VetScan HM2 Hematology System

Transcription

1 VetScan HM2 Hematology System For Veterinary Use Only Customer and Technical Support November 2006 PN: Rev. A Assy PN: Rev. A Text 2006, Abaxis, Inc. Union City, CA 94587

2 IMPORTANT: READ BEFORE USING THE VETSCAN HM2 ANALYZER FOR THE FIRST TIME To get started quickly, please refer to the Quick Reference Guide in the pocket of this Operator s Manual. Fill in this information for future reference Serial number (from the back of the unit):... Date of installation:... Distributor name and address: Abaxis sales representative name:... Phone: THANK YOU FOR PURCHASING THE VETSCAN HM2! Please fill out the Abaxis Product Warranty and Registration Card located in the front pocket of this Operator's Manual and send it to Abaxis soon to get your benefits started. November 2006 PN: Rev. A 2006, Abaxis, Inc. Union City, CA 94587

3 Table of Contents Section 1: General Information Introduction Customer and Technical Support Symbols Used in Labeling and Hazard Identification Section 2: System Overview and Installation VetScan HM2 System Overview Unpacking the System Selecting a Location Installing the System Turning the Analyzer On and Off Initializing the VetScan HM Standby Mode Section 3: Configuring the VetScan HM Printer Settings Operational Settings Date and Time Settings Fluid Sensor Settings Section 4: Test Procedure and Interpreting Results Collecting and Preparing Samples Before Performing an Analysis Analyzing a Sample Adjusting the Needle Height Adjusting the Lyse Volume Interpreting Results Printing and Exporting Results Combining Chemistry and Hematology Results Using Prediluted Mode Interpreting CBC Parameters Section 5: Calibration and Quality Control Calibration Performing Quality Control Section 6: Managing the Database Database Overview Database Table Navigating the Database System Table of Contents TOC-1

4 Section 7: Maintenance & Service Periodic Maintenance Cleaning the Aperture Bleach-Cleaning Shut-Down Procedures Changing the Reagent Pack Running the Self Test Replacing the Peristaltic Pump Tube Assembly Section 8: Special Functions Viewing the Software Version Viewing Status Information Updating the Software Customizing the User-Defined Species Profile Customizing and Printing the Normal Range Section 9: Troubleshooting Warning Indicators Error Messages Evaluating Unexpected Results Preparing the Analyzer for Shipment Section 10: Specifications VetScan HM2 Specifications Linearity Ranges Section A: Introduction to Veterinary Hematology A-1 A.1 Function of Blood A-2 A.2 Composition of Blood A-3 A.3 Blood Cell Parameters A-4 A.4 Normal Hematology Ranges (Dog, Cat, Horse, and Others) A-7 A.5 Veterinary Hematology References A-8 Section B: Operating Principles B-1 B.1 Complete Blood Count (CBC) B-2 B.2 Measurement Methods B-2 B.3 Hemoglobin Determination B-5 B.4 Measured and Calculated Values B-6 B.5 Measured and Calculated Values B-7 TOC-2 Table of Contents

5 Section C: Potential Sample Interferences C-1 Section D: Veterinary Case Studies D-1 D.1 Human Control D-2 D.2 Dogs D-3 D.3 Cats D-10 D.4 Horses D-15 Section E: Glossary E-1 Index I-1 VetScan HM2 Updates following Index Table of Contents TOC-3

6 TOC-4 Table of Contents

7 Section 1 General Information This section provides general information about the Abaxis VetScan HM2 Hematology System. Section Contents 1.1 Introduction Customer and Technical Support Symbols Used in Labeling and Hazard Identification General Information 1-1

8 1.1 Introduction The VetScan HM2 Hematology System is an impedance-based hematology system designed specifically for veterinary in vitro diagnostic use. The VetScan HM2 provides a fully automated complete blood count (CBC) analysis of white blood cells, red blood cells, and platelets in whole blood in a veterinary setting. The system offers an 18-parameter CBC analysis, including a three-part WBC differential, and three cellular histograms. The VetScan HM2 is calibrated to analyze multiple veterinary species. Check the list of species on the HM2 itself (see page 4-8), or contact Abaxis Technical Support for the full list of available species. Additional species may be added to the analyzer software from time to time by means of a simple CD-ROM upgrade, provided to HM2 customers at no charge. The HM2 system features the following components: the hematology analyzer itself, including a keypad, display, and internal printer an external keyboard that can be connected to the analyzer if needed a reagent pack, including a diluent, lyse agent, cleaner, and rinse solution, each in its own container. The diluent container can also be used as a waste container for the next reagent pack. For added convenience, the HM2 can be connected to a VetScan or VetScan2 Chemistry Analyzer so that results from both instruments can be consolidated and printed on a single, standard-size page. In addition, the two analyzers can be connected in tandem to a computer for use with veterinary data management systems. The HM2 alone will also interface directly with an external computer. 1.2 Customer and Technical Support Abaxis Technical Support personnel can answer your questions regarding the VetScan HM2 Hematology Analyzer, or the combined VetScan HM2/VS2 system. Telephone: Beginning January 1, 2007, Abaxis Technical Support will be available seven days a week from 5:00 a.m. to 5:00 p.m. (PST), except on major U.S. holidays. Prior to January 1, 2007, Abaxis Technical Support is available 5:00 a.m. to 5:00 p.m. (PST) Monday through Friday, and 7:00 a.m. to 1:00 p.m. Saturday. technicalsupport@abaxis.com Web: Fax: General Information

9 Note: Abaxis contact information is also available in the HM2 s software. To view the information, press the Utilities key on the instrument s front panel, then press 6 to select Service. 1.3 Symbols Used in Labeling and Hazard Identification The VetScan HM2 uses safety features to protect the operator from injury, prevent damage to the HM2, and to alert the operator of problems with results. Symbols used in labeling and through this manual are shown below. Computer connection. Use only a recommended computer interface cable. Fragile. Use caution when handling. Do not drop. Keyboard connection. Use the keyboard provided. Up. Always store with the arrow facing up. Printer connection. Use only printers recommended and validated by Abaxis. Use only the recommended printer cable. Warning: Alerts the operator of hazardous conditions that can cause injury or damage the analyzer. Read all warning statements carefully, and proceed with caution. Sunlight. Keep out of direct sunlight. Caution: Procedures that must be followed to avoid damaging the analyzer. Read all caution statements carefully. The Product Warranty can be voided if caution statements are not followed. Storage temperature. Store only at temperatures within the indicated range. Note: Important, helpful information General Information 1-3

10 1-4 General Information

11 Section 2 System Overview and Installation This section includes an overview of the VetScan HM2: its features, functionality, main components, reagent pack, and available accessories. The section also provides complete installation instructions. Section Contents 2.1 VetScan HM2 System Overview Features Main Components Control Panel Reagents Accessories Subsystems Menu/Command Listing Unpacking the System Selecting a Location Space Requirements Environmental Requirements Electrical Requirements Installing the System Connecting an External Keyboard (Optional) Connecting the Power Supply Installing Paper into the Built-In Printer Connecting an External Printer (Optional) Connecting a VetScan Chemistry Analyzer (Optional) Connecting the HM2 to a Computer Connecting the Reagent Pack Turning the Analyzer On and Off Turning the Analyzer On Turning the Analyzer Off Initializing the VetScan HM Standby Mode System Overview and Installation 2-1

12 2.1 VetScan HM2 System Overview The following pages provide an overview of the VetScan HM Features Small sample size requirement: consumes approximately 25 µl of EDTA-preserved whole blood per CBC measurement. Rapid test turnaround: only 2 3 minutes to results. Advanced, integrated self-cleaning system for optimized performance with very minimal maintenance expanded automatic aperture cleaning after every run. Simple, intuitive, and easy-to-use software interface makes the analyzer very userfriendly, and requires only a short learning curve. Accurate results for out-of-range values: automatic calculations are provided for external 1:5 dilutions in cases of extremely high values or very small sample volumes. Simple, flexible database with large storage capacity: Automatically saves up to 1,000 test records. Data can be downloaded to a USB flash drive or a compatible data-management system. Quality control (QC) results (including Levy-Jennings charts) are stored in the instrument s QC database for viewing and management. User reminders for replacing the reagent pack and performing simple maintenance tasks. Integrated laboratory results: the HM2 can be connected to a VetScan Chemistry Analyzer for integrated results reporting. Advanced user features: Multi user mode, customizable normal range, and adjustable lysing strength. 2-2 System Overview and Installation

13 2.1.2 Main Components The following pages show the main functional components of the VetScan HM2 analyzer. Front View Control panel with high-contrast liquid 2 CD-ROM drive crystal display (LCD) see page USB Type A port 4 Built-in thermal paper printer see page Interchangeable sample tube adapter 6 Sampling rotor System Overview and Installation 2-3

14 Back View Reagent tubing connections 2 Power On/Off switch 3 12 VDC external power supply input 4 USB Type A port 5 USB Type A port 6 Serial (RS-232) port 7 USB Type B port 8 PS/2 keyboard port Built-in Thermal Paper Printer The analyzer includes an internal printer that outputs test results on thermal paper. 2-4 System Overview and Installation

15 2.1.3 Control Panel The analyzer s control panel features an LCD display, a keypad, six hard-function keys, six soft-function keys, a Start button, and a status LED indicator. Hardfunction keys LCD display Soft-function keys Keypad Status LED Start button Start button Press and release to begin an analysis cycle. Status LED Status indicator Start button This LED (above the Start button) indicates the instrument s status, as follows. Table 2-1: Status LED Color Indications LED color Green Red Orange HM2 status The instrument is in ready state (analysis can begin). The instrument is performing an analysis. No new analysis can be started. The instrument is performing a maintenance process, or is in power-saving standby mode (see Standby Mode on page 2-32). System Overview and Installation 2-5

16 Keypad The analyzer s control keypad includes the following: Alpha-numeric keys for entering data and selecting menu items. Decimal point (. ) key. del key for deleting characters. Cursor control keys: and for moving between items displayed on the screen, and and for moving between items on the screen or menu levels. OK key for confirming data, and for moving through items on the screen. Hard-function keys single-function keys on the instrument s front panel above the LCD display, each of which opens a menu of commands for that function. Table 2-2: Hard-Function Keys Press this key... To use this function... Information/Help Measurement/Analysis press to begin analysis. Database press to view stored results. Utilities press to perform quality control, calibration, maintenance, or setting functions, or to check reagent status. Print Exit press to shut down the instrument. Note: See Menu/Command Listing on page 2-12 for a list of the menus and commands that are available through the hard-function keys. 2-6 System Overview and Installation

17 Soft-function keys a row of unlabeled keys just below the LCD display: The functions of these keys depend on the current menu, and are displayed on the screen immediately above the keys, as shown in this example. To use a displayed function, press the soft-function key below it. The following table lists the various soft-function keys that appear on the instrument. Table 2-3: Soft-Function Keys ACCEPT Accepts the results or changes made. CANCEL Leaves the current menu without saving changes. EXIT Exits the current menu or action. GRAPH Displays histograms of the selected result. TABLE Displays table of values for the selected result. MENU Opens the local menu for the functions displayed. PT.ID Enters or modifies patient data. REPEAT Repeats the previous action. PREVIOUS NEXT Move through results displayed on multiple pages. PGUP PGDN Move through screens with multiple pages NEW OLD STOP CHANGE Change the scaling of Levey-Jennings chart (16 or 64 days). When completing a bleach-cleaning, select NEW to use a new reagent pack, OLD to continue using the current reagent pack, or to STOP to shut down the instrument. Resets the status of the reagent pack. LIMITS Displays reference ranges for selected species. CHKSTA Checks the printer status if an error occurs. CONTINUE Attempts to print again after a printer error has been corrected. RESET Resets the printer if an error occurs. ABORT Stops the current process. System Overview and Installation 2-7

18 2.1.4 Reagents VetScan HM2 Reagent Pack The HM2 reagent pack (PN ) consists of bottles containing diluent, cleaner, lyse, and rinse solutions. Note: To ensure accurate test results, use only the reagents supplied by Abaxis. The following table lists the reagents volumes and bottle sizes in the reagent pack. Table 2-4: Reagents and Containers Reagents Descriptions Color Code Volume Bottle Size Diluent Isotonic saline solution used to dilute whole blood specimens, and to rinse the analyzer s fluidic system between analyses. Green 6200 ml 8 liters Rinse Cleaner Lyse Used with diluent to prevent salt built-up on aperture. Used in the fluidic system cleaning process. Used to create hemolysate for three-part WBC differential, and for total WBC and HGB. White 500 ml 500 ml Blue 250 ml 250 ml Yellow 270 ml 500 ml Cat, Dog, and Human Controls Before you can perform analyses, the instrument must first be calibrated using the Human Calibrator 1, and quality control procedures must then be performed using Cat and Dog Control materials. These control materials are not included with the analyzer, but can be ordered from your distributor or directly from Abaxis. If you order from Abaxis, please use these part numbers: , Dog Control, 2 cc vial (1) , Cat Control, 2 cc vial (1) , Human Calibrator, 2 cc vial (1) 1. The Human Calibrator is derived from human sources. Observe universal safety precautions when handling the Calibrator. See Calibration and Quality Control on page System Overview and Installation

19 2.1.5 Accessories This section describes the accessories that are included with the analyzer. To order replacements, contact your distributor or Abaxis. External Mini-Keyboard You can connect the external mini-keyboard to the instrument through its PS/2 keyboard port or one of its USB ports (see Back View on page 2-4). The mini-keyboard provides a convenient way to enter patient and clinic information. The mini-keyboard s function keys F1 through F6 function as equivalent to the analyzer s softfunction keys (from left to right: see page 2-7), and keys F7 through F12 function as equivalent to the analyzer s hard-function keys (see page 2-6): F7 Information/Help key F8 Measurement/Analysis key F9 Database key F10 Utilities key F11 Print key F12 Exit key Power Supply and Power Cord The analyzer uses an external 12 VDC power supply that can operate from a 230 or 110 V main outlet. The power supply s input socket is a standard power cable connection, and its output is a special locking socket. CAUTION: Abaxis recommends using the analyzer with a surge protector designed for a computer. In addition, an uninterruptable power supply (UPS) is strongly recommended if the HM2 will be used in an area prone to electrical surges or power outages. An appropriate power supply is vital for the integrity of the system. System Overview and Installation 2-9

20 Reagent Tubing Kit The reagent tubing kit (Abaxis Part No ) includes the color-coded reagent tubes used to connect the HM2 with the containers in the reagent pack. The kit also include the reagent bottle caps with connectors and drop-down tubes, as shown. Cleaning Tube Kit The cleaning tube kit is needed for bleach-cleaning the instrument, as described in Bleach-Cleaning on page 7-5. Sample Tube Adapters The instrument includes the following sample tube adapters, for use as shown in Analyzing a Sample on page 4-7. Table 2-5: Sample Tube Adapters Tube Adapter Part Number Sample Tube Size 3 5 ml Tube Adapter ml Microtainer Tube Adapter ml Vacutainer Adapter µl Control Vial Adapter ml control vial Peristaltic Pump Tube Assembly As part of routine yearly preventive maintenance, the HM2 s peristaltic pump tube must be inspected, and replaced if needed (see Periodic Maintenance on page 7-2). The HM2 is shipped with one replacement peristaltic pump tube System Overview and Installation

21 Thermal Paper Roll Two thermal paper rolls are included for the instrument s internal printer, to provide quick, convenient printed results. Note: Thermal paper printouts only last about a year. If you need to keep a record for longer than a year, make a photocopy of the printout Subsystems The VetScan HM2 has two primary subsystems. Fluidic system: Performs sampling, diluting, mixing, lysing, and rinsing functions. Generates the regulated vacuum used for moving cells through the aperture during the counting process. Diluent Lyse Cleaner Waste Rinse reagent Diluent solution Lyse solution Cleaner solution Waste container Rinse reagent Data processing system: Counts, measures, and calculates blood parameters, generates and stores numerical results and histograms. System Overview and Installation 2-11

22 2.1.7 Menu/Command Listing The following charts outline the instrument s menu functions and commands. Information / Help Displays help for the current screen. Analysis / Measurement MENU PT.ID 1 Repeat last sample Repeats last sample tested (HM2 assigns new sample ID, all other patient information is unchanged). 2 Measure blank Performs blank measurement. 3 Prediluted mode Enters automatic calculation for 1:5 externally diluted sample ( 1:5 appears in upper RH screen corner). 4 WBC only Analyzes WBC only with next test. 5 Needle Height Setting Adjusts sampling depth. 6 Lyse Volume Adjusts lyse reagent volume by +/ 0.2 ml. Opens dialog for entering patient information. Sample ID Doctor Species Patient ID Name Age Sex HM2 assigns automatically (cannot be modified). LIMITS Displays reference ranges for selected species (can be modified) System Overview and Installation

