Tyramide signal amplification method in multiple-label immunofluorescence confocal microscopy.

Important note: Information in this article was accurate in 2000. The state of the art may have changed since the publication date.

Tyramide signal amplification method in multiple-label immunofluorescence confocal microscopy.

Methods. 1999 Aug;18(4):459-64. Unique Identifier : AIDSLINE MED/99423778
Wang G; Achim CL; Hamilton RL; Wiley CA; Soontornniyomkij V; Department of Pathology (Neuropathology), University of; Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213,; USA.

Abstract: The tyramide signal amplification (TSA) method has recently been introduced to improve the detection sensitivity of immunohistochemistry. We present three examples of applying this method to immunofluorescence confocal laser microscopy: (1) single labeling for CD54 in frozen mouse brain tissue; (2) double labeling with two unconjugated primary antibodies raised in the same host species (human immunodeficiency virus type 1 p24 and CD68) in paraffin-biopsied human lymphoid tissue; and (3) triple labeling for brain-derived neurotrophic factor, glial fibrillary acidic protein, and HLA-DR in paraffin-autopsied human brain tissue. The TSA method, when properly optimized to individual tissues and primary antibodies, is an important tool for immunofluorescence microscopy. Furthermore, the TSA method and enzyme pretreatment can be complementary to achieve a high detection sensitivity, particularly in formalin-fixed paraffin-embedded archival tissues. Using multiple-label immunofluorescence confocal microscopy to characterize the cellular localization of antigens, the TSA method can be critical for double labeling with unconjugated primary antibodies raised in the same host species. Copyright 1999 Academic Press.

Keywords: JOURNAL ARTICLE Acquired Immunodeficiency Syndrome/PATHOLOGY Alzheimer Disease/PATHOLOGY Animal Antibodies Antigens, CD/ANALYSIS/IMMUNOLOGY Antigens, Differentiation, Myelomonocytic/ANALYSIS/IMMUNOLOGY *Brain Chemistry Fluorescent Antibody Technique Giant Cells/CHEMISTRY Glial Fibrillary Acidic Protein/ANALYSIS/IMMUNOLOGY Human HIV Core Protein p24/ANALYSIS/IMMUNOLOGY HIV-1/ISOLATION & PURIF HLA-DR Antigens/ANALYSIS Intercellular Adhesion Molecule-1/ANALYSIS/IMMUNOLOGY Lymphoid Tissue/CHEMISTRY Mice Microscopy, Confocal/*METHODS Microscopy, Fluorescence/METHODS Senile Plaques/CHEMISTRY Sensitivity and Specificity Support, U.S. Gov't, P.H.S. Tyramine/*ANALYSIS/IMMUNOLOGY

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