Trypan Blue Staining - (Feb/24/2015 )

I am trying to test the vitality of my cultures.

Usualy i detach my cells (3t3 for example) with trypsin from the flask, afterwards the staining and the vitality test is no porblem.

Now i want to know if the results are still reliable if i do not detach the cells ? Of course i cannot count them anymore, but just to get visual impression of how many cells still live.

Or is there maybe another method to test vitality of attached cells.

My picture shows the result:

.... my protocol:

Washing with PBS

1:2 dilution of trypan blue with PBS

5 minutes incubattion time

rinsing the well with PBS again.

You can use neutral red, which will only be taken up by living cells.