Photobleaching of arterial autofluorescence for immunofluorescence applications

BioTechniques
K KingsleyG E Plopper

Abstract

Immunohistochemical localization of low-level antigens in the arterial vasculature is complicated by the presence of complex molecules such as collagen, elastin, cholesterol, and fluorescent lipids that exhibit autofluorescence over a wide spectrum of wavelengths. UV irradiation of arterial vasculature has remained ineffective in preparing samples for immunofluorescent staining because of the recovery of the endogenous fluorescence within a short time following treatment. Therefore, we sought to further enhance the signal-to-noise ratio in arteries by optimizing the photobleaching of this tissue. We report here that the use of filtered sunlight significantly reduces arterial autofluorescence compared to standard UV shortwave and longwave irradiation and maintains multiple antigen epitopes suitable for immunohistochemical analysis. Using this method, we localized low-level laminin-5 isoform expression in situ, which was previously indistinguishable from endogenous autofluorescence.

Citations

Oct 15, 2005·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·Dardo E FerraraW Robert Taylor
May 1, 2017·Histochemistry and Cell Biology·Douglas J Taatjes, Jürgen Roth
Sep 29, 2019·Clinical Hemorheology and Microcirculation·Jie ZouAndreas Lendlein
Sep 27, 2008·Cardiovascular Research·Luis Rodriguez-MenocalRoberto I Vazquez-Padron
Dec 3, 2004·Molecular Biology of the Cell·Robert F KleesGeorge E Plopper
Sep 12, 2019·PloS One·Magdalena BlaszkiewiczKristy L Townsend
Jun 8, 2002·Biochemical and Biophysical Research Communications·K KingsleyG E Plopper
Jun 21, 2002·Biochemical and Biophysical Research Communications·K KingsleyG E Plopper
Jul 12, 2002·Kidney International·Jose Luis ReyesLorenza González-Mariscal
Jun 12, 2003·Experimental Cell Research·Kimi C Nishikawa, Albert J T Millis
Jul 5, 2021·Journal of Immunological Methods·Shahid NazirPriscilla F Gerber
Dec 2, 2006·Matrix Biology : Journal of the International Society for Matrix Biology·Robert F KleesGeorge E Plopper

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