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Genetically Encoded Phase Contrast Agents for Digital Holographic Microscopy

Farhadi, Arash and Bedrossian, Manuel and Lee, Justin and Ho, Gabrielle H. and Shapiro, Mikhail G. and Nadeau, Jay L. (2020) Genetically Encoded Phase Contrast Agents for Digital Holographic Microscopy. Nano Letters, 20 (11). pp. 8127-8134. ISSN 1530-6984. https://resolver.caltech.edu/CaltechAUTHORS:20191113-111723156

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Abstract

Quantitative phase imaging and digital holographic microscopy have shown great promise for visualizing the motion, structure, and physiology of microorganisms and mammalian cells in three dimensions. However, these imaging techniques currently lack molecular contrast agents analogous to the fluorescent dyes and proteins that have revolutionized fluorescence microscopy. Here we introduce the first genetically encodable phase contrast agents based on gas vesicles. The relatively low index of refraction of the air-filled core of gas vesicles results in optical phase advancement relative to aqueous media, making them a “positive” phase contrast agent easily distinguished from organelles, dyes, or microminerals. We demonstrate this capability by identifying and tracking the motion of gas vesicles and gas vesicle-expressing bacteria using digital holographic microscopy, and by imaging the uptake of engineered gas vesicles by mammalian cells. These results give phase imaging a biomolecular contrast agent, expanding the capabilities of this powerful technology for three-dimensional biological imaging.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1021/acs.nanolett.0c03159DOIArticle
https://doi.org/10.1101/833830DOIDiscussion Paper
ORCID:
AuthorORCID
Farhadi, Arash0000-0001-9137-8559
Lee, Justin0000-0002-3657-4386
Ho, Gabrielle H.0000-0002-8511-5549
Shapiro, Mikhail G.0000-0002-0291-4215
Nadeau, Jay L.0000-0001-5258-0076
Additional Information:© 2020 American Chemical Society. Received: July 31, 2020; Revised: October 21, 2020; Published: October 29, 2020. A.F. was supported by the NSERC graduate fellowship. Research in the Shapiro lab was supported by the National Institutes of Health (R01EB018975), the Packard Fellowship in Science and Engineering, the Pew Scholarship in the Biomedical Sciences, and the Heritage Medical Research Institute. The Nadeau lab was supported by the Gordon and Betty Moore Foundation (Grant 4038) and the National Science Foundation (NSF) 1828793. Portions of this work were supported under a contract from the Jet Propulsion Laboratory (JPL), California Institute of Technology, under a contract with the National Aeronautics and Space Administration (NASA). The authors thank Raymond Bourdeau for assistance with initial experiments, Kurt Liewer for the hologram simulation code, and Dina Malounda for providing some of the samples. Author Contributions: A.F. and M.B. contributed equally. The authors declare no competing financial interest.
Group:Heritage Medical Research Institute
Funders:
Funding AgencyGrant Number
Natural Sciences and Engineering Research Council of Canada (NSERC)UNSPECIFIED
NIHR01EB018975
David and Lucile Packard FoundationUNSPECIFIED
Pew Charitable TrustUNSPECIFIED
Heritage Medical Research InstituteUNSPECIFIED
Gordon and Betty Moore Foundation4038
NSFPHY-1828793
NASA/JPL/CaltechUNSPECIFIED
Subject Keywords:Quantitative Phase Imaging (QPI), Off-Axis Digital Holographic Microscopy, Volumetric Imaging, Phase Contrast Agent, Gas Vesicles, Reporter Genes, 3D Particle Tracking
Issue or Number:11
Record Number:CaltechAUTHORS:20191113-111723156
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20191113-111723156
Official Citation:Genetically Encoded Phase Contrast Agents for Digital Holographic Microscopy Arash Farhadi, Manuel Bedrossian, Justin Lee, Gabrielle H. Ho, Mikhail G. Shapiro, and Jay L. Nadeau. Nano Letters 2020 20 (11), 8127-8134; DOI: 10.1021/acs.nanolett.0c03159
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:99821
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:13 Nov 2019 19:41
Last Modified:19 Nov 2020 18:15

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