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Related Experiment Videos

Technique and staining optimization leucoconcentration.

J Pierrez1, A Guerci, O Guerci

  • 1Laboratoire de Cytométrie en Flux, CHU Nancy-Brabois, 54500 Vandoeuvre les Nancy, France.

Cytometry
|September 1, 1987
PubMed
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A modified leucoconcentration technique rapidly prepares cell suspensions for flow cytometry. This method, using Saponin lysis and Mithramycin staining, enables precise cell cycle determination with minimal cytological alteration.

Area of Science:

  • Clinical Cytometry
  • Cellular Analysis
  • Biomedical Research

Background:

  • Rapid preparation of cell suspensions with minimal cytological alteration is crucial for flow cytometric clinical applications.
  • Existing methods may cause cell shifts or alter tinctorial affinity, impacting analysis accuracy.

Purpose of the Study:

  • To develop and validate a modified leucoconcentration technique for preparing cell suspensions for flow cytometric analysis.
  • To optimize staining conditions using Mithramycin for DNA analysis.
  • To enable high-resolution cell cycle determination.

Main Methods:

  • Modification of the Herbeuval leucoconcentration technique for cytometry.
  • Fixation of peripheral blood or bone marrow samples with picric acid alcohol solution.

Related Experiment Videos

  • Saponin lysis of red blood cells to obtain cell suspensions.
  • Staining of cells with Mithramycin for DNA analysis.
  • Analysis of parameters defined by Crissman to optimize staining.

    • Main Results:

    • The modified leucoconcentration technique successfully prepares cell suspensions with preserved cell morphology and tinctorial affinity.
    • Optimized Mithramycin staining conditions were established.
    • The combined technique allows for fine-resolution determination of the cell cycle.

    Conclusions:

    • The leucoconcentration technique combined with Mithramycin-DNA staining provides a rapid and reliable method for preparing cell suspensions for flow cytometry.
    • This approach minimizes cytological alteration and enhances the accuracy of cell cycle analysis.