VetScan HM5 Hematology System

Transcription

1 VetScan HM5 Hematology System For Veterinary Use Only Customer and Technical Support August 2009 PN: Rev. A Assy PN: Rev. E Text 2009, Abaxis, Inc. Union City, CA 94587

2 IMPORTANT: READ BEFORE USING THE VETSCAN HM5 HEMATOLOGY ANALYZER FOR THE FIRST TIME To get started quickly, please see 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 HM5 Hematology Analyzer! 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. VetScan is a registered trademark of Abaxis, Inc. August 2009 PN: Rev. E 2009, 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 HM5 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 Running the Stress 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 HM5 Specifications Linearity Ranges Precision 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-9 A.5 Veterinary Hematology References A-10 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-7 B.4 Measured and Calculated Values B-8 B.5 Measured and Calculated Values B-9 Section C: Potential Sample Interferences C-1 Section D: Veterinary Case Studies D-1 D.1 Normal Level Control D-2 D.2 Dogs D-3 D.3 Cats D-9 D.4 Horses D-12 Section E: Glossary E-1 Index TOC-2 Table of Contents

5 Section 1 General Information This section provides general information about the Abaxis VetScan HM5 Hematology System. Section Contents 1.1 Introduction Customer and Technical Support Symbols Used in Labeling and Hazard Identification General Information 1-1

6 1.1 Introduction The VetScan HM5 hematology analyzer is a compact, fully automated cell counter for in-vitro diagnostic use in veterinary clinics, research laboratories, point-of-care centers, and pharmaceutical/biotech companies. The VetScan HM5 can process samples per hour, and is designed to determine the following 22 hematology parameters from a 50µl (2x25µl) whole blood sample: WBC: LYM#, MON#, NEU#, EOS#, BAS# LYM%, MON%, NEU%, EOS%, BAS% (five-part WBC differential) HGB, RBC, HCT, MCV, RDW, MCH, MCHC PLT, MPV, PCT, PDW The VetScan HM5 is calibrated to analyze multiple veterinary species. Check the list of species on the analyzer 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 customers free of charge. The VetScan HM5 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, Lyse2, 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 VetScan HM5 can be connected to a VetScan Classic or VetScan VS2 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 VetScan HM5 alone will also interface directly with an external computer. 1-2 General Information

7 1.2 Customer and Technical Support Abaxis Technical Support personnel can answer your questions regarding the VetScan HM5 Hematology Analyzer, or the combined VetScan HM5/VS2 system. Telephone: Abaxis Technical Support is available 24 hours a day, seven days a week. vetsupport@abaxis.com Web: Fax: Note: Contact information for Abaxis is also available in the VetScan HM5 s software. To view the information, press the Utilities key on the instrument s front panel, then press 6 to select Service. General Information 1-3

8 1.3 Symbols Used in Labeling and Hazard Identification The VetScan HM5 uses safety features to protect the operator from injury, prevent damage to the analyzer, 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 1-4 General Information

9 Section 2 System Overview and Installation This section includes an overview of the VetScan HM5: its features, functionality, main components, reagent pack, and available accessories. The section also provides complete installation instructions. Section Contents 2.1 VetScan HM5 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 VetScan HM5 to a Computer Connecting the Reagent Pack Connecting the Waste Bottle 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

10 2.1 VetScan HM5 System Overview The VetScan HM5, a fully-automated, 5-part cell counter designed specifically for veterinary applications, offers a comprehensive 22-parameter complete blood count with cellular histograms in just minutes. Its superior performance, elegant design, ease of use, true database management capability, and minimal maintenance make it the optimal hematology system for veterinary clinics, research laboratories, and pharmaceutical/biotech companies. The following pages provide an overview of the VetScan HM Features Small sample size requirement: consumes approximately 50 µl of potassium EDTA-preserved whole blood per CBC measurement. Rapid test turnaround: less than 4 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. Reliable results, including out of range values for 3-part and 5-part modes: 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 2,000 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 VetScan HM5 can be connected to a VetScan Classic or VetScan VS2 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

11 Benefits 10 species available now Small footprint Easy to use Reliable results, including out-of-range values Rapid test turn-around with minimal hands-on time Customizable reference ranges Optimized performance with minimal maintenance Automatically saves up to 2000 test records All downloadable through USB pen drive and integrated data management system Allows software updates and future instrument upgrades Integrated result reporting and comprehensive laboratory analysis Software tracks reagent usage and notifies you when replacement is needed Eco-friendly reagents System Overview and Installation 2-3

12 2.1.2 Main Components The following pages show the main components of the VetScan HM5 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 Sampling rotor 6 Interchangeable sample tube adapter 2-4 System Overview and Installation

13 Back View Reagent tubing connections 2 Power On/Off switch 3 12v DC 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. System Overview and Installation 2-5

14 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 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 2-33). 2-6 System Overview and Installation

15 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 2-13 for a list of the menus and commands that are available through the hard-function keys. System Overview and Installation 2-7

16 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. GRAPHS 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. VALUES Displays the results. 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 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. 2-8 System Overview and Installation

17 2.1.4 Reagents VetScan HM5 Reagent Pack The VetScan HM5 reagent pack (PN ) consists of bottles containing diluent, cleaner, lyse, Lyse2, 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 Diluent Isotonic saline solution used to dilute whole blood specimens, and to rinse the analyzer s fluidic system between analyses. Green 9 liters Rinse Used with diluent to prevent salt built-up on White 500 ml aperture. Cleaner Used in the fluidic system cleaning process. Blue 300 ml Lyse Used to create hemolysate for three-part WBC differential, and for total WBC and HGB. Yellow 300 ml Lyse2 Used to dilute whole blood and differentially hemolyse white blood cells to separate eosinophil granulocytes from other WBC by volume. Suitable for determining EOS, EOS%, BAS, and BAS% parameters. White with orange dot 800 ml Controls and Calibrator Before you can perform analyses, the instrument must first be calibrated using the normal control/ calibrator 1, and quality control procedures must then be performed using high, normal or low controls. 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 refer to your VetScan HM5 price list for part numbers. High-level control Normal-level control Low-level control 1. The VetScan HM5 controls are derived from human sources. Observe universal safety precautions when handling the controls. See Calibration and Quality Control on page 5-1. System Overview and Installation 2-9