23 Database MENU 1 Go to specified record Selects one record by time/date, sample ID, and/or patient ID (more criteria narrows search). 2 Selection 1 Select by date, Selects one or more records by time and ID range of times/dates, sample IDs, and/or patient IDs. 2 Select all Selects all records. 3 Deselect all Deselects all records. 3 Change sort order 1 Unsorted Lists records in the order entered (most recent to oldest). 2 Sort by time Lists records by measurement time (oldest to most recent). 3 Sort by sample ID Lists records by sample ID (highest to lowest). 4 Sort by patient ID Lists records by patient ID (highest to lowest). 4 Manage selected records 1 Send selected Sends selected records to a connected records computer. 2 Delete selected Deletes selected records from the records instrument s database. 3 Backup selected Saves selected records to a USB drive. records GRAPH TABLE 5 View external Reads stored records from USB drive (insert a USB drive containing saved HM2 data into one of the instrument s USB ports). 6 Backup one day Backs up all results from a specified date to a USB drive (insert a USB drive with sufficient space into one of the instrument s USB ports). Toggle between graph and table display of selected results. PT.ID Opens dialog for viewing/modifying patient information (sample ID and patient type cannot be changed). Sample ID Doctor Species Patient ID Name Age Sex System Overview and Installation 2-13

24 Utilities 1 Maintenance 2 Calibration 3 Quality control 1 Cleaning 1 Auto self-cleaning Runs cleaning cycle to remove buildup from instrument s aperture. 2 Clean the wash head Moves wash head to cleaning position, provides cleaning instructions. 2 Priming Primes reagent lines (use after replacing reagent). 1 Prime diluent Primes diluent line (green connector). 2 Prime lyse Primes lyse line (yellow connector). 3 Prime rinse Primes rinse line (blue connector). 4 Prime cleaner Primes cleaner line (white connector). 5 Prime all Primes all reagent lines. 3 Drain chamber Drains aperture chamber (use before moving the instrument). 4 Reagent status Displays reagent installation date, usage, and remaining life. 5 Bleaching Bleaches internal lines (perform with each change of reagent pack). 6 Software upgrade Prompts for software upgrade disk and restarts system. 1 Run Calibrator Performs calibration. 1 Whole blood Calibrates using Abaxis calibrator material. 2 Prediluted blood (1:5) Calibrates using externally diluted Abaxis calibrator material to ensure proper calibration of prediluted samples. 2 View Calibration History Displays previous calibrations. 1 Set QC reference ranges Updates QC ranges for current QC lot number. 2 Run QC Runs QC analysis. 3 View table of QC measures Displays previous QC measures. 4 View QC diagram Displays previous QC measures as Levey-Jennings curve. 5 Select QC type (continued) 1 Dog 2 Cat 3 Human 2-14 System Overview and Installation

25 Utilities (continued) 4 Diagnostics 1 Device information Shows model, S/N, software version. 2 Statistics Displays operation statistics. 3 Self test Tests the analyzer s internal systems. 5 Settings 1 Printer settings Dialog for viewing/modifying printer settings. 2 Customize 1 General Settings 2 Units 3 Laboratory 4 User modes Dialog for setting screensaver delay, VetScan date format, and printing combined VetScan/HM2 results. Dialog for setting unit preferences. Dialog for laboratory information (used in report header). Selects Single or Multi user mode. 1 Single user mode 2 Multi user mode Each user has unique user name and password. 3 Date and time Dialog for setting date/time & format. 4 Fluid sensors Enables and disables reagent sensors. 1 Diluent & Cleaner 2 Lyse 3 Rinse 4 Calibrate sensors Calibrates reagent sensors. 6 Service Displays contact information for technical service. Printing Prints selected test results. Exit Information/Help function. 1 Shut down Shuts off the HM2 (use if the unit will not be used for over 72 hours). 2 Preparing for shipment Drains and shuts off the HM2 (use if the unit will be unused for more than 10 days, or if it will be shipped). System Overview and Installation 2-15

26 2.2 Unpacking the System Follow these directions for unpacking the HM2 analyzer from its shipping container. Note: Save the shipping box, accessory box, and packaging materials in case you need to use them later. Refer to the diagram at right. 1. Open the shipping carton box, and remove the Thank-You Letter (A) and Unpacking Instructions (B). Follow the instructions on the Unpacking Instructions (B). 2. Slide the accessory box (D) located at the back of the packaging box up and out of the shipping carton. 3. Pull the Foam Cap (C) off of the analyzer. 4. Place both hands in the box on opposite sides of the analyzer, and carefully lift it out of the carton as shown. 5. Place the instrument temporarily on a clean, level surface that is free of animal hair, dust, and other contaminants. CAUTION: Place the analyzer in upright position only. Do not place the unit on its back, side, or top, or you could cause severe damage System Overview and Installation

27 6. Use the latch to open the side door on the unit, as shown. 7. Inspect the unit for any visible signs of damage. 8. Remove the foam above the needle XYZ moving assembly, and remove the tape from the top of the counting chamber, as shown. 9. Open the accessory box, and use this list to make sure you received all components of the HM2 system: VetScan HM2 Analyzer HM2 Operator's Manual (this document) HM2 Quick Reference Guide HM2 analyzer external power supply and power cord HM2 mini-keyboard Four sample tube adapters HM2 reagent tubing kit (with color-coded connectors) Caps for reagent containers (with color-coded connectors) HM2 cleaning tube kit Thermal paper rolls (two) Warranty card (inside the HM2 Operator's Manual) Spare peristaltic pump tube assembly (store in a convenient location) System Overview and Installation 2-17

28 Note: The HM2 requires a reagent pack to operate. A reagent pack (Abaxis Part No ) must be ordered for installation. Note: To start the warranty on the HM2, make sure to complete the warranty card and mail it to Abaxis, Inc. within 10 days of system installation. Customers who submit the warranty card are automatically placed on the Abaxis customer list, and are entitled free of charge to all the benefits that Abaxis offers, such as software upgrades, on-line training, education materials, and promotions. 2.3 Selecting a Location WARNING: MAKE SURE THE ANALYZER AND ALL ACCESSORIES ARE PROPERLY GROUNDED. IMPROPER GROUNDING CAN CAUSE INJURY, AND WILL VOID THE WARRANTY. The analyzer must be installed in a suitable location. A poor location can adversely affect its performance. To ensure the accuracy and precision of the instrument, and to maintain a high level of operational safety for lab personnel, the environmental and electrical requirements in this section must be met. Be sure to thoroughly consider all the following requirements in selecting a permanent location for the analyzer Space Requirements The location should be flat, level, sturdy, vibration-free, and as dust-free as possible. Leave at least 20 inches (about 0.5 m) of clearance on all sides of the instrument to allow for adequate airflow and access to connectors. Maintain a minimum of 8 inches (0.2 m) behind the analyzer s rear panel to allow for heat dissipation and tube clearance. Leave enough space for the reagent pack (see Connecting the Reagent Pack on page 2-25 for details) System Overview and Installation

29 2.3.2 Environmental Requirements CAUTION: Make sure the location stays at an ambient temperature of F (15 30 C), and a relative humidity of 65% ± 20%. The optimal operating temperature is 77 F (25 C). CAUTION: Make sure the area is not exposed to open windows, heat sources, air conditioners, temperature extremes, or direct sunlight. Make sure the location is well ventilated. Make sure the location is away from devices that can emit radio frequencies, such as a radio or TV, radar, centrifuge, X-ray device, or fan. Such devices can interfere with the HM2 s operation. Operating the HM2 at an altitude above 10,000 ft (3000 m) is not recommended Electrical Requirements The HM2 is powered by a standard wall outlet, and requires a power supply of VAC, Hz, 2 A (this is provided with the analyzer). To avoid power surges or drain, DO NOT plug the analyzer s power supply (provided) into a circuit that includes a centrifuge or other high-current device. CAUTION: Abaxis recommends using the analyzer with a surge protector designed for a computer. In addition, an uninterruptable power supply (UPS) is strongly recommended if the analyzer will be used in an area prone to electrical surges or power outages. An appropriate power supply is vital for the integrity of the system. System Overview and Installation 2-19

30 2.4 Installing the System Once you ve selected a location, install the analyzer system as follows. CAUTION: Make sure all settings are in the off position before proceeding or before connecting the analyzer to the power supply or to any ancillary devices (such as an external printer, external keyboard, or computer) Connecting an External Keyboard (Optional) You can use an external keyboard with either a PS/2 connector or a USB connector. Connect either cable to the correct port on the rear of the analyzer Connecting the Power Supply WARNING: USE ONLY THE POWER SUPPLY PROVIDED WITH THE HM2. USING ANY OTHER POWER SUPPLY CAN DAMAGE THE INSTRUMENT, AND WILL VOID THE WARRANTY. 1. Plug the power cord from the power supply into the connector on the analyzer s rear panel. 2. Plug the other end of the cord into a properly grounded AC outlet ( VAC) Installing Paper into the Built-In Printer Note: Store thermal paper away from direct light and humidity. If you plan to print results using the analyzer s built-in printer, install the thermal paper roll as follows. 1. Remove any tape from the roll of thermal paper, and cut the end of the roll to a point System Overview and Installation

31 2. Open the paper compartment by pressing the button on top of the analyzer. 3. Insert the paper roll into the compartment so that the paper unrolls beneath the roll and toward the front of the instrument. paper must unroll in this direction 4. Make sure the free end of the roll extends out of the front of the compartment. press to open compartment 5. Press the compartment door closed. Note: When the roll is running out, a red mark appears on the paper Connecting an External Printer (Optional) 1. Set up the external printer according to the instructions included with it. Note: Any diskettes or CDs included with the printer are not required for use with the analyzer, but should be saved in case they are needed another time. 2. Attach the USB printer cable to a USB port on the rear of the analyzer, and to the printer. 3. Plug the printer s power cord into a grounded outlet. 4. Make sure the analyzer is configured for use with the printer see Configuring Printer Settings on page Turn the printer on before turning on the analyzer. System Overview and Installation 2-21

32 2.4.5 Connecting a VetScan Chemistry Analyzer (Optional) Connecting the VetScan HM2 with the VetScan Classic Chemistry Analyzer 1. Make sure the HM2 and the VetScan Classic are both turned off. 2. Connect an RS-232 null modem cable (Abaxis Part No ) to the serial port on the back of the HM2, and to the serial (1) port on the back of the VetScan Classic. 3. Turn on the HM2 and the VetScan Classic. serial port RS-232 cable serial port HM2 VetScan Classic 2-22 System Overview and Installation

33 Connecting the VetScan HM2 with the VetScan VS2 Analyzer 1. Make sure the HM2 and the VS2 are both turned off. 2. Connect a USB Type A-B cable (Abaxis Part No ) to a USB Type A port on the back of HM2, and to a USB Type B port on the back of the VS2. 3. Turn on the HM2 and the VS2. HM2 USB cable VetScan VS2 USB ports (Type A on HM2, Type B on VS2) Note: To print VetScan Chemistry Analyzer results on the HM2, the HM2 must be configured to receive data from the VetScan analyzer. For instructions, see Combining Chemistry and Hematology Results on page Connecting the HM2 to a Computer The HM2 can be connected to a computer using either a serial cable or USB Type A-B cable. This enables you to send information from the analyzer for use by a practice management application or other applications on the computer. Using a Serial Cable Connect the analyzer to a serial port on the computer using an RS-232 null modem cable, or a serial cable and a null modem adapter. System Overview and Installation 2-23

34 Using a USB Type A-B Cable The analyzer can be connected to a computer using the USB Type B port on the back of the analyzer. A software driver must also be installed on the computer to enable communication with the analyzer. Note: Do not connect the USB cable until directed to do so. The following are required to enable the analyzer to communicate with a computer: PC computer with Windows XP and a USB port USB Type A-B cable (Abaxis Part No ) Abaxis VetScan HM2 software CD, version 2.74 or higher (includes the USB driver) Note: For current information on the availability of the USB driver for other operating systems, contact Abaxis Technical Support see Customer and Technical Support on page 1-2. Step 1 Installing the HM2 USB Driver 1. Insert the VetScan HM2 software CD (version 2.74 or higher) into the computer s CD drive. The installation program then starts automatically. 2. Click Next on the first dialog box that appears on screen, then click Finish on the second dialog. Driver installation is then complete. Step 2 Connecting the USB Type A-B Cable Connect a USB Type A-B cable to the USB Type B port on the back of the analyzer, and to a USB Type A port on the computer. Step 3 Configuring the USB Driver The analyzer appears on the computer as a virtual communications port. You will need to use the computer s Device Manager to redefine that port s properties. 1. Click Start > Control Panel. 2. Double-click System. 3. Click the Hardware tab, then click Device Manager. 4. Click the + mark next to Ports (COM and LPT). 5. Right-click USB Serial Port (COMx), then select Properties System Overview and Installation

35 6. Click the Port Settings tab, then click Advanced. 7. Select the appropriate COM port from the drop-down list. You may want to make sure this port matches the COM port used by or selected in your practice management application or other software. 8. Click OK. 9. If a warning states that this modification may interfere with current port settings, consult Abaxis Technical Support or your system administrator before proceeding. If no devices are connected to the relevant port on the computer, you can confirm this message. Otherwise, make sure this won't interfere with your computer settings. Step 4 Configuring the Analyzer to Communicate with a PC You can now configure the analyzer to communicate with an interfaced practice management software application that runs on the computer. Make sure the selected port for the application matches the port selected in the above procedure. For detailed configuration information, see General Settings on page Connecting the Reagent Pack Note: During transportation, microbubbles can form in the reagent solutions. These bubbles will affect test results. Before connecting a reagent pack to the analyzer, allow the pack to stand undisturbed for 24 to 36 hours to eliminate any bubbles. CAUTION: If the analyzer has been kept at a temperature below 50 ºF (10 ºC), allow it to sit for an hour at the correct operating temperature (59 86 F, C) before using it. When installing the analyzer in its permanent location, place the reagent pack according to these guidelines. Place the reagent pack near the analyzer, either beside or behind the instrument. For best results, place the reagent pack at the same level as the analyzer. If you have to place the reagent pack on a lower level, make sure that it is no more than 30 inches (0.75 m) below the level of the analyzer s reagent inlets, as measured from the bottom of the reagent pack to the bottom of the analyzer. System Overview and Installation 2-25

36 WARNING: DO NOT PLACE THE REAGENT PACK ABOVE THE ANALYZER (SUCH AS ON A SHELF). DO NOT DROP THE REAGENT PACK THIS CAN CAUSE MICROBUB- BLES TO FORM. CAUTION: When working with the reagent tubing, make sure the tubing does not become pinched or kinked and is not trapped between or beneath objects. The reagents must be able to flow through the tubes freely and without obstruction, or the instrument will not operate properly. Connect the reagent pack as follows. 1. Open the plastic bag located in the accessory box, and take out the five color-coded reagent tubes and caps with drop-down tubes. 2. Locate the color-coded reagent intake valves on the back of the instrument, as shown. Green = Diluent Yellow = Lyse Blue: = Cleaner Red = Waste White = Rinse 3. Remove the protective caps from the intake valves. Save the caps for later use. Note: The protective valve caps will be needed whenever the analyzer is moved, to prevent reagents from leaking. 4. Insert the capped ends of the reagent tubes into the corresponding color-coded reagent intake valves. Finger-tighten the colored caps securely. CAUTION: Do not overtighten the caps, or you could damage the threads on the connectors. This will cause the reagents to leak and damage the instrument System Overview and Installation

37 5. Open the reagent pack box. Open each reagent container (lyse, rinse, cleaner, and diluent) by unscrewing the container caps. 6. Re-cap each container using the bottle cap with the appropriate color-coded connector and drop-down tube. Note: Be sure to save the original reagent caps so you can re-cap the containers when the reagent pack is used up. 7. Using the color codes as guides, attach the free end of each reagent tube to the bottle cap connector on the appropriate container. Press each tube onto its connector as far as it will go. Note: The analyzer is shipped with a flexible waste container, to be used with the first reagent pack. The diluent bottle included with each reagent pack is designed to be used as the waste container for the subsequent reagent pack. CAUTION: Make sure the small air vent on each container cap is not blocked, and that free airflow is maintained at all times. Do not touch the drop-down tubes with your hands, or you could contaminate the reagents. If you must handle the drop-down tubes, be sure to wear gloves. CAUTION: Make sure each tube runs from the reagent intake valve to its corresponding reagent bottle. Use the color codes as guides. If the tubes are not connected correctly, the instrument will not produce accurate results. If you let the reagent tubes pass through the openings on the reagent pack, make sure the top of the reagent pack does not kink or pinch the tubing. System Overview and Installation 2-27

38 2.5 Turning the Analyzer On and Off Turning the Analyzer On CAUTION: If the analyzer has been kept at a temperature below 50 ºF (10 ºC), allow it to sit for an hour at the correct operating temperature (59 86 F, C) before using it. 1. If you have an external printer or a computer connected to the analyzer, turn on the power to the printer/computer before starting the analyzer. 2. Turn on the analyzer on using the power switch on the upper left of its rear panel. The on position is marked by the I symbol. The analyzer then displays several screens while it runs its software start-up procedure. Note: To initialize the analyzer, see Initializing the VetScan HM2 on page Turning the Analyzer Off CAUTION: Never switch the analyzer by simply pressing the power switch on the rear panel, unless an emergency exists, or if you will turn the instrument on again in only a few minutes. CAUTION: When the analyzer is shut down properly as described below, it rinses its fluidic system. Since the analyzer uses isotonic saline solution, the salt from the solution may not be rinsed out properly if you simply turn off the power. This could lead to salt build-up in the system. Therefore, always follow the instructions in this section when turning off the analyzer. CAUTION: If an emergency occurs, turn off the analyzer using the power switch on the back of the instrument, and unplug the power cord from its outlet System Overview and Installation

