Optimization of visibility and quantification of aberrant crypt foci in colonic mucosa in Wistar rats.

PURPOSE Test immersion of microscopy samples in water as an aid to visualizing and quantifying aberrant crypt foci (ACF) in rat colon mucosa. METHODS Carcinogenesis was induced with azoxymethane in Wistar rats kept on a conventional diet or a hypercaloric diet containing unsaturated fat. Fifteen weeks after induction, colon samples were retrieved and fixated in a 10% formaldehyde solution. The samples were divided into segments (distal, middle, proximal) and stained with 1% toluidine blue. The technique tested in the study consisted of immersing microscopy samples in distilled water in order to eliminate the problem of light reflection known from conventional microscopy. RESULTS When samples were immersed in water during microscopy, significantly more ACF could be visualized in all colon segments than with the conventional method proposed by Bird. CONCLUSION Immersing microscopy samples in water aids the visualization and quantification of aberrant crypt foci in rat colon mucosa fixed in formaldehyde.

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