Team:EPF-Lausanne/Protocol/TrypanBlue

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Protocol: Trypan Blue Method

Sampling

Cell Density (10^6/ml) Dilution PBS (µl) Cells (µl) Trypan Blue (µl)
1 - 2 4 100 50 50
2 - 4.5 8 125 25 50
4.5 - 7 12 120 15 45
> 7 16 137.5 12.5 50


  1. Take x µL of PBS in a 96-well plate.
  2. Add the required volume of cell culture. Mix once.
  3. Bring the plate back to the microscope, add Trypan Blue to the PBS + cell mixture just before counting the sample.

Trypan Blue is toxic to cells. If left too long with it, even healthy cells will die.


Calculation of LCD :

LCD = Cell Count/(100*4)*Dilution

Tips :

  1. Mix cells before sampling.
  2. Take the cell sample from the top of the liquid.
  3. Mix Trypan Blue in the PBS + cell solution slowly before loading.