Abstract
Test immersion of microscopy samples in water as an aid to visualizing and quantifying aberrant crypt foci (ACF) in rat colon mucosa. 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. 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. Immersing microscopy samples in water aids the visualization and quantification of aberrant crypt foci in rat colon mucosa fixed in formaldehyde.
Highlights
Colon carcinogenesis has been investigated widely by administrating carcinogenic substances to experimental animals[1]
aberrant crypt foci (ACF) in colonic mucosa stained with toluidine blue is usually darker and larger than normal crypt foci and tend to occur in isolation, in pairs or in small clusters
Eight weeks into the study period, carcinogenesis was induced with 15 mg/kg azoxymethane administered peritoneally once a week for two weeks
Summary
Colon carcinogenesis has been investigated widely by administrating carcinogenic substances to experimental animals[1]. The emergence of aberrant crypt foci (ACF) is one of the most important outcome parameters in such models[2]. ACF in colonic mucosa stained with toluidine blue is usually darker and larger than normal crypt foci and tend to occur in isolation, in pairs or in small clusters. Bird in 1987 proposed a method for visualizing ACF in samples of rat colon using light microscopy at 40X. Many researchers have since employed the method in study protocols, clinical practice and colonoscopic screening for pre-neoplastic lesions[3,4,5,6]. The objective of this study was to test immersion of microscopy samples in water as an aid to visualizing and quantifying aberrant crypt foci (ACF) in rat colon mucosa
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