Non-reversible tissue fixation retains extracellular vesicles for in situ imaging
Gupta, Mrinali P.; Tandalam, Sangeetha; Ostrager, Shariss; Lever, Alexander S.; Fung, Angus R.; Hurley, David D.; Alegre, Gemstonn B.; Espinal, Jasmin E.; Remmel, H. Lawrence; Mukherjee, Sushmita; Levine, Benjamin M.; Robins, Russell P.; Molina, Henrik; Dill, Brian D.; Kenific, Candia M.; Tuschl, Thomas; Lyden, David; D’Amico, Donald J.; Pena, John T.G.
(2019) Nature Methods, volume 16, issue 12, pp. 1269 - 1273
(Article)
Abstract
Extracellular vesicles (EVs) are secreted nanosized particles with many biological functions and pathological associations. The inability to image EVs in fixed tissues has been a major limitation to understanding their role in healthy and diseased tissue microenvironments. Here, we show that crosslinking mammalian tissues with formaldehyde results in significant EV
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loss, which can be prevented by additional fixation with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) for visualization of EVs in a range of normal and cancer tissues.
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Keywords: Biotechnology, Biochemistry, Molecular Biology, Cell Biology
ISSN: 1548-7091
Publisher: Nature Research
Note: Funding Information: We thank J. Moore for his technical support for this manuscript. This work was performed in part at the Weill Cornell Medicine Imaging Core (for Electron Microscopy), and Genomics Resources Core, Cornell NanoScale Facility, a member of the National Nanotechnology Coordinated Infrastructure (NNCI), which is supported by the National Science Foundation (Grant NNCI-1542081). The Heed Foundation supported M.G. and J.T.G.P. The Rockefeller University Proteomics Resource Center acknowledges funding from the Leona M. and Harry B. Helmsley Charitable Trust for mass spectrometer instrumentation. T.T. was supported by an Extracellular RNA Communication grants U19CA179564 from the NIH. D.L. was supported by NCI R35 CA232093 Outstanding Investigator Award from the NIH. J.T.G.P. supported by Daedalus Fund at Weill Cornell Medicine, The Shulsky Foundation, Research to Prevent Blindness Foundation and Knights Templar Eye Foundation Inc. Publisher Copyright: © 2019, The Author(s), under exclusive licence to Springer Nature America, Inc.
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