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Whole Mount Immunohistochemistry
Sponsored,vendor-submitted protocol    Published in November 2009 (p.20) DOI: 10.2144/000113015

Whole mount staining is the staining of small pieces of tissue, usually embryos, without sectioning onto slides first. This is often used on embryos by stem cell and embryonic development researchers and also neuroscientists who are able to stain the whole embryos at various stages to follow the expression of target proteins through the development of the animal. Whole mount staining is very similar to immunocytochemistry (ICC) or staining of cryosections. If an antibody has been used successfully on cryosections then the antibody should work for a whole mount embryo. The difference being that the sample being stained is much larger, and thicker, than a normal section on a slide. Therefore, incubations for fixative, blocking buffer, antibody, wash buffer, permeabilization, and substrate color development will need to be much longer to allow for permeabilization right into the center of the sample. Researchers use different times, but the details in these procedures provide a guideline for optimizing the experiment at these stages if necessary. The full protocol is for chick or mouse whole mount immunohistochemistry.


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