How to Count Cells An Overview of Cell Counting Methods
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- The microscope is then focused on an area of the counting chamber and the cells are counted using a tally counter. This is typically done using the 1 mm2, 100 nl area of the counting chamber and a 4x or 10x objective, but the precise area and objective used will depend on the size of your cells and their density in suspension. This process is generally repeated using four different 1 mm2 areas and the results are averaged. If determining cell viability, separate counts should be made for live and dead cells, with the dead cells appearing blue due to the permeability of their damaged membranes to trypan blue.
- Fig 1 Schematic representation of a hemocytometer grid.
- Automated cell counters were designed to be a faster, easier, automated alternative to manual counting. They use the same principles of operation as hemocytometers; they perform multiple counts of cells within a known area and average out the results. They also can discern live cells from dead cells using dye exclusion methods (such as trypan blue). Automated cell counters may either operate as standalone devices or require a connection to a computer. In addition to a cell count, most counters also provide statistical information on cell size. Benefits & Drawbacks: Manual cell counting is the least expensive method of determining cell count and cell density, however it is also the slowest and most tedious. It can also be one of the least reliable due to the possibility for human error, particularly if performing many cell counts sequentially, and performing a large number of cell counts can also cause eye strain. If a lab's needs to count cells is sporadic or infrequent, and if a high degree of precision in the cell count is not required, then hemocytometers can be a good choice. For more frequent use, when greater accuracy is required, or in higher-throughput applications, counting chambers fall short. Additionally, human counters are often poor at discerning between multiple types of cells in a suspension unless the differences in size and / or shape between the various cell types are extreme.
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