CASY Model TT E L E C T R I C A L C U R R E N T EXCLUSION ECE BASED CELL ANALYSIS

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1 CASY Model TT Cell Counter + AnalyZer E L E C T R I C A L C U R R E N T EXCLUSION ECE BASED CELL ANALYSIS

2 ELECTRICAL CURRENT EXCLUSION CASY Model TT Cell Counter + Analyzer R&D Easy and fast adaptation to an unlimited number of applications combined with a non-invasive measurement principle, are key features that make the CASY Model TT an indispensable tool for cell culture analysis in R&D. Different precision measurement capillaries, together with cell-specific setups, allow for the measurement of all types of mammalian cells, as well as bacteria, yeast, algae, parasites, pollen and sperm. MS-Excel bases data transfer software provides complete documentation and flexible data evaluation to follow the metabolic state of cells during any type of cell culture experiment. P r o c e s s Development Product Description The CASY Cell Counter + Analyzer System Model TT determines not only the cell number but also the size distribution and viability of a cell sample quickly and reliably. In addition, the aggregation level in the cell sample is determined and automatically included in the determination of the cell concentration. The basic results provided are: Cell concentration Cell viability Cell volume Cell aggregation Cell debris Improving cell culture processes requires fast and accurate data about the quality of the cells. In addition to cell concentration, CASY Model TT monitors cell viability, cell size and aggregation. Also, cell debris has become an increasingly important quality control parameter in process control. With a measurement time of only 10 seconds for a single measurement, different parameters that indicate the state of the cell culture can be thoroughly tested in a short time. CASY Model TT Measuring Results Example

3 ECE -BASED CELL ANALYSIS Benefits Electrical Current Exclusion (ECE ) is the only well-accepted method for the dye-free determination of cell viability. Viability Measurement by Electrical Current Exclusion, ECE Together with the short measurement time and the low sample volume, CASY - Technology offers a unique combination of features for quality control of cell cultures that makes it fast, easy to use, flexible and precise. Comparison of Count and Volume Distribution of CHO Cells with an Aggregation Factor of 1.19 Simultaneous quantification of small debris, dead cells, viable cells and aggregates. Accurate and reproducible results that can be directly compared from user to user, day to day and CASY system to CASY system based on the factory certified calibration. Non-invasive method for determination of cell viability, avoiding the cytotoxic side effects of dyes. Highly precise measurement of cell volume. Correct quantification of even heavily aggregated cell samples. Low sample volume (5 100 µl). Low measurement time (10 sec). Fully qualified for use in clean rooms. Low running cost per sample. Nearly maintenance-free. Calibration identical for all devices and stable for the entire service life. Quality of Factory Certified Calibration Size Distribution of 10 µm Test Beads

4 ELECTRICAL CURRENT EXCLUSION CASY Model TT Cell Counter + Analyzer H i g h R e s o l u t i o n Size Distribution The high resolution size distribution offered by CASY -Technology ensures that the size of every individual cell or particle passing through the measuring pore can be detected. A clear-cut differentiation between cell debris, dead cells, viable cells and cell aggregates is provided, and even volume differences of a fraction of a femtoliter can be detected. Cell volume is a very sensitive parameter that can be used to detect changes in a cell culture. Therefore, a visualized finger-print of the metabolic state of the cell culture is provided. Typical CASY High Resolution Size Distribution Technology CASY Model TT is based on the international accepted reference method for cell counting, ISO In addition, the system is equipped with an advanced method for digital pulse processing called Pulse Area Analysis. This technique is used to get complete information from the signals created when a cell or particle passes through the high precision measuring pore in a low voltage field. Each single object is scanned with a frequency of one million measurements per second, resulting in up to hundreds of readings per cell. Thus, the complete time course of the signals is taken into account when analyzing samples. Pulse Area Analysis provides a unique, high resolution recording of the cell size distribution and an outstanding dynamic measurement range. It is the basis for the dye-free, non-invasive determination of cell viability. Pulse Area Analysis How it works

5 ECE -BASED CELL ANALYSIS Product Number CASY Modell TT Product No Data Management The data transfer program, CASY excell, is an easy-to-use MS-Excel based tool for saving and viewing measurement date acquired with the CASY Model TT Besides easy scrolling of all measurements saved in the same data file it allows visual comparison of the size distributions with a control measurement and the alternatively display of the size distribution on a count or a volume scale. Technical Data Measurement principle Electronic pulse area analysis with 1 Mio. measurements / sec. Viability determination Electrical Current Exclusion (ECE ) Dynamic range in volume > 1: in diameter > 1:40 Measured size channels Displayed size channels 400 Measurement range µm Resolution 1 in Typical analysis time 10 sec. Graphic User Interface of CASY excell Typical sample volume µl Interfaces RS 232 (DB9), Parallel (DB25)

