Vanderbilt Summer Science Academy

Epitope Stability in Unstained Slides for Immunohistochemistry

Immunohistochemistry is used to detect the location of proteins in intact tissue, which is critical in many research studies that interrogate the progression of disease. It utilizes antibodies that bind to specific epitopes. These antibodies are usually conjugated to a specific enzyme that catalyzes a reaction to produce a certain colored stain. After tissues from research subjects are collected and fixed they are then embedded in paraffin wax and sectioned onto microscope slides. Usually, these slides are stored before the immunostaining process, which might lead to degradation of the epitopes needed for staining.  The goal of this project is to determine is the best method for preserving the epitopes sectioned tissue mounted on glass slides. Mouse spleen and four different markers will be used: CD3, CD4, CD8, and F4/80. These are common leukocyte markers used in immunology research. Four conditions for slide storage will be tested: room temperature slides dipped in paraffin wax, room temperature non-dipped, -4°C non-dipped, and -4°C dipped slides. We will examine the effect of these different parameters on tissue cut 2.5 weeks before, 1.5 weeks before, and 1 week before the staining process. These will be compared to tissues sectioned the night before the staining process. After examining the stained slides we will see what is the proper epitope preservation technique and hope to apply that to the overall process in the future.