39 If the Analyzer Will Not Be Used for 72 Hours or More Shut down the instrument as follows. 1. Press the Exit key on the front panel. 2. Select Shut down, then press the YES softfunction key when asked to confirm. The analyzer displays a message and sounds a tone when it s safe to shut it off. 3. Turn off the analyzer using the power switch on the rear panel. The off position is marked by the O symbol. Note: If you perform fewer than ten CBC samples a week, Abaxis strongly recommends that you shut down the analyzer once a week. This will prevent salt from building up in the system, and increase the instrument s efficiency. If the Analyzer Will Not Be Used for 10 Days Or More, or Will Be Shipped Shut down and prepare the instrument as follows. You will need the cleaning tube kit (shown on page 2-10) and 100 ml of distilled water for this procedure. 1. Press the Exit key on the front panel. 2. Select Preparing for shipment, then press the YES soft-function key when asked to confirm. The display then shows Pneumatic system is initializing. Please wait. 3. Follow the instructions that appear on the display. When prompted, remove the four reagent tubing connectors (green, yellow, blue, and white) from the reagent tubing intake valves, but leave the waste tubing connected. System Overview and Installation 2-29

40 4. When prompted, connect the cleaning tube kit to the reagent connectors, as shown. 5. Submerge the free end in a bottle containing at least 100ml of distilled water. 6. Press ACCEPT. The analyzer then flushes any remaining reagents into the waste container. 7. When prompted, remove the cleaning tube kit. Press ACCEPT. 8. When the analyzer displays Now it is safe to turn off the instrument, turn off the analyzer using the power switch on its rear panel. 9. Disconnect the waste tubing. The analyzer is now ready for shipment or extended nonuse. 10. If you need to prepare the analyzer for shipment, continue with the instructions in Preparing the Analyzer for Shipment on page System Overview and Installation

41 2.6 Initializing the VetScan HM2 Use this procedure to initialize the HM2 after installation, or after it has been turned back on after an extended shut-down. 1. Press the Measurement/Analysis key. The analyzer then initializes its pneumatic system and runs a blank measurement. When the process is complete, the instrument displays the result of the blank measurement. 2. When the blank measurement is OK, press ACCEPT to accept the results. The analyzer then displays a sample measurement screen, as shown, and is now ready to perform an analysis. Note: You will be notified if the blank does not fall within specifications (these are listed on page 4-6). If this occurs, run one to three cleaning cycles (see Cleaning the Aperture on page 7-5), and re-run the blank as needed until it falls in range (no flags, and Blank OK is displayed on the screen). If the problem persists, call Abaxis Technical Support see Customer and Technical Support on page Verify the HM2 s system settings, and make any needed changes see Configuring the VetScan HM2 on page If you are installing a new system, or have moved the analyzer or its reagent pack to new locations, recalibrate the fluid sensors to maximize the number of tests performed per reagent pack on your system see Fluid Sensor Settings on page Allow the analyzer and its reagents to fully reach room temperature (usually about five minutes) before beginning an analysis. This will avoid damage from condensation caused by rapid temperature changes, as well as interference and background noise caused by microbubbles in the reagents during installation. System Overview and Installation 2-31

42 2.7 Standby Mode The analyzer automatically enters a power-saving standby mode after a period of standing idle. When in standby mode, the analyzer displays a Ready screen as shown at right, with rotating information at the bottom of the screen showing available user functions. The analyzer can remain in standby for up to 72 hours. To bring the analyzer out of standby, simply press any key System Overview and Installation

43 Section 3 Configuring the VetScan HM2 This section describes how to configure the VetScan HM2 for optimal performance and to meet your particular lab requirements. Section Contents 3.1 Printer Settings Configuring Printer Settings Printout Examples Troubleshooting Printer Settings Operational Settings General Settings Unit Settings Laboratory Information Settings User Mode Settings Date and Time Settings Fluid Sensor Settings Configuring the VetScan HM2 3-1

44 3.1 Printer Settings Configuring Printer Settings Before using a printer, whether the analyzer s built-in printer or an external printer, you must configure the analyzer for the printer and its requirements, as follows. 1. Press the key, then select Settings. 2. Select Printer settings. 3. Use the and keys to set Printer to the printer you ll use (the default is Seiko LPTH245 Built in). 4. Press the ACCEPT soft key to accept the printer settings. The following table lists the available printer selections, along with the printer language and specific printer models corresponding to each. Table 3-1: Compatible Printers Printer Selection Printer Language Supported Printer Models Seiko LPTH245 Built in Special printer language HM2 built-in thermal printer Epson ESC/P Raster Epson ESC/P Raster Epson Stylus series (select models only a ) Canon BJC in 9-pin Canon BJC Canon BJ and BJC series (select models only a ) mode, Canon BJC Canon i 70 Canon Canon i series (select models only a ) Seiko DPU414 Special printer language Seiko DPU414 printer Epson 24-pin in 9-pin mode, Epson 24-pin ESC/P2 Epson 24-pin dot matrix series (LQ-580, Epson 440, etc.) Epson 9-pin ESC/P Epson 9-pin dot matrix series (FX-980, etc.) HP LaserJet PCL4 HP LaserJet series and compatibles (LJ1100, etc.) HP DeskJet PCL4 HP DeskJet series (select models only a ) a. For more information, contact Abaxis Technical Support see Customer and Technical Support on page Configuring the VetScan HM2

45 Note: Although the HM2 can be connected to any printer compatible with any of the above printer languages, printout sizes and fonts may vary depending on manufacturer and printer settings. Abaxis carries printers that have been tested and validated specifically for use with the HM2. For best results, Abaxis highly recommends using only printers validated by Abaxis. For a current list and further information, contact Abaxis Technical Support see Customer and Technical Support on page Configure the remaining settings as follows. Use the and keys to change the settings, and the keypad or external keyboard to enter text or numbers. Use the PGUP and PGDN soft keys and the and keys to move through the settings. Use PGUP and PGDN to move between page of settings. When using the analyzer's built-in printer, these settings are recommended: Printout format: Narrow full (with histograms) Table format: Normal or Narrow Printer cable: COM2 Paper: RollPaper Units: inch or cm Paper size: 2.24 x Left margin: 0.00 Top margin: 0.00 Printing mode: Normal Physical margin: Normal One result per page: No Rollpaper: Yes Vertical spacing: 0.50 Configuring the VetScan HM2 3-3

46 Autoprint: Yes (Yes prints results automatically after each analysis; No requires you to press the Print key.) Limits format: L Graph H Print flags: Yes Print warning flags: Yes Clogging report: No Serial number on result: Yes When using an external printer, the following default settings are recommended: Printout format: Full with histograms Printer cable: USB Paper: select as appropriate for your printer (Note: Never select Rollpaper for an HP LaserJet.) Left and top margins: 1.00 Rollpaper: No (especially for HP LaserJets) 6. Configure any other settings as needed. See Table 3-2 for a complete list. Table 3-2: Printer Settings Setting Description Printer Printer used with the HM2 see Table 3-1 on page 3-2. Printout format Overall format used for printouts. Available selections depend on the selected Printer. (See page 3-6 for examples.) Table format Printed width of tables included in the printouts. Printer cable Interface used for an external printer: USB. Paper Paper type used in the selected Printer. Units Paper size Left margin Top margin Printing mode Physical margin Default units of measure, paper size, and print margins for the selected Paper. These may be customized when using a non-standard paper. Left and top margins control the position of printed results on the page. Overall printout size (Mini, Small, Normal, Enhanced, or Large), width (Wide or Narrow), and print speed (Fast). The available modes depend on the selected Printer. Margin correction for printers that have smaller printable areas than usual. Use Normal unless the right edge of the printed text is missing or appears in the next line. 3-4 Configuring the VetScan HM2

47 Table 3-2: Printer Settings (Continued) Setting One result per page Rollpaper Vertical spacing Autoprint Limits format Print flags Print warning flags Clogging report Serial number on result Description Select Yes to print each analysis on a separate page. Select Yes if using the built-in printer. The vertical distance between printed results when multiple reports are printed on each page. Select Yes to print all results automatically after each analysis. The format for printing reference ranges: To print numeric values, select the displayed range (for example, [ ]). None prints no range values. Graphical prints a graph showing where the results falls within the range. L-H Graph prints the low and high limits of the range, followed by a graph showing where the result falls within the range. Graph L-H prints a graph showing where the result falls within the range, followed by the low and high limits of the range. L Graph H prints the low limit of the range, a graph showing where the result falls within the range, and the high limit of the range. Select Yes to include in the printout any measured values that fell outside the reference range, as well as any errors during analysis. Select Yes to include all warning flags in the printouts. Select Yes to include information on aperture clogging in all printouts. Select Yes to include the analyzer s serial number in all printouts. 7. Press the ACCEPT soft key to accept the settings. Configuring the VetScan HM2 3-5

48 3.1.2 Printout Examples Thermal Paper Printout Report header Patient information Patient species Report date Test date & time (dates and times assigned automatically) HM2 serial number Sample ID (assigned automatically) Reference ranges Test parameters shows location of test results within reference range limits lower limit upper limit Test results Test result units WBC histogram RBC histogram PLT histogram 3-6 Configuring the VetScan HM2

49 8 x 11.5 Letter Printout Report header Sample ID (assigned automatically) Patient information Patient species Patient information Report date (assigned automatically) and HM2 serial number WBC histogram RBC histogram Test parameters Test results Reference ranges PLT histogram Test result units shows location of test results within reference range limits lower limit upper limit Configuring the VetScan HM2 3-7

50 3.1.3 Troubleshooting Printer Settings Use this table to solve problems related to printer settings. Table 3-3: Troubleshooting Printer Settings Problem Incorrect characters appear on the printout. Right side of the printed report is missing or appears on the next line. Printed report is too small, with excess space on the paper. End of the printout appears on the next page. Another patient report could fit on the page. Printed result is not centered horizontally. Printed result is not centered vertically. Results are printed too close together or too far apart. Possible Causes / Solutions Selected printer type does not match your printer. Select the correct type in Printer. Decrease the Left margin and Top margin. Try using a smaller Printout format. Try using a larger Printout format. Enter the correct Paper size. Try decreasing the Left margin and Top margin. Enter the correct Paper size. Try increasing the Left margin, Top margin, and Vertical spacing. Modify the Left margin. Modify the Top margin. Modify the Vertical spacing. 3-8 Configuring the VetScan HM2

51 3.2 Operational Settings Use the Customize menu to set the language and date format used by the HM2, along with the screensaver delay, and the date range used for printing combined results from the HM2 and a connected VetScan analyzer General Settings 1. Press the Utilities key. 2. Select Settings. 3. Select Customize. 4. Select General Settings. Configuring the VetScan HM2 3-9

52 5. Configure the settings as needed. Use the and keys to move through the settings. Use the and keys to change the settings, and the keypad or an external keyboard to enter text or numbers. Use the on-screen PGUP and PGDN soft keys to move between pages of settings. Table 3-4 describes the available settings. Table 3-4: General Settings Screensaver wait time Language PC Link Serial Protocol Version Serial s Decimal Dot VSx Link Date format of VetScan 0 to 30 minutes (0 = screensaver off). English. International customers: please contact your distributor for information about available languages. Offline, USB, Baud rate (150 to ). Protocol used by the analyzer to communicate with a connected computer. If the analyzer is not connected to a computer, choose Offline. If the analyzer is connected to a computer using a USB cable, choose USB. If the analyzer is connected to a computer through serial cable, 9600 is correct for most cases, or you can contact Abaxis Technical Support for the correct setting see Customer and Technical Support on page , 1.7, 2.20, 2.23, 3.0, or 3.1. Protocol versions used by practice management software to interface with the analyzer. Contact Abaxis Technical Support (see page 1-2) or your practice management software provider for the proper selection. (Newer versions have higher numbers.) Dot (period), slash, comma, or lang def (as defined by language). Offline, USB, or RS232. Protocol used by the HM2 to communicate with a connected VetScan Classic or VS2 for combining results. If the HM2 is not connected to a VetScan Classic or VS2, choose Offline. If the HM2 is connected to a VetScan VS2 through a USB cable, choose USB. If the HM2 is connected to VetScan Classic through a serial cable, choose RS232. When printing combined HM2 and VetScan results, the date format used by the VetScan: MM/DD/YY or DD/MM/YY (DD = date, MM = month, YY = year). Set both instruments to the same format Configuring the VetScan HM2

53 Table 3-4: General Settings (Continued) VetScan-HM2 combining within 365, 90, 60, 30, 15, 8, 5, 3, 1 day, Same day, Don t care. When printing combined results from the HM2 and VetScan Classic or VS2, defines the time range within which HM2 and VetScan hematology results will be paired: HM2 Patient ID number is paired with the Patient Number on the VetScan Classic, or with the Sample ID on the VS2. For example, if the clinic uses the same patient ID numbers each day, so that a given number may be assigned to more than one patient, select Same day to avoid inadvertently pairing results from different patients. Chemistry and hematology results using the same patient ID number will then only be paired if run in the same day. If this is not a requirement, use Don t care. The HM2 then matches the most recent results on the VetScan Classic or VS2. 6. Press the ACCEPT soft key to accept the settings Unit Settings Use the Units menu to set the measurement units the HM2 will use. 1. Press the Utilities key. 2. Select Settings. 3. Select Customize. 4. Select Units. Configuring the VetScan HM2 3-11

54 5. Set the units as needed: Use the and keys to select a unit. Use the and keys to select a setting for that unit. Table 3-5 lists the available unit settings. Table 3-5: Unit Settings Count unit HGB unit PCT, HCT unit RDW, PDW mode cells/liter (cells/l), cells/µl (cells/µl). Prec. displays a more precise value. grams/liter (g/l), grams/deciliter (g/dl), millimols/liter (mmol/l) percentage (%), absolute (abs) standard deviation (sd), coefficient of variation (cv) 6. Press the ACCEPT soft-key to save the settings Laboratory Information Settings Enter your clinic or laboratory information as follows, to be printed automatically on report headers. You can also use this procedure to edit or change this information. 1. Press the Utilities key. 2. Select Settings. 3. Select Customize Configuring the VetScan HM2

55 4. Select Laboratory. 5. Enter the name and address of your clinic or laboratory. Press OK to move to the next line. You can enter up to 40 characters in each line. 6. Press ACCEPT to accept the setting User Mode Settings If you want to require users to log in and enter passwords to use the HM2, and to be able to track individual usage of the unit, you can enable the HM2 s Multi user mode. This will require each user to have a unique user ID and password. Note: In general, Single user mode is recommended unless the specific features enabled by Multi user mode are required. 1. Press the Utilities key. 2. Select Settings. Configuring the VetScan HM2 3-13

56 3. Select Customize. 4. Select User modes. 5. Follow the on-screen instructions to enable Multi user mode, to add or modify users, or to change a user s password. CAUTION: Write down user names and passwords and store them in a secure location. If this information is lost, you will need to contact Abaxis Technical Support (see page 1-2) to reset the HM Configuring the VetScan HM2

57 3.3 Date and Time Settings The date and time of each analysis is stored with the results. Use the Date and Time menu to set the HM2 s built-in clock and calendar, and the format used to display the date. 1. Press the Utilities key, then select Settings. 2. Select Date and time. 3. Select Set date and time. 4. Type in the date and time (use the and keys to select AM or PM), then press ACCEPT. 5. Select formats for displaying the date (item 2, 3, or 4) and time (item 5 or 6), then press EXIT. 6. Press ACCEPT to accept the settings. Configuring the VetScan HM2 3-15

58 3.4 Fluid Sensor Settings The fluid sensors allow the HM2 to monitor reagent consumption correctly. To maximize the life of the reagent pack, recalibrate the fluid sensors in the following situations: on installation when the instrument is moved to a new location if the reagent pack is moved relative to the instrument (changed distance or height of the pack relative to the instrument) Recalibrate the fluid sensors as follows. 1. Press the Utilities key, then select Settings. 2. Select Fluid Sensors. 3. Make sure all fluid sensors are on (checked). To turn on a sensor, select it and press OK so that its box is checked. 4. If a message indicates that one or more fluid sensors are disabled, turn those sensors on, then select Calibrate sensors Configuring the VetScan HM2

59 Section 4 Test Procedure and Interpreting Results This section describes how to prepare and analyze samples with the HM2, as well as how to interpret and print results. Section Contents 4.1 Collecting and Preparing Samples Sample Quality Storing Samples Before Performing an Analysis Daily Cleaning Check Reagent Status and Tubing Run a Blank Measurement Analyzing a Sample Adjusting the Needle Height Adjusting the Lyse Volume Interpreting Results CBC Parameters Histograms Printing and Exporting Results Printing Saved Results Exporting Results Combining Chemistry and Hematology Results Combining Previously Saved Results Combining Results During Testing Printout Examples Using Prediluted Mode Interpreting CBC Parameters Test Procedure and Interpreting Results 4-1

