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The use of quenching agents to enable immunofluorescent examination of collagen-based biomaterials showing glutaraldehyde-derived autofluorescence

Authors
Journal
Clinical Materials
0267-6605
Publisher
Elsevier
Publication Date
Volume
6
Issue
1
Identifiers
DOI: 10.1016/0267-6605(90)90040-3

Abstract

Abstract Examination of glutaraldehyde-treated collagen-based vascular prostheses by immunofluorescence methods indicated that a high degree of autofluorescence was present. This background autofluorescence, which resulted from the glutaraldehyde used in the manufacture of the prostheses, made it very difficult to visualise specific antibody-associated fluorescence. The use of different fluorochromes with a variety of filter blocks did not lead to acceptable conditions, since the autofluorescence showed a broad emission spectrum at various excitation wavelengths. An alternative which proved satisfactory was the use of quenching agents. In particular, 1% Evans Blue led to a shift in the autofluorescent spectrum to longer wavelengths. Specific monoclonal antibody detection of collagen was then possible, with the use of additional filters to mask out the longer autofluorescence wavelengths.

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