A Method for Serial Tissue Processing and Parallel Analysis of Aberrant Crypt Morphology,Mucin Depletion,and Beta-Catenin Staining in an Experimental Model of Colon Carcinogenesis |
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Authors: | John N McGinley Matthew D Thompson Henry J Thompson |
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Institution: | (1) Cancer Prevention Laboratory, Colorado State University, 1173 Campus Delivery, Fort Collins, CO 80523, USA; |
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Abstract: | The use of architectural and morphological characteristics of cells for establishing prognostic indicators by which individual
pathologies are assigned grade and stage is a well-accepted practice. Advances in automated micro- and macroscopic image acquisition
and digital image analysis have created new opportunities in the field of prognostic assessment; but, one area in experimental
pathology, animal models for colon cancer, has not taken advantage of these opportunities. This situation is primarily due
to the methods available to evaluate the colon of the rodent for the presence of premalignant and malignant pathologies. We
report a new method for the excision and processing of the entire colon of the rat and illustrate how this procedure permitted
the quantitative assessment of aberrant crypt foci (ACF), a premalignant colon pathology, for characteristics consistent with
progression to malignancy. ACF were detected by methylene blue staining and subjected to quantitative morphometric analysis.
Colons were then restained with high iron diamine–alcian blue for assessment of mucin depletion using an image overlay to
associate morphometric data with mucin depletion. The subsequent evaluation of ACF for beta-catenin staining is also demonstrated.
The methods described are particularly relevant to the screening of compounds for cancer chemopreventive activity. |
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