60 4.1 Collecting and Preparing Samples Because sample integrity is essential for accurate test results, always follow the optimal sample collection and proper sample handling techniques described in this section. Note: For multiple tube draws, always fill in this order: 1) red top, 2) green top, 3) lavender top. CAUTION: Use tubes containing EDTA only for CBC analysis Sample Quality The quality of the blood sample will directly affect the quality of the analysis results. Follow the recommendations below for optimal results. Selecting an anti-coagulant: Use manufactured EDTA lavender-top tubes for blood storage. Discarding the first portion of the blood sample: To prevent hair and skin pieces from getting into the sample, discard the first few drops of blood after collection. Filling blood tubes correctly: Each tube must be filled at least half full to avoid inaccuracy due to improper dilution of EDTA. When using homemade containers pre-dosed with EDTA, the final concentration of EDTA in the blood should not exceed 3 mg/ml. Mixing samples thoroughly: Even the most minute blood clots can adversely affect results. To avoid clotting, thoroughly mix each sample immediately after filling the tube by gently inverting the filled EDTA sample tube by hand 10 to 15 times. If testing is delayed and the sample was placed on a rocker, be sure to mix it thoroughly again before analysis by gently inverting the tube 10 to 15 times. This ensures the homogeneity of the sample. Discarding clotted samples: Before analysis, visually check all samples for clots while rocking the sample tube in your fingers. This is especially important for difficult sample draws. If you suspect a clot, swirl a wooden applicator stick through the sample: if you see clots or fibrin strands, discard the sample, and collect a fresh sample. 4-2 Test Procedure and Interpreting Results

61 Note: Do not shake samples! Doing so can damage the blood cells, and can form microbubbles that will cause inaccurate results. Note: Never use a rocker for samples smaller than 1.0 ml. Note: Rockers do not mix samples. Each sample must be inverted by hand 10 to 15 times immediately after drawing, and then 10 to 15 times again before running the sample. Note: Feline samples frequently demonstrate platelet aggregation (clumps). Vortex mixing for up to 30 seconds can disaggregate these clumps with no adverse effect on the sample. Some practitioners find that collecting blood from the medial saphenous vein using a vacutainer minimizes platelet clumping Storing Samples Analysis is best performed with a fresh, anti-coagulated sample. Always use proper sample handling technique, and make sure the sample is well mixed. If analysis is to take place more than 4 hours after the sample is obtained, refrigerate the sample at ºF (2 8 ºC), and test the sample within 8 hours. Note: When analyzing a refrigerated sample, be sure to warm the sample to room temperature before analysis. Roll the sample tube between your palms to speed up warming. This also helps mix the sample. If the sample is not mixed correctly, PLTs and WBCs can form clumps, causing erroneous results. Mix the sample thoroughly again before analysis. Test Procedure and Interpreting Results 4-3

62 4.2 Before Performing an Analysis CAUTION: If the analyzer has been kept at a temperature below 50 F (10 C), such as during shipment or a power failure, allow it to stand for an hour at the correct operating temperature before turning it on see Environmental Requirements on page Note: Each day, before analyzing samples, perform the procedures in this section to make sure the instrument is in proper operating condition. The HM2 automatically goes into standby mode after a period of standing idle. Press any button to bring the analyzer out of standby Daily Cleaning 1. Wipe up any spills on the sample rotor. 2. Keep the instrument and immediate surroundings as clean as possible. Sample tube adapter Sample rotor Check Reagent Status and Tubing 1. Check the status of the reagent pack to make sure enough reagent is available for sample analysis for the day see Viewing Status Information on page 8-2. If any reagent is at a level of 10% or less, change the reagent pack soon to ensure reliable results. For instructions, see Changing the Reagent Pack on page Inspect the reagent tubes and connections to make sure the reagents can flow freely. Make sure the tubes are not pinched, kinked, or trapped between or beneath objects. 4-4 Test Procedure and Interpreting Results

63 3. Check the reagent lines (except the red waste line) for bubbles or air gaps. If any exist, prime the affected tubes as follows. a. Press the Utilities key, then select Maintenance. b. Select Priming. c. Select the line to prime. d. Repeat as needed until all air gaps are removed. Note: Air gaps in the waste line (red) are normal Run a Blank Measurement A blank measurement checks the cleanliness of the HM2 s fluidic system, and is used to establish a baseline for sample measurements. The results of a blank are used to determine if the background will affect the test results, and whether the HM2 needs cleaning or maintenance. Note: A blank must be run every 12 hours. Each blank measurement remains valid for 12 hours of continuous operation, after which the HM2 displays BLANK needed, and a new blank must be run. 1. If BLANK needed appears on the screensaver, press the Measurement/Analysis key. The HM2 then runs a blank measurement, and displays the results. Test Procedure and Interpreting Results 4-5

64 2. Check the displayed results, and make sure the following CBC parameters fall within the ranges shown for an acceptable blank: Table 4-1: Acceptable Blank Parameters Measurement WBC RBC PLT HGB Allowed range 0.0 to 0.5x10 3 cells/µl 0.0 to 0.05x10 6 cells/µl 0 to 25x10 3 cells/µl 0 to 10 g/l If the values are within the allowed ranges: proceed to step 3, below. If the values are not within the allowed ranges (the message UNSUCCESSFUL BLANK MEASURE is displayed): a. Repeat the measurement by pressing the REPEAT soft key. b. If the new results are still above the acceptable ranges, run one to three cleaning cycles (see Cleaning the Aperture on page 7-5), and re-run the blank (up to several times). c. If the results still do not fall within the acceptable range, run a bleach-cleaning see Bleach-Cleaning on page 7-5. d. If the problem persists, call Abaxis Technical Support see Customer and Technical Support on page When the blank measurement is OK, press ACCEPT to accept the results. The HM2 is then ready for analysis. Note: To run a blank measurement other than at startup (such as for troubleshooting), press the Measurement/Analysis key, then press the MENU soft key, then select Measure blank. 4-6 Test Procedure and Interpreting Results

65 4.3 Analyzing a Sample Use this general procedure to analyze samples with the HM2. 1. Prepare a well-mixed, EDTA-preserved sample see Collecting and Preparing Samples on page 4-2 for details. 2. Select the appropriate sample tube adapter (see below), and place the adapter into the sampling rotor. Tube adapter (3 ml) Microtube adapter Controls/calibrator adapter Vacutainer adapter Note: Sponge disks are provided for the microtube adapter. If you have a very short sample, you can put the sponge disk into the microtube adapter to raise the tube height. Sponge plate for microtainer adapter 3. Press the Measurement/Analysis key, and select PT.ID. Test Procedure and Interpreting Results 4-7

66 4. Enter the species: view the available profiles using the and keys. Press the keypad OK key to move to the next field. 5. Enter other patient identification data as indicated. Use the OK key and the and keys to move through the settings. Use the and keys to change the settings, and the keypad or external keyboard to enter text or numbers. Note: When using the HM2 in conjunction with the VetScan Classic, make sure the Patient ID number on the HM2 is the same as the Patient Number on the VetScan Classic. When using the HM2 with the VetScan VS2, make sure the Patient ID number on the HM2 is the same as the Sample ID on the VS2. Otherwise, the results will not print together. Press the ACCEPT soft key to accept the settings. Note: Make sure the species for the sample displayed in the upper right corner of the screen is correct. The sample will be processed and tested according to the species selected. 6. Gently mix the sample by inverting the tube 10 to 15 times. Then remove the cap from the tube and place the tube into the sample adaptor as shown at right. 7. Begin analysis by pressing the Start button. Analysis is complete in less than three minutes. All results and histograms are automatically saved in the HM2 s database. 4-8 Test Procedure and Interpreting Results

67 8. When analysis is complete, a results screen displays all measured and calculated parameters as well as the WBC, RBC, and PLT histograms: You can view the results as follows: Use the PREV and NEXT soft keys to move through the results. If the HM2 is not set to print results automatically, press the Print key to print the results. (See Printing and Exporting Results on page 4-15 for details.) Results falling outside of the specified species reference range are marked as follows: Results higher than the species reference range are highlighted in black and marked with a plus sign (+). Results falling below the reference range are highlighted and marked with a minus sign (-). Warnings and errors flags are indicated using upper- and lower-case letters at the bottom. See Troubleshooting on page 9-1 for details about identifying and resolving any identified flags. For details on results and interpretation, see Interpreting Results on page Test Procedure and Interpreting Results 4-9

68 4.4 Adjusting the Needle Height Adjust the HM2 s needle height when you switch from one tube type to another, when you run a control, or when the sample volume is low enough to require it. 1. Press the Measurement/Analysis key, then press the MENU soft key. 2. Press Needle Height Setting repeatedly to cycle through the available needle heights: A:-2mm, B: 0mm, C: 5mm, D: 10mm, and E: 15mm. 3. When the correct needle height is displayed, press ACCEPT. Use this table and diagram as a guide to appropriate needle height settings.. Table 5: Needle Heights Setting and Needle Height Suitable Situations A:-2mm Measuring control or short sample B: 0mm Default setting C: 5mm Using a false-bottom tube D: 10mm Have plenty of blood sample E: 15mm Have plenty of blood sample ~3 4 mm E: 15 mm D: 10 mm C: 5 mm B: 0 mm A: 2 mm 4-10 Test Procedure and Interpreting Results

69 4.5 Adjusting the Lyse Volume The lyse volume generally only needs adjustment if a histogram indicates over- or under-lysing (this is relatively rare). If you receive such a histogram, contact Abaxis Technical Support before adjusting the lyse volume see Customer and Technical Support on page 1-2. When needed, adjust the lyse volume as follows. 1. Press the Measurement/Analysis key, then press the MENU soft key. 2. Press Lyse Volume or OK on the keypad repeatedly to cycle through the available volumes: 0.50 ml ml ml ml ml 3. When the correct volume is displayed, press ACCEPT. Test Procedure and Interpreting Results 4-11

70 4.6 Interpreting Results The HM2 produces a printed report containing the patient ID, measurement data, numeric results with flags (if any), and histograms showing the three different cell populations. The following graphic shows a set of typical results and histograms: 4-12 Test Procedure and Interpreting Results

71 4.6.1 CBC Parameters The Complete Blood Count (CBC) parameters included in the results are useful for assessing the overall health of the patient, as well as identifying and monitoring certain disease states. Note: Always check whether the results include any warning flags see Warning Indicators on page 9-2. For details about CBC parameters and associated clinical indications, see Interpreting CBC Parameters on page Histograms Histograms display population distributions of each cell type: leukocytes (white blood cells WBC), erythrocytes (red blood cells RBC) and thrombocytes (platelets PLT). The histograms show the relative frequency (percentage) of cells on the vertical (Y) axis, and cell volume in femtoliters (fl) on the horizontal (X) axis. Histograms enable you to quickly scan results for abnormalities, and also allow the versed practitioner to derive more information about the sample than is displayed by the values alone. The following pages describe each of the three histograms (WBC, RBC, and PLT), and show a typical example of each with an explanation. White Blood Cell Histogram (WBC) The WBC histogram shows white blood cell populations sorted by size. Cells larger than discriminator 1 are counted as WBCs. Blood includes three WBC populations: discriminator 1 discriminator 2 discriminator 3 Lymphocytes (LYM), shown by the first peak in the histogram. Monocytes (MON), indicated by the area between the second and third discriminators (although the MON region does demonstrate a distinctive peak of its own, this peak is not always clear in histogram form). LYM GRA MON (fl) Granulocytes (GRA), indicated by the peak to the right of the third discriminator. Test Procedure and Interpreting Results 4-13

72 The histogram on the previous page shows the lymphoctye cell population (LYM in the CBC parameters) to be in the range fl. The two other discriminators define a MON population of fl, and a GRA population of > 102 fl (above discriminator 3). Neutrophils normally make up the vast majority of the granulocytes, although considerable increases in eosinophils/basophils from an allergic or parasitic condition may be identified by the presence of an additional peak between the monocyte and primary granulocyte populations. Note: Measurements detected to the left of or near the first discriminator can include nucleated RBCs, and giant or clumped platelets. Red Blood Cell Histogram (RBC) The distribution of red blood cells normally appears as a single, steep bell curve. The presence of distinctive cell populations of various sizes, such as occurs in anemia, can be identified by the presence of more than one peak in the RBC histogram. This RBC histogram follows a normal distribution, with the RDW value within the normal range, and with the histogram normalized (100% fit) to the RBC peak. (fl) Platelet Histogram (PLT) The PLT histogram is a magnified portion of the beginning of the RBC histogram. The example PLT histogram at right follows a lognormal distribution, with a good separation from RBCs. The most commonly identified anomaly in platelet histograms results from aggregated (clumped) platelets. This appears as a flattened, lumpy histogram. Note: Feline samples may demonstrate platelet aggregation (clumps). Careful sample collection and vortex mixing for up to 30 seconds can avoid or disaggregate clumps with no adverse effect on the sample. See Veterinary Case Studies on page D-1 for a variety of sample test results and interpretations Test Procedure and Interpreting Results

73 4.7 Printing and Exporting Results The analyzer can print a wide range of results, statistics, and settings, and prints analysis results automatically if the Autoprint function is selected (see Table 3-2 on page 3-4). If Autoprint is not selected, you can print by pressing the Print key. Note: Printouts from the integrated (thermal) printer last approximately one year. To ensure the results remain legible for longer, photocopy the printout. Do not expose the thermal paper printout to heat extremes. Heat causes the printout to deteriorate more rapidly. To print any displayed screen, press the Print key. For instructions on connecting and setting up an external printer, see Printer Settings on page Printing Saved Results The HM2 automatically stores the most recent 1,000 test results (record 1,001 overwrites record 1, record 1,002 overwrites record 2, and so on). For more information about saved results, see Database Table on page 6-2. To print a single saved result, press the Database key. Use the and keys to highlight the record, then press the Print key. To print multiple saved results, press the Database key, then press the OK key to select the records to print (as indicated by filled boxes in the SmpID column). When the records to print are selected, press the Print key Exporting Results You can export results through the HM2 s serial port or USB Type B port, using a special software interface developed specially to communicate with the VetScan HM2. For availability, check with Abaxis Technical Support (see page 1-2) or your data management system (DMS) provider. To export a saved result, press the Database key, select the result, then select the Send Selected Records function in the Database menu. Note: Abaxis does not create interface programs for data management software, but can initiate the process with your current software provider. For further information, please contact Abaxis Technical Support see Customer and Technical Support on page 1-2. Test Procedure and Interpreting Results 4-15

74 4.8 Combining Chemistry and Hematology Results The HM2 can print results from a connected VetScan Classic or VetScan VS2 analyzer. The HM2 can print the VetScan results only, or the combined VetScan and HM2 results for a given patient ID. (For connection instructions, see Connecting a VetScan Chemistry Analyzer (Optional) on page 2-22.) Printing VetScan results requires that the HM2 be configured to receive data from the VetScan (see General Settings on page 3-9). This is normally done on installation. If you have questions, contact Abaxis Technical Support see Customer and Technical Support on page Combining Previously Saved Results To export combined VetScan and HM2 data, first display the CBC results to be combined on the HM2, then do the following on the VetScan: 1. Press the RECALL key. The following appears: Press ENTER to Recall Results MENU for next option EXIT to leave. 2. Press ENTER. The following appears: 3. Press ENTER to select patient results. The following appears: Recall:1 Results 1-Patient results 2-Patient flags ENTER to accept. Recall:1 Single 1-Single result card 2-All patients ENTER to accept. 4. Press ENTER to select a single result. The following appears: Select:1 Print 1-Print result card 2-Transmit patient ENTER to accept. 5. Press 2 and ENTER to transmit. If the Patient IDs match, both results will be transmitted Test Procedure and Interpreting Results

75 4.8.2 Combining Results During Testing 1. Use a USB cable to connect the printer to the HM2. 2. Turn on the printer, then turn on the HM2 and the VetScan. 3. Make sure the HM2 uses the same date format as the VS2 see Date and Time Settings on page Configure the HM2 s printer settings as follows (see Printer Settings on page 3-2 for instructions): Set Printer to HP 5650, Canon i455, or SeikoLPTH245 Built in. HP 5650 only: set Printer cable to USB (this setting is made automatically for the Canon i455 and built-in printers). Set Autoprint to No. 5. Configure the HM2 s general settings as follows (see General Settings on page 3-9): Set PC Link to Offline. Set VSx Link to USB. Set VetScan-HM2 combining within as needed. 6. To print a VetScan result only: Note: Be sure to set VetScan-HM2 combining within the time frame so that no matching record will exist in the HM2. For example, if a VetScan analysis was run for a given patient 5 days ago, you would set VetScan-HM2 combining within to 4 days or less to exclude any earlier records. As long as no matching VetScan record exists, the VS2 results will be printed. Note: If you run VetScan and VS2 analyses for the same patient on the same day and want to print only the VetScan result, you can change the patient ID on the HM2 so that there will be no matching record. Test Procedure and Interpreting Results 4-17

