Document Detail

A novel method for the detection of viable human pancreatic beta cells by flow cytometry using fluorophores that selectively detect labile zinc, mitochondrial membrane potential and protein thiols.
MedLine Citation:
PMID:  18412218     Owner:  NLM     Status:  MEDLINE    
Improvement over current methods of beta cell viability assessment is highly warranted in order to efficiently predict the viability and function of beta cells prior to transplantation into type 1 diabetes patients. Dispersed human islet cells were stained with the cell-permeable zinc-selective dye, FluoZin-3-AM, along with the mitochondrial membrane potential indicator [(tetramethylrhodamine ethylester (TMRE)] and the thiol-binding dye, monochlorobimane (mBcl), and analyzed by flow cytometry. Islets were subjected to various experimental conditions to validate the usefulness of this method to accurately determine the viability and function of beta cells. Staining with FluoZin-3 revealed the presence of higher amounts of chelatable zinc ions in beta cells than in lymphoid cells and fibroblasts. An intracellular zinc chelator competitively inhibited the binding of FluoZin-3 to zinc ions. Mitochondrial depolarization or oxidative stress minimally affected the binding of mBcl and FluoZin-3, respectively, to thiols and zinc ions. The combination of FluoZin-3, TMRE, and mBcl was sufficient and necessary for the determination of the viability and function of beta cells. The data demonstrate the usefulness of the zinc-specific dye and the indicators of mitochondrial function and thiol levels, to accurately estimate the beta cell viability and function. This novel flow cytometry method has implications for islet transplantation in type 1 diabetes patients.
Sundararajan Jayaraman
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cytometry. Part A : the journal of the International Society for Analytical Cytology     Volume:  73     ISSN:  1552-4930     ISO Abbreviation:  Cytometry A     Publication Date:  2008 Jul 
Date Detail:
Created Date:  2008-06-19     Completed Date:  2008-08-07     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101235694     Medline TA:  Cytometry A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  615-25     Citation Subset:  IM    
Copyright Information:
(c) 2008 International Society for Analytical Cytology
Department of Surgery, College of Medicine, University of Illinois at Chicago, Chicago, Illinois 60612, USA.
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MeSH Terms
Cell Line
Cell Separation
Flow Cytometry / methods*
Fluorescent Dyes / pharmacology
Insulin-Secreting Cells / cytology*
Membrane Potential, Mitochondrial
Mitochondrial Membranes / metabolism*
Proteins / chemistry*
Sulfhydryl Compounds / chemistry*
Zinc / chemistry*
Reg. No./Substance:
0/Fluorescent Dyes; 0/Proteins; 0/Sulfhydryl Compounds; 7440-66-6/Zinc

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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