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Trypan blue viability test

Trypan blue viability test


Trypan blue will stain dead or dying cells. Viable cells are able to repell the dye and do not stain. Note: Trypan blue has a greater affinity for serum proteins than for cellular protein. If the background is too dark, cells should be pelleted and resuspended in protein-free medium or salt solution prior to counting.

Supplies & Equipment:



  1. Prepare a cell suspension in HBSS
  2. Transfer into Eppendorf tube:
  3. Allow to stand for 5 to 15 minutes
    Note: after prolonged incubation, viable cells start to take up dye as well.
  4. Pipet 10Ál of this mix into cover-slipped chambers of hemocytometer
    Note: avoid cell clusters by pipetting up and down.
  5. Count viable and non-viable cells
    Note: for optimal results, adjust cell density to 20-50 cells / square.
  6. Calculations: