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Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy (Conpokal) on Live Cells

Techniques available for micro- and nano-scale mechanical characterization have exploded in the last few decades. From further development of the scanning and transmission electron microscope, to the invention of atomic force microscopy, and advances in fluorescent imaging, there have been substanti...

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Autores principales: Sandin, Joree N., Aryal, Surya P., Wilkop, Thomas, Richards, Christopher I., Grady, Martha E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7680637/
https://www.ncbi.nlm.nih.gov/pubmed/32865532
http://dx.doi.org/10.3791/61433
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author Sandin, Joree N.
Aryal, Surya P.
Wilkop, Thomas
Richards, Christopher I.
Grady, Martha E.
author_facet Sandin, Joree N.
Aryal, Surya P.
Wilkop, Thomas
Richards, Christopher I.
Grady, Martha E.
author_sort Sandin, Joree N.
collection PubMed
description Techniques available for micro- and nano-scale mechanical characterization have exploded in the last few decades. From further development of the scanning and transmission electron microscope, to the invention of atomic force microscopy, and advances in fluorescent imaging, there have been substantial gains in technologies that enable the study of small materials. Conpokal is a portmanteau that combines confocal microscopy with atomic force microscopy (AFM), where a probe “pokes” the surface. Although each technique is extremely effective for the qualitative and/or quantitative image collection on their own, Conpokal provides the capability to test with blended fluorescence imaging and mechanical characterization. Designed for near simultaneous confocal imaging and atomic force probing, Conpokal facilitates experimentation on live microbiological samples. The added insight from paired instrumentation provides co-localization of measured mechanical properties (e.g., elastic modulus, adhesion, surface roughness) by AFM with subcellular components or activity observable through confocal microscopy. This work provides a step by step protocol for the operation of laser scanning confocal and atomic force microscopy, simultaneously, to achieve same cell, same region, confocal imaging, and mechanical characterization.
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spelling pubmed-76806372020-11-22 Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy (Conpokal) on Live Cells Sandin, Joree N. Aryal, Surya P. Wilkop, Thomas Richards, Christopher I. Grady, Martha E. J Vis Exp Article Techniques available for micro- and nano-scale mechanical characterization have exploded in the last few decades. From further development of the scanning and transmission electron microscope, to the invention of atomic force microscopy, and advances in fluorescent imaging, there have been substantial gains in technologies that enable the study of small materials. Conpokal is a portmanteau that combines confocal microscopy with atomic force microscopy (AFM), where a probe “pokes” the surface. Although each technique is extremely effective for the qualitative and/or quantitative image collection on their own, Conpokal provides the capability to test with blended fluorescence imaging and mechanical characterization. Designed for near simultaneous confocal imaging and atomic force probing, Conpokal facilitates experimentation on live microbiological samples. The added insight from paired instrumentation provides co-localization of measured mechanical properties (e.g., elastic modulus, adhesion, surface roughness) by AFM with subcellular components or activity observable through confocal microscopy. This work provides a step by step protocol for the operation of laser scanning confocal and atomic force microscopy, simultaneously, to achieve same cell, same region, confocal imaging, and mechanical characterization. 2020-08-11 /pmc/articles/PMC7680637/ /pubmed/32865532 http://dx.doi.org/10.3791/61433 Text en http://creativecommons.org/licenses/by-nc-nd/3.0/ JoVE Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License
spellingShingle Article
Sandin, Joree N.
Aryal, Surya P.
Wilkop, Thomas
Richards, Christopher I.
Grady, Martha E.
Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy (Conpokal) on Live Cells
title Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy (Conpokal) on Live Cells
title_full Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy (Conpokal) on Live Cells
title_fullStr Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy (Conpokal) on Live Cells
title_full_unstemmed Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy (Conpokal) on Live Cells
title_short Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy (Conpokal) on Live Cells
title_sort near simultaneous laser scanning confocal and atomic force microscopy (conpokal) on live cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7680637/
https://www.ncbi.nlm.nih.gov/pubmed/32865532
http://dx.doi.org/10.3791/61433
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