76 7. To print combined HM2 and VS2 results: Note: The Patient ID of the HM2 record must match the Sample ID field of the VS2 record. Start the CBC first, and then run the chemistry rotor. When the rotor finishes, the VS2 automatically transmits the results to the HM2, and the HM2 prints out the combined results Printout Examples Thermal Paper Printout VetScan Chemistry Results Patient information Patient species VetScan information Test results Test parameters Test result units Test quality indicators (rotor QC, interfering substances) Reference ranges 4-18 Test Procedure and Interpreting Results

77 Thermal Paper Printout HM2 + VetScan Chemistry Results Report header Patient information Report date Test date & time (dates and times assigned automatically) Patient species HM2 serial number Sample ID (assigned automatically) Reference ranges Test parameters Test results lower limit Test result units upper limit location of test results within reference range limits WBC histogram RBC histogram PLT histogram VetScan results Test Procedure and Interpreting Results 4-19

78 8 x 11.5 Letter Printout HM2 + VetScan Chemistry Results Report header Sample ID (assigned automatically) Patient information Patient species Patient information Report date (assigned automatically) and HM2 serial number WBC histogram Test parameters Test results and unit Reference ranges RBC histogram PLT histogram VetScan results lower limit shows location of test results within reference range limits upper limit 4-20 Test Procedure and Interpreting Results

79 4.9 Using Prediluted Mode The HM2 software provides a special predilution function for use in the following situations: if an insufficient amount of blood is available for a routine CBC if the CBC results are abnormally high (non-linear), resulting in an m, M, or N flag (see Warning Indicators on page 9-2) Note: Before using prediluted mode, always call Abaxis Technical Support see Customer and Technical Support on page 1-2. Perform an external predilution of the sample using HM2 reagent diluent, or an isotonic saline solution. Dilute the sample by a 1:6 ratio (1 part sample to 5 parts diluent) for example, 50 μl sample to 250 μl diluent or saline. Mix well. Note: Prediluted mode must be calibrated before use. The HM2 is calibrated at installation, and can be recalibrated as described in Calibration on page 5-2 (select Prediluted blood (1:5) as the calibrator). After the dilution, perform a QC run for the 1:5 diluted quality control blood. See Performing Quality Control on page 5-6 for instructions. Perform the analysis as follows. 1. Press the Measurement/Analysis key, then press the MENU soft key. 2. Select Prediluted mode. (The accompanying checkbox is filled in.) Test Procedure and Interpreting Results 4-21

80 3. Press the ACCEPT soft key. Notice that 1:5 now appears in the upper left corner of the screen. 4. If you are processing a new sample: press the PT.ID soft key, enter the patient information, then press ACCEPT. If you are processing a sample that has already been tested as non-prediluted: select Repeat Last Sample to avoid re-entering the patient information. 5. Press the Start button to begin the analysis. The HM2 will automatically calculate the results with the 1:5 predilution factor. Note: The HM2 automatically returns to normal mode after the prediluted analysis completes Test Procedure and Interpreting Results

81 4.10 Interpreting CBC Parameters Complete Blood Count (CBC) parameters are useful in assessing the overall wellness of a patient, as well as for identifying and monitoring certain disease states. The following tables outline the various CBC parameters and associated clinical indications. Table 4-1: White Blood Cells and Associated Indications White Blood Cell (Leukocyte) Role Increase in Disease State Decrease in Disease State Non-Granulocytic Lymphocytes B-cells: humoral immunity (antibody synthesis) T-cells: cellular immunity Monocytes Immature macrophages phagocytosis of debris/foreign material, killer-cell activation Granulocytes Neutrophils Left shift: Increased numbers of immature neutrophils/band cells. regenerative: < 10% bands, neutrophilia, absence of myelocytes/metamyelocytes degenerative: >10% bands, depressed total neutropenia, presence of myelocytes/metamyelocytes (poor prognostic indicator) Phagocytize/kill microorganisms, initiate and modify inflammatory process, cytotoxic Chronic inflammation Acute infection/recovery Lymphocytic anemia Hypoadrenocorticism Necrotic, malignant, hemorrhagic, or immune-mediated disease Inflammation Neoplasm Stress Exercise/excitement Acute/severe disease Viral disease Endotoxemia Hyperadrenocorticism Stress-related corticosteroid response Rare, no known significance Bacterial infection Viral infection Drug-induced (bone marrow depression) Right shift: Increased number of hypermature (hyper-segmented) neutrophils, usually seen with noninfectious inflammatory process (e.g. malignancy) Eosinophils Parasiticidal Cytotoxic Phagocytic Basophils Initiate inflammation Prevent coagulation Activate lipoprotein lipase Parasitic infection Allergic responses Hypoadrenocorticism Allergic reactions Parasitic infection Neoplasia Stress Hyperadrenocorticism ACTH therapy No known significance Test Procedure and Interpreting Results 4-23

82 Table 4-2: Red Blood Cell Parameters and Associated Indications Parameter Definition Diagnostic Consideration Hematocrit (HCT) Percentage of total cellular constituents (primarily red blood cells) in a unit of whole blood Anemia exists when the HCT falls below the reference range for the species. Increase in red cell count or polycythemia is most commonly the result of dehydration. Hematocrit will normally have a value of approximately three times (3X) the hemoglobin value. Hemoglobin (HGB) RBC Indices Anemia Characterization MCV Mean Corpuscular Hemoglobin MCH Mean Corpuscular Hemoglobin MCHC Mean Corpuscular Hemoglobin Concentration RDW Red Cell Distribution Width The oxygen-carrying component of red blood cells; allows for the calculation of MCH and MCHC Measure of the volume of an average RBC: MCV = (HCT x 10) / RBC Calculated HGB concentration of an average RBC MCH = (HGB x 10) / RBC (in picograms) Calculated HGB concentration in an average RBC MCHC = MCH / MCV (in grams of HGB per 100 ml RBCs) Measure of red blood cell anisocytosis (cell size variation) Hemoglobin normally falls in the range of one third (1/3) of the hematocrit value. Increase: most commonly associated with reticulocytes/regenerative anemia Decrease: iron-deficiency anemia Increase: most commonly the result of hemolysis Decrease: hypochromasia common in irondeficiency anemia and reticulocytosis Increase: most commonly the result of hemolysis Decrease: hypochromasia common in irondeficiency anemia and reticulocytosis Increase: indicates increased production of large immature cells. RDW increases even before the MCV exceeds the normal range: early indicator of regenerative anemia. RDW also increases with the production of small RBCs in iron-deficiency anemia. Table 4-3: Platelet Parameters and Associated Indications Parameter Increase in Disease State Decrease in Disease State Total Platelet Count Thrombocytosis is present with excess Disseminated intravascular coagulation bleeding, iron deficiency anemia and Bone marrow depression myeloproliferative syndromes. Autoimmune hemolytic anemia Severe hemorrhage MPV Mean Platelet Volume PCT Platelet Hematocrit PDW Platelet Distribution Width Indirect evidence of increased (bone marrow) megakaryocyte response. Volume of platelets expressed as a percentage of whole blood (used as a research tool). Increased measure of platelet anisocytosis (platelet size variation) indicative of active platelet release. Not an accurate predictor of decreased megakaryocyte response. Volume of platelets expressed as a percentage of whole blood (used as a research tool). No known clinical significance Test Procedure and Interpreting Results

83 Section 5 Calibration and Quality Control This section describes the calibration and quality control procedures for the VetScan HM2. Section Contents 5.1 Calibration When to Calibrate Required Calibration Materials and Handling Calibration Procedure Viewing Calibration History Performing Quality Control Required Quality Control Materials Quality Control Procedure Monitoring Quality Control Values Over Time Viewing the Accepted QC Database Calibration and Quality Control 5-1

84 5.1 Calibration When to Calibrate The HM2 is factory-calibrated for optimal performance. In the following situations, a fine-tuning calibration can be done (using three measures of calibrator material): on initial installation when quality control measurements show a systematic error (bias the curve is shifted upward or downward) or are repeatedly outside predefined limits after service that replaces any component related to the process of dilution or measurement after relocating the instrument after software upgrades (as directed by Abaxis) before using prediluted mode Required Calibration Materials and Handling The calibration procedure requires the Hematology Calibrator, Abaxis Part No WARNING: THE CALIBRATION PROCEDURE REQUIRES A SPECIAL CALI- BRATOR THAT IS DERIVED FROM HUMAN SOURCES. OBSERVE UNIVERSAL SAFETY PRECAUTIONS WHEN HAN- DLING THE CALIBRATOR SAMPLE. 5-2 Calibration and Quality Control

85 5.1.3 Calibration Procedure WARNING: IT IS STRONGLY RECOMMENDED THAT LATEX GLOVES BE WORN DURING THIS PROCEDURE. Note: Make sure to bring the calibrator up to room temperature before beginning this procedure. 1. Press the Utilities key, then select Calibration. 2. Select Run Calibrator. 3. Select the calibrator: Whole blood or Prediluted blood (1:5). Select Prediluted blood (1:5) only if you are using prediluted mode. 4. Enter the assay values for each parameter from the calibrator package insert. Use the PGUP and PGDN soft keys to move between pages. 5. When all parameters are set, press the soft key. ACCEPT Calibration and Quality Control 5-3

86 Note: Needle height must be at 2 mm. To adjust, press MENU, select 3: Needle Height, press OK repeatedly until needle height = 2mm, then press ACCEPT. The display shows 1/3 Calib at top center, and Calib at top right. 6. Mix the calibrator by gently inverting the tube between thumb and forefinger 10 to 15 times. Remove the tube cap. 7. Place the calibrator into the control tube adapter. 8. Press the Start button. 9. When analysis is complete and the display shows the results, press ACCEPT. The display then shows 2/3 Calib at the top. 10. Remove the tube from the adapter, and mix by gently inverting the tube between thumb and forefinger 10 to 15 times. 11. Replace the tube in the adapter, and press the Start button. 5-4 Calibration and Quality Control

87 12. When the second analysis is complete and the results are displayed, press ACCEPT. The display then shows 3/3 Calib at top. 13. Remove the tube from the adapter, and mix by gently inverting the tube between thumb and forefinger 10 to 15 times. 14. Replace the tube in the adapter, and press Start. 15. When the third analysis is complete, press ACCEPT. The new calibration factors are then displayed. Use the NEXT and PREV soft keys to move between pages. Calibration and Quality Control 5-5

88 16. Press ACCEPT to accept the new calibration factors. The instrument is now calibrated. 17. Press EXIT. Note: To complete the calibration, you need to press the ACCEPT soft key after each calibration measurement. When all three measurements are accepted, the instrument then calculates and displays the new calibration factors. Do not change any of these displayed calibration factors Viewing Calibration History All calibration times and the settings used are logged and saved, and can be viewed as follows: 1. Press the Utilities key, then select Calibration. 2. Select View Calibration History. If the message 1 or more of your factors are out of range appears, call Abaxis Technical Support see Customer and Technical Support on page Performing Quality Control The HM2 is pre-programmed to monitor three different quality control (QC) types: Dog, Cat, and Human. You should perform QC determinations regularly to verify continued optimal performance. We recommend testing control material (dog control or cat control) each time the reagent pack is changed, or otherwise approximately once per month. Also, perform a quality control measure after each service. The analyzer stores all accepted QC results in an internal database Required Quality Control Materials The quality control procedure requires the following materials: Dog Control, Abaxis Part No Cat Control, Abaxis Part No Human Calibrator, Abaxis Part No Calibration and Quality Control

89 5.2.2 Quality Control Procedure Note: Before performing this procedure, be sure to bring the quality control materials up to room temperature. Perform a quality control (QC) check as follows. 1. Press the Utilities key, then select Quality control. 2. Select Select QC type, then select the QC material you want to use: Dog, Cat, or Human. 3. Select Set QC reference ranges. 4. From the package insert, enter the lot no., exp. date, and assay and gap values for each test parameter. To disable QC of a parameter, set that parameter to 0.0. Use the PGDN and PGUP soft keys to view and enter additional parameters. 5. Press ACCEPT to accept the data, then press ACCEPT again to confirm. 6. Mix the control vial by gently inverting the tube between thumb and forefinger 10 to 15 times, and remove its cap. 7. Put the control vial onto the control vial adapter on the sample rotor. Calibration and Quality Control 5-7

90 8. Select Run QC to start the analysis. Press the Start button. The results screen displays Quality Control as ID. 9. Press ACCEPT to accept the results and save them in the QC database. Note: Any values that are out of range are highlighted. If you don't accept the QC results, they will not be saved in the instrument s database. 10. Abaxis strongly recommends performing three successful QC runs after calibrating the instrument Monitoring Quality Control Values Over Time Many laboratories monitor the recovery values of quality control material over time to identify trends and potential performance drift. The HM2 allows you to do this easily using a database table and in graphic form Viewing the Accepted QC Database You can view all of the accepted QC results from the instrument s database in table or graphic (Levy-Jennings) form. The QC measurement results are assigned sequential ID numbers. 1. Press the Utilities key, then select Quality control. 2. Select View table of QC measures. Use the and keys to select entries, and the and keys to move among parameters. To print a selected QC result, press GRAPH, then press the Print key. 5-8 Calibration and Quality Control

91 Section 6 Managing the Database This section describes how to use the analyzer s database. Section Contents 6.1 Database Overview Database Table Navigating the Database System Managing the Database 6-1

92 6.1 Database Overview The analyzer can store up to 1,000 complete test results in its internal database. Results are stored chronologically by date and time. (When more than 1,000 measurements are stored, the most recent file replaces the oldest file.) You can retrieve these results anytime. 6.2 Database Table To access the Database table, press the Database key on the front panel. The first screen shows a table of the most recently saved tests, as shown at right. You can view the results in this screen as follows: Use the and keys to view results that extend past the edge of the screen. Each test result includes the following: Sample ID Warnings (if any) Mode Date Name Doctor Patient ID Age Lyse 18 CBC parameters Sex Clogging report Use the and keys to move through individual results. Select the highlighted patient results by pressing the OK key on the keypad. (The check box next to the patient name is then filled.) To display the WBC, RBC, and PLT histograms, press the histogram GRAPH soft key. To view the second panel, containing PLT results, press the NEXT soft key. To view sample information, press PT.ID. To view the results in table format, press TABLE to return to the previous screen. To go to the Local Menu, press MENU. Print selected patient results by pressing the Print key. The screen then displays: Print as table select this to print the selected results in table format. Print result by result select this to print each result with its sample information, CBC values, and histograms. To print an individual result, highlight the result, press GRAPH, then press Print. 6-2 Managing the Database

93 6.3 Navigating the Database System Use the Database local menu to access and manage the records stored in the analyzer s database. From the Database table screen, enter the Database local menu by pressing the MENU key. The menu contains the following items: Go to specified record: Jumps to a particular sample record. Enter the date and time, sample ID, and patient ID of the sample you want to view, and press the ACCEPT key. The first sample meeting your parameters is then displayed. If Patient ID is blank (0), records are searched by date/time only. Selection: Selects all records in memory, or all those with a specific date, time, and ID. Select by date, time and ID selects a range of records by date, time, and/or ID number. Select all selects all records, and Deselect all deselects all records. Entering 0 as an ID searches by date only. Corresponding results are marked with a filled box. Change sort order: Changes the order in which results are displayed: Unsorted, Sort by time, Sort by sample ID, or Sort by patient ID. Managing the Database 6-3

94 Manage selected records: Enables you to Send selected records to a PC, Delete selected records, or Backup selected records to a USB flash drive. (Before selecting Backup selected records, insert a USB flash drive into the USB port on the analyzer s front-panel.) View external: Views previously saved data from a USB flash drive inserted into the USB port on the analyzer s front-panel. Backup one day: Backs up all records from a specified day to a USB flash drive. 1. Select the date to back up, and confirm by pressing the ACCEPT soft key. 2. Insert a flash drive in the USB port on the front of the analyzer, then press ACCEPT. The records are then copied to the flash drive. 6-4 Managing the Database

95 Section 7 Maintenance & Service This section details maintenance and service procedures for the analyzer. Section Contents 7.1 Periodic Maintenance Daily Maintenance Weekly Maintenance Monthly Preventive Maintenance Cleaning the Aperture Bleach-Cleaning Shut-Down Procedures Changing the Reagent Pack Running the Self Test Replacing the Peristaltic Pump Tube Assembly Maintenance & Service 7-1

96 7.1 Periodic Maintenance Performing the simple and easy maintenance procedures in this section will help keep your analyzer in optimal operating condition, thus ensuring peak performance, high-quality results, and a good return on your investment. Note: Pay particular attention to the weekly maintenance Daily Maintenance The analyzer s automatic standby mode is designed to keep the system s internal workings in optimal condition, and so reduces the amount of maintenance needed. Always keep the analyzer and its immediate surroundings as clean as possible to prevent debris from getting into the system. Check all reagent lines for kinks, pinches, abnormalities, and leaks or spills. Clean up any standing fluid near the analyzer. Wipe up any spills on the sample rotor Weekly Maintenance Perform the following tasks once a week. Clean the Wash Head The wash head cleans the outer surface of the sample needle with a saline diluent. Salt can build up around the wash head if it is not cleaned regularly. As a reminder, once a week the analyzer displays this message on the bottom of its screensaver: It is time to clean the needle wash head. Go to instructions., Maintenance, Cleaning for When this message appears, follow the instructions below to clean the wash head and prevent salt accumulation. The process takes only a few seconds to complete. CAUTION: Do not touch any components inside the instrument except as specifically directed. 7-2 Maintenance & Service

