Exclusion The following procedure will enable you to accurately determine the cell viability. Cell viability is calculated as the number of viable cells divided by the total number of cells within the grids on the hemacytometer.

**Trypan Blue:**

Exclusion
The following procedure will enable you to accurately determine the cell viability.
Cell viability is calculated as the number of viable cells divided by the total
number of cells within the grids on the hemacytometer. If cells take up trypan
blue, they are considered non-viable.

1.
Determine the cell density of your cell
line suspension using a hemacytometer.

2.
Prepare a 0.4% solution of trypan blue
in buffered isotonic salt solution, pH 7.2 to 7.3 (i.e., phosphate-buffered
saline).

3.
Add 0.1 mL of trypan blue stock solution
to 1 mL of cells.

4.
Load a hemacytometer and examine immediately
under a microscope at low magnification.

5.
Count the number of blue staining cells
and the number of total cells. Cell viability should be at least 95% for
healthy log-phase cultures. Remember to correct for the dilution factor.

To
calculate the number of viable cells per mL of culture, use the formula below.

Live
cell count/ Total cell count =Viability

Determine
total viable cell yield using the formula below.

Viable
cell count/ Quadrants counted x Dilution factor x Hemocytometer factor x
Current volume (mL) = Viable cell yield.

Study Material, Lecturing Notes, Assignment, Reference, Wiki description explanation, brief detail

**Related Topics **

Copyright © 2018-2020 BrainKart.com; All Rights Reserved. Developed by Therithal info, Chennai.