Flow Cytometry Calculator
Enter the name and event count for each gated population. The calculator will give percentages of parent and total events.
| Population Name | Events in Gate | |
|---|---|---|
Calculate absolute cell counts using counting beads or known sample volume.
Calculate how much antibody to add per sample based on the recommended dilution and number of cells.
Estimate spillover and compensation requirements for a multi-color panel. Enter the fluorochrome name and its spillover percentage into neighbouring channels.
Results
How to Use the Flow Cytometry Calculator
This calculator has four modules. Population % takes your total acquired events, parent gate (e.g. live cells), and individual gated populations and returns the percentage of parent and percentage of total for each. Absolute Count converts percentage data into real cell numbers per mL using either counting beads or a known sample volume. Antibody Volume calculates how much antibody to add to each staining tube based on your recommended dilution, staining volume, and number of samples. Compensation estimates the controls needed and their cell requirements for multi-color panels.
Key Formulas
Compensation Controls
Compensation corrects for spectral overlap between fluorochromes. For each fluorochrome in your panel, you need a single-colour control — either antibody-capture beads stained with that antibody, or cells known to be positive for that marker. Each control tube should have the same number of cells as your experimental samples. An unstained control is also required as a baseline reference.
Tips for Accurate Flow Data
- Always include a viability dye to exclude dead cells from analysis — dead cells autofluoresce and bind antibodies non-specifically.
- Use fluorescence-minus-one (FMO) controls to set gates accurately in multi-color panels.
- Run compensation controls on the same day as the experiment using the same instrument settings.
- Aim for at least 10,000 events in the parent gate for statistically robust population analysis.
- Keep cells on ice and process quickly — surface markers internalise and degrade at room temperature.