Combined Atomic Force Microscope and Volumetric Light Sheet System for Correlative Force and Fluorescence Mechanobiology Studies

Abstract The central goals of mechanobiology are to understand how cells generate force and how they respond to environmental mechanical stimuli. A full picture of these processes requires high-resolution, volumetric imaging with time-correlated force measurements. Here we present an instrument that...

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Autores principales:	E. Nelsen, C. M. Hobson, M. E. Kern, J. P. Hsiao, E. T. O’Brien III, T. Watanabe, B. M. Condon, M. Boyce, S. Grinstein, K. M. Hahn, M. R. Falvo, R. Superfine
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b2465c36f5de475da8a8f1ee5b8006ac
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spelling	oai:doaj.org-article:b2465c36f5de475da8a8f1ee5b8006ac2021-12-02T14:59:09ZCombined Atomic Force Microscope and Volumetric Light Sheet System for Correlative Force and Fluorescence Mechanobiology Studies10.1038/s41598-020-65205-82045-2322https://doaj.org/article/b2465c36f5de475da8a8f1ee5b8006ac2020-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-65205-8https://doaj.org/toc/2045-2322Abstract The central goals of mechanobiology are to understand how cells generate force and how they respond to environmental mechanical stimuli. A full picture of these processes requires high-resolution, volumetric imaging with time-correlated force measurements. Here we present an instrument that combines an open-top, single-objective light sheet fluorescence microscope with an atomic force microscope (AFM), providing simultaneous volumetric imaging with high spatiotemporal resolution and high dynamic range force capability (10 pN – 100 nN). With this system we have captured lysosome trafficking, vimentin nuclear caging, and actin dynamics on the order of one second per single-cell volume. To showcase the unique advantages of combining Line Bessel light sheet imaging with AFM, we measured the forces exerted by a macrophage during FcɣR-mediated phagocytosis while performing both sequential two-color, fixed plane and volumetric imaging of F-actin. This unique instrument allows for a myriad of novel studies investigating the coupling of cellular dynamics and mechanical forces.E. NelsenC. M. HobsonM. E. KernJ. P. HsiaoE. T. O’Brien IIIT. WatanabeB. M. CondonM. BoyceS. GrinsteinK. M. HahnM. R. FalvoR. SuperfineNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-12 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q E. Nelsen C. M. Hobson M. E. Kern J. P. Hsiao E. T. O’Brien III T. Watanabe B. M. Condon M. Boyce S. Grinstein K. M. Hahn M. R. Falvo R. Superfine Combined Atomic Force Microscope and Volumetric Light Sheet System for Correlative Force and Fluorescence Mechanobiology Studies
description	Abstract The central goals of mechanobiology are to understand how cells generate force and how they respond to environmental mechanical stimuli. A full picture of these processes requires high-resolution, volumetric imaging with time-correlated force measurements. Here we present an instrument that combines an open-top, single-objective light sheet fluorescence microscope with an atomic force microscope (AFM), providing simultaneous volumetric imaging with high spatiotemporal resolution and high dynamic range force capability (10 pN – 100 nN). With this system we have captured lysosome trafficking, vimentin nuclear caging, and actin dynamics on the order of one second per single-cell volume. To showcase the unique advantages of combining Line Bessel light sheet imaging with AFM, we measured the forces exerted by a macrophage during FcɣR-mediated phagocytosis while performing both sequential two-color, fixed plane and volumetric imaging of F-actin. This unique instrument allows for a myriad of novel studies investigating the coupling of cellular dynamics and mechanical forces.
format	article
author	E. Nelsen C. M. Hobson M. E. Kern J. P. Hsiao E. T. O’Brien III T. Watanabe B. M. Condon M. Boyce S. Grinstein K. M. Hahn M. R. Falvo R. Superfine
author_facet	E. Nelsen C. M. Hobson M. E. Kern J. P. Hsiao E. T. O’Brien III T. Watanabe B. M. Condon M. Boyce S. Grinstein K. M. Hahn M. R. Falvo R. Superfine
author_sort	E. Nelsen
title	Combined Atomic Force Microscope and Volumetric Light Sheet System for Correlative Force and Fluorescence Mechanobiology Studies
title_short	Combined Atomic Force Microscope and Volumetric Light Sheet System for Correlative Force and Fluorescence Mechanobiology Studies
title_full	Combined Atomic Force Microscope and Volumetric Light Sheet System for Correlative Force and Fluorescence Mechanobiology Studies
title_fullStr	Combined Atomic Force Microscope and Volumetric Light Sheet System for Correlative Force and Fluorescence Mechanobiology Studies
title_full_unstemmed	Combined Atomic Force Microscope and Volumetric Light Sheet System for Correlative Force and Fluorescence Mechanobiology Studies
title_sort	combined atomic force microscope and volumetric light sheet system for correlative force and fluorescence mechanobiology studies
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/b2465c36f5de475da8a8f1ee5b8006ac
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Combined Atomic Force Microscope and Volumetric Light Sheet System for Correlative Force and Fluorescence Mechanobiology Studies

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