97 1. Press the Utilities key, then select Maintenance. 2. Select Clean the wash head. 3. Open the door on the right side of the instrument. The wash head s location is shown at right. 4. Use a soft, lint-free cloth and warm tap water to gently rub the lower surface of the wash head to remove any salt buildup. 5. When finished, close the side door, then press the ACCEPT soft key. Once this procedure is complete, the screensaver reminder will disappear, and will reappear in one week. Maintenance & Service 7-3

98 7.1.3 Monthly Preventive Maintenance Once a month, clean the analyzer as follows. 1. Turn off and (if possible) unplug the analyzer. 2. Use a slightly damp cloth to clean the analyzer s cover, front panel, and sampling rotor. 3. Open the side and rear doors, and clean the internal metal chassis underlay if necessary. CAUTION: Do not use chemical cleaners such as bleach, and do not use an excessively wet cloth. Water can cause the instrument s electronics to malfunction. Avoid touching internal components. 4. Clean the entire pneumatic system (tubes, valves, chamber, and aperture) with a Prime all cycle: a. Press the Utilities key, then select Maintenance. b. Select Priming. c. Select Prime all. The analyzer then performs its priming process, including cleaning. Note: Step 4 can be skipped if the reagent pack is changed monthly and the instrument is shut down weekly. 7-4 Maintenance & Service

99 7.2 Cleaning the Aperture The analyzer automatically performs a three-step cleaning process after each analysis. In addition, the analyzer provides a software-driven process that you can use to clear the measuring aperture of any debris when needed (such as when directed to perform as a part of troubleshooting). 1. Press the Utilities key, select Maintenance, then select Cleaning. 2. Select Automatic self-cleaning. The analyzer then performs its cleaning process, and returns to the Maintenance menu when finished. 7.3 Bleach-Cleaning Use this procedure to bleach-clean the analyzer s internal tubing each time you change the reagent pack. You will need the following materials: cleaning tube kit (shown on page 2-10) household-strength chlorine bleach at least 5 ml distilled water at least 195 ml Run the bleach-cleaning process as follows: 1. Prepare a 5% bleach solution using 5 ml of bleach and 95 ml of distilled water. In addition to the bleach solution, you will need at least 100 ml of distilled water for rinsing. Maintenance & Service 7-5

100 2. Press the Utilities key, then select Maintenance. 3. Select Bleaching. 4. When asked to confirm, press ACCEPT. 5. When prompted, disconnect the reagent tubing from the reagent inlets (diluent, lyse, cleaner, and rinse) on the back of the analyzer, but leave the waste tubing connected. 6. Press ACCEPT to continue. Note: If you do not have the cleaning tubing, leave the reagent lines connected to the analyzer, and instead disconnect the rinse, lyse, diluent, and cleaner bottle caps, and submerge all four lines into the bleach solution see step When prompted, connect the cleaning tube kit to the reagent inputs as shown. 7-6 Maintenance & Service

101 8. Submerge the free end of the cleaning tube in the bleach solution. 9. Press ACCEPT to continue. 10. When prompted, disconnect the cleaning tube (leaving the waste tubing connected), then press ACCEPT. 11. When prompted, connect the cleaning tube kit to the reagent inputs as shown in step 7, and submerge the free end of the cleaning tube in a container of at least 100 ml of distilled water, as shown in step Press ACCEPT to continue. 13. When prompted, disconnect the cleaning tube (leaving the waste tubing connected), then press ACCEPT. 14. When prompted, select from the following options as needed: To install a new reagent pack, press NEW. To reconnect the old reagents, press OLD. To turn off the analyzer, press STOP. 7.4 Shut-Down Procedures Note: Proper use of the analyzer s shut-down procedure is critical for maintaining the instrument in good working condition. If you will not use the instrument for more than 72 hours, shut it down as described in Turning the Analyzer Off on page Maintenance & Service 7-7

102 7.5 Changing the Reagent Pack Follow this procedure to change the reagent pack. 1. Disconnect the reagent tubing from the reagent inlets (diluent, lyse, cleaner, and rinse) on the back of the analyzer, but leave the waste tubing connected. 2. Perform a bleach cleaning see Bleach-Cleaning on page Reconnect the reagent tubing to the inlets on the back of the analyzer. 4. Remove the caps (with tubing still attached) from the old reagent pack (including the waste container). 5. Connect a new reagent pack, using the same tubing and reagent cap connectors. 6. Re-cap the used reagents with their original caps, and dispose of them in an environmentally appropriate manner. 7. Prime all reagent lines: a. Press the Utilities key, then select Maintenance. b. Select Priming, then select Prime All. 8. Reset the reagent status: a. Press the Utilities key, then select Maintenance. b. Select Reagent Status. 7-8 Maintenance & Service

103 c. Press the CHANGE soft key, then press ACCEPT to confirm. The analyzer updates the installation date, reagent lifetime, and the amount of reagent in each container. d. Press EXIT when finished. Note: If you selected NEW at the last step of the bleach-cleaning, the reagent status will be automatically reset for a new pack (100% for each reagent). 9. If you place the new reagent pack in a different location relative to the instrument (compared to the old pack), recalibrate the fluid sensors to obtain the maximum number of tests per pack. (Calibration is not necessary if the new pack is placed in the same location as the old one.) If needed, calibrate the fluid sensors see Fluid Sensor Settings on page Run a quality control procedure using a dog or cat sample see Performing Quality Control on page 5-6. Maintenance & Service 7-9

104 7.6 Running the Self Test The analyzer includes an automated Self test that verifies proper operation of essential components of the instrument. Run the Self test at these times: at installation after replacing any component after any extended time out of use if you suspect the analyzer is not working properly 1. Press the Utilities key, and select Diagnostics. 2. Select Self test. The analyzer then lists and checks off subsystems as it tests each. 3. When the test is finished, the analyzer displays a summary of the results. Press PGDN and PGUP to move through the pages and view details. Note: If the Overall result displayed is Errors, call Abaxis Technical Support see Customer and Technical Support on page Maintenance & Service

105 7.7 Replacing the Peristaltic Pump Tube Assembly The analyzer s peristaltic pump is designed to be maintenance-free. However, after extended, longterm use (normally longer than a year), the pump tubing may eventually need to be replaced. The analyzer includes a spare peristaltic pump tube set, with two plastic connectors and two metal spring clips. Though replacement is a simple process, Abaxis strongly recommends that you call Abaxis Technical Support before beginning, so that they can verify the need for pump tube replacement and guide you through the process if necessary. See Customer and Technical Support on page 1-2. Replace the peristaltic pump tube assembly as follows. WARNING: MAKE SURE THE INSTRUMENT IS POWERED OFF BEFORE YOU BEGIN THIS PROCEDURE. Note: Wear gloves while performing this procedure. Keep paper towels on hand in case any liquid spills from the tubes. 1. Open the analyzer s rear door, and locate the peristaltic pump at the lower left of the rear compartment. Disconnect the color-coded tubes attached to the base of the pump housing, as shown. 2. Locate the locking mechanism on the underside of the pump housing. Use the middle finger and thumb of your right hand (as shown) to twist the fixing lock at the base of the pump counterclockwise to loosen the tube case. turn counterclockwise Maintenance & Service 7-11

106 3. Lift the pump tube case upward and remove it. 4. Remove the tubing from inside the case and replace it with the new tubing provided. Make sure the two plastic connectors at the ends of the tubing are properly placed into the slots of the tube case, and that the metal clips face inward. Otherwise, the tube case cannot be installed properly. 5. Fit the tube case back onto the pump housing from the top, and firmly press both sides of the case. 6. Twist the fixing lock at the base of the pump clockwise until it clicks. The pump tube is then properly installed. Make sure the tubing case fits properly and tightly in the correct position. 7. Reconnect the color-coded tubes to their proper connectors: white to white, green to green. 8. Wipe up any spilled fluids from the base of the instrument. Close the rear door Maintenance & Service

107 Section 8 Special Functions This section describes special functions you can use in operating the instrument. Section Contents 8.1 Viewing the Software Version Viewing Status Information Updating the Software Customizing the User-Defined Species Profile Customizing and Printing the Normal Range Special Functions 8-1

108 8.1 Viewing the Software Version To view the software version number, press the Utilities key, select Diagnostics, then select Device information. 8.2 Viewing Status Information To view device statistics number of analyses performed, clogs, and errors, and other information that may be useful for technical service press the Utilities key, select Diagnostics, then select Statistics. To view the status of the reagent pack, press the Utilities key, select Maintenance, then select Reagent status. Note: Do not press the CHANGE soft key unless you are changing the reagent pack. Otherwise, the analyzer will not display the true reagent status. 8-2 Special Functions

109 8.3 Updating the Software From time to time, Abaxis will send you an updated version of the analyzer software on a CD. Use the following procedure to install the update from the CD. 1. Press the Utilities key, and select Maintenance. 2. Select Software upgrade. 3. Open the analyzer s CD-ROM drive by pressing the button on the drive door. 4. Insert the update CD in the drive, then close the drive. 5. Press ACCEPT. The system then restarts. 6. When the Upgrade/Install menu appears, select Upgrade existing version. 7. When the CD-ROM drive opens, remove the CD from the drive and store it in a safe place. 8. Restart the analyzer by turning it off and then on again using the power switch on the back of the instrument The analyzer then automatically functions with the upgraded software version. 8.4 Customizing the User-Defined Species Profile The analyzer includes a User-Defined Species Profile that enables you to define your own species for testing. This profile can only be activated by Abaxis Technical Support personnel. If you need to create custom species, please contact Abaxis Technical Support see Customer and Technical Support on page 1-2. They will guide you through activating the profile, and help you to customize it to suit your requirements. Special Functions 8-3

110 8.5 Customizing and Printing the Normal Range If you need to print out the normal range for particular species, do so as follows. 1. Press the Measurement/Analysis key, then press the PT.ID soft key. 2. Select the species you want, then press the ACCEPT key. The species you selected are shown at the upper right corner of the next screen. 3. Press LIMITS. The normal ranges of the selected species are then displayed. 4. Change any limits as needed, then press ACCEPT. 5. Press the Print key. The normal range of the selected species then prints. 8-4 Special Functions

111 Section 9 Troubleshooting Use the information in this section to help diagnose and solve problems with the analyzer. Section Contents 9.1 Warning Indicators Blank Flags Measurement conditions Out-of-Range Results Results Flags Error Messages Evaluating Unexpected Results Examples Preparing the Analyzer for Shipment Troubleshooting 9-1

112 9.1 Warning Indicators This section lists warning indicators (flags) that can appear in test results, along with possible solutions for each Blank Flags If analysis errors occur or the blank measurement is too high, an E error flag appears along with the affected parameter, and --- is displayed instead of results. In this situation, perform a cleaning see Cleaning the Aperture on page 7-5. If the problem persists, contact Abaxis Technical Support see Customer and Technical Support on page Measurement conditions Warning flags are grouped according to measurement conditions and problems with the sample. c, C, q, and Q flags are related to aperture clogging. Perform three to five cleanings as needed (see Cleaning the Aperture on page 7-5), and repeat the blank measurement (see Run a Blank Measurement on page 4-5). b, p, E, H, and B are generally due to reagent contamination or lack of sample. Check the sample (needle height, etc.). If contamination is suspected, change the reagent pack (see Changing the Reagent Pack on page 7-8). m, M, and N are caused by too high a cell concentration. Perform an external 1:5 dilution and repeat the measurement (see Using Prediluted Mode on page 4-21). Run a quality control procedure if needed (see Performing Quality Control on page 5-6). An asterisk (*) near a parameter indicates uncertainty. A variety of causes are possible. If a result warning flag is present, refer to Table 9-1 on page 9-3 for possible causes and solutions Out-of-Range Results If a parameter is outside the normal reference range, it is flagged with if below the normal range, or + if above the normal range. Reference ranges can be edited and customized in the Limits menu. A setting of 0 for a range limit results in no reference range flagging Results Flags If a warning occurs, a * flag appears before the result. Specific warning flags are displayed on the results screen, and are grouped according to measurement conditions and the problems relating to the blood sample: Uppercase warning flags (E, H, B, C, Q) are related to WBC or HGB measures. Lowercase flags (p, b, c, q) are related to RBC or PLT measures. 9-2 Troubleshooting

113 The following table summarizes these flags. Table 9-1: Result Warning Flags Flag Meaning Description / Recommended action E Empty sample. Needle may not reach sample. Check needle height and adjust to 2 mm if needed. H B No WBC 3-part differential (unable to report a WBC differential) HGB blank is high, or no HGB blank WBC blank is high, or no WBC blank M, N Linearity error, or WBC coincidence is too high May occur in pathological lymphocytosis. Review slide if WBC count is outside normal limits. Possible lyse problem. Prime lyse and repeat the blank. Possible interference. Make sure the side door is closed, and repeat the blank measurement. Possible bubbles in the WBC chamber. Check the connections of the reagent inputs and perform a cleaning. Repeat the blank. Check to see if wash head needs cleaning. If cleaning is needed, perform 3 to 5 cleaning cycles and repeat the blank measurement. If the blank is too high, perform 3 to 5 cleaning cycles and repeat the blank measurement. Possible lyse problem. Prime lyse and repeat the blank. Possible lyse contamination. Check lyse container and line for contamination. Repeat the blank. Possible electrical noise interference. Be sure to use an uninterruptable power supply (UPS). Repeat the blank. The results are out of the linearity range. Make a dilution with an external method to achieve 1:5 dilution rate, and run again. (1:5 calibration should be current.) Prime lyse and repeat the blank. C, Q WBC clogging Aperture clogging. Perform 3 to 5 cleaning cycles and repeat the measurement. If the problem persists, contact Abaxis Technical Support see page 1-2. Low-temperature reagents (mainly diluent) can cause this. Be sure to allow reagents to reach room temperature. p b m PLT blank is high, or no PLT blank RBC blank is high, or no RBC blank Linearity error, or RBC/PLT coincidence is too high Check to see if wash head needs cleaning. If cleaning is needed, perform 3 to 5 cleaning cycles and repeat the blank measurement. Possible reagent contamination or system cleanliness problem. If the problem persists, change the reagent pack, perform a cleaning, and repeat. If the problem still remains, contact Abaxis Technical Support see page 1-2. Check to see if wash head needs cleaning. If cleaning is needed, perform 3 to 5 cleaning cycles and repeat the blank measurement. Possible reagent contamination or system cleanliness problem. If the problem persists, change the reagent pack, perform a cleaning, and repeat. If the problem still remains, contact Abaxis Technical Support see page 1-2. Results are out of linearity range. contact Abaxis Technical Support see page 1-2. Run a control cycle. c RBC/PLT clogging Low-temperature reagents (mainly diluent) can cause this. Be sure to allow reagents to reach room temperature. Aperture clogging. Run cleaning and repeat the measurement. If the problem persists, contact Abaxis Technical Support see page 1-2. Troubleshooting 9-3

114 9.2 Error Messages Below are a few very common error messages and solutions to the errors. Error message: Out of diluent, lyse, cleaner, or rinse Solutions: Visually check the reagent containers for reagent. If needed, change the reagent pack see Changing the Reagent Pack on page 7-8. Check for bubbles in the reagent tubing. If bubbles are present, check the connections at the back of the instrument, at the top of the corresponding reagent container, and under the container cap to make sure they are tight. Make sure the reagent sensor is on if it is not, there will be an S in the top left corner of the display. In this case, calibrate the fluid sensor see Fluid Sensor Settings on page Error message: Pressure or vacuum error Solutions: Check for loose, detached, or kinked tubing inside and outside the instrument. Make sure there are no leaks. Check the tubing inside the instrument for clots. Check for bubbles in the reagent tubing. If bubbles are present, check the connections at the back of the instrument, at the top of the corresponding reagent container, and under the container cap to make sure they are tight. Visually check the reagent containers for reagent. If needed, change the reagent pack see Changing the Reagent Pack on page Troubleshooting

115 9.3 Evaluating Unexpected Results This section presents several histograms demonstrating unexpected results. These examples will assist you in identifying various cell populations, their position, and their ratio as indicated by their respective histograms. The figure at right shows a full-spectrum histogram. The first two discriminators in this example show a clean valley between 29 and 34, which is ideal. The first discriminator indicates the end of the platelet population (and is not normally displayed in the PLT histogram), and the second indicates the start of the WBC category. High peaks or lack of a valley here indicate one or more of the following: Discriminators existence of nucleated RBCs or reticulocytes existence of resistive RBCs (bigger pieces of hemolysed RBCs in WBC solution) contaminated reagents, or other interference such as electrical noise, etc. Note that this illustration shows a full-spectrum histogram (2 400 fl) following the RBC lysing process, whereby the entire RBC population is ideally lysed for accurate counting of white blood cells and platelets. In the above histogram, the third and fourth discriminators establish the WBC differential: The area (cells) between the second and third discriminators is classified as lymphocytes. The area between the third and fourth discriminators is classified as monocytes. The area to the right of the fourth discriminator is classified as granulocytes. Troubleshooting 9-5

