Team:EPF-Lausanne/Protocol/TrypanBlue

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| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''Sampling'''
| align="center" style="background:#f0f0f0;"|'''Sampling'''
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#Take xµL ofPBS in 96 well plate
#Take xµL ofPBS in 96 well plate

Revision as of 14:24, 13 August 2012

Protocol: Trypan Blue Method

' Sampling ' ' ' '
DilutionsCell Density ( 10^6/ml)DilutionPBS (µL)Cells (µL)Trypan Blue(µL)
1-241005050
2- 4.581252550
4.5 - 7121201545
>716137.512.550
  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