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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 |
- Take x µL of PBS in a 96-well plate.
- Add the required volume of cell culture. Mix once.
- 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 :
- Mix cells before sampling.
- Take the cell sample from the top of the liquid.
- Mix Trypan Blue in the PBS + cell solution slowly before loading.
