Analysis of Adiposity in Mice: Method Development

Abstract

<p>Purpose: To setup a method to measure area and diameter of fat cells in white adipose tissue from mice. The plan is to use the best method for the study of fat metabolism in a murine model of diabetes. Methods: We tested two methods: 1} Oil Red to stain the lipid in fat cells in frozen tissue sections and 2} Paraformaldehyde fixed and paraffin embedded adipose tissue stained with hematoxylin and eosin (H&E). Oil Red is a lipophilic dye which penetrates the cells, outlines the cell membrane and stains the nucleus a blue color. Results: Results showed that the Oil Red stain did not work well with the frozen tissue sections. The temperature required for sectioning fat was below the range of the cryostat. Histological examination of the tissue showed that the cell membranes were deformed, allowing the oil red stained fat to leak out. In contrast, the results with the H&E stained, fixed tissue was much better. The adipose tissue was clearly preserved showing a network of fat cells, resembling a wire mesh. Using this method, it will be possible to measure cell size and number, combining photomicroscopy with computer imaging methods. Future Work: The next step will be to conduct studies in an animal model of diet induced diabetes, examining the changes in adiposity.</p><p>This presentation occurred at the Wright State University Campus-Wide Celebration of Research, Scholarship and Creative Activities on April 16, 2010</p>