18 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 2-5). 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 2-8), and keys F7 through F12 function as equivalent to the analyzer s hard-function keys (see page 2-7): 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 12v DC power supply that can operate from a 230 or 110 volt 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 VetScan HM5 will be used in an area prone to electrical surges or power outages. An appropriate power supply is vital for system integrity System Overview and Installation

19 Reagent Tubing Kit The reagent tubing kit (Abaxis Part No ) includes the color-coded reagent tubes used to connect the VetScan HM5 with the containers in the reagent pack. The kit also includes the reagent bottle caps with connectors, drop-down tubes, and cleaning tube 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 3 ml Control Adapter ml control tube Peristaltic Pump Tube Assembly As part of routine yearly preventive maintenance, the analyzer s peristaltic pump tube must be inspected, and replaced if necessary (see Periodic Maintenance on page 7-2). The analyzer is shipped with one replacement peristaltic pump tube. System Overview and Installation 2-11

20 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 HM5 has two primary subsystems. Fluidic system: Performs sampling, diluting, mixing, lysing, and rinsing func- Diluent Lyse Cleaner Lyse2 tions. Generates the regu- Waste Rinse lated vacuum used for moving cells through the aperture during the counting process. Diluent solution Lyse Cleaner Waste container Rinse Lyse2 Data processing system: Counts, measures, and calculates blood parameters, generates and stores numerical results and histograms System Overview and Installation

21 2.1.7 Menu/Command Listing The following charts outline the VetScan HM5 s menu functions. Information / Help Displays help for the current screen. Analysis / Measurement MENU 1 Repeat last sample Repeats last sample tested (analyzer assigns new sample ID, all other patient information is unchanged). 2 Measure blank Performs blank measurement. 3 Predilution mode Enters automatic calculation for 1:5 externally diluted sample ( 1:5 appears in upper RH screen corner). (Available for 3-part and 5-part modes.) 4 Needle Height Setting Adjusts sampling depth of needle. 5 Lyse volume Adjusts lyse reagent volume by +/ 0.1 ml. PT.ID Opens dialog for entering patient information. Sample ID Patient ID Species Doctor Name Age Sex Analyzer assigns automatically cannot be modified). This field can be entered for each sample. GRAPHS VALUES Displays details about the sample. System Overview and Installation 2-13

22 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, 2 Select all and/or patient IDs. Selects all records. 3 Deselect all Deselects all records. 3 Change sort order 1 Unsorted Lists records in the order entered 2 Sort by time (most recent to oldest). Lists records by time of measurement (oldest to most recent). 3 Sort by sample ID Lists records by sample ID (highest 4 Sort by patient ID to lowest). Lists records by patient ID (highest to lowest). 4 Manage selected records 1 Send selected Sends selected records to a computer records 2 Delete selected through a serial connection. Deletes selected records from the records analyzer s database. 3 Backup selected Saves selected records to a USB drive. records 5 View external Reads stored records from a USB drive. 6 Backup one day Backs up all results from a specified date to an inserted USB drive. DETAILS Toggle between graph and table display of selected results. PT.ID Opens dialog for viewing/modifying patient information (Sample ID and Patient ID cannot be changed). Sample ID Patient ID Species Doctor Name Age Sex 2-14 System Overview and Installation

23 Utilities 1 Maintenance 1 Cleaning 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 (white connector). 4 Prime cleaner Primes cleaner line (blue connector). 5 Prime Lyse2 Primes Lyse2 line (orange connector). 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. 2 Calibration 3 Quality control 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. 3 Enzymatic Cleaning Cleans analyzer with enzymatic cleaner. 6 Prime all Primes all reagent lines. 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. 3 Reset Calibration For troubleshooting calibration issues. 1 Set QC reference ranges Updates QC ranges for 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. Selected QC type: 3-5 Part Control Only option for QC control material. 1 5-Part Control (continued) System Overview and Installation 2-15

24 Utilities (continued) 4 Diagnostics 5 Settings 1 Device information Shows model, S/N, software version. 2 Statistics Displays operation statistics. 3 Self test Tests the analyzer s internal systems. 4 Stress test 10 cycles Higher pressure test of fluidics and measurement systems. 5 Log File Viewer Displays history of analyzer actions. 1 Printer settings Dialog for viewing/modifying 2 Customize printer settings. 1 General Settings 2 Units 3 Laboratory 4 User modes 1 Single user mode 2 Multi-user mode Dialog for setting screensaver delay, analyzer date format, and printing combined results. Dialog for setting unit preferences. Dialog for laboratory information (used in report header). Selects Single or Multi-user mode. Each user has unique user name and password. 3 Date and time Sets date/time and format. 4 Fluid sensors Enables and disables reagent sensors. 1 Diluent & Cleaner 2 Lyse 3 Lyse2 4 Rinse 5 Calibrate sensors Calibrates reagent sensors (use if analyzer incorrectly reports missing reagents). 6 Service Displays contact information for technical service. Printer Prints selected test results. Exit 1 Shut down Shuts off the analyzer (use if the unit will not be used for over 72 hours). 2 Preparing for shipment Drains and shuts off the analyzer (use if the unit will be shipped, or unused for more than 2 weeks) System Overview and Installation

25 2.2 Unpacking the System Follow these directions for unpacking the VetScan HM5 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) 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 2-17

26 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. foam tape 9. Open the accessory box, and use this list to make sure you received all components of the VetScan HM5 system: VetScan HM5 Analyzer VetScan HM5 Operator's Manual (this document) VetScan HM5 Quick Reference Guide External power supply and power cord Mini-keyboard Five sample tube adapters Reagent tubing kit, including caps for reagent containers (with color-coded connectors) and reagent tubing Cleaning tube Thermal paper rolls (two) Warranty card (inside the Operator's Manual) Spare peristaltic pump tube assembly (store in a convenient location) 2-18 System Overview and Installation

27 Note: The VetScan HM5 requires a reagent pack to operate. A reagent pack (Abaxis Part No ) must be ordered for installation. Note: To start the warranty on the VetScan HM5, 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 2-26 for details). System Overview and Installation 2-19

28 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 analyzer s operation. Operating the analyzer at an altitude above 10,000 ft (3000 m) is not recommended Electrical Requirements The VetScan HM5 is powered by a standard wall outlet, and requires a power supply of v AC, Hz, 2 A (this is provided with the analyzer). To avoid power surges or drain, DO NOT plug the analyzer s power supply 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

29 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 VETSCAN HM5. 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 ( v AC) 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. System Overview and Installation 2-21

