Chapter 34
Viability of Cells
Subprotocol 1
Vital Staining Method
The following is the method of Melnick and Opton (1956) to stain living
cells with neutral red.
Materials
- Regular biologic microscope
- Incubator
- Slide glasses
- Cover glasses
- Pasteur pipettes
- 0.01% neutral red
Procedure
1. Suspend the cells to be examined in the culture medium.
2. Add 0.05 ml neutral red solution to 1 ml cell suspension.
3. Incubate at 30°C for 30 min.
4. Observe the cells under a microscope.
Results
Viable cells are stained red.
Subprotocol 2
Dye-Exclusion Method
Living cells extrude dye after the dye enters the cells. Using this characteristic, cell viability can be measured. Several kinds of dye are used to
measure cell viability with the dye-exclusion method.
Trypan Blue
This method was originally devised by McLimans et al. (1957) for mammalian cells.
PROTOCOL
Equipment
Solutions
Viability of Cells
Subprotocol 1
Vital Staining Method
The following is the method of Melnick and Opton (1956) to stain living
cells with neutral red.
Materials
- Regular biologic microscope
- Incubator
- Slide glasses
- Cover glasses
- Pasteur pipettes
- 0.01% neutral red
Procedure
1. Suspend the cells to be examined in the culture medium.
2. Add 0.05 ml neutral red solution to 1 ml cell suspension.
3. Incubate at 30°C for 30 min.
4. Observe the cells under a microscope.
Results
Viable cells are stained red.
Subprotocol 2
Dye-Exclusion Method
Living cells extrude dye after the dye enters the cells. Using this characteristic, cell viability can be measured. Several kinds of dye are used to
measure cell viability with the dye-exclusion method.
Trypan Blue
This method was originally devised by McLimans et al. (1957) for mammalian cells.
PROTOCOL
Equipment
Solutions