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Related Techniques
Materials
Equipment - Regular biologic microscope
- Slide glasses
- Cover glasses
- Pasteur pipettes
Solutions - 0.5% Trypan blue solution
Procedure
1. Suspend the cells to be examined in the culture medium.
2. Add 0.5 ml of 0.5% trypan blue solution to 1 ml cell suspension.
3. Observe the cells immediately under a microscope.
Comments
Trypan blue is toxic to cells, and, therefore, cells will die if the cell suspension is left in contact with the trypan blue for too long.
Results
Only dead cells are stained blue.
Erythrosin B
The following procedure is based on the methods of Phillips and
Terryberry (1957).
Materials
Equipment - The equipment used is the same as given for the trypan blue protocol
Solutions - Erythrosin B solution (20 mg erythrosin BIIOO ml balanced salt solution or culture medium)
Procedure
1. Suspend the cells to be examined in the culture medium.
2. Centrifuge the cell suspension at 150 g for 5 min.
3. Suspend the cell pellet in erythrosin B solution.
4. Observe the cells after 5-15 min, and finish counting the cells within
2h.
Results
Only dead cells are stained red.