BioDeviceHub

Cells per cm² Seeding Density Calculator

Convert a seeding density in cells per cm² into the number of cells for a flask, dish or well, using nominal growth areas for common culture vessels.

Formula

Nvessel=Agrowth×ρseedN_{\text{vessel}} = A_{\text{growth}} \times \rho_{\text{seed}}
Vsuspension=NvesselcstockV_{\text{suspension}} = \dfrac{N_{\text{vessel}}}{c_{\text{stock}}}
AgrowthA_{\text{growth}}
surface available to the cells, in cm²
ρseed\rho_{\text{seed}}
cells per cm² at the time of seeding

How it works

Adherent cells respond to how crowded the surface is, not to the volume of medium, so seeding is specified per unit area. Multiplying the density by the growth area of the vessel gives the cells to add, and the same density then transfers between vessel sizes.

The growth areas offered are nominal values. They differ slightly between manufacturers, so the field can be edited to match your plasticware.

Worked example

Seed three T-75 flasks at 10⁴ cells/cm² from a suspension at 1.06 × 10⁶ cells/mL.

  1. Cells per flask = 75 cm² × 10⁴ = 7.5 × 10⁵.
  2. All three = 2.25 × 10⁶ cells.
  3. Suspension per flask = 7.5 × 10⁵ / 1.06 × 10⁶ per mL = 707.5 µL.

Add 7.5 × 10⁵ cells, about 708 µL of suspension, to each flask and top up with medium.

These are the values the calculator opens with, so you can check its output against this example.

Assumptions

  • Cells attach and spread evenly over the whole growth area.
  • The nominal area matches your vessel.

Common mistakes

  • Using the total area of a multi-well plate instead of the area of one well.
  • Carrying a cells-per-well number from one plate format to another. Convert through cells per cm².
  • Not rocking the vessel after seeding, which leaves cells concentrated in the centre.

Related tools

Cell Seeding CalculatorSuspension and medium for a plate of wellsOpen calculatorCell Confluence CalculatorEstimated confluence and time to reach a targetOpen calculatorSplit Ratio CalculatorSuspension and medium per vessel when passagingOpen calculatorHemocytometer Cell Counting CalculatorCells per mL from a hemocytometer countOpen calculator

Related equipment

Service documentation, failure modes and parts for the instruments this calculation is used with.