30 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

31 2.4.5 Connecting a VetScan Chemistry Analyzer (Optional) Connecting the VetScan HM5 with the VetScan Classic Chemistry Analyzer 1. Make sure the VetScan HM5 and the VetScan Classic are both turned off. 2. Connect a serial cable (Abaxis Part No ) to the serial port on the back of the VetScan HM5, and to the serial (1) port on the back of the VetScan Classic. 3. Turn on the VetScan HM5 and the VetScan Classic. serial port serial cable serial port VetScan HM5 VetScan Classic System Overview and Installation 2-23

32 Connecting the VetScan HM5 with the VetScan VS2 Analyzer 1. Make sure both analyzers are turned off. 2. Connect a USB Type A-B cable (Abaxis Part No ) to a USB Type A port on the back of VetScan HM5, and to a USB Type B port on the back of the VS2. 3. Turn on the VetScan HM5. VetScan HM5 USB cable VetScan VS2 4. When the VetScan HM5 has completed its start-up, turn on the VetScan VS2. USB ports (Type A on VetScan HM5, Type B on VS2) Note: To print VetScan Classic or VetScan VS2 Chemistry Analyzer results on the VetScan HM5, the VetScan HM5 must be configured properly. For instructions, see Combining Chemistry and Hematology Results on page Connecting the VetScan HM5 to a Computer The VetScan HM5 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 female-female null modem cable System Overview and Installation

33 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 HM5 software CD, version 1.05 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 1-3. Step 1 Installing the VetScan HM5 USB Driver 1. Insert the VetScan HM5 software CD (version 1.05 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 2-25

34 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, micro-bubbles 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 at least 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 18 inches (0.45 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

35 WARNING: DO NOT PLACE THE REAGENT PACK ABOVE THE ANALYZER (SUCH AS ON A SHELF). DO NOT DROP THE REAGENT PACK THIS CAN CAUSE MICRO- BUBBLES 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 six 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 Orange = Lyse2 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 2-27

36 5. Open the reagent pack box. Open each reagent container (lyse, Lyse2, 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. 8. Connect the tube that has no colored band to the Lyse2 bottle cap. Note: 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 Connecting the Waste Bottle 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. A temporary waste bottle is provided with your new VetScan HM5. Be sure to empty and then discard this container after using the first pack of reagents System Overview and Installation

37 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 HM5 on page Turning the Analyzer Off CAUTION: Never switch the analyzer off 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 2-29

38 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 soft-function 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 2-11) 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 five reagent tubing connectors (green, yellow, blue, white, and orange) from the reagent tubing intake valves, but leave the waste tubing (red) connected System Overview and Installation

39 4. When prompted, connect the cleaning tube kit to the reagent connectors, as shown. 5. When prompted, submerge the free end in a bottle containing at least 100 ml 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 2-31

40 2.6 Initializing the VetScan HM5 Use this procedure to initialize the VetScan HM5 after installation, or after it has been turned back on after an extended shut-down. 1. Press the Measurement key. 2. 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. 3. 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 analyzer s system settings, and make any needed changes see Configuring the VetScan HM5 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 System Overview and Installation

41 6. 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 micro-bubbles in the reagents during installation. 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 2-33

42 2-34 System Overview and Installation

43 Section 3 Configuring the VetScan HM5 This section describes how to configure the VetScan HM5 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 HM5 3-1

44 3.1 Printer Settings The following table summarizes the print settings available for the analyzer. These settings are usually configured whenever the analyzer is first set up, or when another printer is to be used. (Configuration procedures begin on page 3-2.) Table 3-1: Printer Settings Setting Printer Printout Mode Margins Font Size Autoprint Print warning flags Unit Color Print Histograms Limits Technical Info Description Auto-detects a variety of Hewlett-Packard printers connected to the HM5 by a USB cable. Normal or Fast. Default print margins. These can be customized to control the position of printed results on the page. Font size for the printout: Normal or Large. Select Yes to print all results automatically after each analysis. Select Yes to include all warning flags in the printouts. inch or cm. Select Yes to print in color. Select Yes to include histograms in results. Select Yes to print reference ranges and range bars for the parameters. Select Yes to print analyzer settings (probe voltages, lyse volume settings for the sample, and software version) in sample reports Configuring Printer Settings Before using the analyzer s built-in printer or an external printer, you must configure the analyzer for the printer as follows. 1. Press the key, then select Settings. 2. Select Printer settings. 3. Use the and keys to select the Printer you ll use (the default is Built in). 4. Press the ACCEPT soft key to accept the printer settings. 3-2 Configuring the VetScan HM5

45 The following lists available printer selections, along with language and models for each. Table 3-2: Compatible Printers Printer Selection Printer Language Supported Printer Models Seiko LPTH245 Built in Special printer language VetScan HM5 built-in thermal printer HP LaserJet PCL4 HP LaserJet series and compatibles (LJ1100, etc.) HP DeskJet PCL4 HP DeskJet series (select models only) Note: Other HP printers may function with the HM5, but have not been validated. For current 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: Margins: Left 0.00, Top 0.00 Font size: Normal VSx Same Page: Yes Autoprint: Yes (prints automatically after analysis; No requires you to press the Print key ) Print warning flags: Yes When using an external printer, the following default settings are recommended (settings are available only in software version 1 or greater): Left and Top margins: 0.39 (optimum setting when combining with VetScan chemistry results is 0.35) Font size: Normal Autoprint: Yes Print Warning Flags: Yes Histograms: Yes 6. Configure any other settings as needed. See the following table for a complete list. 7. Press the ACCEPT soft key to accept the settings. Configuring the VetScan HM5 3-3

46 3.1.2 Printout Examples Thermal Paper Printout Report header Patient information Sample ID (assigned automatically) Patient species Test date & time (assigned automatically) VetScan HM5 serial number Report date Test parameters Results falling out of range high or low are shown in boxes Reference ranges heavy line shows location of test results relative to reference range lower limit upper limit Test results WBC histogram Test result units inner two lines indicate upper and lower limits of normal EOS histogram RBC histogram PLT histogram 3-4 Configuring the VetScan HM5

47 8.5 x 11 Letter Printout Patient information Report header Patient species Report time and date (assigned automatically) and analyzer serial number WBC histogram Results falling out of range high or low are shown in boxes EOS histogram RBC histogram Test parameters PLT histogram Test results Test result units Reference ranges heavy line shows location of test results relative to reference range lower limit upper limit inner two lines indicate upper and lower limits of normal Configuring the VetScan HM5 3-5

