Confocal microscopy

From: Oberdan Leo (oleo@dbm.ulb.ac.be)
Date: Fri Oct 12 2001 - 08:54:30 EST


Hi everyone,
although not strictly a flow question, my query relates to staining
cells with fluorescent antibodies so...
We are trying to look at freshly isolated murine lymphocytes by confocal
microscopy. We would like to look at both cell-surface and intracellular
antigens, and need some advice. Let's say we want to stain for CD3 :
1. Should you stain the cells in a tube, fix, and then try to make the
cells stick to a coverslip ?
2. Or have the cells adhere first, stain and fix ?
3. What type of mounting media / coverslip ?
Hope someone is able to help us

Greetings,

Oberdan





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