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Characterization and Quenching of Autofluorescence in Piglet Testis Tissue and Cells
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 effecti...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi Publishing Corporation
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3437279/ https://www.ncbi.nlm.nih.gov/pubmed/22970375 http://dx.doi.org/10.1155/2012/820120 |
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author | Yang, Yanfei Honaramooz, Ali |
author_facet | Yang, Yanfei Honaramooz, Ali |
author_sort | Yang, Yanfei |
collection | PubMed |
description | 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 fluorescence labeling of gonocytes with lectin Dolichos biflorus agglutinin (DBA). We speculate that the lipofuscin or lipofuscin-like pigments within Leydig cell granules were mainly responsible for the observed intrinsic fluorescence in piglet testes. The method described in the present study can facilitate the identification and characterization of piglet gonocytes using fluorescence microscopy. |
format | Online Article Text |
id | pubmed-3437279 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Hindawi Publishing Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-34372792012-09-11 Characterization and Quenching of Autofluorescence in Piglet Testis Tissue and Cells Yang, Yanfei Honaramooz, Ali Anat Res Int Research Article 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 fluorescence labeling of gonocytes with lectin Dolichos biflorus agglutinin (DBA). We speculate that the lipofuscin or lipofuscin-like pigments within Leydig cell granules were mainly responsible for the observed intrinsic fluorescence in piglet testes. The method described in the present study can facilitate the identification and characterization of piglet gonocytes using fluorescence microscopy. Hindawi Publishing Corporation 2012 2012-08-30 /pmc/articles/PMC3437279/ /pubmed/22970375 http://dx.doi.org/10.1155/2012/820120 Text en Copyright © 2012 Y. Yang and A. Honaramooz. https://creativecommons.org/licenses/by/3.0/ 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. |
spellingShingle | Research Article Yang, Yanfei Honaramooz, Ali Characterization and Quenching of Autofluorescence in Piglet Testis Tissue and Cells |
title | Characterization and Quenching of Autofluorescence in Piglet Testis Tissue and Cells |
title_full | Characterization and Quenching of Autofluorescence in Piglet Testis Tissue and Cells |
title_fullStr | Characterization and Quenching of Autofluorescence in Piglet Testis Tissue and Cells |
title_full_unstemmed | Characterization and Quenching of Autofluorescence in Piglet Testis Tissue and Cells |
title_short | Characterization and Quenching of Autofluorescence in Piglet Testis Tissue and Cells |
title_sort | characterization and quenching of autofluorescence in piglet testis tissue and cells |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3437279/ https://www.ncbi.nlm.nih.gov/pubmed/22970375 http://dx.doi.org/10.1155/2012/820120 |
work_keys_str_mv | AT yangyanfei characterizationandquenchingofautofluorescenceinpiglettestistissueandcells AT honaramoozali characterizationandquenchingofautofluorescenceinpiglettestistissueandcells |