I am trying to isolate THP-1 cells derived macrophages by stimulation of PHA. To check live and dead population to take appropriate numbers in further assay i am getting problem to get actual picture of tryphan blue stained cells control.
If anybody have the same please share it.
What is the issue you are experiencing? This is a very routine assay. Could you describe your situation more?
Thanking you to trying me to solve my issue.
In my case when i stained with trypan blue to check viability and enumaration of cells whole cells as well some time nucleus seen blue colour without stain with cytoplasm. It is very difficult to judge live or dead and enumerate to put specific number of cells in next assay. If i gate know picture or experience it will be help ful to me to resolve the same.
DEAR Nawaj Shaikh,
Trypan blue is a vital stain used to selectively color dead tissues or cells blue. so only the dead cell or cells with any defects in their wall will be stained and appear blue under microscope with very clear projections on the cells walls, while living cells are stainless with the intact center and just by adjusting the eye lens you can also clearly find the wall of the intact cells.
regards
regards