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Related Concept Videos

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Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
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Related Experiment Video

Updated: Jun 12, 2026

Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes
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Published on: December 13, 2012

Amine-reactive dyes for dead cell discrimination in fixed samples.

Stephen P Perfetto1, Pratip K Chattopadhyay, Laurie Lamoreaux

  • 1Vaccine Research Center, NIAID, NIH, Bethesda, Maryland, USA.

Current Protocols in Cytometry
|June 26, 2010
PubMed
Summary

Amine-reactive dyes are fluorescent viability stains that irreversibly label dead cells after fixation. These stains enable accurate dead cell identification using flow cytometry, even after permeabilization.

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Area of Science:

  • Cell Biology
  • Immunology
  • Biotechnology

Background:

  • Cell viability assays are crucial for experimental accuracy.
  • Traditional viability dyes can be lost during fixation and permeabilization.
  • Amine-reactive dyes offer a solution for identifying dead cells in fixed samples.

Purpose of the Study:

  • To describe the use of amine-reactive dyes for dead cell identification.
  • To provide protocols for using ViViD and Aqua Blue amine-reactive dyes.
  • To offer troubleshooting guidance for these dead cell stains.

Main Methods:

  • Utilizing amine-reactive dyes (ViViD, Aqua Blue) that cross membranes of dead cells.
  • Staining dead cells via reaction with cytoplasmic free amines.
  • Analyzing stained cells using fluorescence and flow cytometry.

Main Results:

  • Amine-reactive dyes irreversibly bind to dead cells.
  • Live cells exclude the dyes due to intact cell membranes.
  • Bound dye remains associated with dead cells after fixation and permeabilization.

Conclusions:

  • Amine-reactive dyes are effective for identifying dead cells in fixed samples.
  • These dyes are compatible with intracellular staining procedures.
  • ViViD and Aqua Blue are reliable tools for dead cell discrimination in flow cytometry.