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A versatile, multi-laser twin-microscope system for light-sheet imaging

Light-sheet microscopy offers faster imaging and reduced phototoxicity in comparison to conventional point-scanning microscopy, making it a preferred technique for imaging biological dynamics for durations of hours or days. Such extended imaging sessions pose a challenge, as it reduces the number of...

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Detalles Bibliográficos
Autores principales: Keomanee-Dizon, Kevin, Fraser, Scott E., Truong, Thai V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AIP Publishing LLC 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7255815/
https://www.ncbi.nlm.nih.gov/pubmed/32486724
http://dx.doi.org/10.1063/1.5144487
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author Keomanee-Dizon, Kevin
Fraser, Scott E.
Truong, Thai V.
author_facet Keomanee-Dizon, Kevin
Fraser, Scott E.
Truong, Thai V.
author_sort Keomanee-Dizon, Kevin
collection PubMed
description Light-sheet microscopy offers faster imaging and reduced phototoxicity in comparison to conventional point-scanning microscopy, making it a preferred technique for imaging biological dynamics for durations of hours or days. Such extended imaging sessions pose a challenge, as it reduces the number of specimens that can be imaged in a given day. Here, we present a versatile light-sheet imaging instrument that combines two independently controlled microscope-twins, built so that they can share an ultrafast near-infrared laser and a bank of continuous-wave visible lasers, increasing the throughput and decreasing the cost. To permit a wide variety of specimens to be imaged, each microscope-twin provides flexible imaging parameters, including (i) operation in one-photon and/or two-photon excitation modes, (ii) delivery of one to three light-sheets via a trio of orthogonal excitation arms, (iii) sub-micron to micron imaging resolution, (iv) multicolor compatibility, and (v) upright (with provision for inverted) detection geometry. We offer a detailed description of the twin-microscope design to aid instrument builders who wish to construct and use similar systems. We demonstrate the instrument’s versatility for biological investigation by performing fast imaging of the beating heart in an intact zebrafish embryo, deep imaging of thick patient-derived tumor organoids, and gentle whole-brain imaging of neural activity in behaving larval zebrafish.
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spelling pubmed-72558152020-06-03 A versatile, multi-laser twin-microscope system for light-sheet imaging Keomanee-Dizon, Kevin Fraser, Scott E. Truong, Thai V. Rev Sci Instrum ARTICLES Light-sheet microscopy offers faster imaging and reduced phototoxicity in comparison to conventional point-scanning microscopy, making it a preferred technique for imaging biological dynamics for durations of hours or days. Such extended imaging sessions pose a challenge, as it reduces the number of specimens that can be imaged in a given day. Here, we present a versatile light-sheet imaging instrument that combines two independently controlled microscope-twins, built so that they can share an ultrafast near-infrared laser and a bank of continuous-wave visible lasers, increasing the throughput and decreasing the cost. To permit a wide variety of specimens to be imaged, each microscope-twin provides flexible imaging parameters, including (i) operation in one-photon and/or two-photon excitation modes, (ii) delivery of one to three light-sheets via a trio of orthogonal excitation arms, (iii) sub-micron to micron imaging resolution, (iv) multicolor compatibility, and (v) upright (with provision for inverted) detection geometry. We offer a detailed description of the twin-microscope design to aid instrument builders who wish to construct and use similar systems. We demonstrate the instrument’s versatility for biological investigation by performing fast imaging of the beating heart in an intact zebrafish embryo, deep imaging of thick patient-derived tumor organoids, and gentle whole-brain imaging of neural activity in behaving larval zebrafish. AIP Publishing LLC 2020-05-01 2020-05-27 /pmc/articles/PMC7255815/ /pubmed/32486724 http://dx.doi.org/10.1063/1.5144487 Text en © 2020 Author(s). 0034-6748/2020/91(5)/053703/14/$0.00 All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle ARTICLES
Keomanee-Dizon, Kevin
Fraser, Scott E.
Truong, Thai V.
A versatile, multi-laser twin-microscope system for light-sheet imaging
title A versatile, multi-laser twin-microscope system for light-sheet imaging
title_full A versatile, multi-laser twin-microscope system for light-sheet imaging
title_fullStr A versatile, multi-laser twin-microscope system for light-sheet imaging
title_full_unstemmed A versatile, multi-laser twin-microscope system for light-sheet imaging
title_short A versatile, multi-laser twin-microscope system for light-sheet imaging
title_sort versatile, multi-laser twin-microscope system for light-sheet imaging
topic ARTICLES
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7255815/
https://www.ncbi.nlm.nih.gov/pubmed/32486724
http://dx.doi.org/10.1063/1.5144487
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