34 Viability of Cells
339
Eosin Y
Hanks and Wallace (1958) devised a method to distinguish between dead
and living cells based on the action of living cells to exclude eosin.
Materials
- Regular biologic microscope
- Slide glasses
- Cover glasses
- Pasteur pipettes
- A platinum line with I-mm diameter loop at the tip
- Petri dish
- 1% or 0.15% eosin Y solution
Procedure
l. Insert the platinum loop of Imm diameter into the 1 % eosin solution.
2. Spot eosin solution onto a slide glass using the platinum loop holding
eosin solution.
3. Air dry the slide glass, and store until use at room temperature.
4. Place wet filter paper on the bottom of a petri dish, and place the slide
glass with eosin spots on the filter paper.
5. Place the cell suspension onto the eosin spots using the platinum loop
of 2mm diameter.
6. Place a lid over the petri dish, and leave the cells for 2 min.
7. Observe the cells under a microscope.
Alternative Method
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 0.15% eosin solution.
4. Observe the cells under a microscope.
Results
Only dead cells stain red. If the alternative procedure is performed under sterile conditions, unstained living cells can be cultured further.
Nigrosin
This method was devised by Kaltenbach et al. (1958).
Materials
Equipment
Solutions
The equipment used is the same as that given for the trypan blue proto- Equipment
col.
339
Eosin Y
Hanks and Wallace (1958) devised a method to distinguish between dead
and living cells based on the action of living cells to exclude eosin.
Materials
- Regular biologic microscope
- Slide glasses
- Cover glasses
- Pasteur pipettes
- A platinum line with I-mm diameter loop at the tip
- Petri dish
- 1% or 0.15% eosin Y solution
Procedure
l. Insert the platinum loop of Imm diameter into the 1 % eosin solution.
2. Spot eosin solution onto a slide glass using the platinum loop holding
eosin solution.
3. Air dry the slide glass, and store until use at room temperature.
4. Place wet filter paper on the bottom of a petri dish, and place the slide
glass with eosin spots on the filter paper.
5. Place the cell suspension onto the eosin spots using the platinum loop
of 2mm diameter.
6. Place a lid over the petri dish, and leave the cells for 2 min.
7. Observe the cells under a microscope.
Alternative Method
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 0.15% eosin solution.
4. Observe the cells under a microscope.
Results
Only dead cells stain red. If the alternative procedure is performed under sterile conditions, unstained living cells can be cultured further.
Nigrosin
This method was devised by Kaltenbach et al. (1958).
Materials
Equipment
Solutions
The equipment used is the same as that given for the trypan blue proto- Equipment
col.