Seeding Density Calculator
Select Vessel Format
c/mL
mL
c/mL
Please enter a target seeding density and media volume.
Know how many cells you added? Find the resulting seeding density for any vessel.
mL
Please enter cells added and media volume.
Estimate when cells will reach confluency based on seeding density and doubling time.
c/mL
hrs
%
Please fill in seeding density, doubling time, and confluent density.
🌱
Results
How to Use the Seeding Density Calculator
Select Seed Vessel to calculate how many cells and what volume of suspension to add to a vessel based on your target density and stock concentration. Select Find Density from Count to work backwards — enter how many cells you added to find the resulting density. The Confluency Estimator predicts when cells will reach your target confluency given a seeding density and doubling time.
Seeding Density Formula
Cells Needed = Target Density (cells/mL) × Media Volume (mL)
Volume from Stock = Cells Needed ÷ Stock Concentration (cells/mL)
Cells/cm² = Cells Added ÷ Growth Area (cm²)
Standard Vessel Growth Areas and Media Volumes
| Vessel | Growth Area (cm²) | Recommended Media | Typical Seeding (cells/cm²) |
|---|---|---|---|
| 96-well | 0.32 | 0.1–0.2 mL | 5,000–20,000 |
| 24-well | 1.9 | 0.5–1 mL | 10,000–50,000 |
| 12-well | 3.8 | 1–2 mL | 20,000–80,000 |
| 6-well | 9.5 | 2–3 mL | 50,000–200,000 |
| 60mm dish | 20 | 4–5 mL | 100,000–400,000 |
| 100mm dish | 57 | 8–12 mL | 300,000–1,000,000 |
| T-25 | 25 | 5–7 mL | 100,000–500,000 |
| T-75 | 75 | 12–20 mL | 300,000–1,500,000 |
Tips for Consistent Seeding
- Always count cells immediately before seeding — density changes within minutes after trypsinisation.
- Mix cell suspension thoroughly before taking aliquots to ensure uniform distribution.
- For multi-well plates, prepare a master mix and dispense equal volumes per well for consistency.
- Rock the plate gently in a cross pattern (not circular) to distribute cells evenly across the well.
- Let plates sit flat on the bench for 30 seconds before placing in the incubator to prevent cells pooling in the centre.
- For transfection experiments, seed at 70–80% of the confluency density to ensure cells are still dividing at transfection.