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Trypan Blue if cells cultured in 96 well plate

trypan blue flow culture

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#1 H. Aly

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Posted 22 November 2014 - 02:45 AM

I am new to cell culture and I am going to measure cell viability with trypan blue but some one told me that it can't be done if i had my cells plated in 96 well plate. Is that true? 

 

And if I am going to use flow cytometry for direct staining, do i have to culture my cells in different plates for it or if i can make the cell suspension and use some of it for viability and some for flow?



#2 science noob

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Posted 22 November 2014 - 08:29 AM

You will have to trypsinize your cultured cells and count them using trypan blue and a haemocytometer. You won't get alot of cells from a 96-well plate. That said, what is your aim of measuring cell viability? Are you treating your cells with a chemical to see if they undergo apoptosis?

 

For your second question, you will need more cells to run a flow cytometry experiment. Most people use between 100,000 - 1,000,000 cells to stain and take 5,000-10,000 cells to measure the fluorescence positivity. What cells are you working with?

 

Help yourself by referring to trypan blue protocols online. There are alot of information you can get with a simple Google search. Here is a good overview of the technique: http://www.ncbi.nlm....pubmed/18432654



#3 H. Aly

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Posted 22 November 2014 - 09:46 AM

Thanks science noob,

Yes, I am treating my cells to check the apoptosis, and I am working with MDA-MB-231.







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