Protocol for Whole mount staining of intestinal tissue
Whole Mount X-gal Staining
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posted 13 Sep, 2011
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mouse tissue with LacZ
β-gal staining of whole mount intestine.
After staining the intestines for β-gal, the number of positive villi score under a Leica MZ6 dissecting microscope at 20X power, a 25mm2 field of view. The field of view represented 1/28 of each strip of small intestine, thus 1/168 of the entire small intestine (each strip was 1/6 of the entire small intestine). The 25mm2 field of view represented 1200 villi, which extrapolates to 201,600 villi in the entire small intestine. The number of β-gal+ villi were counted in each field of view and at least 20 β-gal+ foci were counted for each third of the small intestine. Nearby β-gal+ foci were considered independent if not arising from the same crypt and surrounded by non-staining crypts. Adenomas, which involved multiple villi, were scored by whole mount and in cross sections. Microadenomas, which involved a single villus, were determined by scoring cross sections.