Trypan blue cell death assay on cardiac myocytes - (Aug/17/2006 )

Hi! though it is a simple procedure, I am having some concerns/problems...
can anybody tell me the method of assessing the cell viability on cardiac myocytes by trypan blue method if anybody has done it.
-Its difficult to use Haemocytometer bcz of much bigger cell size or can we?
-time to stain... 15 mins or less???
so, just to look upon a area under microscope and count???
any other precaution....

thanks a lot.

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After adding trypsin blue, count the cells immediately.
You do not need to use haemocytometer, you can put the cells on a glass slide and count 100 cells. This will give you the percentage of viable cells.

I hope this may help.

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yeah! did that and it worked well. but even after 15 minutes, some of the myocytes are not stained though they were round (which indicates that they are in contracture) shall I count them as dead or live??? I count them as dead generally.
thanks.

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Its OK to leave ur cells in trypan blue for some minutes as long as its not over 30 minutes as its toxic to the cells and cause death to them.

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