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I'm never going to run these type of experiments but I do need to have a good idea of their timescale. After I fix my cells, if I stained my cells with histology stains like DAPI, Fluorescein, and various labeled antibodies, how long do I have to wait before I can examine my cells with microscopy?

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up vote 6 down vote accepted

For fluorescence immunocytochemistry, there are 3 main incubations that determine duration of the procedure. Generally it takes 2 days to stain your cells before you can visualize them. Once cells or tissue are fixed onto glass, you first incubate with a primary antibody to directly target a specific antigen(s), usually overnight. After primary labelling, a secondary antibody is applied to target the primary antibody, which usually takes a few hours. These secondary antibodies come conjugated to some fluorescent dye (Cy5, TRITC, FITC, AlexaFluors, etc). A nuclear counterstain is really quick, only 5 minutes. After all the labelling and nuclear stain, you can mount the coverslip with mounting media and once it cures you can visualize them.

Abcam has one of many quick reference guides.

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The temperature plays an important role, too: incubating at 4°C or at RT makes a big difference. – nico Apr 7 '12 at 6:49
Certainly. I also left out blocking which is arguably most critical to the process, but on the time scale of the whole procedure, is relatively quick (30-60 minutes). @bobthejoe was simply asking for an idea of total time. – user560 Apr 7 '12 at 23:31
And another factor is what you are staining: times are shorter for cells in culture compared to tissue slices (i.e.: ICC vs IHC), as the antibody will need time to deeply penetrate in the tissue. – nico Apr 8 '12 at 7:30

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