High Resolution Light Microscopy and Immunocytochemistry Using Glycol Methacrylate-Embedded Sections and Immunogold-Silver Staining
Abstract
The immunogold silver technique was modified so that 1-2 micron thick glycol methacrylate sections could routinely be used for high resolution immunocytochemistry. A rabbit anti-human hemoglobin antibody was used to demonstrate the distribution of exogenous hemoglobin in mouse tissues. Inclusion of 0.1% glutaraldehyde in the primary formalin fixation improved tissue appearance. Glutaraldehyde treatment after colloidal gold antibody application increased the staining intensity, and gold chloride intensification of the silver-enhanced colloidal gold further improved immunosensitivity. Thus a more accurate analysis of hemoglobin distribution in tissue was obtained. The benefit of these enhancements was also demonstrated for localizing myelin basic protein in brain and for determining the proliferative status of tissue by localization of bromodeoxyuridine incorporation into nuclei. The efficiency of manual capillary-action staining performed during all steps in the immunogold-silver staining protocol was also demonstrated. This greatly simplified the procedure, reduced technician time, and produced better staining consistency. The immunogold-silver staining technique is simple, sensitive, economical, and relatively nontoxic.
Document Details
- Document Type
- Technical Report
- Publication Date
- Dec 30, 1991
- Accession Number
- ADA245163
Entities
People
- David E. Tacha
- Luann Mckinney
- Phillip D. Bowman
Organizations
- Letterman Army Hospital