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Frequently Asked QuestionsCellometer Frequently Asked Questions

Frequently Asked Questions about Cellometer Auto T4

Q: What is the principle behind the Cellometer® automated cell counter?

A: The basic principle of Cellometer® automated cell counters is image-based cytometry. It is the same as the hemacytometer in terms of spreading cells into a thin layer of liquid, as in the CHT4. The total counting area for the Auto T4 model, including area A and B, is equivalent to all four corner squares of the traditional hemacytometer.

Cellometer automates the cell counting by taking images of cells in the counting chamber, analyzing the number of cells in each image using image analysis software, and converting the number of cells into cell concentration.

Q: Does Cellometer measure viability?

A: Yes, when trypan blue is added to the sample, live cell concentration and viability are both measured.

Q: What concentration of trypan blue should be used?

A: A stock solution of 0.2% trypan blue in PBS is recommended

Q: What are the limits of cell concentration (minimum and maximum) for accurate analysis?

A: The limits of cell concentration depend on the cell size to certain degree. Large cells have a lower minimum, and small cells have a higher maximum. The common range for reliable counting is from (0.25 - 10) x 106cells/ml. For small cells, the maximum can go up to 15 x 106cells/ml.

Q: What is the size range for Cellometer Auto T4?

A: 5 microns to 60 microns.

Q: Does Cellometer Auto T4 count trypan blue positive and negative cells at the same time in the same sample?

A: Yes, if trypan blue stained cells are loaded into the counting chamber and the "Test Viability" option is selected, both trypan blue positive and negative cells are measured at the same time, from the same sample.

Q: Can Cellometer Auto T4 measure heterogeneous cell populations?

A: Yes, cells are spread into a thin layer within the disposable counting chamber. The same sample can be measured multiple times using different parameters.

Q: How does Cellometer Auto T4 count clumpy cells?

A: Cellometer cell counting software recognizes the bright centers of viable cells within clumps. Clumps can be de-clustered to count individual cells or remain counted as one, depending on the user's selection.

Q: Can Cellometer® Auto T4 count cells based on size?

A: Yes, Cellometer® Auto T4 is an image-based cell counter. The user can set minimum and maximum size for counting from 5 microns to 60 microns.

Q: Will cell debris be counted?

A: No. In a typical cell culture, cell debris is smaller in size and different in morphology than live cells. Cellometer counting software excludes debris.

Q: What is the minimum quantity of sample required for counting?

A: The disposable counting chamber requires 20 microliters of sample.

Q: How reproducible are the results?

A: The CV for various cell types tested is less than 10%, including sampling variability.

Q: How many types of cells can the Auto T4 count?

A: Auto T4 uses size and morphology for counting. Currently, more than 300 types of cells have been counted. The list is growing each day.

Q: Does Cellometer® Auto T4 measure irregular shaped cells?

A: Yes. Cellometer® software can be set to count cells of irregular shapes.

Q: Do I need any special buffer or reagent for cell counting?

A: No special buffer is required. Cells suspended in growth media or PBS are routinely counted. Trypan blue is used to stain dead cells.

Q: How does Cellometer® Auto T4 count different size cells?

A: Through cell type parameters, the user defines the size range for counting. Depending on the application, different sized cells can be included or excluded. The same cell image can be analyzed for different cell types.

Q: How do I set up parameters for a new cell type?

A: There are three options for setting up a new cell type:

Option 1: Input from "Cell Library." Currently, the "Cell Library" in the Cellometer software consists of more than 300 cell types.
Option 2: Use the "New Cell Type Wizard." The Cellometer software will guide you through an automatic set-up process.
Option 3: Save the cell image and upload it to the Nexcelom web site. Our technical support team will analyze cell images, set up the cell type, and e-mail it back to you.

Q: Does Cellometer Auto T4 measure cell size?

A: Yes. Cell size is measured from the image. Mean cell diameter is included in the cell counting results. Diameters of all counted cells and a cell size histogram are included in the "Analysis" function.