48 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. 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 Font Size. Try using a larger Font Size. Enter the correct Paper size. Try decreasing the Left margin and Top margin. Enter the correct Paper size. Try increasing the Left margin and Top margin. Modify the Left margin. Modify the Top margin. 3-6 Configuring the VetScan HM5

49 3.2 Operational Settings Use the Customize menu to set the language and date format used by the VetScan HM5, along with the screensaver delay, and the date range used for printing combined results General Settings 1. Press the Utilities key. 2. Select Settings. 3. Select Customize. 4. Select General Settings. Configuring the VetScan HM5 3-7

50 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. The following table lists 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 Protocol version used by practice management software to interface with the analyzer. Contact Abaxis Technical Support (see page 1-3) 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 VetScan HM5 to communicate with a connected VetScan Classic or VetScan VS2 for combining results. If the VetScan HM5 is not connected to a VetScan Classic or VetScan VS2, choose Offline. If the VetScan HM5 is connected to a VetScan VS2 through a USB cable, choose USB. If the VetScan HM5 is connected to VetScan Classic through a serial cable, choose RS232. When printing combined results, the date format used: MM/DD/YY or DD/MM/YY (DD = date, MM = month, YY = year). Set both instruments to the same format. 3-8 Configuring the VetScan HM5

51 Table 3-4: General Settings (Continued) VetScan-HM5 combining within 365, 90, 60, 30, 15, 8, 5, 3, 1 day, Same day, Don t care. When printing combined results from the VetScan HM5 and VetScan Classic or VetScan VS2, defines the time range within which results will be paired: VetScan HM5 Patient ID number is paired with the Patient Number on the VetScan Classic, or with Patient ID on the VetScan 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 VetScan HM5 then matches the most recent results on the VetScan Classic or VetScan VS2. 6. Press the ACCEPT soft key to accept the settings Unit Settings Use the Units menu to set the measurement units the VetScan HM5 will use. 1. Press the Utilities key. 2. Select Settings. 3. Select Customize. 4. Select Units. Configuring the VetScan HM5 3-9

52 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/microliter (cells/µl), cells/liter prec. (cells/l prec.), cells/microliter prec. (cells/ul prec.) 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 HM5

53 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 VetScan HM5, and to be able to track individual usage of the unit, you can enable the analyzer s Multi-user mode. This requires 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 HM5 3-11

54 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 1-3) to reset the VetScan HM Configuring the VetScan HM5

55 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 VetScan HM5 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 the date format by pressing the desired number. 4. Select Set date and time. 5. Type in the date and time (use the and keys to select AM or PM), then press ACCEPT. 6. Select formats for displaying the date (item 2, 3, or 4) and time (item 5 or 6), then press EXIT. 7. Press ACCEPT to accept the settings. Configuring the VetScan HM5 3-13

56 3.4 Fluid Sensor Settings The fluid sensors allow the VetScan HM5 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. FLUID SENSORS1 4. If a message indicates that one or more fluid sensors are disabled, turn those sensors on, then select Calibrate sensors. FLUID SENSORS Configuring the VetScan HM5

57 Section 4 Test Procedure and Interpreting Results This section describes how to prepare and analyze samples with the VetScan HM5, 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

58 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 or tiger top, 2) green top, 3) lavender top. CAUTION: Use tubes containing potassium 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. Remove needle from syringe before dispensing blood into the EDTA tube. Do not use tuberculin syringes to draw blood samples. Filling blood tubes correctly: Each tube must be filled at least half full to avoid inaccuracy due to improper dilution of potassium EDTA. When using homemade containers pre-dosed with potassium EDTA, the final concentration of potassium 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 potassium 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

59 Note: Do not shake samples! Doing so can damage the blood cells, and can form micro-bubbles 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 10 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. Slowly 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

60 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 VetScan HM5 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 or kinked, or trapped between or beneath objects. 4-4 Test Procedure and Interpreting Results

61 3. Check the reagent lines (except the red waste line) for bubbles or air gaps. If any bubbles or gaps are present, 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 VetScan HM5 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 analyzer 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 analyzer displays BLANK needed, and a new blank must be run. 1. If BLANK needed appears on the screensaver, press the Measurement/Analysis key. The analyzer then runs a blank measurement, and displays the results. Test Procedure and Interpreting Results 4-5

62 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 EOS 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 0 to 0.1x10 3 cells/µl 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 VetScan HM5 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

63 4.3 Analyzing a Sample Use this general procedure to analyze samples with the VetScan HM5. 1. Prepare a well-mixed, potassium EDTA-preserved sample see Collecting and Preparing Samples on page 4-2 for details. 2. Select the appropriate sample tube adapter see below. Tube adapter (3 ml) Microtube adapter Controls/calibrator adapter Vacutainer adapter 3 ml control adapter 3. Place the adapter into the sampling rotor. 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 4. Press the Measurement/Analysis key, and select PT.ID. Test Procedure and Interpreting Results 4-7

64 5. Enter the species: view the available profiles using the and keys. Press the keypad OK key to move to the next field. 6. Enter the species and 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 VetScan HM5 in conjunction with the VetScan Classic, make sure the Patient ID number on the VetScan HM5 is the same as the Patient Number on the VetScan Classic. When using the VetScan HM5 with the VetScan VS2, make sure the Patient ID number on the VetScan HM5 is the same as the Patient ID on the VetScan 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. 7. 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. 8. Begin analysis by pressing the START button. Analysis is complete in less than four minutes. All results and histograms are automatically saved in the analyzer s database. 4-8 Test Procedure and Interpreting Results

65 9. When analysis is complete, a results screen displays all measured and calculated parameters as well as the WBC, EOS, RBC, and PLT histograms: You can view the results as follows: Use the VALUES and GRAPHS soft keys to move through the results. If the analyzer 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

66 4.4 Adjusting the Needle Height Adjust the VetScan HM5 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 desired needle height is displayed, press ACCEPT. Note: For most situations, a needle height of 0 mm is appropriate. Use the following 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

67 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 1-3. 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: n ml n+0.1 ml n+0.2 ml n-0.1 ml n-0.2 ml where the number n depends on the selected species profile. 3. When the correct volume is displayed, press ACCEPT. Test Procedure and Interpreting Results 4-11

68 4.6 Interpreting Results The VetScan HM5 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 shows a set of typical results and histograms: 4-12 Test Procedure and Interpreting Results

