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Two-layer gradient separation of granulocytes for functional tests.

A Naess1, H Sjursen, I Talstad

  • 1Medical Department B, University of Bergen, Haukeland Hospital, Norway.

APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica
|December 1, 1990
PubMed
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Two gradient separation methods efficiently isolate pure granulocytes from blood, showing comparable functionality to traditional techniques for research applications.

Area of Science:

  • Immunology
  • Cell Biology
  • Hematology

Background:

  • Granulocyte separation is crucial for immunological and cellular studies.
  • Traditional dextran sedimentation methods can be time-consuming.
  • Optimizing granulocyte isolation is essential for accurate functional assays.

Purpose of the Study:

  • To compare two gradient separation techniques with dextran sedimentation for granulocyte isolation.
  • To evaluate the purity, yield, and functional integrity of isolated granulocytes.
  • To determine the suitability of gradient methods for preparing pure granulocyte suspensions.

Main Methods:

  • Comparison of two gradient separation techniques against dextran sedimentation.
  • Assessment of granulocyte purity and yield.

Related Experiment Videos

  • Flow cytometry analysis of surface membrane markers.
  • Functional testing including random migration, chemotaxis, and chemiluminescence.

    • Main Results:

    • Both gradient techniques provided adequate yield and excellent purity of granulocytes.
    • No evidence of subset selection was observed via flow cytometry.
    • Separated granulocytes from all methods exhibited comparable functional performance.
    • Gradient methods are rapid and efficient for high-purity granulocyte preparation.

    Conclusions:

    • Gradient separation techniques offer a rapid and efficient alternative for isolating pure granulocytes.
    • These methods preserve granulocyte functionality for various research applications.
    • The techniques are suitable for preparing near 100% pure granulocyte suspensions for functional studies.