Journal
NATURE PROTOCOLS
Volume 15, Issue 5, Pages 1649-1672Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/s41596-020-0300-1
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Funding
- Schmidt Futures
- MIT Media Lab
- Chan Zuckerberg Initiative
- NIH [U01MH114819, 1U19MH114821, 1R01NS102727, 1R01EB024261, 1R01MH110932, 1RM1HG008525]
- Ludwig Foundation
- Open Philanthropy project
- HHMI-Simons Faculty Scholars Program
- US Army Research Laboratory [W911NF1510548]
- US Army Research Office [W911NF1510548]
- Ludwig Center at Harvard
- Harvard Catalyst (the Harvard Clinical and Translational Science Center (National Center for Research Resources, National Institutes of Health)) [UL1 TR001102]
- Harvard Catalyst (the Harvard Clinical and Translational Science Center (National Center for Advancing Translational Sciences, National Institutes of Health)) [UL1 TR001102]
- Carnegie Mellon University
- NIH Director's New Innovator Award [DP2 OD025926-01]
- U.S. Department of Defense (DOD) [W911NF1510548] Funding Source: U.S. Department of Defense (DOD)
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In pathology, microscopy is an important tool for the analysis of human tissues, both for the scientific study of disease states and for diagnosis. However, the microscopes commonly used in pathology are limited in resolution by diffraction. Recently, we discovered that it was possible, through a chemical process, to isotropically expand preserved cells and tissues by 4-5x in linear dimension. We call this process expansion microscopy (ExM). ExM enables nanoscale resolution imaging on conventional microscopes. Here we describe protocols for the simple and effective physical expansion of a variety of human tissues and clinical specimens, including paraffin-embedded, fresh frozen and chemically stained human tissues. These protocols require only inexpensive, commercially available reagents and hardware commonly found in a routine pathology laboratory. Our protocols are written for researchers and pathologists experienced in conventional fluorescence microscopy. The conventional protocol, expansion pathology, can be completed in similar to 1 d with immunostained tissue sections and 2 d with unstained specimens. We also include a new, fast variant, rapid expansion pathology, that can be performed on <5-mu m-thick tissue sections, taking <4 h with immunostained tissue sections and <8 h with unstained specimens.
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