37.5
cells/square
750,000
cells/mL
750,000
cells/mL
750,000
cells
750,000
cells
37.5
cells/square
750,000
cells/mL
750,000
cells/mL
750,000
cells
750,000
cells
The Hemocytometer Calculator converts your raw cell counts from a hemocytometer (Neubauer chamber) into a concentration in cells per milliliter. This counting chamber is one of the most widely used tools in cell biology laboratories for determining cell density before experiments, passages, or cryopreservation.
Enter the total number of cells you counted, the number of large squares examined, and any dilution factor applied (such as Trypan Blue), and the calculator will give you the cell concentration instantly.
Each large square on a standard hemocytometer has a volume of 0.1 µL (0.0001 mL). The formula is:
Cells/mL = (Total cells counted / Number of squares) × Dilution factor × 10,000
The factor of 10,000 converts from the 0.1 µL square volume to 1 mL. The dilution factor accounts for any dilution performed before loading (e.g., 1:2 with Trypan Blue gives a dilution factor of 2).
Inputs
Results
Counting 150 cells across 4 squares with a 1:2 Trypan Blue dilution gives 7.5×10⁵ cells/mL. The average of 37.5 cells per square is in the ideal range of 20-50.
Inputs
Results
With a 1:10 dilution and 500 cells across 5 squares, the original concentration is 1×10⁷ cells/mL. The high per-square average suggests a greater dilution would improve accuracy.
Count at least 4 of the large corner squares (each 1 mm²) for reliable results. If cell density is low, count all 5 large squares (4 corners + center). Aim for 100-200 total cells counted for good statistical accuracy. If you count fewer than 100, increase the number of squares or reduce dilution.
The ideal range is 20 to 50 cells per large square. If you see fewer than 20, your sample may need less dilution or more counting squares. If you see more than 50, dilute further before loading the hemocytometer to improve counting accuracy.
Each large square on a Neubauer hemocytometer has dimensions of 1 mm × 1 mm × 0.1 mm depth, giving a volume of 0.1 µL or 1×10⁻⁴ mL. Multiplying by 10,000 (10⁴) converts from this 0.0001 mL chamber volume to cells per 1 mL.
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