Counterstaining Improves Visualization of the Myenteric Plexus in Immunolabelled Whole-Mount Preparations

• DOI: 10.1007/s10895-006-0120-x
• Publication Date: 2006-09-11T17:19:03Z
• Authors: Parkash Mandhan; Bao Quan Qi; Jacqueline I. Keenan; Salim Ismail; Spencer W. Beasley; Michael J. Sullivan
• Keywords: 0301 basic medicine; Staining and Labeling; Myenteric Plexus; Immunohistochemistry; Rats; 3. Good health; Rats, Sprague-Dawley; 03 medical and health sciences; Fetus; Pregnancy; Phosphopyruvate Hydratase; Animals; Female; Fluorescent Antibody Technique, Indirect; Fluorescent Dyes; Vasoactive Intestinal Peptide

Immunocytochemistry has emerged as a powerful research tool in neurobiology. One of the widely used methods is an indirect fluorescence technique that uses FITC- conjugated IgG to visualise protein expression within tissues, but a major drawback of this technique is the high background fluorescence due to non-specific antibody binding. Gut innervation is complex and best visualized in three-dimensions in whole mount preparations. We describe a simple and easy to use counterstaining procedure in conjunction with an indirect immunofluorescence technique in gut whole mount preparations that largely eliminates background fluorescence and creates a contrasting background against the bright antigen-antibody complexes. Furthermore, this technique allows the detailed qualitative and quantitative study of myenteric plexuses in whole-mount preparations.