Aug 21, 2020

Public workspace6: Protocol optimization for SABER-FISH in tissues

This protocol is a draft, published without a DOI.
  • Jocelyn Y. Kishi1,2,3,
  • Sylvain W. Lapan4,3,
  • Brian J Beliveau1,2,5,3,6,
  • Emma R. West4,3,
  • Allen Zhu1,2,
  • Hiroshi M. Sasaki1,2,
  • Sinem K Saka1,2,
  • Yu Wang1,2,
  • Constance L Cepko4,7,6,
  • Peng Yin1,2,6
  • 1Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA;
  • 2Department of Systems Biology, Harvard Medical School, Boston, MA, USA;
  • 3These authors contributed equally;
  • 4Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, MA, USA;
  • 5Present address: Department of Genome Sciences, University of Washington, Seattle, WA, USA;
  • 6Correspondence: py@hms.harvard.edu (P.Y.), cepko@genetics.med.harvard.edu (C.L.C.), beliveau@uw.edu (B. J. B.);
  • 7Howard Hughes Medical Institute, Chevy Chase, MD, USA
  • Human Cell Atlas Method Development Community
Jocelynkishi
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External link: http://saber.fish/
Protocol Citation: Jocelyn Y. Kishi, Sylvain W. Lapan, Brian J Beliveau, Emma R. West, Allen Zhu, Hiroshi M. Sasaki, Sinem K Saka, Yu Wang, Constance L Cepko, Peng Yin 2020. 6: Protocol optimization for SABER-FISH in tissues. protocols.io https://protocols.io/view/6-protocol-optimization-for-saber-fish-in-tissues-bh9vj966
Manuscript citation:
Kishi, J.Y., Lapan, S.W., Beliveau, B.J. et al. SABER amplifies FISH: enhanced multiplexed imaging of RNA and DNA in cells and tissues. Nat Methods 16, 533–544 (2019). https://doi.org/10.1038/s41592-019-0404-0
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
Created: July 06, 2020
Last Modified: August 21, 2020
Protocol Integer ID: 38933
Keywords: tissue, SABER-FISH,
Abstract
This protocol summarizes several tested variations to simplify the SABER-FISH protocol in tissues; In most cases robust signal was found.



Note
This protocol is part of the SABER-FISH collection.

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SABER amplifies FISH...
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For hazard information and safety warnings, please refer to the SDS (Safety Data Sheet).
Figure S3: Testing variations in the RNA FISH staining protocol. a, Overview of main steps in the RNA FISH protocol. The most efficient condition tested in this experiment is shown below each step. b, Table of eight conditions tested in side-byside comparison. See Methods for additional details. c, Quantification of signal intensity for conditions tested, with each line representing a replicate (N=2 retinal sections per condition). Each condition was tested using two-color FISH for Vsx1 and Grik1. d, Representative images of conditions tested. Scale bar is 10 µm. All sections 40 µM, from P25 animals.