116 9.3.1 Examples 1 High Blank, Flag p The blank measurement shown below indicates a high PLT background, most likely due to a contaminated system. Blank is optimal if PLT < 26. any cells in this area may reflect the presence of aggregated/giant platelets, nrbcs, retics, etc. Evaluation The above histograms indicate the following (see the arrows): WBC normal: shows no traces of particles RBC shows some background noise PLT shows a p flag, indicating background noise (such as chamber contamination) Solution 1. Perform three to five cleanings as needed see Cleaning the Aperture on page Repeat the blank measurement see Run a Blank Measurement on page If the problem persists, contact Abaxis Technical Support see Customer and Technical Support on page Troubleshooting

117 2 No results due to having accepted a high blank flags p, b, and B A report similar to that shown below results from measuring a sample without first generating an acceptable blank. The p (high PLT), b (high RBC), and B (high WBC) flags show that the previous blank measurement had high background values. Although histograms are shown, the values are not reported (see the arrows in the values listing). Evaluation The above histograms indicate the following (see the arrows): WBC B flag, high WBC background RBC b flag, high RBC background PLT p flag, high WBC background Solution 1. Perform three to five cleanings as needed see Cleaning the Aperture on page Repeat the blank measurement see Run a Blank Measurement on page Repeat the sample run after obtaining an acceptable blank. 4. If the problem persists, contact Abaxis Technical Support see Customer and Technical Support on page 1-2. Troubleshooting 9-7

118 3 WBC clogging, flag C In cases of aperture clogging, the histogram represents apparent large cells, since clogging results in a smaller effective aperture, which makes the particles passing through appear relatively larger. The histograms are drawn, but WBC results are not available. Note: In cases of RBC clogging, MCV (and consequently HCT) are artificially larger. Evaluation The above histograms indicate the following (see the arrows and highlight): WBC flag C shows WBC aperture clogging HGB the result is unbelievably high (resulting in erroneous MCH/MCHC results) RBC normal PLT normal Solution 1. Perform three to five cleanings as needed see Cleaning the Aperture on page Repeat the measurement. 3. If the problem persists, contact Abaxis Technical Support see Customer and Technical Support on page Troubleshooting

119 4 WBC non-linear, flag M This particular sample shows a typical lymphocytosis (WBC count beyond linearity limits). Evaluation The above histograms indicate the following (see the arrow and highlight): WBC WBC is very high, indicating leukocytosis. Flag M means that the WBC results are out of the linearity range. The absolute and percentage values are marked with asterisks (*), due to overlapping cell populations (cell discriminators may not be accurate). RBC normal PLT normal Solution Contact Abaxis Technical Support see Customer and Technical Support on page 1-2. Troubleshooting 9-9

120 9.4 Preparing the Analyzer for Shipment If Abaxis Technical Support determines that your instrument needs to be sent in for service, Abaxis will send you a loaner unit for use in the meanwhile. After you receive the loaner unit, prepare your analyzer for shipping as follows. 1. Switch off the instrument as described on page Remove the waste connector from the instrument. 3. Put the cap on each reagent intake valve on the instrument to prevent reagent solutions from leaking out. 4. Remove the thermal printer paper roll from the built-in printer. 5. Remove the sample adapter from the sample rotor. 6. Unplug the external power supply, printer cable, and mini-keyboard from the instrument. 7. Open the instrument side door. Wipe up any liquid left inside the instrument. 8. Insert the foam piece in the position shown to prevent sample needles from sliding. 9. If you received a loaner instrument, transfer the tape from the loaner instrument's counting chamber onto the top of your instrument s counting chamber to protect it from dust. If you did not receive a loaner instrument, proceed to the next step. CAUTION: DO NOT use any other type of tape, as the tape s adhesive may damage the counting chamber. 10. Close the side door. Wipe up any liquid left outside of the instrument. 11. Place the analyzer into its original plastic bag and place it inside its shipping box. Make sure that the forms and instrument fit perfectly inside of the box. 12. Place the power supply and power cord properly into the shipping box. 13. Close the box and seal the box using a strong tape. The instrument is now ready for shipping Troubleshooting

121 Section 10 Specifications This appendix contains technical specifications for the HM2 system, and lists its linearity ranges. Section Contents 10.1 VetScan HM2 Specifications Linearity Ranges Specifications 10-1

122 10.1 VetScan HM2 Specifications Sample volume 25 µl whole blood (50 µl in prediluted mode) Aperture diameter 80 µm Throughput Approximately 25 tests/hour Characteristics Sampling method Clog prevention Cleaning procedure Quality control Calibration Multi user feature User interface Languages available Data capacity Host computer interface Data back-up method Software upgrade method External printer interface Built-in printer Display Keypad External keyboard Power supply Power supply (input) Operating temperature Dimensions (W x D x H) Net weight Accuracy (max. deviance from expected) Reproducibility (CV) Carry-over between samples Test range WBC 3% < 3% < 1% RBC 3% < 3% < 1% HCT 3% < 3% < 1% MCV 2% < 1% N/A HGB 2% < 2% < 1% 9 16 PLT 4% < 4% < 3% HCT 3% < 3% < 1% Open tube system with automatic sample rotor High-voltage pulse and chemical cleaning of the aperture in each analysis cycle High-voltage burst of the aperture, high-pressure back-flush, chemical cleaning of the aperture 3 QC options. QC parameters include: mean, ± range, SD and CV for all measured and calculated parameters, 16- and 64-day Levy-Jennings charts, separate QC database Automatic based on 3 measurements (default method). Predilution method must be calibrated independently. Three-level Multi user operation with selective privilege levels, user identification with ID and password Easy-to-use, menu-driven user interface with 6 hard-function buttons, graphic icons, and on-screen help English. For other languages, contact Abaxis Technical Support see page results, including histograms on-board. Data can be saved to USB flash drive or downloaded to computer. Serial (RS-232) computer link and USB USB flash drive USB flash drive or CD-ROM USB Easy Paper Operation built-in thermal printer 240x128-dot, high-contrast, CCFL, backlit, graphics liquid crystal diode 24 foil keys + START button Standard PS/2 or USB-compatible keyboard External 12 VDC, 8 A power module 100/240 V, Hz, 10 W stand by, 80 W operating F (15 30 C). Optimal temperature is 77 F (25 C) x 10.2 x 14.4 in (320 x 260 x 365 mm) 26.4 lbs (12 kg) 10-2 Specifications

123 10.2 Linearity Ranges The HM2 is guaranteed to provide specified accuracies within its linearity range. Beyond this range, results may still be displayed, but accuracy is no longer guaranteed. If the value is over the maximum range of guaranteed linearity, the instrument cannot measure it, and the result will be marked with an E, m, M, or N flag. To measure a sample whose parameters exceed the maximum linear value indicated in the table below, predilution is recommended see Using Prediluted Mode on page The following tables list the linearity ranges for primary parameters in normal measuring mode and prediluted mode. Table 10-1: Linearity Ranges in Normal Measuring Mode Parameter Linearity Ranges Maximum Unit WBC cells/liter RBC cells/liter PLT cells/liter HGB g/l HCT % MCV Fl MPV Fl Table 10-2: Linearity Ranges for Prediluted Mode Parameter Linearity Ranges Maximum Unit WBC cells/liter RBC cells/liter PLT cells/liter Specifications 10-3

124 10-4 Specifications

125 Appendix A Introduction to Veterinary Hematology This section introduces several fundamental concepts of veterinary hematology. Having a basic knowledge of these concepts will help you better understand the results from the analyzer. Appendix Contents A.1 Function of Blood A-2 A.2 Composition of Blood A-3 A.3 Blood Cell Parameters A-4 A.3.1 Red Blood Cells, Hemoglobin A-4 A.3.2 White Blood Cells A-5 A.3.3 Automated WBC Classification A-5 A.3.4 Platelets A-6 A.4 Normal Hematology Ranges (Dog, Cat, Horse, and Others). A-7 A.5 Veterinary Hematology References A-8 Introduction to Veterinary Hematology A-1

126 A.1 Function of Blood Blood circulates in the body and acts as a transport medium that carries oxygen, essential nutrients, and other materials to the cells of the body. It also serves to transport waste products for disposal. Neurons, muscle cells, connective tissue cells, and epithelial cells draw their nourishment from interstitial spaces, and respond to the glucose and oxygen content of that environment. Fresh supplies of oxygen and glucose are exchanged with the blood circulating in the capillaries. Blood receives its oxygen from lungs and glucose from the intestines and liver. Normal cell function depends on the rapid removal of toxic metabolic products (CO 2 and NH 3 ) from the interstitial fluid environment. These waste products are taken up by the plasma and red blood cells, and eliminated as the blood passes through the kidneys and lungs. Blood also delivers hormones, lipids, amino acids, salts, and vitamins, and removes urea and conjugated acids. Blood distributes the heat generated by metabolizing body cells, so that body temperature is maintained at a constant level. In the event of vascular injury, blood platelets and plasma coagulation mechanisms prevent blood loss by aggregating with other platelets to form large hemostatic plugs (clots). White blood cells protect against infections by identifying and killing invasive bacteria. The number, size and distribution of blood cells provide important information for clinical diagnosis and therapy. The aim of hematology is to collect this information. A-2 Introduction to Veterinary Hematology

127 A.2 Composition of Blood Whole blood contains three cellular components: Red blood cells (erythrocytes, RBC) White blood cells (leukocytes, WBC) Platelets (thrombocytes, PLT) Table A-1: Cell Composition of Whole Blood* Red Blood Cells White Blood Cells Platelets Normal density 5 12 x cells/l 6 15 x 10 9 cells/l x10 9 cells/l Nucleated? No ** Yes No Sub-populations and their percentages Shape, size RBC: 99.9% NRBC ** (Nucleated RBC) 0.1% Biconcave (donut) shape diameter: 5 7 µm thickness: µm GRAnulocytes: 65% NEUtrophil: 60% EOSinophil: 4% BASophil: 1% LYMphocytes: 32% MONocytes: 3% GRAnulocytes: µm LYMphocytes: 8 15 µm MONocytes: µm Fragments with a diameter of 2-4 µm Volume of cell fl fl 5 15 fl Volumetric% in whole blood 40 45% 0.1% 0.3% * Data generated using whole human blood. **Mature mammalian RBCs are non-nucleated. Introduction to Veterinary Hematology A-3

128 A.3 Blood Cell Parameters A.3.1 Red Blood Cells, Hemoglobin Red blood cells RBC are formed in the bone marrow. A mature human red blood cell is nonnucleated, and has a mean corpuscular volume (MCV) of approx. 70 fl. RBCs are the most numerous cell type in blood. There are approximately 5 10 x cells/l in the blood of a healthy dog. RBC and MCV are directly measured primary parameters. Hematocrit HCT is the proportion of RBCs to plasma (liquid portion) in blood. HCT is the most accurate and simplest way to measure the degree of anemia, and is calculated from the RBC and MCV values: HCT percent = RBC x MCV / 10 HCT absolute = HCT percent / 10 Typically, HCT = 0.45 = 45% Red Blood Cell Distribution Width RDW is a measure of RBC anisocytosis, the degree of cell size variation. In a healthy sample, RBCs demonstrate a normal (Gaussian) distribution (bell curve), as shown at right. RDW can be characterized by a standard deviation (RDW-SD) or a coefficient of variation (RDW-CV) represented as a percentage. (fl) Hemoglobin HGB is the main component of RBCs. It is a conjugated protein (with Fe), and its main function is to transport oxygen from the lungs to tissues and carbon dioxide from the tissues back to the lungs. Normal HGB concentration of samples is approximately 14 g/dl or 140 g/l or 87 mmol/l. Mean Corpuscular Hemoglobin MCH is the average hemoglobin content of RBCs, and is calculated from RBC and HGB values: MCH = HGB / RBC x 10, reported in picograms or fmol Mean Corpuscular Hemoglobin Concentration MCHC is the concentration of HGB in an average RBC, calculated from the HGB and HCT values: MCHC = HGB / HCT absolute, reported in g/dl, g/l or mmol/l A-4 Introduction to Veterinary Hematology

129 A.3.2 White Blood Cells White Blood Cells WBC are formed in the bone marrow. During their maturation sequence they differentiate from the parent cell into five sub-populations. WBCs are nucleated and classified as granulocytes (neutrophils, eosinophils, and basophils), lymphocytes, and monocytes. WBCs are equipped with all cell organelles necessary to perform vital protective functions in the body. The normal WBC is in the range of 7 x 10 9 cells/l, a fraction of the RBC population. In pathological conditions, the WBC count can increase dramatically (up to 300 x 10 9 cells/l in extreme leukemia). In these cases, predilution of the sample is recommended for the most accurate results (see Diluting Whole Blood on page B-3). Three-part differential histograms (volume distribution curves) of WBCs can be used as a simple, visual evaluation of the number and relative percentage of lymphocytes (LYM, LYM%), monocytes (MON, MON%), and granulocytes (GRA, GRA%). WBC-related parameters are defined as follows: WBC = LYM + MON + GRA LYM% = LYM / WBC MON% = MON / WBC GRA% = GRA / WBC A.3.3 Automated WBC Classification The analyzer evaluates each sample as a unique population, using dynamic cellular discriminators to assess the cellular distribution most accurately. To determine WBC sub-populations, the analyzer first sets discriminator 1 at the limit of hemolysed RBCs + PLTs (on the left) and LYM population, then fits normal distribution curves to the remaining WBC histogram (shown below in different shades of grey). Discriminators: RBC/PLT region GRA population LYM population MID population Introduction to Veterinary Hematology A-5

130 Eosinophilia is often depicted as a peak between the MON and GRA classifications. As with any automated system, good laboratory practice requires that all abnormal results be verified by slide (blood smear) review. A.3.4 Platelets Platelets PLT are non-nucleated fragments of the megakaryocyte. Note that platelets are formed by cellular fragmentation and not by a so-called maturation sequence. Therefore, the platelet histogram normally has a logarithmic shape on the left side, and a normal shape on the right side ( log-normal distribution). Normal PLT concentrations range from x 10 9 cells/l (for dogs), depending on the mean platelet volume (MPV), but can vary from x10 9 cells/l under certain circumstances. PLTs are relatively small compared to RBCs. The mean platelet volume MPV is approximately 10 fl, so PLTs can effectively be separated from RBCs by their size. Platelet aggregation is common, particularly in feline species, and is depicted by a flattened, lumpy histogram, as shown at right. This effect can be minimized with proper sample collection and vortex mixing of the sample (up to 30 seconds) before analysis. The analyzer calculates the volumetric ratio of PLTs in whole blood as follows: PCT percent = PLT x MPV/10%, PCT absolute = PCT percent / 10 Typically, PCT = = 0.3% Platelet Distribution Width PDW is a measure of platelet anisocytosis, the degree of size variation. In a healthy sample, platelets demonstrate a normal (Gaussian) distribution (bell curve). PDW can be characterized by a standard deviation (PDW-SD) or a coefficient of variation (PDW-CV) represented as a percentage. A-6 Introduction to Veterinary Hematology

131 A.4 Normal Hematology Ranges (Dog, Cat, Horse, and Others) The following table summarizes normal ranges of blood cell parameters. Keep in mind that normal values vary from population to population. Parameter Unit Dog Cat Horse Bovine Pig Mouse WBC 10 9 cells/l LYM% % MID% % GRA% % RBC cells/l HCT % MCV fl RDWcv % HGB g/dl MCH pg MCHC g/dl PLT 10 9 cells/l MPV fl Parameter Unit Dog Cat Horse Bovine Pig Mouse WBC 10 9 cells/l LYM% % MID% % GRA% % RBC cells/l HCT % MCV fl RDWcv % HGB g/dl MCH pg MCHC g/dl PLT 10 9 cells/l MPV fl Introduction to Veterinary Hematology A-7

132 A.5 Veterinary Hematology References Schalm s Veterinary Hematology, 5th ed., Feldman, Bernard, et al, Lippincott Williams & Wilkins, Veterinary Hematology, Atlas of Common Domestic Species, Reagan, William, et al, Iowa State Press, Veterinary Laboratory Medicine, Interpretation & Diagnosis, 3rd ed., Meyer, Denny & Harvey, John, Elsevier Press, A Guide to Hematology in Dogs & Cats, Rebar, Alan, et al, Teton New Media, Automated Blood Counts & Differentials, A Practical Guide, Bessman, J David, Johns Hopkins University Press, Hematology Techniques & Concepts for Veterinary Technicians, Voigt, Gregg, Blackwell Publishing, A-8 Introduction to Veterinary Hematology

133 Appendix B Operating Principles This appendix explains the basic operating principles of the analyzer. Appendix Contents B.1 Complete Blood Count (CBC) B-2 B.2 Measurement Methods B-2 B.2.1 Volumetric Impedance Method B-2 B.2.2 Diluting Whole Blood B-3 B.2.3 Three-Part WBC Differential Method B-4 B.3 Hemoglobin Determination B-5 B.3.1 Hemoglobin Determination by Photometry B-5 B.4 Measured and Calculated Values B-6 Operating Principles B-1