Q: Does Cellometer® Auto T4 need calibration?

A: No. The counted volume is determined by the chamber height and area.

Q: Does Cellometer® Auto T4 need maintenance?

A: No. There is no liquid handling system inside the Auto T4. Required maintenance is similar to a microscope.

Q: How long does the light source last?

A: The light source is a light-emitting diode which has a very long lifetime. It should last for the life of the instrument.

Q: How do I verify the counting results?

A: After counting, you can verify results by showing counted cells. Green circles indicates live cells, while the red circles indicate dead cells.

Q: Do I need a computer to operate the Cellometer Auto T4?

A: Yes. Cellometer Auto T4 is connected to a PC via USB cable. An existing PC in your lab running other instruments can be used. (Computer checking procedure). A lap-top PC with factory-loaded Cellometer software is also available for purchase.

Q: What are the Computer Requirements?

A: Computer Recommendations:
Windows XP (SP3), Vista, or Windows 7
1.6 GHz or higher speed processor
2GB RAM recommended
USB 2.0 port (required)
1024 x 768 pixel display (or higher)

Q: Is the disposable cell counting chamber special?

A: Cellometer® counting chamber is specially designed as an integral part of the counting system.

Q: Can I save cell concentration data?

A: Cell counting data is saved in the file indicated in the box located in the lower left corner of the software. Each time you click "Save Data", the just counted data is saved in the file indicated in the box. The data can be reviewed and printed or exported to Excel for further analysis.

Q: Can I save cell images?

A: Yes, cell images can be saved for reference. They are also used in our technical support. When you send us your image, we can observe and analyze the sample data.

Q: How fast can I count one sample?

A: Counting time depends on the number of cells and morphology of the sample, as well as the computer speed. Typically, cell lines require less than 30 seconds per sample.

Q: Does it automatically mix the sample with trypan blue?

A: No, the trypan blue mixing is performed by the user prior to loading the disposable counting chamber. This is the same procedure that is used to count cells with a manual hemacytometer.

Q: Does it count Primary cells and Cell lines?

A: Most mammalian cell lines and cultured primary cells can be counted. Please refer to cell list.

Q: What is the advantage of Cellometer over traditional, flow based automatic cell counting systems?

A: Cellometer requires only 20 microliters of sample. There is no liquid handling, no clogging, and no daily maintenance. The counting time is less than 30 seconds for a typical sample. Images can be viewed for visual verification and archived.

Q: What is the advantage of Cellometer over hand-held counters and other image-based automatic cell counters?

A: Cellometer Auto T4 can measure viability based on the trypan blue dye exclusion method. The Cellometer pattern-recognition software is superior at handling cell clumps and irregular-shaped cells. The Cellometer software is designed to differentiate and count cells based on size.

Q: Can Cellometer count platelets?

A: A special model Cellometer Auto M10, which is designed with a higher optical magnification than the Auto T4, can be used to count platelets.

Q: Can Cellometer count PBMC?

A: The Cellometer Auto 2000 is recommended for analysis of PBMCs from heterogeneous samples. Cultured PBMCs can be analyzed with the Cellometer Auto T4.

Q: Can Cellometer count yeast?

A: Yeast cells are typically smaller than 5 microns and are best counted using the Cellometer Auto M10, Auto X4 (10x), or Vision (10x). The Auto M10 is recommended for simple yeast counting. The Auto X4 (10x) is recommended for yeast concentration and viability in clean samples. For yeast concentration and viability in more complex samples, the Cellometer Vision (10x) is recommended.

Q: Can Cellometer count algae?

A: The Cellometer Vision CBA (10x) is recommended for algae.

Q: Can Cellometer count bacteria?

A: Bacteria are too small to be counted by Cellometer Auto T4 cell counters. Some bacteria may be counted with the Cellometer Vision (10x) Cell Analyzer. Please contact Nexcelom for more information.