69 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 In 5-part differential mode, histograms display population distributions of each cell type: leukocytes (white blood cells WBC), eosinophils (EOS), 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 four histograms (WBC, EOS, 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: Lymphocytes (LYM), shown by the first peak in the histogram. WBC discriminator 1 discriminator 2 discriminator 3 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) (fl) LYM GRA MON Granulocytes (GRA), indicated by the peak to the right of the third discriminator. Test Procedure and Interpreting Results 4-13

70 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. Eosinophil Histogram (EOS) The distribution of eosinophils is shown by the second peak in the histogram. The first peak (dotted line) is the RBC ghost and other WBCs. Red Blood Cell Histogram (RBC) The distribution of red blood cells normally appears as a single, steep, bell-shaped 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. 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 Test Procedure and Interpreting Results

71 The most commonly identified anomaly in platelet histograms results from aggregated (clumped) platelets. This appears as a flattened, lumpy histogram. Note: Feline samples frequently demonstrate platelet aggregation (clumps). Careful sample collection and vortex mixing for up to 10 seconds can avoid or disaggregate clumps with no adverse effect on the sample. 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 3-1 on page 3-2). 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 VetScan HM5 automatically saves the most recent 2,000 test results (record 2,001 overwrites record 1, record 2,002 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. Test Procedure and Interpreting Results 4-15

72 4.7.2 Exporting Results You can export results through the VetScan HM5 s serial port or USB Type B port, using a software interface developed specially to communicate with the analyzer. Results can also be exported to AbaxisLab software, a software package for archiving and printing results from Abaxis instruments. For availability, check with Abaxis Technical Support (see page 1-3) 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 Combining Chemistry and Hematology Results The VetScan HM5 can print results from a connected VetScan Classic or VetScan VS2 analyzer. The VetScan HM5 can print the results from the other analyzer only, or the combined results for a given patient ID. (For connection instructions, see Connecting a VetScan Chemistry Analyzer (Optional) on page 2-23.) Printing VetScan Classic or VetScan VS2 results requires that the VetScan HM5 be properly configured see General Settings on page 3-7. 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 data, first display the CBC results to be combined on the VetScan HM5, then do the following on the VetScan Classic: 1. Press the RECALL key. The following appears: Press ENTER to Recall Results MENU for next option EXIT to leave Test Procedure and Interpreting Results

73 2. Press ENTER. The following appears: Recall:1 Results 1-Patient results 2-Patient flags ENTER to accept. 3. Press ENTER to select patient results. The following appears: 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. Transmitting Results from the VS2 to the HM5 When transmitting results from the VS2 to the HM5, be sure to select the VS2 analyzer printer and remove the VS2 printer paper from its holder to avoid printer errors. To transmit, uncheck Print and check Xmit in the Print Result screen. For details, see the VS2 user's manual or contact Abaxis Technical Support see Customer and Technical Support on page Combining Results During Testing 1. Use a USB cable to connect the printer to the VetScan HM5. 2. Turn on the printer, then turn on the analyzers. 3. Make sure the VetScan HM5 uses the same date format as the VetScan VS2 see Date and Time Settings on page Configure the VetScan HM5 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). Test Procedure and Interpreting Results 4-17

74 Set Autoprint to No. 5. Configure the VetScan HM5 s general settings as follows (see General Settings on page 3-7): Set PC Link to Offline. Set VSx Link to USB. Set VetScan-HM5 combining within as needed. 6. To print a VetScan VS2 result only: Note: Be sure to set VetScan-HM5 combining within the time frame so that no matching record will exist in the VetScan HM5. For example, if a VetScan analysis was run for a given patient 5 days ago, you would set VetScan-HM5 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 VetScan HM5 so that there will be no matching record. 7. To print combined VetScan HM5 and VS2 results: Note: The Patient ID of the VetScan HM5 record must match the Patient 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 VetScan HM5, and the VetScan HM5 prints out the combined results Test Procedure and Interpreting Results

75 4.8.3 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 Test Procedure and Interpreting Results 4-19

76 Thermal Paper Printout VetScan HM5 + VetScan Chemistry Results Report header Patient information Report date Test date & time (dates and times assigned automatically) Results falling out of range are shown in boxes Test parameters Test results WBC histogram Patient species VetScan HM5 serial number Sample ID (assigned automatically) Reference ranges Test result units heavy line shows location of test results relative to reference range lower limit upper limit inner two lines indicate upper and lower limits of normal EOS histogram RBC histogram PLT histogram VetScan results 4-20 Test Procedure and Interpreting Results

77 8 x 11.5 Letter Printout VetScan HM5 + VetScan Chemistry Results Report header Sample ID (assigned automatically) Patient information Patient species Report date (assigned automatically) and analyzer serial number Test parameters WBC histogram EOS histogram RBC histogram Test results and unit Reference ranges PLT histogram VetScan results Test Procedure and Interpreting Results 4-21

78 4.9 Using Prediluted Mode The VetScan HM5 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, call Abaxis Technical Support see Customer and Technical Support on page 1-3. Perform an external predilution of the sample using VetScan HM5 reagent diluent, or an isotonic saline solution. Make sure the diluent or saline is at room temperature before performing the dilution. Dilute the sample by a 1:6 ratio (1 part sample to 5 parts diluent) for example, 50 l sample to 250 l diluent or saline. Mix well. Note: The VetScan HM5 must be calibrated for prediluted mode before analyzing samples. The analyzer 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. Use diluent or normal saline to dilute the control blood. See Performing Quality Control on page 5-7 for instructions Test Procedure and Interpreting Results

79 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 then filled in.) 3. Press the ACCEPT soft key. 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. Test Procedure and Interpreting Results 4-23

80 5. Press the START button to begin the analysis. The VetScan HM5 automatically calculates the results with the 1:5 predilution factor. Note: The VetScan HM5 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: up to 50% 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 leukemia 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, often 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 4-25

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. 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 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 Normal MCV is consistent with non-regenerative anemia, often due to chronic disease. MCV should always be interpreted in light of other clinical data. Increase: most commonly the result of hemolysis Decrease: hypochromasia common in irondeficiency anemia and reticulocytosis In the anemic state, normal MCHC (with normal MCV) is consistent with non-regenerative anemia due to chronic disease. Decrease: hypochromasia common in irondeficiency anemia and reticulocytosis. Elevated RDW is typically indicative of anisocytosis. In the anemic state, increased RDW with an associated increase in MCV can indicate increased levels of immature RBCs. 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 Liver disease Parasites 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 Test Procedure and Interpreting Results 4-27

