Immunofluorescence is a widely used technique to visualize the localization of proteins of interest. Nucleoside analogs, such as 5-ethynyl-2'-deoxyuridine (EdU), are incorporated into newly synthesized DNA and enable permanent labeling of newly divided cells. Both these techniques can be applied to long-term organotypic culture of in a fashion similar to that already described for tissue sections. We report here our optimized method for immunofluorescence and EdU staining of organotypic slices.
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