Overview
The TC20™ automated cell counter has two counting modes. Depending on the sample type, the user-defined gates can be either enabled or disabled.
Samples with Multiple Cell Populations (stem cells and other primary cells)
User-defined gates must be enabled. Once the user-defined gates are enabled, a histogram will be displayed at the beginning of each count and the user can select the population of interest by adjusting the position of the cell size gates. Only objects within the designated range will be analyzed as cells; those with diameters outside of the size range will be excluded from the cell count.
If the user-defined gates are disabled, the cell counting algorithm will automatically determine the population to count without any user input. The algorithm will count the cells of the most predominate cell type that typically comprise the largest peak, which may not be the population the user wants to count.
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| Gating setup window. | Cell size gating window. |
To make counting of such samples easier, when counting multiple sample replicates, the TC20 cell counter can save the positions of gates and apply them to subsequent counts (choose Yes in the “Use saved gates” field in Gating setup).
If the Image preview is On, an image of cells will be displayed during gating. In the image, only cells with diameters between the selected size gates will be marked with a yellow dot.
Immortal Cell Lines
When counting immortal cell lines or other samples that are composed of cells with similar cell size, the user-defined gates can be disabled. The cell counting algorithm then automatically identifies the cell population of interest without requiring user input.
The TC20 automated cell counter can accurately count a broad range of cell types:
- Immortal cell lines
- Primary cells from tissue or blood (peripheral blood mononuclear cells, primary lymphocytes)
- Stem cells
The TC20 counter can count cells with a round shape after trypsinization and a cell diameter between 6 and 50 µm.
These cells were counted using the TC20 automated cell counter either by Bio-Rad scientists or customers. If you do not find your cell name in this list, please contact us at 1-800-424-6723 (Technical Support).
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Launch the Gallery |
The data below show the TC20™ automated cell counter is compatible with a broad range of cell types, such as cell lines, primary cells (from tissue or blood), and stem cells. The TC20 cell counter can count cells with a 6–50 µm cell diameter and within a broad concentration range of 5 x 104–1 x 107 cells/ml, which eliminates the need to dilute cells, thus reducing the error associated with sample dilutions prior to counting.
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| The TC20 automated cell counter demonstrates accurate cell counts across a range of cell sizes. Small (PBMC, Jurkat), medium (HeLa), and large (MEF) cells were counted with a hemocytometer, a TC20 automated cell counter, and a competitor's image-based automated cell counter. The TC20 counter and hemocytometer cell counts showed no statistically significant differences. Precision is indicated by the standard deviations; error bars represent average standard deviations. Cell counts on the TC20 counter were performed on one instrument with four sample replicates. |
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| The TC20 automated cell counter demonstrates accurate counts of viable cells. Pan T cells mixed with trypan blue (1:1) were counted with a hemocytometer, a TC20 automated cell counter, and a competitor’s image-based automated cell counter. The TC20 counter and hemocytometer cell counts showed no statistically significant differences. Precision is indicated by the standard deviations; error bars represent average standard deviations. Cell counts on the TC20 counter were performed on four different instruments with five sample replicates. |
The TC20™ automated cell counter uses microscopy with auto-focus that analyzes multiple focal planes to identify the best plane. The cell counting algorithm then uses the image acquired from the best focal plane to identify cells and exclude debris, thereby calculating the total cell count.
Using auto-focus instead of subjective manual focusing is especially important when assessing cell viability because an incorrectly selected focal plane will lead to inaccurate results.
If trypan blue is detected along with the total cell count, the TC20 counter assesses cell viability. The conventional method of analyzing viability using a single focal plane can lead to inaccurate conclusions because light scattering and the alignment of cells at different heights in a counting chamber can change the appearance of cells — live cells may appear to be dead and vice versa. To determine if cells are viable, the TC20 counter analyzes each cell using images acquired from multiple focal planes during the focusing step.
| Effect of multifocal plane analysis on live/dead count accuracy for HeLa cells. | ||
| Viability Assessment | Live Cells, % | Dead Cells, % |
| Single best focal plane* | 35 | 65 |
| Multiple focal planes* | 65 | 35 |
View, Export, and Analyze Your Data
| After viewing cell count results, users can choose to view an image of the counted cells on the TC20™ automated cell counter with the ability to zoom in on an image with the click of a button. |
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After a count, a JPEG file of the cell image is available for immediate export to a USB flash drive or, to save every cell image, automatic data export can be enabled. Each cell image contains a metatag with the count data and can be further analyzed using TC20 data analyzer software under Download Tab. This optional software can be used to review single images, perform multi-file comparative analyses, and generate reports. |
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Results from 100 counts are stored in the TC20 cell counter so you can always go back and recover counts based on their acquisition date and time stamp. Previous count results can be exported via the USB port and opened in a Microsoft Excel spreadsheet, allowing easy comparison of results between experiments. |
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You can assign a custom sample name that will be associated with the count result and the JPEG image. To save time when working with multiple sample replicates, enable the automatic name serialization option to populate numerically serialized names. |
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The optional thermal label printer can be connected to the TC20 counter to print results onto labels that can be placed into a laboratory notebook for traceability of the count results. |
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Users can access the onboard dilution calculator to determine volume adjustments required to achieve the cell concentration needed for the next experiment. |
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Mammalian tissue or cell culture techniques were developed in the early 1900s as a method of maintaining animal organs outside of the body. During the 1940s and 1950s cell culture technology emerged as an essential tool to support virology research by enabling vaccine production through viral purification. Today, cell culture is used worldwide as a model for a variety of studies, including intra- and intercellular communication, tissue/organ development or organization, and different disease states, such as cancer, Parkinson's disease, and diabetes.
In this guide the most common cell types used in cell biology are described.
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Mammalian tissue or cell culture techniques were developed in the early 1900s as a method of maintaining animal organs outside of the body. During the 1940s and 1950s cell culture technology emerged as an essential tool to support virology research by enabling vaccine production through viral purification. Today, cell culture is used worldwide as a model for a variety of studies, including intra- and intercellular communication, tissue/organ development or organization, and different disease states, such as cancer, Parkinson's disease, and diabetes.
This presentation provides an introduction to growing and maintaining mammalian cells in culture.
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Basic Cell Culture Introduction |
