Flow Cytometry Sample Concentration Planner

What This Tool Does

This planner helps you estimate the cell concentration and dilution needed to achieve a target number of events per sample in flow cytometry. It relates:

• Desired events per sample to the required cell concentration
• Starting concentration to a simple dilution plan (stock + diluent)
• Optional flow rate to estimated acquisition time and event rate

Core Relationship

The fundamental relationship between events and concentration is:

Rearranging to find the required concentration:

Dilution Factor

The dilution factor indicates how much to dilute your starting sample:

• Dilution factor ≥ 1: You can dilute the stock to reach the target
• Dilution factor < 1: Required concentration is higher than starting; you would need to concentrate the sample

Volume Calculations

For each prepared sample:

• Prep Volume: Acquisition volume × (1 + overage%)
• Stock Volume: Prep volume ÷ Dilution factor
• Diluent Volume: Prep volume − Stock volume

The overage ensures you prepare enough volume to account for pipetting losses.

Flow Rate & Acquisition Time

If you provide the instrument flow rate (µL/min), the tool estimates:

• Acquisition time: Acquisition volume ÷ Flow rate
• Event rate: Achievable events ÷ Acquisition time (in seconds)

These are rough estimates; actual rates depend on instrument settings and sample quality.

Limitations

This tool provides simple concentration and dilution math. It does NOT:

• Recommend markers, antibody panels, or fluorophore combinations
• Design gating strategies or compensation matrices
• Set voltages, thresholds, or instrument parameters
• Account for doublets, debris, or dead cells
• Interpret flow cytometry data or suggest clinical cutoffs

Key Assumptions

• Each recorded event corresponds approximately to a single cell
• Cell suspension is homogeneous and accurately counted
• No significant cell loss during sample handling
• Acquisition volume equals the volume actually processed by the cytometer