A polymer 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:10.1039/D0LC01233E

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

Xiaofei Han,

‡*^ab Yijun Su,‡*^ac Hamilton White,

‡*^de Kate M. O'Neill,

^fg Nicole Y. Morgan,^h Ryan Christensen,^a Deepika Potarazu,^a Harshad D. Vishwasrao,^c Stephen Xu,^a Yilun Sun,ⁱ Shar-yin Huang,ⁱ Mark W. Moyle,^j Qionghai Dai,^b Yves Pommier,ⁱ Edward Giniger,^f Dirk R. Albrecht,

‡^dk Roland Probst

‡^l and Hari Shroff ‡^acm

Author affiliations

* Corresponding authors

^a Laboratory of High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA
E-mail: suy4nih@gmail.com

^b Department of Automation, Tsinghua University, Beijing, China
E-mail: hxf16@mails.tsinghua.edu.cn

^c Advanced Imaging and Microscopy Resource, National Institutes of Health, Bethesda, MD, USA

^d Department of Biomedical Engineering, Worcester Polytechnic Institute, 100 Institute Road, Worcester, MA, USA
E-mail: hawhite@wpi.edu

^e Department of Neurobiology, University of Massachusetts Medical School, Worcester, MA, USA

^f National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA

^g Institute for Physical Science and Technology, University of Maryland College Park, College Park, MD, USA

^h National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA

ⁱ Developmental Therapeutics Branch and Laboratory of Molecular Pharmacology, Center for Cancer Research, National Institutes of Health, Bethesda, MD, USA

^j Department of Neuroscience and Department of Cell Biology, Yale University School of Medicine, New Haven, CT, USA

^k Department of Biology and Biotechnology, Worcester Polytechnic Institute, 100 Institute Road, Worcester, MA, USA

^l ACUITYnano, Innovation in Biomedical Imaging, North Bethesda, MD, USA

^m Marine Biological Laboratory Fellows Program, Woods Hole, MA, USA

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.

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

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

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