Team:EPF-Lausanne/Protocol/TrypanBlue

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Protocol: Trypan Blue Method



Sampling Dilutions {|class="wikitable" style="text-align: center; color: black;" |Cell Density |Dilution |PBS ( µL) | Cells( µL) |Trypan Blue

|- | ||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 xµL ofPBS in 96 well plate
  2. Add required volume of cell culture. Mix once
  3. Bring the plate back to 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 trypan blue, even healthy cells will die.


Calculation of LCD :

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

Tips :

  1. Mix cells before sampling
  2. Take cell sample from top of the liquid
  3. Mix trepan blue into the PBS + cel solution slowly and well before loading