Mar 14, 2024

Public workspaceCell Viability Assessment

DOI
dx.doi.org/10.17504/protocols.io.n92ldmd9ol5b/v1
  • Bo Am Seo1,2,3,4,5,6,
  • Valina L. Dawson1,2,7,8,9,10,
  • Ted M. Dawson1,2,7,8,9,10,11,
  • Hanseok Ko1,2,7
  • 1Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA;
  • 2Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA;
  • 3Department of Convergence Medicine, Yonsei University Wonju College of Medicine, Wonju-si, Gangwon-do, Republic of Korea;
  • 4Department of Global Medical Science, Yonsei University Wonju College of Medicine, Wonju-si, Gangwon-do, Republic of Korea;
  • 5Mitohormesis Research Center, Yonsei University Wonju College of Medicine, Wonju-si, Gangwon-do, Republic of Korea;
  • 6Natural Product Research Center, Korea Institute of Science and Technology, Gangneung Gangwon-do, Republic of Korea;
  • 7Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA;
  • 8Adrienne Helis Malvin Medical Research Foundation, New Orleans, LA 70130-2685, USA;
  • 9Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205 USA;
  • 10Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA;
  • 11Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA
Eileen Ruth Torres
Open access
DOI: dx.doi.org/10.17504/protocols.io.n92ldmd9ol5b/v1
Protocol Citation: Bo Am Seo, Valina L. Dawson, Ted M. Dawson, Hanseok Ko 2024. Cell Viability Assessment. protocols.io https://dx.doi.org/10.17504/protocols.io.n92ldmd9ol5b/v1
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
We use this protocol and it's working
Created: February 01, 2024
Last Modified: May 31, 2024
Protocol Integer ID: 94559
Keywords: ASAPCRN, cell viability
Abstract
Assess neuronal viability.
Materials
  • Hoechst 33342 (7 µM)
  • propidium iodide (PI) (2 µM) (Invitrogen)
  • Axiovision 4.6 software (Carl Zeiss)
Stain neurons with Hoechst 33342 (Concentration7 micromolar (µM) ) and propidium iodide (PI) (Concentration2 micromolar (µM) ) (Invitrogen) at Temperature37 °C for Duration00:05:00 .

5m
Capture images by a microscope and dead cells are automatically counted by Axiovision 4.6 software (Carl Zeiss).
Subject background signals of the control group intensity.
Calculate the neuronal death percentage by dividing the PI signal by the Hoechst 33342 signal.
Protocol references
Yun SP, Kam TI, Panicker N, Kim S, Oh Y, Park JS, Kwon SH, Park YJ, Karuppagounder SS, Park H, Kim S, Oh N, Kim NA, Lee S, Brahmachari S, Mao X, Lee JH, Kumar M, An D, Kang SU, Lee Y, Lee KC, Na DH, Kim D, Lee SH, Roschke VV, Liddelow SA, Mari Z, Barres BA, Dawson VL, Lee S, Dawson TM, Ko HS. Block of A1 astrocyte conversion by microglia is neuroprotective in models of Parkinson's disease. Nat Med. 2018 Jul;24(7):931-938. doi: 10.1038/s41591-018-0051-5. Epub 2018 Jun 11. PMID: 29892066; PMCID: PMC6039259.