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Related Experiment Videos

A rapid preparation technique for leucocytes.

D A McCarthy1, J Bernhagen, Y C Liu

  • 1School of Biological Sciences, Queen Mary College, London.

Journal of Microscopy
|April 1, 1990
PubMed
Summary
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This study presents a new method for preparing white blood cells (leukocytes) from blood samples. The technique allows for the identification of polarized polymorphonuclear leukocytes (PMN) in circulation, aiding in inflammatory disease research.

Area of Science:

  • Hematology
  • Cell Biology
  • Microscopy Techniques

Background:

  • Accurate identification of leukocyte subtypes is crucial for diagnosing and understanding inflammatory diseases.
  • Existing methods for preparing leukocytes for microscopy can be time-consuming and may alter cell morphology.
  • Quantifying circulating leukocyte polarization offers potential insights into inflammatory processes.

Purpose of the Study:

  • To describe a novel ex vivo procedure for preparing fixed leukocyte samples from blood.
  • To validate the use of this method for identifying and quantifying polarized polymorphonuclear leukocytes (PMN).
  • To assess the prevalence of circulating polarized PMN in healthy individuals.

Main Methods:

  • Blood samples were fixed ex vivo using buffered formaldehyde.

Related Experiment Videos

  • Erythrocytes and platelets were removed via lysis and differential centrifugation.
  • Fixed leukocyte preparations were processed for light microscopy, identifying lymphocytes, monocytes, and PMN by nuclear morphology.
  • Polarized PMN were identified by their non-spherical shape.

    • Main Results:

    • The procedure successfully yielded identifiable fixed leukocyte preparations.
    • Polarized PMN were readily distinguished from spherical PMN based on morphology.
    • In normal subjects, 84.0% of circulating PMN were spherical, with 14.3% showing slight polarization.

    Conclusions:

    • The described method provides a reliable means for preparing and analyzing fixed leukocytes.
    • This technique enables the detection and quantification of circulating polarized PMN.
    • The findings suggest a low prevalence of significantly polarized PMN in healthy individuals, providing a baseline for future studies in inflammatory conditions.