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Protocol: Trypan Blue Method
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| align="center" style="background:#f0f0f0;"|Sampling
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| Dilutions||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
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| ||4.5 - 7||12||120||15||45
|-
| ||>7||16||137.5||12.5||50
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- Take xµL ofPBS in 96 well plate
- Add required volume of cell culture. Mix once
- 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 :
- Mix cells before sampling
- Take cell sample from top of the liquid
- Mix trepan blue into the PBS + cel solution slowly and well before loading