6 ELECTRICAL CURRENT EXCLUSION CASY Model TT Cell Counter + Analyzer Applications With the CASY Model TTC there are no limitations concerning the type of cell and the type of application. The system allows to follow each type of cell culture experiment and is recommended to be used for the quality control, the improvement and the longterm-monitoring of biological processes. Typical applications include: Counting of all types of cells Control of cell seeding Success or failure of many cell culture experiments depend on seeding the cells in the correct concentration and in good shape at the very beginning. Precise determination of the cell concentration and the state of the cells therefore is indispensable for cell seeding control. Determination of cell proliferation Cell proliferation strongly depends on the conditions used to grow up the cells. Even minimal changes in the composition of the medium or the addition of stimu lating or inhibiting substances can effect the rate of cell proliferation considerably. Therefore the measurement of cell proliferation is a standard method to optimize cell culture processes. Determination of cytotoxic effects on cells 125 Measuring of Cytotoxicity: IC50 - Determination With a size range of 0.7 to 160 µm, nearly all types of cells can be measured. This includes bacteria, yeast cells, algae cells, protozoa and all types of mammalian cells. The CASY high resolution size distribution also provides important information about the metabolic state of the cells, in addition to counting cells. Decrease in Cell Viability (% of Control) WST-8 Neutral Red Trypan Blue CASY-ECE 0,001 0,01 0, Concentration of Cytotoxic Agent (mm, log) The sensitivity of any cytotoxicity assay depends on the efficiency of quantification of the reproductive integrity of cells (inhibition of cell proliferation) and the structural integrity of cells (cell death). Based on the high resolution size distribution, as well as Electrical Current Exclusion, CASY Model TT determines both effects of toxic substances on cells independently.

7 ECE -BASED CELL ANALYSIS Application Support Our application support team has 20 years of experience with all kinds of cell culture applications. A huge application data base helps to answer all questions coming up fast and allows to provide you with typical measurement examples from our comprehensive cell library. Based on this information our team is well prepared to assist you in establishing new applications and to develop adapted data evaluation solutions. Monitoring of virus infections of cells Consumables Further information see page 88/89. Baculovirus infections of insect cells are used to produce large amounts of protein in infected cells. During infections, cell volume increases dramatically. Precise volume determination allows the user to monitor the process and calculate the degree of infection (MOI).

8 ELECTRICAL CURRENT EXCLUSION CASY Models TT Cell Counter + Analyzer Consumables CASY ton CASY ton is a ready-to-use isotonic dilution liquid for cell cultures especially developed for use with CASY Cell Counter + Analyzer Systems. The particle purity of the liquid guarantees a low background to the measurements. CASY ton is manufactured according to the strict standards of the quality management system ISO 9000 ff. Properties CASY ton reduces aggregation of cells in the measuring cup and stabilizes the cells for the duration of the measurement time. The defined conductivity guarantees consistent measurements. Physiological ph and osmolarity ensure the preservation of cell size. Principle CASY supplies are especially designed for standardized CASY equipment. Only the exclusive and regular use of CASY supplies guarantees long-term consistency of measurement results, as well as long-term stability and optimal functionality of the equipment. Description Packaging CASY ton is available directly from innovatis AG in convenient 10 liter containers, including a plastic dosage tap. Shelf-life and Storage The best-before date is indicated on the product packaging. CASY ton can be stored at room temperature. CASY ton is an isotonic, particle-pure, odorless and colorless saline solution. Waste Disposal Waste must be disposed of in accordance with local cell culture laboratory regulations. Product Number Product No

