characterization and quenching of autofluorescence in piglet testis tissue and cells表征和自发荧光猝灭小猪睾丸组织和细胞.pdfVIP

Hindawi Publishing Corporation Anatomy Research International Volume 2012, Article ID 820120, 10 pages doi:10.1155/2012/820120 Research Article Characterization and Quenching of Autofluorescence in Piglet Testis Tissue and Cells Yanfei Yang and Ali Honaramooz Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon, SK, Canada S7N 5B4 Correspondence should be addressed to Ali Honaramooz, ali.honaramooz@usask.ca Received 11 June 2012; Accepted 11 July 2012 Academic Editor: Udo Schumacher Copyright © 2012 Y. Yang and A. Honaramooz. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Significant intrinsic fluorescence in tissues and in disassociated cells can interfere with fluorescence identification of target cells. The objectives of the present study were (1) to examine an intrinsic fluorescence we observed in both the piglet testis tissue and cells and (2) to test an effective method to block the autofluorescence. We observed that a number of granules within the testis interstitial cells were inherently fluorescent, detectable using epifluorescence microscopy, confocal laser scanning microscopy, and flow cytometry. The emission wavelength of the autofluorescent substance ranged from 425 to 700 nm, a range sufficiently broad that could potentially interfere with fluorescence techniques. When we treated the samples with Sudan Black B for different incubation times, the intrinsic fluorescence was completely masked after treatment for 10–15 min of the testis tissue sections or for 8 min of the testis cells, without compromising specific