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Measuring cell viability with membrane impermeable zinc fluorescent indicator.

Christian J Stork1, Yang V Li

  • 1Department of Biomedical Sciences, Molecular and Cellular Biology Program, Ohio University, Athens, OH 45701, USA.

Journal of Neuroscience Methods
|February 10, 2006
PubMed
Summary

Cytoplasmic zinc accumulation is key in cell death. Cell-impermeable zinc fluorescent indicators can reliably detect unhealthy cells in tissue, correlating with cell death.

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Area of Science:

  • Cell Biology
  • Neuroscience
  • Biochemistry

Background:

  • Cytoplasmic zinc accumulation ([Zn2+]i) is increasingly recognized as a critical factor in the cell death cascade.
  • Oxidative stress can trigger the release of intracellular zinc, contributing to cell demise.

Purpose of the Study:

  • To evaluate the efficacy of cell-impermeable zinc fluorescent indicators for assessing cell viability in tissue slices.
  • To establish a correlation between intracellular zinc accumulation and neuronal death.

Main Methods:

  • Utilized Newport Green (NG), a non-toxic, membrane-impermeable zinc fluorescent indicator.
  • Employed confocal microscopy to visualize NG fluorescence in conjunction with propidium iodide (PI) staining.
  • Tested additional zinc dyes, Fluozin-3 and Zinpyr-4, for their ability to label dying cells.

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Main Results:

  • Newport Green (NG) successfully labeled unhealthy cells in tissue slices, similar to propidium iodide (PI).
  • PI-labeled nuclei colocalized with NG fluorescence, indicating that compromised cell membranes allowed dye entry.
  • Increased NG fluorescence was observed in damaged/dead cells, and other impermeable zinc dyes also detected cytosolic zinc in PI-labeled cells.
  • Intracellular zinc accumulation was demonstrated to correlate with neuronal death.

Conclusions:

  • Cell-impermeable zinc fluorescent indicators offer a fast, simple, non-toxic, and reliable method for visualizing cell viability in vitro.
  • Intracellular zinc ([Zn2+]i) dynamics are a significant indicator of cell death, particularly in neuronal tissues.