9 ECE -BASED CELL ANALYSIS ceramus Dispenser 2 10 ml The ceramus Dispenser, made by Hirschmann Laborgeräte, Eberstadt, Germany, is the digitally adjustable top bottle dispenser for the dosage of liquids that innovatis recommends for use with CASY ton. Principle Packaging The ceramus Dispenser is delivered with an individual certificate of performance. The package contains several adapters for different types of bottle necks. One of the adapters fits the inlet of the CASY ton container. A manual explains all of the necessary aspects of installation, handling and cleaning. The ceramus Dispenser allows CASY users to easily adjust the dose of CASY ton, in steps of 0.25 ml. The delivered discharge tube unit is equipped with a Luer lock adapter. Therefore it is possible to rinse each portion of CASY ton through a particle filter with a defined pore size compatible to the capillary used with the CASY system. Additional Features The ceramus Dispenser can be autoclaved (refer to the manual for instructions). The recirculation valve allows for priming in order to avoid the accumulation of air bubbles in the CASY ton. Properties The permanent use of the ceramus dispenser allows for a highly reproducible adjustment of the liquid volume of CASY ton needed for every measurement. In combination with a connected particle filter, this guarantees optimum quality in purity and volume precision. Operation Conditions The ceramus dispenser is suitable for operating temperatures between +4 C and +50 C. Product Number Product No

10 ELECTRICAL CURRENT EXCLUSION CASY Models TT Cell Counter + Analyzer Consumables CASY clean CASY clean is a ready-to-use cleaner for the CASY Cell Counter + Analyzer Systems. Principle CASY supplies are especially designed for standardized CASY equipment. Only the exclusive and regular use of CASY supplies guarantees long-term consistency of measurement results, as well as long-term stability and optimal functionality of the equipment. Properties Regular use protects the device and measurement capillaries from biological deposits. Through its emulsifying effect, CASY clean prevents blockages and encrustations of the internal liquid container system and measurement capillaries. Packaging CASY clean is available directly from innovatis AG as a package of 3 x 500 ml bottles. Description CASY clean is a foam-free, alkaline, odorless and colorless cleaning solution for the system and measurement capillaries. CASY clean is non-toxic and non-flammable. Shelf-life and Storage The best-before date is indicated on the product packaging. CASY clean should be stored in a cool, dry place and should be protected from light. Waste Disposal Waste must be disposed of in accordance with local cell culture laboratory regulations. Product Number Product No

11 ECE -BASED CELL ANALYSIS CASY blue CASY blue creates a standardized control sample of dead cells. Principle CASY supplies are especially designed for use with standardized CASY equipment. Only the use of CASY blue guarantees a standardized viability determination with the CASY Cell Counter + Analyser Systems of innovatis AG. Description CASY blue is a ready-to-use, blue, alcoholic dilution for the determination of the cell matrix volume. CASY blue acts highly specific and is used together with the CASY Cell Counter + Analyzer Systems of innovatis AG for establishing cell specific setups for viability determination and for the ongoing quality control of cells. Utilization Use CASY blue on a regular basis to check the cursor settings defined for viability determination by ECE and for the on-going quality control of your cells to guarantee cell identity. CASY blue covers all TQC requirements of cell culture labs in research and production. Please find the detailed standard operating procedure (SOP) for using the CASY blue enclosed. Packaging CASY blue is available directly from innovatis as a packaging unit of 4 x 30 ml bottles. Shelf-life and Storage The best-before date is indicated on the product packaging. CASY blue should be stored in a cool, dry place, protected from light. Close bottle tightly after use. Waste Disposal Waste must be disposed of in accordance with local cell culture laboratory regulations. Product Number Product No

12 ELECTRICAL CURRENT EXCLUSION CASY Models TT Cell Counter + Analyzer Consumables CASY cups CASY cups are measurement cups with lids designed for use with CASY Cell Counter + Analyzer Systems. The material properties, size and volume of CASY cups are optimally adapted for cell culture sample preparation. CASY cups are disposable and designed for single use. Repeated use can have an effect on measurement results. Principle CASY supplies are especially designed for use with standardized CASY equipment. Only the exclusive and regular use of CASY supplies guarantees long-term consistency of measurement results, as well as long-term stability and optimal functionality of the equipment. Description Packaging CASY cups with lids are available directly from innovatis in packages of 1,800 cups. Shelf-life and Storage CASY cups must be stored at room temperature in a dry, dust-free location. The shelf-life is unlimited. CASY cups cups measure 22 x 65 mm with a filling volume of 15 ml. They are made of Polypropylene (PPN), and are transparent and milky in color. CASY cups are particle-pure and biologically inert. The material prevents cells from adhering to the cup walls. The cups are heat-resistant up to 80 C and slowly inflammable. CASY rack for 24 pcs CASY cups. Waste Disposal Waste must be disposed of in accordance with local cell culture laboratory regulations. Product Number CASY cups CASY rack

13 ECE -BASED CELL ANALYSIS

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