84 4-28 Test Procedure and Interpreting Results

85 Section 5 Calibration and Quality Control This section describes the calibration and quality control procedures for the VetScan HM5. Section Contents 5.1 Calibration When to Calibrate Required Calibration Materials and Handling Calibration Procedure Viewing Calibration History Resetting Calibration 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

86 5.1 Calibration When to Calibrate The VetScan HM5 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 normal control. WARNING: THE CALIBRATION PROCEDURE REQUIRES A SPECIAL CON- TROL MATERIAL THAT ARE DERIVED FROM HUMAN SOURCES. OBSERVE UNIVERSAL SAFETY PRECAUTIONS WHEN HANDLING THE CONTROL. 5-2 Calibration and Quality Control

87 5.1.3 Calibration Procedure WARNING: WE RECOMMEND WEARING LATEX GLOVES DURING THIS PROCEDURE. 1 Note: Make sure to bring the normal control up to room temperature before beginning this procedure. 1. Press the Utilities key, then select Calibration Select Run Calibrator. 3. Select the calibrator: Whole blood or Prediluted blood (1:5). Select Whole Blood if samples will be analyzed as undiluted samples. Select Prediluted blood (1:5) only if you are using prediluted mode. Note: You must recalibrate the analyzer if you switch between whole blood and prediluted modes. 1. The VetScan HM5 controls are derived from human sources. Observe universal safety precautions when handling the controls. See Calibration and Quality Control on page 5-1. Calibration and Quality Control 5-3

88 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 Note: Needle height must be at 0 mm. To adjust, press MENU, select Needle Height Setting, press OK repeatedly until needle height = 0 mm, then press ACCEPT. The display shows Calib (1/3) at upper left. 6. Mix the normal control by gently inverting the tube between thumb and forefinger 10 to 15 times. Remove the tube cap. 7. Place the normal control into the control tube adapter. 8. Press the START button. 9. When analysis is complete and the display shows the results, press ACCEPT. 5-4 Calibration and Quality Control

89 The display then shows Calib (2/3) 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. 12. When the second analysis is complete and the results displayed, press ACCEPT. The display then shows Calib (3/3) 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 and the results displayed, press ACCEPT. Calibration and Quality Control 5-5

90 The analyzer then displays the new calibration factors. Use the NEXT and PREV soft keys to move between pages. 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 Calibration and Quality Control

91 5.1.5 Resetting Calibration If calibration fails repeatedly, use the Reset Calibration function to bring the analyzer s calibration closer to factory settings. 1. Press the Utilities key, then select Calibration. 2. Select Reset Calibration. Press Yes to confirm. 5.2 Performing Quality Control The VetScan HM5 is programmed to monitor a single type of quality control (QC) material in three levels: high, normal and low. You should perform QC determinations regularly to verify continued optimal performance. We recommend testing control material 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 at least one of the following materials: High control Low control Normal control 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 Set QC reference ranges. Calibration and Quality Control 5-7

92 3. From the package insert, enter the lot no., expiration date, and assay and gap values for each test parameter. Assay values precede +-. Gap values follow +-. 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. 4. Press ACCEPT to accept the data, then press ACCEPT again to confirm. 5. Mix the control vial by gently inverting the tube between thumb and forefinger 10 to 15 times, and remove its cap. 6. Put the control vial onto the control vial adapter on the sample rotor. 7. Select Run QC to start the analysis. Press the START button. The results screen displays Quality Control as ID. 8. 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. 9. Abaxis strongly recommends performing three successful QC runs after calibrating the instrument. 5-8 Calibration and Quality Control

93 5.2.3 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 VetScan HM5 allows you to do this easily using a database table and histograms 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 GRAPHS, then press the Print key. \Calibration and Quality Control 5-9

94 5-10 Calibration and Quality Control

95 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

96 6.1 Database Overview The analyzer can store up to 2,000 complete test results in its internal database. Results are stored chronologically by date and time. (When more than 2,000 measurements are stored, the most recent result replaces the oldest result.) 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 22 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 results of a patient record, press the DETAILS soft key. To view the WBC, EOS, RBC and PLT histograms, press the GRAPH 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 DETAILS, then press Print. 6-2 Managing the Database

97 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

98 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

99 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 Running the Stress Test Replacing the Peristaltic Pump Tube Assembly Maintenance & Service 7-1

100 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

101 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 distilled water to gently rub the lower surface of the wash head to remove any salt build-up. 5. When finished, close the side door, then press the ACCEPT soft key. Once this procedure is complete, the screensaver reminder disappears, and reappears after one week. Maintenance & Service 7-3

102 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

103 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: VetScan HM5 cleaning tube (shown on page 2-11) household-strength chlorine bleach (typically 5 6% hypocholorite) 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 (for a final concentration of 0.25% hypocholorite). In addition to the bleach solution, you will need at least 100 ml of distilled water for rinsing. Maintenance & Service 7-5

104 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, Lyse2, 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, Lyse2, 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

105 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

106 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, Lyse2, 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. Loosen the caps on the containers in the new reagent pack. 5. Loosen the caps (with tubing still attached) on the containers in the old reagent pack (including the waste container). 6. Use a lint-free wipe moistened with distilled water to clean and connect each reagent drop-down tube as follows: a. Grasp the cap containing the reagent tube, and pull it up and out of the old reagent bottle. b. Use the damp lint-free wipe to remove any lint or dirt from the drop-down tube connected to the inside of the cap. c. Place the tube into the appropriate color-coded bottle in the new reagent pack. 7. Re-cap the used reagents with their original caps, and dispose of them in an environmentally appropriate manner. Save the used diluent container to use as the waste container. 8. Prime all reagent lines: a. Press the Utilities key, then select Maintenance. b. Select Priming, then select Prime all. 9. Reset the reagent status: a. Press the Utilities key, then select Maintenance. b. Select Reagent Status. 7-8 Maintenance & Service

107 c. Press the CHANGE soft key, then press ACCEPT to confirm. The analyzer updates the installation date, reagent lifetime, and 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). 10. 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 normal control see Performing Quality Control on page 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. Maintenance & Service 7-9

