Label-free determination of hemodynamic parameters in the microcirculaton with third harmonic generation microscopy.

Determination of blood flow velocity and related hemodynamic parameters is an important aspect of physiological studies which in many settings requires fluorescent labeling. Here we show that Third Harmonic Generation (THG) microscopy is a suitable tool for label-free intravital investigations of th...

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Autores principales:	Steffen Dietzel, Joachim Pircher, A Katharina Nekolla, Mazhar Gull, André W Brändli, Ulrich Pohl, Markus Rehberg
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/832d8d5c1b2e4f1495074f95fdf9353e
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spelling	oai:doaj.org-article:832d8d5c1b2e4f1495074f95fdf9353e2021-11-18T08:15:28ZLabel-free determination of hemodynamic parameters in the microcirculaton with third harmonic generation microscopy.1932-620310.1371/journal.pone.0099615https://doaj.org/article/832d8d5c1b2e4f1495074f95fdf9353e2014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24933027/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Determination of blood flow velocity and related hemodynamic parameters is an important aspect of physiological studies which in many settings requires fluorescent labeling. Here we show that Third Harmonic Generation (THG) microscopy is a suitable tool for label-free intravital investigations of the microcirculation in widely-used physiological model systems. THG microscopy is a non-fluorescent multi-photon scanning technique combining the advantages of label-free imaging with restriction of signal generation to a focal spot. Blood flow was visualized and its velocity was measured in adult mouse cremaster muscle vessels, non-invasively in mouse ear vessels and in Xenopus tadpoles. In arterioles, THG line scanning allowed determination of the flow pulse velocity curve and hence the heart rate. By relocating the scan line we obtained velocity profiles through vessel diameters, allowing shear rate calculations. The cell free layer containing the glycocalyx was also visualized. Comparison of the current microscopic resolution with theoretical, diffraction limited resolution let us conclude that an about sixty-fold THG signal intensity increase may be possible with future improved optics, optimized for 1200-1300 nm excitation. THG microscopy is compatible with simultaneous two-photon excited fluorescence detection. It thus also provides the opportunity to determine important hemodynamic parameters in parallel to common fluorescent observations without additional label.Steffen DietzelJoachim PircherA Katharina NekollaMazhar GullAndré W BrändliUlrich PohlMarkus RehbergPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 6, p e99615 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Steffen Dietzel Joachim Pircher A Katharina Nekolla Mazhar Gull André W Brändli Ulrich Pohl Markus Rehberg Label-free determination of hemodynamic parameters in the microcirculaton with third harmonic generation microscopy.
description	Determination of blood flow velocity and related hemodynamic parameters is an important aspect of physiological studies which in many settings requires fluorescent labeling. Here we show that Third Harmonic Generation (THG) microscopy is a suitable tool for label-free intravital investigations of the microcirculation in widely-used physiological model systems. THG microscopy is a non-fluorescent multi-photon scanning technique combining the advantages of label-free imaging with restriction of signal generation to a focal spot. Blood flow was visualized and its velocity was measured in adult mouse cremaster muscle vessels, non-invasively in mouse ear vessels and in Xenopus tadpoles. In arterioles, THG line scanning allowed determination of the flow pulse velocity curve and hence the heart rate. By relocating the scan line we obtained velocity profiles through vessel diameters, allowing shear rate calculations. The cell free layer containing the glycocalyx was also visualized. Comparison of the current microscopic resolution with theoretical, diffraction limited resolution let us conclude that an about sixty-fold THG signal intensity increase may be possible with future improved optics, optimized for 1200-1300 nm excitation. THG microscopy is compatible with simultaneous two-photon excited fluorescence detection. It thus also provides the opportunity to determine important hemodynamic parameters in parallel to common fluorescent observations without additional label.
format	article
author	Steffen Dietzel Joachim Pircher A Katharina Nekolla Mazhar Gull André W Brändli Ulrich Pohl Markus Rehberg
author_facet	Steffen Dietzel Joachim Pircher A Katharina Nekolla Mazhar Gull André W Brändli Ulrich Pohl Markus Rehberg
author_sort	Steffen Dietzel
title	Label-free determination of hemodynamic parameters in the microcirculaton with third harmonic generation microscopy.
title_short	Label-free determination of hemodynamic parameters in the microcirculaton with third harmonic generation microscopy.
title_full	Label-free determination of hemodynamic parameters in the microcirculaton with third harmonic generation microscopy.
title_fullStr	Label-free determination of hemodynamic parameters in the microcirculaton with third harmonic generation microscopy.
title_full_unstemmed	Label-free determination of hemodynamic parameters in the microcirculaton with third harmonic generation microscopy.
title_sort	label-free determination of hemodynamic parameters in the microcirculaton with third harmonic generation microscopy.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/832d8d5c1b2e4f1495074f95fdf9353e
work_keys_str_mv	AT steffendietzel labelfreedeterminationofhemodynamicparametersinthemicrocirculatonwiththirdharmonicgenerationmicroscopy AT joachimpircher labelfreedeterminationofhemodynamicparametersinthemicrocirculatonwiththirdharmonicgenerationmicroscopy AT akatharinanekolla labelfreedeterminationofhemodynamicparametersinthemicrocirculatonwiththirdharmonicgenerationmicroscopy AT mazhargull labelfreedeterminationofhemodynamicparametersinthemicrocirculatonwiththirdharmonicgenerationmicroscopy AT andrewbrandli labelfreedeterminationofhemodynamicparametersinthemicrocirculatonwiththirdharmonicgenerationmicroscopy AT ulrichpohl labelfreedeterminationofhemodynamicparametersinthemicrocirculatonwiththirdharmonicgenerationmicroscopy AT markusrehberg labelfreedeterminationofhemodynamicparametersinthemicrocirculatonwiththirdharmonicgenerationmicroscopy
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Label-free determination of hemodynamic parameters in the microcirculaton with third harmonic generation microscopy.

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