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The electrophysical properties of the human polymorphonuclear leukocyte
Dissertation

The electrophysical properties of the human polymorphonuclear leukocyte

Michael Anthony DellaVecchia
Doctor of Philosophy (Ph.D.), Drexel University
1984
DOI:
https://doi.org/10.17918/00009752
pdf
DellaVecchia_Michael_198431.41 MB
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Abstract

Leucocytes Neutrophils
The polymorphonuclear leukocyte is the primier hematologic cellular component of acute inflammation and is responsible for many concomitant and subsequent physiological and pathological processes in the human body. In this treatise the electrophysiological properties of the human polymorphonuclear leukocyte with emphasis on its dielectric relaxation mechanisms and cataphoretic behavior is presented. In order to guaranty specificity and viability of testing samples, new techniques were developed by the author for the isolation of the human polymorphonuclear leuykocyte from human peripheral circulation and for assessing its viability. The leukocytes are isolated from human peripheral circulation via Dextran centrifugation gradient. A 99% isolation viability was determined as tested by microscopically counted polystyrene microsphere phagocytosis. This technique compared favorably to non-vital trypan blue viability testing. Polystyrene microsphere phagocytosis testing along with transmission and scanning electron microscopy was employed to determine that measurement parameters did not significantly alter test sample viability. Electrophoretic measurements of the polymorph suspended in M15 phosphate buffered solution undertaken on a Northrup-Kunitz cataphorometer yielded a mobility of 0.47 +/- 0.02 mu-cm/volt-sec. From this data was subsequently calculated a zeta potential of 6.26 mV; a surface charge density of 5.49 x 10('-7)Coulomb/cm('2); a net charge of 1.82 x 10('7) effective charges per polymorphonuclear leukocyte, and a fractional ionic area of 0.3%. Dielectric measurements were taken over the frequency spectrum from .01 megahertz to 0.1 megahertz using a Wayne Kerr Universal B221 Mk III bridge, from 0.1 to 3 megahertz using the Wayne Kerr B201 transformer-ratio arm bridge, and from 0.5 to 180 megahertz using the Boonton General Radio 250 RX bridge. Relaxation was observed in the lower frequency ranges consistent with beta frequency membrane dispersion exhibiting a relatively constant relaxation time, a dispersion which increases with increasing volume fraction and a membrane capacitance of 0.44 +/- 0.023 uf/cm('2). Recommendations are given for advancement in measurement techniques and further studies on the physiological and pathological variants of the human polymorphonuclear leukocyte and suspendable cytological samples.

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