Worthington Tissue Dissociation Guide

Optimization Techniques: Measure of Viability

One of the simplest methods to approximate cell viability is the dye exclusion technique. This method utilizes an indicator dye to demonstrate cell membrane damage. Cells which absorb the dye become stained and are considered non-viable. Dyes such as trypan blue, erythrosin, and nigrosin are commonly used with trypan blue being the most common in preliminary cell isolation procedures.

This procedure can be performed along with the cell counting procedure but cell density may require adjustment in order to obtain approximately 106 cells per milliliter.

  1. Mix 1 drop of trypan blue with one drop of the cell suspension and allow 1 - 2 minutes for absorption
  2. Prepare hemocytometer and load chambers as described in "Cell Quantitation".
  3. Count both the total number of cells and the number of stained (dark) cells.
Percent Viability = 
    Total Cells Counted - Stained Cells x 100
 Total Cells Counted

Total Cells / 1 mm2 = 182
Stained Cells = 24
% Viability =182 - 24=158 x 100
 182 182 
  =86.8% Viability
Note: Dye exclusion viability procedures tend to give high estimates of cell viability when compared to cell attachment or metabolic assays, but for optimization of cell isolation procedures trypan blue does provide a rapid estimate of dissociation performance in conjunction with yield quantitation.

Next: Tissue Culture Glossary

Tissue Tables (references, grouped by tissue type and species)

Adipose/Fat Adrenal Bone Brain
Cartilage Colon Endothelial Epithelial
Eye Heart Intestine Kidney
Liver Lung Lymph nodes Mammary
Miscellaneous Muscle Neural Pancreas
Parotid Pituitary Prostate Reproductive
Scales Skin Spleen Stem
Thymus Thyroid/Parathyroid Tonsil Tumor

Note: We have not limited the references listed to only those papers using Worthington enzymes. Generally speaking, the tissue dissociation enzymes offered by Worthington can be used interchangeably for most preparations cited.