Colonoscopic Guided Murine Wound Model: A Method to Isolate Biopsy Induced Wound Bed from Colon to Understand the Healing Process
Transcrição
Colonoscopic-guided biopsies in murine models induce wound sites in the colon. These wound beds are isolated for molecular and histological studies to understand the process of wound healing in inflammatory diseases of the gastrointestinal, or GI tract.
To begin wound bed isolation, place a euthanized experimental mouse with a colonoscopic-induced wound in the supine position. Incise the abdominal skin and underlying muscles to expose the body cavity.
Place closed scissors under the colon and lift it. Cut the colon at its midpoint, and at the anus to separate it from the body. Flush the colon with an appropriate buffer to remove fecal content.
Place the colon on filter paper and cut it open longitudinally, with the mesenteric side facing down on the filter paper. Now, add methylene blue dye over the colon and observe under the microscope.
The dye stains the dead and granulated tissue in the wound bed, making it appear darker than the surrounding healthy tissue, allowing for its easy identification. Preserve the isolated wound bed for gene expression studies.
