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...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24933027/pdf/?tool=EBI |
| _version_ | 1819180970116382720 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-22T22:22:48Z |
| format | Article |
| id | doaj.art-832d8d5c1b2e4f1495074f95fdf9353e |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-22T22:22:48Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-832d8d5c1b2e4f1495074f95fdf9353e2022-12-21T18:10:38ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0196e9961510.1371/journal.pone.0099615Label-free determination of hemodynamic parameters in the microcirculaton with third harmonic generation microscopy.Steffen DietzelJoachim PircherA Katharina NekollaMazhar GullAndré W BrändliUlrich PohlMarkus RehbergDetermination 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.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24933027/pdf/?tool=EBI |
| spellingShingle | 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. PLoS ONE |
| title | 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_short | 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 |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24933027/pdf/?tool=EBI |
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