134 B.1 Complete Blood Count (CBC) The analyzer utilizes impedance technology whereby electrically neutral blood cells pass through an electrically charged aperture thereby generating a pulse. Cell counts are determined by the number of pulses measured in a given volume of blood over a set period of time. The decrease in electrical conductance (degree of intensity) as measured is directly proportional to the cell volume. This size discrimination, along with susceptibility to various lysing agents distinguish the basic cell types (red, whites, platelets). B.2 Measurement Methods This section provides an overview of the hematology measurement methods used by the analyzer. B.2.1 Volumetric Impedance Method The analyzer uses a volumetric impedance method of counting blood cells. The following figure illustrates this method. The principle of this method is that blood diluted with an isotonic solution (diluent) conducts electric current by ionic conduction. A counting chamber made of an insulating material (plastic) holds this diluted blood, while a small circular hole (aperture) in this chamber allows the flow of diluted blood. (The analyzer aperture diameter and length is 80 µm the optimal size for veterinary hematology.) Placing two electrodes on the two sides of this aperture and applying constant electric current causes the isotonic solution to conduct electricity, and allows a voltage to be measured on the aperture. B-2 Operating Principles

135 Applying pressure to the diluted sample causes it to flow through the aperture. When a cell is passing the aperture, a small change in electric impedance occurs, so that the voltage rises somewhat and a small electric pulse occurs. The amplitude of this pulse is proportional to the ratio of the cell volume (size) and the aperture volume: the bigger the cell, the higher the pulse. Proper counting (or differentiation) of cells requires passing of only one cell through the aperture at a time. To help ensure this, the blood samples must be diluted, since cell concentrations are otherwise too high. Although diluted blood is used, in cases of extremely high concentrations (such as leukemia) WBC density can be 100x higher than normal, causing two or more cells to pass through the aperture at a time, generating one pulse instead of two (or more). This is called coincidence, and results in non-linear counting of cells. Flags m, M, and N appear in this case. The WBC linearity range is 100 x 10 9 cells/l. In cases of cell counts beyond the linear range, an external predilution of the sample is recommended see Using Prediluted Mode on page B.2.2 Diluting Whole Blood The analyzer uses the following procedure to dilute the blood sample for processing: Operating Principles B-3

136 In this procedure, 25 µl of whole blood is diluted with 4 ml of diluent to form a 1:160 first dilution. 25 µl of this dilution is then mixed with 0.5 ml of lyse to form an overall 1:180 dilution, suitable for measuring WBC and HGB. After WBC-HGB measurement, 25 µl of the first dilution is mixed with 5 ml of diluent to form an overall 1:32000 dilution, suitable for measuring RBC and PLT. B.2.3 Three-Part WBC Differential Method Since RBCs are typically 1000 times more concentrated in normal blood than are WBCs, they would interfere with WBC counting if left intact. In addition, the analyzer directly measures HGB, which is inside the RBCs and so must be extracted. Therefore, a hemolysing reagent (lyse) is used to dissolve cellular membranes, thus destroying RBCs, and creating a complex solution suitable for photometry of HGB and counting WBCs. The following figure shows the changes in blood cell characteristics that occur during three-part differential hemolysis. The membranes of the WBCs become selectively permeable, so that they begin to shrink down to their nuclei in the slightly hypertonic lyse solution. Effectively hemolysed samples contain WBC particles in the fl region (for veterinary species). B-4 Operating Principles

137 The figure below shows a typical three-part differential WBC histogram of selective hemolysis (dog). LYM MON GRA B.3 Hemoglobin Determination Hemoglobin is measured directly by means of the traditionally used cyanomethoglobin reaction, but using cyanide-free substances to reach the same endpoint: Fe 2+ converted to Fe 3+ methemoglobin + KCN cyanmethemoglobin Hemoglobin concentration is measured photometrically. B.3.1 Hemoglobin Determination by Photometry HGB determination is one of the most important hematology parameters, as it relates to the oxygen carrying capacity of blood. The analyzer uses a cyanide-free lysing reagent to minimize environmental impact. The effect of cyanide-free lyse is very similar to that of lyse containing cyanide, but the chemical reaction is slightly different. The figure below illustrates the HGB measurement method. HGB is measured by passing light (540 nm) through the WBC dilution, and measuring the transmitted light with a photo detector. The light intensity (I) of the sample liquid is logarithmically proportional to the concentration of HGB: HGB log (I reference / I sample ) Operating Principles B-5

138 B.4 Measured and Calculated Values Each sample is analyzed to produce a complete, 18-parameter blood count (CBC), including the following measured or calculated parameters: WBC total white blood cell count LYM lymphocytes count MON monocyte count* GRA granulocytes count LYM% lymphocyte percentage MO% monocyte* percentage GRA% granulocytes percentage RBC red blood cell count HGB hemoglobin HCT hematocrit MCV mean corpuscular volume MCH mean corpuscular hemoglobin MCHC mean corpuscular hemoglobin concentration RDWc red cell distribution width PLT platelet count PCT platelet percentage MPV mean platelet volume PDWc platelet distribution width * The monocyte category consists primarily of monocytes. Impedance counters categorize white blood cell types (differential) according to size, and therefore a certain percentage of eosinophils may have a mass that falls in the normal range for monocytes (the exact percentage depends on the individual animal and is generally inconsequential due to the very low numbers of eosinophils in a healthy animal). Eosinophilias, however, can typically be visualized as a distinct peak between the monocyte range and the granulocyte peak on a histogram. B-6 Operating Principles

139 B.5 Measured and Calculated Values The HM2 measures and calculates the following values from tested blood samples. Table B-1: Measured and Calculated Values Values White Blood Cells WBC (reportable as: cells/l, cells/µl) Red Blood Cells RBC (reportable as: cells/l, cells/µl) Hemoglobin concentration HGB (reportable as: g/dl, g/l, mmol/l) Mean Corpuscular Volume MCV (fl) Hematocrit HCT (reportable as: percentage, absolute) Mean Corpuscular Hemoglobin MCH (reportable as: picogram, fmol) Mean Corpuscular Hemoglobin Concentration MCHC (reportable as: g/dl, g/l, mmol/l) Red Cell Distribution Width (reportable as: RDW-SD [fl], RDW-CV [absolute]) Definitions Total number of leukocytes (white blood cells). WBC = WBC cal x (cells/l, cells/µl) Total number of erythrocytes (red blood cells). RBC = RBC cal x (cells/l, cells/µl) Measured photometrically at 540 nm (see page B-2 for details). HGB = HGB cal x (HGB measured HGB blank ) Average volume of individual erythrocytes derived from the RBC histogram. Also known as Packed Cell Volume (PCV). Calculated from the RBC and MCV values: HCT percentage = RBC x MCV / 10 HCT absolute = RBC x MCV Average hemoglobin content of erythrocytes, calculated from RBC and HGB values: MCH = HGB / RBC Calculated from the HGB and HCT values: MCHC = HGB / HCT absolute Measure of the degree of RBC anisocytosis. Calculated using the distribution width of the erythrocyte or platelet population derived from the histogram at 20% of peak: Platelet PLT (reportable as: cells/l, cells/µl) Mean Platelet Volume MPV (fl) Platelet Distribution Width (reportable as: PDW-SD [fl], PDW-CV [absolute]) Platelet Hematocrit (Thrombocrit) PCT (reportable as: percentage, absolute) Number of thrombocytes (platelets). PLT = PLT cal x (cells/l, cells/µl) Average volume of individual platelets derived from the PLT histogram. Measure of the degree of platelet anisocytosis. Calculated from the PLT and MPV values: PCT percentage = PLT x MPV x 100 PCT absolute = PLT x MPV Operating Principles B-7

140 Values White Blood Cell Differential: LYM, LYM%: lymphocytes MON, MON%: monocytes GRA, GRA%: neutrophil, eosinophil and basophil granulocytes Definitions Absolute values counted in the channels determined by the three WBC discriminators: LYM MON GRA Percentages calculated from the absolute WBC value. B-8 Operating Principles

141 Appendix C Potential Sample Interferences Table C-1 lists situations in which substances in the samples themselves can interfere with accurate analysis, and provides possible solutions. Potential Sample Interferences C-1

142 Table C-1: Intrinsic Substances: Potential Sample-Induced Interferences Sample Condition Indicators Results Affected Causes Solutions Lipemia Cloudy, white plasma. High MCHC. Increased HGB, MCHC, MCH. RBCs may be smudged on blood film. Hemolysis Pink or red plasma. Decreased RBC, PCV Decreased MCHC Clumped platelets Decreased platelet count w/ platelet clumps often visible on blood smear or applicator stick dipped in sample. Rising left side of lymphocyte curve. Small pale clots stick to wooden applicator stick swirled in sample. Giant platelets Right side of platelet histogram runs into RBC histogram. Clotted sample Visible clot in sample. Red clot(s) stick to wooden applicator stick swirled in sample. Platelet clumps may or may not be visible on blood smear. Lyse-resistant RBCs Decreased PLT +/- increased WBC Decreased PLT, decreased MPV Decreased PLT, decreased WBC, and/or decreased RBC (varies with clot size) Rising left side of lymphocyte curve. Increased WBC, increased Lymphs Non-fasted sample Redraw fasted sample. Metabolic disorder a Use RBC, PCV, MCV, and RDW rather than HGB, MCH, MCHC values to assess anemia. Traumatic venipuncture Hemolytic anemia N/A Traumatic venipuncture or feline species Excess EDTA (under-filled tube) Miscellaneous/ idiopathic Thrombopoiesis, feline species Traumatic venipuncture and/ or delayed transfer to anti-coagulant Idiopathic (most commonly with felines) Redraw w/ clean venipuncture. Redraw; collect blood in anti-coagulated syringe or use vacutainer system; invert tube several times immediately after filling; vortex sample immediately before testing. Fill tube at least halfway, or remove portion of EDTA before filling tube. Use alternative anti-coagulant as heparin or citrate; vortex sample immediately prior to sampling. Confirm platelet estimate on smear and/ or manual platelet count. Redraw with clean venipuncture; use vacutainer system or anti-coagulated syringe; mix collection tube by multiple inversions immediately after filling. Clotted sample may clog the sample needle. Lyse volume may be adjusted for the sample (for additional information, contact Abaxis Technical Support see page 1-2). a).diabetes mellitus, nephrotic syndrome, hypothyroidism, lipoprotein lipase deficiency, acute pancreatitis, cholestasis, hyperadrenocorticism, hypercholesterolemia in briards, idiopathic hyperlipidemia of miniature schnauzers. C-2 Potential Sample Interferences

143 Appendix D Veterinary Case Studies The following pages present a variety of veterinary case studies. Appendix Contents D.1 Human Control D-2 D.2 Dogs D-3 D.2.1 Dog: Normal Sample D-3 D.2.2 Dog: High LYM%, Low GRA% D-4 D.2.3 Dog: High PLT, High GRA D-5 D.2.4 Dog: Low PLT, High WBC and GRA D-6 D.2.5 Dog: High RBC and HGB D-7 D.2.6 Dog: Low RBC and PLT, high WBC D-8 D.2.7 Dog: Eosinophilia D-9 D.3 Cats D-10 D.3.1 Cat: Optimal Sample D-10 D.3.2 Cat: Low PLT and RBC D-11 D.3.3 Cat: Clumped/Aggregated Platelets D-12 D.3.4 Cat: High WBC and GRA, Low RBC, HCT, and HGB D-12 D.3.5 Cat: Possible nrbcs and Clumped Platelets D-13 D.4 Horses D-15 D.4.1 Horse: Normal Sample D-15 D.4.2 Horse: Low LYM%, High GRA% D-16 D.4.3 Horse: Low PLT and MPV D-17 D.4.4 Horse: Low RBC and HGB D-18 Veterinary Case Studies D-1

144 D.1 Human Control The following report shows typical histograms of normal human control blood (that is, human control run in QC mode). Note the differences in cell populations: human sample and control blood contains larger cells than does animal blood. D-2 Veterinary Case Studies

145 D.2 Dogs Dog samples show RBC peaks around fl with a good separation of the PLTs from the RBCs. Canine lymphocyte populations (lymphocytes, monocytes, and granulocytes) can overlap as a result of similar cell sizes. D.2.1 Dog: Normal Sample The following shows a normal dog histogram. These histograms indicate the following: WBC All cells larger than 39 fl are counted as WBCs (indicated by discriminator 1). Discriminators 2 and 3 show the MON population. RBC All cells larger than 25 fl are counted as RBCs. The RBC histogram follows a normal distribution. The expected MCV is near 60 fl, and the expected RBC is near 7x10 12 cells/l. PLT PLT cell population is between 2 and 25 fl. The PLT histogram follows a lognormal distribution. Veterinary Case Studies D-3

146 D.2.2 Dog: High LYM%, Low GRA% The following shows a sample that has high LYM%. (LYM% is normally near 25 30% for dogs.) This case shows a lymphocytosis. These histograms indicate the following: WBC High LYM%, low GRA%. RBC Normal. PLT Normal. D-4 Veterinary Case Studies

147 D.2.3 Dog: High PLT, High GRA In this sample, the absolute value of GRA and PLT is high, and the LYM population is much smaller than the GRA population. This case demonstrates a neutrophilia and thrombocytosis. These histograms indicate the following: WBC Three-part differential curve with good separation between populations. RBC Normal. PLT High PLT. (Note that the RBC histogram shows that the peak of the PLT curve, to the left of the RBC discriminator, is higher than normal.) Veterinary Case Studies D-5

148 D.2.4 Dog: Low PLT, High WBC and GRA This sample shows a very low PLT, while the WBC is high due to an extremely high GRA. This case demonstrates a neutropenia and thrombocytosis. These histograms indicate the following: WBC Discriminator 1 is at 53 fl, accurately separating the RBC/PLT region. Discriminators 2 and 3 are placed correctly according to the WBC population. RBC Normal. Note that the PLT curve is very flat. PLT Very low PLT. D-6 Veterinary Case Studies

149 D.2.5 Dog: High RBC and HGB This sample shows a slightly high RBC and HGB, while the WBC and PLT are normal. These histograms indicate the following: WBC Discriminator 1 is at 56 fl, accurately separating the RBC/PLT region. Discriminators 2 and 3 are placed correctly according to the WBC population. RBC Slightly high RBC, normal distribution. PLT Normal, with a perfect discriminator at 25 fl. Veterinary Case Studies D-7

150 D.2.6 Dog: Low RBC and PLT, high WBC This sample shows low RBC, PLT, HGB, and HCT values. The WBC is high due to a high GRA population. This case demonstrates anemia and neutrophilia. These histograms indicate the following: WBC Discriminator 1 is at 45 fl, accurately separating the RBC/PLT region. Discriminators 2 and 3 are placed correctly according to the WBC population. RBC Very low RBC, with a normal distribution, indicating anemia. PLT Low PLT. D-8 Veterinary Case Studies

151 D.2.7 Dog: Eosinophilia This sample demonstrates an eosinophilic canine patient. These histograms indicate the following: WBC The histogram indicates a normal WBC accompanied by an absolute lymphopenia and relative granulocytosis. The presence of the secondary eosinophil peak (indicated by the arrow) indicates a eosinophil count higher than normal, and a need for a manual blood smear. Veterinary Case Studies D-9

152 D.3 Cats An important characteristic of cat blood is that the RBCs are much smaller than those of dogs (20 50 fl, MCV = fl), potentially causing the RBC and PLT histograms to overlap slightly. Cats also commonly demonstrate both platelet aggregation and giant platelets. The analyzer minimizes these effects with a proprietary technology and dynamic discriminator approach to maximize accuracy. A good practice when taking blood from cats is to discard the first few drops of the sample. This can prevent hair and skin pieces from getting into the sample. Some clinics have minimized stressinduced platelet aggregation by collection from the saphenous vein using a vacutainer. Pre-analytical vortex mixing (up to 30 seconds) also helps disaggregate platelets, with no deleterious effects. D.3.1 Cat: Optimal Sample The following shows an optimal normal cat sample. These histograms indicate the following: WBC Normal. RBC Normal. PLT Optimal: the PLT is well separated from the RBC. D-10 Veterinary Case Studies

153 D.3.2 Cat: Low PLT and RBC This sample demonstrates a very low PLT, and a typical feline WBC histogram. These histograms indicate the following: WBC Normal. (Note that the WBC histogram at discriminator 1 is high, indicating the presence of lyse-resistive RBCs.) RBC Slightly low RBC, with a correct distribution. PLT Very low PLT. Veterinary Case Studies D-11

154 D.3.3 Cat: Clumped/Aggregated Platelets The following samples show normal and clumped platelets, respectively: Normal Clumped D.3.4 Cat: High WBC and GRA, Low RBC, HCT, and HGB The results for this cat indicate anemia and leukocytosis. These histograms indicate the following: WBC Discriminators are set correctly, showing a high GRA population. RBC Normal distribution, low RBC. PLT Discriminators are set correctly, showing a slightly low PLT. D-12 Veterinary Case Studies

155 D.3.5 Cat: Possible nrbcs and Clumped Platelets This sample demonstrates a patient that may have nucleated RBCs and clumped platelets. These histograms indicate the following: WBC Rising left side of the histogram may indicate nucleated RBCs, or highly lyseresistant RBCs, which can be present in feline samples. RBC Normal distribution. PLT Flattened, lumpy histogram indicates aggregated (clumped) platelets. Veterinary Case Studies D-13