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Comparison of techniques for the assessment of polymorphonuclear leukocyte polarisation in suspension
Author(s) -
Harkin Damien G,
Gadd Stephen J,
Bignold Leon P
Publication year - 1993
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1016/0248-4900(93)90144-4
Subject(s) - fixative , flow cytometry , biology , cytometry , lysis , suspension culture , isolation (microbiology) , pathology , cytoplasm , microbiology and biotechnology , immunology , cell culture , medicine , genetics
Summry— Polarisation of polymorphonuclear leukocytes (PMN) is suspension was assessed using three techniques: 1) visual classification; 2) computerized morphometry; and 3) flow cytometry. While visual classification detected the formation, polarisation and type of cytoplasmic extensions produced by PMN, morphometry and flow cytometry detected only the formation of extensions. The area, perimeter and ellipticity were, in general, statistically different for each subtype of PMN‐shape identified by visual classification. Furthermore, the magnitude and direction of changes detected by flow cytometry were affected by the use of erythrocyte lysis (during isolation of the cells) and the fixative used prior to analysis. The findings of this study demonstrate that visual classification is a more sensitive, reliable and appropriate assay of PMN polarisation than current morphometric and flow cytometric methods.