Trypan Blue Staining
Trypan Blue is a dye commonly used to assess cell viability.
|Trypan Blue solution, 0.4%|
|Hemocytometer with brightfield/phase microscope or automated cell counter (i.e. Countess™ automated cell counter)|
Note on Trypan Blue
- Trypan Blue is toxic to cells. After adding Trypan Blue dye to cells, move quickly to determine cell viability.
- Add 10 µL of 0.4% Trypan Blue to 10 µL washed cells. Pipette to mix.
- Transfer 10 µL of cell-Trypan Blue mix to a counting slide.
- View under brightfield/phase microscope or cell counter.
- Assess cell morphology and viability and troubleshoot as needed.
- Viable cells are impermeable to Trypan Blue and will exclude the dye (Trypan Blue negative; Figure A).
- Dead cells are permeable to Trypan Blue and will stain Trypan Blue positive (Figure A).
- Isolated nuclei will stain Trypan Blue positive (Figure B).
Figure. (A) Washed K562 cells are mostly viable (bright white and round). A dead cell (blue, Trypan positive) is circled in red. (B) Successful nuclei harvest shows Trypan Blue stained nuclei. An intact cell (bright white, Trypan negative) is circled in red.