Cell isolation from tissue -- how do you tell how many viable cells you get? - (Jan/16/2008 )

I would like to know how many whole cells I am getting from a cell isolation from tissue. How would I measure how many viable cells I am getting?

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The more important questions you should be asking are:

What cells am I isolating and what is the PURITY of those cells ?

Then once they are characterised, how many do I need for my experiments ? .......and what passage number do I go upto ?

Is my marker that I am interested in still there ? Are my cells contaminated with bacteria/fungus/mycoplasma ?

Many questions BEFORE looking at viabilty of cells that could be a mixture of Fibroblasts, epithelial, endothelial, smooth muscle, WBC/RBC's or bacteria.

Once all these have been answered, then look for viabilty of the cells you are interested in.....Trypan blue exclusion, annexin V, MTT assays etc.

Rhombus

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Count cells in a hemocytometer. Add some trypan blue to cell suspension. The dead cells will take up the blue color.

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depends primarily on the type of tissue, how much of it there is and the method used to extract cells... e.g growing tissues such as gut or skin you can easily get lots of cells, but tissues such as brain which are non-proliferative will give very few, if any cells.

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