Document Detail


A platinum-based covalent viability reagent for single-cell mass cytometry.
MedLine Citation:
PMID:  22577098     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In fluorescence-based flow cytometry, cellular viability is determined with membrane-impermeable fluorescent reagents that specifically enter and label plasma membrane-compromised nonviable cells. A recent technological advance in flow cytometry uses antibodies conjugated to elemental metal isotopes, rather than to fluorophores, to allow signal detection by atomic mass spectrometry. Unhampered by the limitations of overlapping emission fluorescence, mass cytometry increases the number of parameters that can be measured in single cells. However, mass cytometry is unable to take advantage of current fluorescent viability dyes. An alternative methodology was therefore developed here in which the platinum-containing chemotherapy drug cisplatin was used to resolve live and dead cells by mass cytometry. In a 1-min incubation step, cisplatin preferentially labeled nonviable cells from both adherent and suspension cultures, resulting in a platinum signal quantifiable by mass cytometry. This protocol was compatible with established sample processing steps for intracellular cytometry. Furthermore, the live/dead ratios were comparable between mass- and fluorescence-based cytometry. Importantly, although cisplatin is a known DNA-damaging agent, a 1-min "pulse" of cisplatin did not induce observable DNA damage or apoptotic responses even within 6-h post-exposure. Cisplatin can therefore be used as a viability reagent for a wide range of mass cytometry protocols.
Authors:
Harris G Fienberg; Erin F Simonds; Wendy J Fantl; Garry P Nolan; Bernd Bodenmiller
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2012-05-10
Journal Detail:
Title:  Cytometry. Part A : the journal of the International Society for Analytical Cytology     Volume:  81     ISSN:  1552-4930     ISO Abbreviation:  Cytometry A     Publication Date:  2012 Jun 
Date Detail:
Created Date:  2012-05-22     Completed Date:  2012-09-14     Revised Date:  2014-03-21    
Medline Journal Info:
Nlm Unique ID:  101235694     Medline TA:  Cytometry A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  467-75     Citation Subset:  IM    
Copyright Information:
Copyright © 2012 International Society for Advancement of Cytometry.
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MeSH Terms
Descriptor/Qualifier:
Cell Adhesion
Cell Membrane Permeability
Cell Survival
Cisplatin / chemistry*
DNA / analysis,  chemistry
Female
Flow Cytometry / methods*
Fluorescent Dyes
Humans
Mass Spectrometry / methods*
Platinum / chemistry*
Single-Cell Analysis
Staining and Labeling / methods*
Tumor Cells, Cultured
Grant Support
ID/Acronym/Agency:
1R01CA130826/CA/NCI NIH HHS; 5U54CA143907/CA/NCI NIH HHS; CA 09-011/CA/NCI NIH HHS; HHSN268201000034C/HL/NHLBI NIH HHS; HHSN272200700038C//PHS HHS; HV-10-05(2)/HV/NHLBI NIH HHS; P01 CA034233/CA/NCI NIH HHS; R01 CA130826/CA/NCI NIH HHS; T32 AI007328/AI/NIAID NIH HHS; U19 AI057229/AI/NIAID NIH HHS; U19 AI057229/AI/NIAID NIH HHS; U19 AI100627/AI/NIAID NIH HHS; U54 CA119367/CA/NCI NIH HHS; U54 CA143907/CA/NCI NIH HHS; U54 CA149145/CA/NCI NIH HHS; U54CA149145/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Fluorescent Dyes; 49DFR088MY/Platinum; 9007-49-2/DNA; Q20Q21Q62J/Cisplatin
Comments/Corrections

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