108 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 Running the Stress Test In Stress Test mode, the analyzer performs measuring cycles continuously without a sample (blank measurements). This verifies the function of the fluidic or pneumatic system, and also provides information about the stability, cleanliness, Hgb readings, and PLT readings as a measure of analyzer function. To pass this test, results should match an acceptable blank run (see page 4-5). Consult technical support if the results do not become acceptable within 5 10 cycles. 1. Press the Utilities key, and select Diagnostics. 2. Select Stress 10 Cycles. To exit the stress test before 10 cycles complete, press the START until the current cycle is completed. or OK button 7-10 Maintenance & Service

109 7.8 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 1-3. 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

110 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

111 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

112 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

113 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 1-3. They will guide you through activating the profile, and help you to customize it to suit your requirements. Special Functions 8-3

114 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 is shown at upper left in 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

115 Section 9 Troubleshooting Use the information in this section to help diagnose and solve problems with the VetScan HM5. 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

116 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, Q, and Y 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, B, and X 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 4-22). Run a quality control procedure if needed (see Performing Quality Control on page 5-7). Z is caused by a possible lysing problem. Make sure there is adequate 5D lyse reagent in the container, and prime if needed (see Check Reagent Status and Tubing on page 4-4). An asterisk (*) near a parameter indicates uncertainty. A variety of causes are possible. If a result warning flag is present, refer to Results Flags 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. 9-2 Troubleshooting

117 9.1.4 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. 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, clean the wash head, 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 1-3. Low-temperature reagents (mainly diluent) can cause this. Be sure to allow reagents to reach room temperature. p PLT blank is high, or no PLT 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 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 1-3. Troubleshooting 9-3

118 b m 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, clean the wash head, 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 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 1-3. Results are out of linearity range. contact Abaxis Technical Support see page 1-3. 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 1-3. L Potential lyse-resistant RBCs or RBC ghosts or pathological condition in canine samples Table 9-1: Result Warning Flags Flag Meaning Description / Recommended action Re-run the sample at a higher lyse volume (call Technical Support) or perform a smear to examine the cells. W Extremely large cells Abnormally high number of very large cells in feline samples. This may indicate clumping of cells or a pathological condition. Vortex sample for 5 10 seconds at high speed and re-run. Perform manual smear to examine cells. X EOS blank is high Check to see if wash head needs cleaning. If cleaning is needed, clean the wash head and repeat the blank measurement. If the blank is too high, perform 3 to 5 cleaning cycles and repeat the blank measurement. Y EOS clogging Lyse2 aperture clogging. Perform 3 to 5 cleaning cycles and repeat the measurement. If the problem persists, contact Abaxis Technical Support see page 1-3. Z Noise in the EOS channel Possible lysing problem. Make sure there is adequate Lyse2 reagent in the container, and prime if needed (see Check Reagent Status and Tubing on page 4-4). 9-4 Troubleshooting

119 9.2 Error Messages Below are a few very common error messages and solutions to the errors. Error message: Check 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. Make sure the tubing in the reagent pack is not kinked. 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 7-8. Troubleshooting 9-5

120 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. The EOS histogram (shown at right) overlaps in size with the WBC histogram, but its results are derived from a different measurement. The EOS histogram displays leukocytes that have not been lysed by the Lyse2 reagent (eosinophils). Lysed WBCs and lysed RBCs appear in the faint peak on the far left. EOS EM fl Histogram 400 fl 9-6 Troubleshooting

121 9.3.1 Examples 1 High Blank, Flag p This flag is seen when the EOS blank is somewhat high, between 0.1 to 0.2. Evaluation The EOS histogram shows a large number of cells that may invalidate this blank measurement. EOS blank values > 0.2 are rejected and prevent blood measurements. 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 1-3. Troubleshooting 9-7

122 2 EOS clogging, flag Y A Y flag indicates a clog or partial clog in the aperture during the EOS measurement. Evaluation The above histograms indicate the following: The clogs are detected through changed in the probe voltages. The probe voltages are displayed on the upper right of the histograms on the graphs screen. Y flag shows at the bottom of the screen. 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 Troubleshooting

123 3 Noisy EOS channel, flag Z The Z flag indicates that the measured EOS value is higher than the measured GRA population from the WBC histogram. This flag can occur if the Lyse2 is contaminated or if there is underlysing in the Lyse2 lysing step. Evaluation The above histograms indicate the following: EOS histogram indicates a very large number of cells. This flag can appear when control blood is run in the wrong profile or the wrong species profile is selected. Troubleshooting 9-9

124 Solution 1. Make sure the correct species profile is selected for the measurement. 2. If the profile is correct, in very rare cases the sample can contain a rare pathology: perform a manual smear to confirm the results. 3. If the WBC histogram is shaped normally, the problem is likely underlysing with Lyse2. Check for bubbles in the Lyse2 tubing and/or sufficient solution in the bottle. Prime Lyse2 if needed. Replace the reagent pack if the Lyse2 fluid is low. 4. If you run a blank and the EOS value is high, the Lyse2 solution could be contaminated. Confirm this by re-running the blank. If the EOS blank does not decrease after several cleaning cycles, replace the reagent pack after bleaching. 5. If the problem persists, contact Abaxis Technical Support see Customer and Technical Support on page Troubleshooting

125 4 Lack of Separation between RBC and LYM peaks, flag L The L flag indicates when there is significant overlap between the RBC and the WBC histogram. Evaluation The above histograms indicate the following: An obvious peak of RBCs intrudes into the left side of the WBC histogram. This flag is often seen when the sample contains lyse-resistant RBCs. Solution 1. Perform a manual smear and examine with a microscope to confirm the cell morphology is normal. 2. Contact Abaxis Technical Support (see Customer and Technical Support on page 1-3), and as directed, increase the lyse volume by 0.1 or 0.2 ml to achieve proper lysis of RBCs for this sample. (Lyse volume returns to default for the next sample.) 3. If the increased lyse volume does not yield a normally shaped WBC histogram, the sample may contain an underlying pathology. Perform a manual smear to check cell morphology and differential. Troubleshooting 9-11

126 5 Unusually High Number of Large Cells, flag W When the WBC histogram indicates a large number of extremely large cells, the sample is flagged W. The flag may indicate large numbers of highly clumped PLT or a pathological condition. Evaluation The above histograms indicate the following: Normal WBC histograms should descend to zero near 200 fl, while a W flagged sample may have a WBC histogram extending to near 400 fl. Solution 1. Vortex the sample at high speed for 5 10 seconds and re-run the sample. Be sure not to vortex too much as the resulting foam can damage cells and distort cell counts. 2. Perform a manual smear to confirm the presence of the large cells Troubleshooting

