Microscopic Cell Counting
Microscopic Cell Counting - Related Videos
10:19
Using a Hemacytometer to Count Cells
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2023
Many biomedical experiments require manipulation of a known quantity of cells, in order to achieve accurate, reproducible, and statistically-relevant data. Therefore, learning how to count cells is a particularly essential technique for any successful biomedical scientist. The most common way to count cells is by using a hemacytometer - an instrument that bears two laser-etched grids, which aid in the enumeration of an aliquot cells under a simple light microscope. This data can then be used to...
05:47
Counting and Determining the Viability of Cultured Cells
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Cited by 19 •
2008
Determining the number of cells in culture is important in standardization of culture conditions and in performing accurate quantitation experiments. In this video, we demonstrate how cells are counted using a hemacytometer.
06:37
Counting Human Neural Stem Cells
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Cited by 14 •
2007
Knowledge of the exact number of viable cells is required for many tissue culture manipulations. This protocol describes how to differentiate between live and dead cells and quantify cells using a hemacytometer. Although it describes counting human neural stem/precursor cells (hNSPCs), it can be used for other cell types.
11:01
Automated Quantification and Analysis of Cell Counting Procedures Using ImageJ Plugins
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Cited by 90 •
2016
This paper describes the quantification of hemocytometer and migration/invasion micrographs through two new open-source ImageJ plugins Cell Concentration Calculator and migration assay Counter. Furthermore, it describes image acquisition and calibration protocols as well as discusses in detail the input requirements of the plugins.
12:25
Counting Proteins in Single Cells with Addressable Droplet Microarrays
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Cited by 2 •
2018
Here we present addressable droplet microarrays (ADMs), a droplet array based method able to determine absolute protein abundance in single cells. We demonstrate the capability of ADMs to characterize the heterogeneity in expression of the tumor suppressor protein p53 in a human cancer cell line.
FAQs
The hemocytometer’s calibrated grid and known chamber depth establish the volume represented by the counted squares. After cells in the defined area are tallied, that count is converted to cells per unit volume using the chamber geometry. This makes the result quantitative rather than simply an estimate of how many cells appear in the microscope field.
A dilution factor is essential when the original suspension cannot be counted directly at a suitable concentration. The observed count applies to the diluted material, so the calculated concentration must be adjusted to represent the starting sample. Recording the dilution preserves the connection between the microscope measurement and the actual cell concentration used in experiments.
Viability dyes add a functional distinction to the numerical count: cells classified as living are separated from those identified as nonviable. Consequently, the measurement can report more than total cell abundance, allowing investigators to assess the viable portion of a suspension. That distinction is particularly relevant when deciding how much culture to use for a reproducible experiment.
A typical workflow begins with a cell suspension, transfers it into the hemocytometer, views the calibrated grid under a microscope, and counts cells in the defined squares. The count is then converted to cells per unit volume, with dilution included when applicable. Keeping these stages linked prevents the raw microscopic tally from being mistaken for the final concentration.
In cell culture maintenance, the resulting concentration helps determine how much suspension is needed for a planned seeding setup. Rather than relying on an unmeasured volume, researchers can use the counted concentration to prepare comparable starting conditions across experiments. This supports reproducibility when cultures are maintained or established for subsequent biological analysis.
Measurements collected during culture growth monitoring can reveal changes in cell abundance over time. Because the method produces a concentration expressed per unit volume, results from samples can be compared quantitatively rather than only described visually. When viability information is included, interpretation can also distinguish changes in total cells from changes in the living fraction.