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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
AIP Publishing LLC
2020
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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. |
format | Online Article Text |
id | pubmed-7255815 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | AIP Publishing LLC |
record_format | MEDLINE/PubMed |
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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