Intravital fluorescence microscopy with negative contrast.

Advances in intravital microscopy (IVM) have enabled the studies of cellular organization and dynamics in the native microenvironment of intact organisms with minimal perturbation. The abilities to track specific cell populations and monitor their interactions have opened up new horizons for visuali...

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Autores principales:	Juwell W Wu, Yookyung Jung, Shu-Chi A Yeh, Yongwan Seo, Judith M Runnels, Christian S Burns, Toshihide Mizoguchi, Keisuke Ito, Joel A Spencer, Charles P Lin
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d4d8b879f7d245b2bc20758c74e82da3
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spelling	oai:doaj.org-article:d4d8b879f7d245b2bc20758c74e82da32021-12-02T20:15:13ZIntravital fluorescence microscopy with negative contrast.1932-620310.1371/journal.pone.0255204https://doaj.org/article/d4d8b879f7d245b2bc20758c74e82da32021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0255204https://doaj.org/toc/1932-6203Advances in intravital microscopy (IVM) have enabled the studies of cellular organization and dynamics in the native microenvironment of intact organisms with minimal perturbation. The abilities to track specific cell populations and monitor their interactions have opened up new horizons for visualizing cell biology in vivo, yet the success of standard fluorescence cell labeling approaches for IVM comes with a "dark side" in that unlabeled cells are invisible, leaving labeled cells or structures to appear isolated in space, devoid of their surroundings and lacking proper biological context. Here we describe a novel method for "filling in the void" by harnessing the ubiquity of extracellular (interstitial) fluid and its ease of fluorescence labelling by commonly used vascular and lymphatic tracers. We show that during routine labeling of the vasculature and lymphatics for IVM, commonly used fluorescent tracers readily perfuse the interstitial spaces of the bone marrow (BM) and the lymph node (LN), outlining the unlabeled cells and forming negative contrast images that complement standard (positive) cell labeling approaches. The method is simple yet powerful, offering a comprehensive view of the cellular landscape such as cell density and spatial distribution, as well as dynamic processes such as cell motility and transmigration across the vascular endothelium. The extracellular localization of the dye and the interstitial flow provide favorable conditions for prolonged Intravital time lapse imaging with minimal toxicity and photobleaching.Juwell W WuYookyung JungShu-Chi A YehYongwan SeoJudith M RunnelsChristian S BurnsToshihide MizoguchiKeisuke ItoJoel A SpencerCharles P LinPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 8, p e0255204 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Juwell W Wu Yookyung Jung Shu-Chi A Yeh Yongwan Seo Judith M Runnels Christian S Burns Toshihide Mizoguchi Keisuke Ito Joel A Spencer Charles P Lin Intravital fluorescence microscopy with negative contrast.
description	Advances in intravital microscopy (IVM) have enabled the studies of cellular organization and dynamics in the native microenvironment of intact organisms with minimal perturbation. The abilities to track specific cell populations and monitor their interactions have opened up new horizons for visualizing cell biology in vivo, yet the success of standard fluorescence cell labeling approaches for IVM comes with a "dark side" in that unlabeled cells are invisible, leaving labeled cells or structures to appear isolated in space, devoid of their surroundings and lacking proper biological context. Here we describe a novel method for "filling in the void" by harnessing the ubiquity of extracellular (interstitial) fluid and its ease of fluorescence labelling by commonly used vascular and lymphatic tracers. We show that during routine labeling of the vasculature and lymphatics for IVM, commonly used fluorescent tracers readily perfuse the interstitial spaces of the bone marrow (BM) and the lymph node (LN), outlining the unlabeled cells and forming negative contrast images that complement standard (positive) cell labeling approaches. The method is simple yet powerful, offering a comprehensive view of the cellular landscape such as cell density and spatial distribution, as well as dynamic processes such as cell motility and transmigration across the vascular endothelium. The extracellular localization of the dye and the interstitial flow provide favorable conditions for prolonged Intravital time lapse imaging with minimal toxicity and photobleaching.
format	article
author	Juwell W Wu Yookyung Jung Shu-Chi A Yeh Yongwan Seo Judith M Runnels Christian S Burns Toshihide Mizoguchi Keisuke Ito Joel A Spencer Charles P Lin
author_facet	Juwell W Wu Yookyung Jung Shu-Chi A Yeh Yongwan Seo Judith M Runnels Christian S Burns Toshihide Mizoguchi Keisuke Ito Joel A Spencer Charles P Lin
author_sort	Juwell W Wu
title	Intravital fluorescence microscopy with negative contrast.
title_short	Intravital fluorescence microscopy with negative contrast.
title_full	Intravital fluorescence microscopy with negative contrast.
title_fullStr	Intravital fluorescence microscopy with negative contrast.
title_full_unstemmed	Intravital fluorescence microscopy with negative contrast.
title_sort	intravital fluorescence microscopy with negative contrast.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/d4d8b879f7d245b2bc20758c74e82da3
work_keys_str_mv	AT juwellwwu intravitalfluorescencemicroscopywithnegativecontrast AT yookyungjung intravitalfluorescencemicroscopywithnegativecontrast AT shuchiayeh intravitalfluorescencemicroscopywithnegativecontrast AT yongwanseo intravitalfluorescencemicroscopywithnegativecontrast AT judithmrunnels intravitalfluorescencemicroscopywithnegativecontrast AT christiansburns intravitalfluorescencemicroscopywithnegativecontrast AT toshihidemizoguchi intravitalfluorescencemicroscopywithnegativecontrast AT keisukeito intravitalfluorescencemicroscopywithnegativecontrast AT joelaspencer intravitalfluorescencemicroscopywithnegativecontrast AT charlesplin intravitalfluorescencemicroscopywithnegativecontrast
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Intravital fluorescence microscopy with negative contrast.

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