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A polymer gel index-matched to water enables diverse applications in fluorescence microscopy

Xiaofei Han, Yijun Su, Hamilton White, Kate M. O’Neill, Nicole Y. Morgan, Ryan Christensen, Deepika Potarazu, Harshad D. Vishwasrao, Stephen Xu, Yilun Sun, Shar-yin Huang, Mark W. Moyle, Qionghai Dai, Yves Pommier, Edward Giniger, Dirk R. Albrecht, Roland Probst, Hari Shroff
doi: https://doi.org/10.1101/2020.10.04.324996
Xiaofei Han
1Laboratory of High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA
2Department of Automation, Tsinghua University, 100084, Beijing, China
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  • For correspondence: hxf16@mails.tsinghua.edu.cn suy4nih@gmail.com hawhite@wpi.edu
Yijun Su
1Laboratory of High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA
3Advanced Imaging and Microscopy Resource, National Institutes of Health, Bethesda, Maryland, USA
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  • For correspondence: hxf16@mails.tsinghua.edu.cn suy4nih@gmail.com hawhite@wpi.edu
Hamilton White
4Department of Biomedical Engineering, Worcester Polytechnic Institute, 100 Institute Road, Worcester, MA, 01609, USA
5Department of Neurobiology, University of Massachusetts Medical School, Worcester, MA 01655, USA
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  • For correspondence: hxf16@mails.tsinghua.edu.cn suy4nih@gmail.com hawhite@wpi.edu
Kate M. O’Neill
6National Institute of Neurological Disorders and Stroke, NIH, Bethesda, MD, 20892, USA
7Institute for Physical Science and Technology, University of Maryland College Park, College Park, Maryland, USA
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Nicole Y. Morgan
8National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA
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Ryan Christensen
1Laboratory of High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA
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Deepika Potarazu
1Laboratory of High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA
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Harshad D. Vishwasrao
3Advanced Imaging and Microscopy Resource, National Institutes of Health, Bethesda, Maryland, USA
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Stephen Xu
1Laboratory of High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA
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Yilun Sun
9Developmental Therapeutics Branch and Laboratory of Molecular Pharmacology, Center for Cancer Research, National Institutes of Health, Bethesda, Maryland, USA
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Shar-yin Huang
9Developmental Therapeutics Branch and Laboratory of Molecular Pharmacology, Center for Cancer Research, National Institutes of Health, Bethesda, Maryland, USA
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Mark W. Moyle
10Department of Neuroscience and Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06536, USA
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Qionghai Dai
2Department of Automation, Tsinghua University, 100084, Beijing, China
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Yves Pommier
9Developmental Therapeutics Branch and Laboratory of Molecular Pharmacology, Center for Cancer Research, National Institutes of Health, Bethesda, Maryland, USA
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Edward Giniger
6National Institute of Neurological Disorders and Stroke, NIH, Bethesda, MD, 20892, USA
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Dirk R. Albrecht
4Department of Biomedical Engineering, Worcester Polytechnic Institute, 100 Institute Road, Worcester, MA, 01609, USA
11Department of Biology and Biotechnology, Worcester Polytechnic Institute, 100 Institute Road, Worcester, MA, 01609, USA
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Roland Probst
12ACUITYnano, Innovation in Biomedical Imaging, North Bethesda, Maryland, USA
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Hari Shroff
1Laboratory of High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA
3Advanced Imaging and Microscopy Resource, National Institutes of Health, Bethesda, Maryland, USA
13Marine Biological Laboratory Fellows Program, Woods Hole, MA, USA
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Abstract

We demonstrate diffraction-limited and super-resolution imaging through thick layers (tens-hundreds of microns) of BIO-133, a biocompatible, UV-curable, commercially available polymer with a refractive index (RI) matched to water. We show that cells can be directly grown on BIO-133 substrates without the need for surface passivation and use this capability to perform extended time-lapse volumetric imaging of cellular dynamics 1) at isotropic resolution using dual-view light-sheet microscopy, and 2) at super-resolution using instant structured illumination microscopy. BIO-133 also enables immobilization of 1) Drosophila tissue, allowing us to track membrane puncta in pioneer neurons, and 2) Caenorhabditis elegans, which allows us to image and inspect fine neural structure and to track pan-neuronal calcium activity over hundreds of volumes. Finally, BIO-133 is compatible with other microfluidic materials, enabling optical and chemical perturbation of immobilized samples, as we demonstrate by performing drug and optogenetic stimulation on cells and C. elegans.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. This article is a US Government work. It is not subject to copyright under 17 USC 105 and is also made available for use under a CC0 license.
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Posted October 11, 2020.
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A polymer gel index-matched to water enables diverse applications in fluorescence microscopy
Xiaofei Han, Yijun Su, Hamilton White, Kate M. O’Neill, Nicole Y. Morgan, Ryan Christensen, Deepika Potarazu, Harshad D. Vishwasrao, Stephen Xu, Yilun Sun, Shar-yin Huang, Mark W. Moyle, Qionghai Dai, Yves Pommier, Edward Giniger, Dirk R. Albrecht, Roland Probst, Hari Shroff
bioRxiv 2020.10.04.324996; doi: https://doi.org/10.1101/2020.10.04.324996
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A polymer gel index-matched to water enables diverse applications in fluorescence microscopy
Xiaofei Han, Yijun Su, Hamilton White, Kate M. O’Neill, Nicole Y. Morgan, Ryan Christensen, Deepika Potarazu, Harshad D. Vishwasrao, Stephen Xu, Yilun Sun, Shar-yin Huang, Mark W. Moyle, Qionghai Dai, Yves Pommier, Edward Giniger, Dirk R. Albrecht, Roland Probst, Hari Shroff
bioRxiv 2020.10.04.324996; doi: https://doi.org/10.1101/2020.10.04.324996

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