High-resolution intravital microscopy.

Cellular communication constitutes a fundamental mechanism of life, for instance by permitting transfer of information through synapses in the nervous system and by leading to activation of cells during the course of immune responses. Monitoring cell-cell interactions within living adult organisms i...

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Autores principales:	Volker Andresen, Karolin Pollok, Jan-Leo Rinnenthal, Laura Oehme, Robert Günther, Heinrich Spiecker, Helena Radbruch, Jenny Gerhard, Anje Sporbert, Zoltan Cseresnyes, Anja E Hauser, Raluca Niesner
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/cf05e5fc5e2444deb4d03774228d10c1
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spelling	oai:doaj.org-article:cf05e5fc5e2444deb4d03774228d10c12021-11-18T08:05:08ZHigh-resolution intravital microscopy.1932-620310.1371/journal.pone.0050915https://doaj.org/article/cf05e5fc5e2444deb4d03774228d10c12012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23251402/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Cellular communication constitutes a fundamental mechanism of life, for instance by permitting transfer of information through synapses in the nervous system and by leading to activation of cells during the course of immune responses. Monitoring cell-cell interactions within living adult organisms is crucial in order to draw conclusions on their behavior with respect to the fate of cells, tissues and organs. Until now, there is no technology available that enables dynamic imaging deep within the tissue of living adult organisms at sub-cellular resolution, i.e. detection at the level of few protein molecules. Here we present a novel approach called multi-beam striped-illumination which applies for the first time the principle and advantages of structured-illumination, spatial modulation of the excitation pattern, to laser-scanning-microscopy. We use this approach in two-photon-microscopy--the most adequate optical deep-tissue imaging-technique. As compared to standard two-photon-microscopy, it achieves significant contrast enhancement and up to 3-fold improved axial resolution (optical sectioning) while photobleaching, photodamage and acquisition speed are similar. Its imaging depth is comparable to multifocal two-photon-microscopy and only slightly less than in standard single-beam two-photon-microscopy. Precisely, our studies within mouse lymph nodes demonstrated 216% improved axial and 23% improved lateral resolutions at a depth of 80 µm below the surface. Thus, we are for the first time able to visualize the dynamic interactions between B cells and immune complex deposits on follicular dendritic cells within germinal centers (GCs) of live mice. These interactions play a decisive role in the process of clonal selection, leading to affinity maturation of the humoral immune response. This novel high-resolution intravital microscopy method has a huge potential for numerous applications in neurosciences, immunology, cancer research and developmental biology. Moreover, our striped-illumination approach is able to improve the resolution of any laser-scanning-microscope, including confocal microscopes, by simply choosing an appropriate detector.Volker AndresenKarolin PollokJan-Leo RinnenthalLaura OehmeRobert GüntherHeinrich SpieckerHelena RadbruchJenny GerhardAnje SporbertZoltan CseresnyesAnja E HauserRaluca NiesnerPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 12, p e50915 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Volker Andresen Karolin Pollok Jan-Leo Rinnenthal Laura Oehme Robert Günther Heinrich Spiecker Helena Radbruch Jenny Gerhard Anje Sporbert Zoltan Cseresnyes Anja E Hauser Raluca Niesner High-resolution intravital microscopy.
description	Cellular communication constitutes a fundamental mechanism of life, for instance by permitting transfer of information through synapses in the nervous system and by leading to activation of cells during the course of immune responses. Monitoring cell-cell interactions within living adult organisms is crucial in order to draw conclusions on their behavior with respect to the fate of cells, tissues and organs. Until now, there is no technology available that enables dynamic imaging deep within the tissue of living adult organisms at sub-cellular resolution, i.e. detection at the level of few protein molecules. Here we present a novel approach called multi-beam striped-illumination which applies for the first time the principle and advantages of structured-illumination, spatial modulation of the excitation pattern, to laser-scanning-microscopy. We use this approach in two-photon-microscopy--the most adequate optical deep-tissue imaging-technique. As compared to standard two-photon-microscopy, it achieves significant contrast enhancement and up to 3-fold improved axial resolution (optical sectioning) while photobleaching, photodamage and acquisition speed are similar. Its imaging depth is comparable to multifocal two-photon-microscopy and only slightly less than in standard single-beam two-photon-microscopy. Precisely, our studies within mouse lymph nodes demonstrated 216% improved axial and 23% improved lateral resolutions at a depth of 80 µm below the surface. Thus, we are for the first time able to visualize the dynamic interactions between B cells and immune complex deposits on follicular dendritic cells within germinal centers (GCs) of live mice. These interactions play a decisive role in the process of clonal selection, leading to affinity maturation of the humoral immune response. This novel high-resolution intravital microscopy method has a huge potential for numerous applications in neurosciences, immunology, cancer research and developmental biology. Moreover, our striped-illumination approach is able to improve the resolution of any laser-scanning-microscope, including confocal microscopes, by simply choosing an appropriate detector.
format	article
author	Volker Andresen Karolin Pollok Jan-Leo Rinnenthal Laura Oehme Robert Günther Heinrich Spiecker Helena Radbruch Jenny Gerhard Anje Sporbert Zoltan Cseresnyes Anja E Hauser Raluca Niesner
author_facet	Volker Andresen Karolin Pollok Jan-Leo Rinnenthal Laura Oehme Robert Günther Heinrich Spiecker Helena Radbruch Jenny Gerhard Anje Sporbert Zoltan Cseresnyes Anja E Hauser Raluca Niesner
author_sort	Volker Andresen
title	High-resolution intravital microscopy.
title_short	High-resolution intravital microscopy.
title_full	High-resolution intravital microscopy.
title_fullStr	High-resolution intravital microscopy.
title_full_unstemmed	High-resolution intravital microscopy.
title_sort	high-resolution intravital microscopy.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/cf05e5fc5e2444deb4d03774228d10c1
work_keys_str_mv	AT volkerandresen highresolutionintravitalmicroscopy AT karolinpollok highresolutionintravitalmicroscopy AT janleorinnenthal highresolutionintravitalmicroscopy AT lauraoehme highresolutionintravitalmicroscopy AT robertgunther highresolutionintravitalmicroscopy AT heinrichspiecker highresolutionintravitalmicroscopy AT helenaradbruch highresolutionintravitalmicroscopy AT jennygerhard highresolutionintravitalmicroscopy AT anjesporbert highresolutionintravitalmicroscopy AT zoltancseresnyes highresolutionintravitalmicroscopy AT anjaehauser highresolutionintravitalmicroscopy AT ralucaniesner highresolutionintravitalmicroscopy
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High-resolution intravital microscopy.

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