Immunofluorescence microscopy fixation

The immunofluorescent labeling procedure described in this section is analogous to that described in Section III,A. It involves fixation and permeabilization of the cells and denaturation of the DNA in order to give the antibodies access to the halo atoms of the incorporated IdU and CldU nucleotides. A very similar dual-labeling protocol, more specifically for flow cytometry rather than for fluorescence microscopy, has been described by Aten et al. (1994). 

Immunolabelling of organelles or stmctiues by fluorescent antibodies in living cells has the advantage, compared to work in ftxed cells, that the dynamics of the labelled structures or molecules can be studied by for example time-lapse microscopy. Also, labelling in living cells does not suffer from possible problems induced by the fixation or permeabilization of the cells as is necessary for immunofluorescence or electron microscopy. 

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