Calculate cell concentration and viability from hemocytometer grid counts using the Trypan Blue exclusion method for accurate cell culture analysis.
Viable Cell Concentration
--
Total Cell Concentration
--
Cell Viability
--
Total Viable Cells
--
Total Cells in Suspension
--
A hemocytometer is a specialized microscope slide with a precise grid pattern used to count cells in suspension. It consists of large and small squares with known volumes, allowing accurate calculation of cell concentration.
The standard dilution factor is 2, achieved by mixing equal volumes of cell suspension and 0.4% Trypan Blue solution (1:1 ratio). This allows viable cells to remain unstained while dead cells appear blue.
Count at least 4 large squares (1mm x 1mm each) for statistical accuracy. If cell density is very low, count 5 squares. For very high density, you may need to dilute further and count fewer squares.
Cell viability above 95% is considered excellent, 90-95% is good, 80-90% is fair, and below 80% is poor. Low viability may indicate cell stress, contamination, or improper handling.
The factor 10,000 converts cells per large square to cells per mL. It accounts for the volume of one large square (0.1 μL) and converts to mL: 1/(0.1 × 10⁻³) = 10,000.
Viable cells have intact membranes and exclude Trypan Blue, appearing bright and refractile. Dead cells have compromised membranes, take up the dye, and appear blue under the microscope.
If counts are too high (>30 cells per square), dilute further. If too low (<10 cells per square), use a higher concentration or count more squares for better statistical accuracy.
Yes, this calculator works for any cell type counted with a standard hemocytometer. Adjust the dilution factor based on your specific staining protocol and cell concentration needs.