Enter the average cells per square, the dilution factor, and the volume of the square to determine the cell density.
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Cell Density Formula
The following formula is used to calculate cell density (cell concentration) from a hemocytometer count.
CD = ACSQ * DF / VC
- Where CD is the cell density (cells per mL, if VC is in mL)
- ACSQ is the average number of cells counted per square
- DF is the dilution factor (this is 1 if the sample has not been diluted at all)
- VC is the volume represented by 1 counted square (mL)
To calculate a cell density, multiply the average number of cells per square by the dilution factor, then divide by the volume represented by one counted square.
If you are using an Improved Neubauer hemocytometer and your “square” is the 1 mm × 1 mm large square with a 0.1 mm chamber depth, then VC = 0.0001 mL and the formula becomes CD = ACSQ × DF × 10,000.
Cell Density Definition
What is cell density?
Cell density is a unit of measure that describes the total number of cells (often reported as viable/living cells) per unit volume.
What is the cell density of human tissue?
There is no single “average” cell density for human tissue because it varies widely by tissue type, cell size, and how the measurement is defined (e.g., nuclei counts vs. intact cells). For context, human blood contains roughly 4–6×109 red blood cells per mL, while many mammalian cell culture suspensions are commonly worked with in the 105–107 cells/mL range.
How to calculate cell density?
The following example outlines how to calculate the cell density of a sample.
First, determine the average number of cells per square under the microscope. This is usually the average of 4 or more squares. In this example, the average number of cells per square is 500.
Next, determine the volume represented by each counted square. For an Improved Neubauer hemocytometer large square (1 mm × 1 mm) with a 0.1 mm depth, the volume per square is 0.0001 mL.
Next, determine the dilution factor. In this problem, there was no dilution, so the dilution factor is 1.
Finally, calculate the cell density using the formula above:
CD = ACSQ * DF / VC
CD = 500 * 1 / 0.0001
CD = 5,000,000 cells / mL.
