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The way to count adherent cells in 96 well - (Feb/24/2009 )

Hi,

I am currently planning to count my endothelial cells in a 96 well plate when they are confluent. My experience with them is that they really sticky and would be hard to trypsinise them. Another problem would be that prolonged exposure to trypsin would surely be harmful to them.

As for the counting, i was planning to use trypan blue exclusion test since it's much easier but would like to hear more about other alternatives in case it does not work.

Thanks in advance!

-timothyl-

There are a number of good competing products to Trypsin which are very safe to cells. I have personally used Accutase for many cell lines which were particularly sensitive to Trypsin and Invitrogen also offers a reliable Trypsin alternative (Tryple).

-jajell-

I would like to stand corrected that the one from Invitrogen is the Tryple Express?

Thanks for the suggestion.

I would like to keep this as soon as possible.

Thanks again

-timothyl-

timothyl on Feb 25 2009, 07:36 AM said:

Hi,

I am currently planning to count my endothelial cells in a 96 well plate when they are confluent. My experience with them is that they really sticky and would be hard to trypsinise them. Another problem would be that prolonged exposure to trypsin would surely be harmful to them.

As for the counting, i was planning to use trypan blue exclusion test since it's much easier but would like to hear more about other alternatives in case it does not work.

Thanks in advance!

Dear Timothyl
If u r not goin to reuse/ reculture the cell from 96 well plate, i will suggest u to use MTT/MTS or other viability assay.
Coz trypan blue exclusion test is not so suitable to count cell in 96 well plate(the cell number is too low and accuracy will be affected)
cheers :wacko:

-CKtong-