127 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. foam tape 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 9-13

128 9-14 Troubleshooting

129 Section 10 Specifications This appendix contains technical specifications for the VetScan HM5 system, and lists its linearity ranges. Section Contents 10.1 VetScan HM5 Specifications Linearity Ranges Precision Specifications 10-1

130 10.1 VetScan HM5 Specifications Chambers Chambers Reagent system Aperture diameter Throughput Sampling method 25 µl of whole blood in 3-part mode, 50 µl of whole blood in 5-part mode mode. 50 µl of whole blood in 5-part prediluted mode of 1:5; 25 µl of prediluted blood in 3-part prediluted mode of 1:5. 1 unified chamber for diluting whole blood and counting. VetScan HM5 Reagent pack. 80 µm tests/hour in 3-part mode, tests/hour in 5-part mode. Open tube system with automatic sample rotor, support for all HM2 and VetScan HM5 test tubes. Sample types 5-part mode: dog, cat, horse, control. 3-part mode: mouse, rat, rabbit, ferret, pig, cattle, monkey (monkey is research only). Clog prevention High-voltage pulse on aperture in each analysis cycle, chemical cleaning and high-pressure back-flush of the aperture using cleaner reagent. Cleaning procedure High-voltage burn of the aperture, high-pressure back-flush, chemical cleaning of the aperture. Quality control Support for 5-part differential control blood. QC parameters include: mean, ± range, SD and CV for all measured and calculated parameters, 16- and 64-day Levy-Jennings charts, separate QC database. Calibration 3-measurement automatic or manual calibration of WBC, HGB, RBC, PLT, MCV, RDW, MPV, and EOS/BAS absolute. Multi-user feature Three-level Multi-user operation with selective privilege levels, user identification with ID and password. User interface Easy-to-use, menu-driven user interface with 6 hard-function buttons, graphic icons, and on-screen help. Languages available English. For other languages, contact Abaxis Technical Support see page 1-3. Data capacity 2000 results, including RBC, PLT, WBC 3-part and 5-part histograms onboard. Data can be saved to USB flash drive or downloaded to computer. Host computer interface 1x USB slave port or 1x RS-232 on real panel. Support for ASCII-based communication protocol only (V3.1). Data back-up method Port for USB mass storage device (flash drive) on front panel. Software upgrade method USB flash drive or CD-ROM. External printer interface USB. Built-in printer Easy Paper Operation built-in thermal printer. Display 240x128-dot, high-contrast, backlit, graphics liquid crystal diode. Keypad 29 foil keys + START button. External keyboard Standard PS/2-compatible keyboard. Power requirement 12v DC, 6 A, 72 W. Power supply unit External, auto-ranging power unit for or VAC, 50 60Hz. Operating temperature F (15 30 C). Optimal temperature is 77 F (25 C). Dimensions (W x D x H) 12.6 x 10.2 x 14.4 in (320 x 260 x 365 mm). Net weight 27 lbs (12.3 kg) Specifications

131 10.2 Linearity Ranges The VetScan HM5 is guaranteed to provide specified accuracies within its linearity range when properly calibrated and maintained. 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 EOS 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 EOS cells/liter RBC cells/liter PLT cells/liter Specifications 10-3

132 10.3 Precision The following precision parameters were established using normal level control on nine instruments over five days, with ten replicate measurements per day. Table 10-3: Control Parameter Precision Parameter Mean SD %CV WBC % RBC % EOS % HCT % HGB % MCV % PLT % MPV % 10-4 Specifications

133 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.3.5 Eosinophils A-8 A.4 Normal Hematology Ranges (Dog, Cat, Horse, and Others). A-9 A.5 Veterinary Hematology References A-10 Introduction to Veterinary Hematology A-1

134 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. A-2 Introduction to Veterinary Hematology

135 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

136 A.3 Blood Cell Parameters A.3.1 Red Blood Cells, Hemoglobin Red blood cells RBC are formed in the bone marrow. A mature dog red blood cell is nonnucleated, and has a mean corpuscular volume (MCV) of approximately 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 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. 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

137 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 Three-Part Differential Method 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 Elevated WBC counts can affect other differential parameters. It is highly recommended that abnormal WBC and other cell differentials be confirmed by a manual blood smear, as is the case with all automated hematology analyzers. 1 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). 1. Bessman, JD. Automated Blood Counts and Differentials. A Practical Guide. (Baltimore: Johns Hopkins University Press, 1986), p Introduction to Veterinary Hematology A-5

138 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% A-6 Introduction to Veterinary Hematology

139 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. Introduction to Veterinary Hematology A-7

140 A.3.5 Eosinophils Eosinophils EOS are granulocytes that are specialized to attack parasites, such as worms and protozoa. They are also the primary effector cells in allergic symptoms. They can be identified morphologically by the presence of eosin (red) -staining granules. Normal EOS concentrations can range from 0 7% of the total WBC count. EOS EM fl Histogram 400 fl A-8 Introduction to Veterinary Hematology

141 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 Cow Pig Mouse WBC 10 9 cells/l LYM% % MID% % GRA% % EOS% % RBC cells/l HCT % MCV fl RDWcv % HGB g/l MCH pg MCHC g/l, g/dl, mmol/l PLT 10 9 cells/l MPV fl Parameter Unit Rat Rabbit Ferret Monkey User_1 User_2 WBC 10 9 cells/l LYM% % MID% % GRA% % RBC cells/l HCT % MCV fl RDWcv % HGB g/l MCH pg MCHC g/l, g/dl, mmol/l PLT 10 9 cells/l MPV fl Introduction to Veterinary Hematology A-9

142 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-10 Introduction to Veterinary Hematology

143 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 Three-Part Differential Method B-3 B.2.3 Five-Part WBC Differential Method B-5 B.3 Hemoglobin Determination B-7 B.3.1 Hemoglobin Determination by Photometry B-7 B.4 Measured and Calculated Values B-8 B.5 Measured and Calculated Values B-9 Operating Principles B-1

144 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

145 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 Three-Part Differential Method To perform a three-part WBC differential count, the RBCs must first be lysed since RBCs are typically 1000 times more concentrated in normal blood than are WBCs and would interfere with WBC counting if left intact. The lysing also serves to release the HGB, which is stored in the RBCs, for direct analysis in the solution. 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. Operating Principles B-3

146 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). Three-Part Differential B-4 Operating Principles