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Real-time volumetric microscopy of in-vivo dynamics and large-scale samples with SCAPE 2.0
The limited per-pixel bandwidth of most microscopy methods requires compromises between field of view, sampling density and imaging speed. This limitation constrains studies involving complex motion or fast cellular signaling, and presents a major bottleneck for high-throughput structural imaging. H...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6885017/ https://www.ncbi.nlm.nih.gov/pubmed/31562489 http://dx.doi.org/10.1038/s41592-019-0579-4 |
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author | Voleti, Venkatakaushik Patel, Kripa B. Li, Wenze Campos, Citlali Perez Bharadwaj, Srinidhi Yu, Hang Ford, Caitlin Casper, Malte J. Yan, Richard Wenwei Liang, Wenxuan Wen, Chentao Kimura, Koutarou D. Targoff, Kimara L. Hillman, Elizabeth M.C. |
author_facet | Voleti, Venkatakaushik Patel, Kripa B. Li, Wenze Campos, Citlali Perez Bharadwaj, Srinidhi Yu, Hang Ford, Caitlin Casper, Malte J. Yan, Richard Wenwei Liang, Wenxuan Wen, Chentao Kimura, Koutarou D. Targoff, Kimara L. Hillman, Elizabeth M.C. |
author_sort | Voleti, Venkatakaushik |
collection | PubMed |
description | The limited per-pixel bandwidth of most microscopy methods requires compromises between field of view, sampling density and imaging speed. This limitation constrains studies involving complex motion or fast cellular signaling, and presents a major bottleneck for high-throughput structural imaging. Here, we combine high-speed intensified camera technology with a versatile, reconfigurable and dramatically improved Swept, Confocally Aligned Planar Excitation (SCAPE) microscope design that can achieve high-resolution volumetric imaging at over 300 volumes-per-second and over 1.2 GHz pixel rates. We demonstrate near-isotropic sampling in freely moving C. elegans, and analyze real-time blood flow and calcium dynamics in the beating zebrafish heart. The same system also permits high-throughput structural imaging of mounted, intact, cleared and expanded samples. SCAPE 2.0’s significantly lower photodamage compared to point-scanning techniques is also confirmed. Our results demonstrate that SCAPE 2.0 is a powerful, yet accessible imaging platform for myriad emerging high-speed dynamic and high-throughput volumetric microscopy applications. |
format | Online Article Text |
id | pubmed-6885017 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
record_format | MEDLINE/PubMed |
spelling | pubmed-68850172020-03-27 Real-time volumetric microscopy of in-vivo dynamics and large-scale samples with SCAPE 2.0 Voleti, Venkatakaushik Patel, Kripa B. Li, Wenze Campos, Citlali Perez Bharadwaj, Srinidhi Yu, Hang Ford, Caitlin Casper, Malte J. Yan, Richard Wenwei Liang, Wenxuan Wen, Chentao Kimura, Koutarou D. Targoff, Kimara L. Hillman, Elizabeth M.C. Nat Methods Article The limited per-pixel bandwidth of most microscopy methods requires compromises between field of view, sampling density and imaging speed. This limitation constrains studies involving complex motion or fast cellular signaling, and presents a major bottleneck for high-throughput structural imaging. Here, we combine high-speed intensified camera technology with a versatile, reconfigurable and dramatically improved Swept, Confocally Aligned Planar Excitation (SCAPE) microscope design that can achieve high-resolution volumetric imaging at over 300 volumes-per-second and over 1.2 GHz pixel rates. We demonstrate near-isotropic sampling in freely moving C. elegans, and analyze real-time blood flow and calcium dynamics in the beating zebrafish heart. The same system also permits high-throughput structural imaging of mounted, intact, cleared and expanded samples. SCAPE 2.0’s significantly lower photodamage compared to point-scanning techniques is also confirmed. Our results demonstrate that SCAPE 2.0 is a powerful, yet accessible imaging platform for myriad emerging high-speed dynamic and high-throughput volumetric microscopy applications. 2019-09-27 2019-10 /pmc/articles/PMC6885017/ /pubmed/31562489 http://dx.doi.org/10.1038/s41592-019-0579-4 Text en Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms |
spellingShingle | Article Voleti, Venkatakaushik Patel, Kripa B. Li, Wenze Campos, Citlali Perez Bharadwaj, Srinidhi Yu, Hang Ford, Caitlin Casper, Malte J. Yan, Richard Wenwei Liang, Wenxuan Wen, Chentao Kimura, Koutarou D. Targoff, Kimara L. Hillman, Elizabeth M.C. Real-time volumetric microscopy of in-vivo dynamics and large-scale samples with SCAPE 2.0 |
title | Real-time volumetric microscopy of in-vivo dynamics and large-scale samples with SCAPE 2.0 |
title_full | Real-time volumetric microscopy of in-vivo dynamics and large-scale samples with SCAPE 2.0 |
title_fullStr | Real-time volumetric microscopy of in-vivo dynamics and large-scale samples with SCAPE 2.0 |
title_full_unstemmed | Real-time volumetric microscopy of in-vivo dynamics and large-scale samples with SCAPE 2.0 |
title_short | Real-time volumetric microscopy of in-vivo dynamics and large-scale samples with SCAPE 2.0 |
title_sort | real-time volumetric microscopy of in-vivo dynamics and large-scale samples with scape 2.0 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6885017/ https://www.ncbi.nlm.nih.gov/pubmed/31562489 http://dx.doi.org/10.1038/s41592-019-0579-4 |
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