Whole Mount Preparation of Mouse Intestine: Using a Simple Gut-incising Device to Prepare Whole Mount
Transcrição
Whole-mount analysis allows the visualization of multi-dimensional tissue cross-sections while preserving the structural interactions between cells and their microenvironment.
To prepare whole mounts of the intestine, begin with a freshly harvested mouse intestine. Remove the mesentery – folds of peritoneal tissue – from the intestine. Flush the intestine with a chilled buffer to remove any fecal matter.
Next, excise the cecum. Dissect the intestine into equal segments. Make small incisions in the intestinal segments to drain the fluid within them.
Gently insert a wet stainless-steel rod into the intestinal segments. Now, position these rods into holding slots at the base of the customized gut-incising device. Fit the lid of the device over its base.
Using angled bars on the lid as the guide, longitudinally incise the intestinal segments. Remove the lid from the device. Moisten the segments. Gently roll the rod sideways to release and spread the segments flat.
Immerse the segments in a bath containing a suitable fixative and incubate for the desired duration. The fixative chemically preserves the cellular structure and tissue architecture for subsequent sampling.
Following fixation, store the tissue in ethanol until further downstream analysis.
