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Mesoscopic Oblique Plane Microscopy via Light-sheet Mirroring

Understanding the intricate interplay and inter-connectivity of biological processes across an entire organism is important in various fields of biology, including cardiovascular research, neuroscience, and developmental biology. Here, we present a mesoscopic oblique plane microscope (OPM) that enab...

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Autores principales: DAETWYLER, STEPHAN, CHANG, BO-JUI, CHEN, BINGYING, ZHOU, FELIX, FIOLKA, RETO
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10441428/
https://www.ncbi.nlm.nih.gov/pubmed/37609162
http://dx.doi.org/10.1101/2023.08.10.552834
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author DAETWYLER, STEPHAN
CHANG, BO-JUI
CHEN, BINGYING
ZHOU, FELIX
FIOLKA, RETO
author_facet DAETWYLER, STEPHAN
CHANG, BO-JUI
CHEN, BINGYING
ZHOU, FELIX
FIOLKA, RETO
author_sort DAETWYLER, STEPHAN
collection PubMed
description Understanding the intricate interplay and inter-connectivity of biological processes across an entire organism is important in various fields of biology, including cardiovascular research, neuroscience, and developmental biology. Here, we present a mesoscopic oblique plane microscope (OPM) that enables whole organism imaging with high speed and subcellular resolution. A microprism underneath the sample enhances the axial resolution and optical sectioning through total internal reflection of the light-sheet. Through rapid refocusing of the light-sheet, the imaging depth is extended up to threefold while keeping the axial resolution constant. Using low magnification objectives with a large field of view, we realize mesoscopic imaging over a volume of 3.7×1.5×1 mm(3) with ~2.3 microns lateral and ~9.2 microns axial resolution. Applying the mesoscopic OPM, we demonstrate in vivo and in toto whole organism imaging of the zebrafish vasculature and its endothelial nuclei, and blood flow dynamics at 12 Hz acquisition rate, resulting in a quantitative map of blood flow across the entire organism.
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spelling pubmed-104414282023-08-22 Mesoscopic Oblique Plane Microscopy via Light-sheet Mirroring DAETWYLER, STEPHAN CHANG, BO-JUI CHEN, BINGYING ZHOU, FELIX FIOLKA, RETO bioRxiv Article Understanding the intricate interplay and inter-connectivity of biological processes across an entire organism is important in various fields of biology, including cardiovascular research, neuroscience, and developmental biology. Here, we present a mesoscopic oblique plane microscope (OPM) that enables whole organism imaging with high speed and subcellular resolution. A microprism underneath the sample enhances the axial resolution and optical sectioning through total internal reflection of the light-sheet. Through rapid refocusing of the light-sheet, the imaging depth is extended up to threefold while keeping the axial resolution constant. Using low magnification objectives with a large field of view, we realize mesoscopic imaging over a volume of 3.7×1.5×1 mm(3) with ~2.3 microns lateral and ~9.2 microns axial resolution. Applying the mesoscopic OPM, we demonstrate in vivo and in toto whole organism imaging of the zebrafish vasculature and its endothelial nuclei, and blood flow dynamics at 12 Hz acquisition rate, resulting in a quantitative map of blood flow across the entire organism. Cold Spring Harbor Laboratory 2023-08-11 /pmc/articles/PMC10441428/ /pubmed/37609162 http://dx.doi.org/10.1101/2023.08.10.552834 Text en https://creativecommons.org/licenses/by-nd/4.0/This work is licensed under a Creative Commons Attribution-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, and only so long as attribution is given to the creator. The license allows for commercial use.
spellingShingle Article
DAETWYLER, STEPHAN
CHANG, BO-JUI
CHEN, BINGYING
ZHOU, FELIX
FIOLKA, RETO
Mesoscopic Oblique Plane Microscopy via Light-sheet Mirroring
title Mesoscopic Oblique Plane Microscopy via Light-sheet Mirroring
title_full Mesoscopic Oblique Plane Microscopy via Light-sheet Mirroring
title_fullStr Mesoscopic Oblique Plane Microscopy via Light-sheet Mirroring
title_full_unstemmed Mesoscopic Oblique Plane Microscopy via Light-sheet Mirroring
title_short Mesoscopic Oblique Plane Microscopy via Light-sheet Mirroring
title_sort mesoscopic oblique plane microscopy via light-sheet mirroring
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10441428/
https://www.ncbi.nlm.nih.gov/pubmed/37609162
http://dx.doi.org/10.1101/2023.08.10.552834
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