Cell seeding density calculator
Maintained by the Talindrew team · Last updated
The number of cells to seed is the target density times the vessel's growth area: cells = cells/cm² × cm². A 6-well plate well is 9.6 cm², so seeding at 2 × 10⁴ cells/cm² takes 1.9 × 10⁵ cells per well; dividing by your stock concentration gives the volume to pipette.
Result
cells = density × growth area. Areas follow Corning's growth-area chart (e.g. 6-well 9.6 cm², T-75 75 cm²): check your vendor's insert, areas differ slightly between brands.
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What it looks like
Formula
cells = density (cells/cm²) × growth area (cm²) · volume = cells / stock (cells/ml)
Worked example
Given: Six wells of a 6-well plate at 2 × 10⁴ cells/cm², stock at 1 × 10⁶ cells/ml
- 1.Per well: 2 × 10⁴ × 9.6 = 1.92 × 10⁵ cells.
- 2.Six wells: 1.15 × 10⁶ cells total.
- 3.Stock volume: 1.15 × 10⁶ / 1 × 10⁶ = 1.15 ml, split across the wells in complete medium.
1.92 × 10⁵ cells per well; 1.15 ml of stock for the plate
How the calculation flows
Units & constants
| 96-well | 0.32 cm² per well |
|---|---|
| 24-well | 1.9 cm² per well |
| 6-well | 9.6 cm² per well |
| 100 mm dish | 56.7 cm² |
| T-25 / T-75 / T-175 | 25 / 75 / 175 cm² |
| Source | Corning growth-area chart; areas vary slightly by vendor |
| Price | Free |
How many cells per well? Seeding chart by vessel
| Vessel | Growth area | 5,000 cells/cm² | 10,000 cells/cm² | 20,000 cells/cm² | 50,000 cells/cm² |
|---|---|---|---|---|---|
| 96-well plate (per well) | 0.32 cm² | 1,600 | 3,200 | 6,400 | 16,000 |
| 48-well plate (per well) | 0.95 cm² | 4,750 | 9,500 | 19,000 | 47,500 |
| 24-well plate (per well) | 1.9 cm² | 9,500 | 19,000 | 38,000 | 95,000 |
| 12-well plate (per well) | 3.8 cm² | 19,000 | 38,000 | 76,000 | 190,000 |
| 6-well plate (per well) | 9.6 cm² | 48,000 | 96,000 | 192,000 | 480,000 |
| 35 mm dish | 8.8 cm² | 44,000 | 88,000 | 176,000 | 440,000 |
| 60 mm dish | 21.5 cm² | 107,500 | 215,000 | 430,000 | 1,075,000 |
| 100 mm dish | 56.7 cm² | 283,500 | 567,000 | 1,134,000 | 2,835,000 |
| 150 mm dish | 152 cm² | 760,000 | 1,520,000 | 3,040,000 | 7,600,000 |
| T-25 flask | 25 cm² | 125,000 | 250,000 | 500,000 | 1,250,000 |
| T-75 flask | 75 cm² | 375,000 | 750,000 | 1,500,000 | 3,750,000 |
| T-175 flask | 175 cm² | 875,000 | 1,750,000 | 3,500,000 | 8,750,000 |
cells = density × growth area. Areas follow Corning's growth-area chart; other brands differ slightly, so check the insert for your plates.
What seeding density should you use?
It depends on the line and the experiment's timeline. Adherent workhorses (HEK293, HeLa, CHO) are commonly seeded at 1–5 × 10⁴ cells/cm² to reach 70–90% confluence in 2–3 days. Seed low for long experiments and clonal growth; seed high (≥ 10⁵/cm²) when you need a monolayer tomorrow. Transfection protocols usually specify the target confluence at transfection time: work backwards from it using the doubling-time calculator.
Why calculate per cm² instead of per well?
Density per area is the transferable number: 2 × 10⁴ cells/cm² behaves the same in a 96-well well and a T-75, while 'cells per well' silently changes meaning between formats. Protocols written per area scale to any vessel with one multiplication, which is exactly what this calculator does.
Frequently asked questions
A 96-well plate well has 0.32 cm² of growth area, so multiply your target density by 0.32: 10,000 cells/cm² is 3,200 cells per well and 30,000 cells/cm² is 9,600. In 100 µl per well, that is a suspension of 32,000 or 96,000 cells/ml.
A T-75 flask has 75 cm² of growth area: 10,000 cells/cm² is 750,000 cells and 20,000 cells/cm² is 1.5 million. The right density depends on the cell line and how many days until you passage.
3,200 per well (0.32 cm² × 10⁴), so 3.1 × 10⁵ cells for a full plate before overage. Add ~10% extra volume for multichannel dead volume.
Close but not identical, a '6-well' well is 9.6 cm² for Corning and 9.0–9.6 cm² elsewhere. For density-sensitive work use the area printed in your plate's certificate.
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