Cell electrophoresis for diagnostic purposes. II. Critical evaluation of conventional cytopherometry

Hoffmann, W.; Kaufmann, R; Steiner, Rudolf; Werner, W

doi:10.1038/bjc.1981.89

Br J Cancer

1981

DOI: 10.1038/bjc.1981.89

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Cell electrophoresis for diagnostic purposes. II. Critical evaluation of conventional cytopherometry

W. Hoffmann¹,

R Kaufmann²,

Rudolf Steiner³

et al.

Abstract: Determination of the electrophoretic mobility of test cells has been widely used in an attempt to detect so-called lymphokines in a laboratory test for cancer, but operational difficulties are inherent in conventional cytopherometers. This study therefore investigates the technical and operational aspects of cell electrophoresis, using the Zeiss cytopherometer; e.g. influence of electro-osmosis, focus uncertainty, movement due to convection and other sources of error. Implications and possible improvements in … Show more

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Cited by 8 publications

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A procedure for stabilization of cytopherometers for immunological applications

1982

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A device commonly used for cell electrophoresis is the cytopherometer. Its application in an immunological tumor test, however, pointed out many shortcomings and instabilities. In order to use cytopherometers in cellular immunology, a procedure was worked out to stabilize this instrument. By enlarging the electrode chambers, using a simple and efficient buffer in all compartments, coating the electrophoresis chamber, and by following a set of electrophoretic parameters (current, temperature, etc.), an intraassay variance of 1.8 % and an interassay variance of approximately 1 % were reached. I 1 Plasma from normal male and female rats was analyzed, after removal of albumin by affinity chromatography, by two-dimensional electrophoresis. The results demonstrated gender differences in two plasma proteins with apparent molecular weights of 70 000 and 84 000 and isoelectric points ranging from 6.2 to 6.4, and 5.1 to 5.5, respectively. Both proteins were found to be more concentrated in the plasma minus albumin (P-alb) fraction of females than males in both Wistar and Sprague-Dawley rats. The 70000 protein is serum albumin which was more completely removed from male plasma than female plasma. A possible mechanism responsible for this difference is discussed. The results support the existence of "maleness" and "femaleness" in blood protein patterns of rats.

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A procedure for stabilization of cytopherometers for immunological applications

1982

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Cell electrophoresis: Automatic measurements by laser light scattering with Lazypher

et al. 1985

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Automated cell electrophoretic measurements with Lazypher are based on the laser Doppler technique. Experiments are performed with high precision (of 1.2 % S. D.) and in short time intervals of 1-3 min. Application to electrophoretic mobility tests with tanned sheep erythrocytes incubated in supernatants from sensitized murine thymus and spleen cells which had been prepared from cultures containing 5 pg purified protein derivative (PPD)/ml caused a reduction of the electrophoretic mobility (EM) of 5 % and 13 %, respectively. Control spleen cell supernatants caused a 4 % decrease in mobility and no effect was observed with supernatants from control thymus cells. Peripheral blood lymphocytes from healthy adults (n = 56) and children with atopic diseases and congenital heart disease (n = 12) are separated into a high mobility peak (p = 1.24 pmcm/Vs for adults, p = 1.19 pmcm/Vs for the child group) corresponding toT-cells and alow mobility peak, p =0.93 pmcm/Vs, representing Bcells. The mobility difference between the adult and child T-cells is highly significant (99 %). Electrophoretic measurements of myeloid cells from healthy donors and patients with chronic myeloid leukemia (CML) and acute myeloid leukemia (AML) show that progressing differentiation within this cell line is accompanied by a decreasing mobility. Blast cells from a patient with acute lymphoblastic leukemia (c-ALL) had an EM, u = 1.09 umcm/Vs. distinct from that of mveloblasts in AML (Y = 1.16 and 1.25 pmcm/Vs) and B-lymphocytes.

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The development of an automated cell electrophoresis analyzer

et al. 1986

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An automated cell-electrophoresis apparatus is described which appears suitable for clinical laboratory applications. A solid state laser is employed; however, the instrument departs from the usual Doppler systems in that coherence is not required and stationary transmission grating optics are used. A dedicated microprocessor performs fast Fourier transforms (FFT), curve fitting for pedestal and low frequency (LF) noise removal, and digital filtering to suppress FFT artifacts. Unique features include fully automated sample handling, and measurements are made at the center of the migration tube in lieu of the "stationary layer" for enhanced signal quality. Experiments show high resolution of spectra from individual and mixed populations of sheep and rabbit erythrocytes. The sample-to-report cycle of 5 or 7.5 min (switchable), high cost/performance ratio and elimination of potential operator errors make the instrument a candidate for clinical use.

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Cell electrophoresis for diagnostic purposes. II. Critical evaluation of conventional cytopherometry

Cited by 8 publications

References 28 publications

A procedure for stabilization of cytopherometers for immunological applications

A procedure for stabilization of cytopherometers for immunological applications

Cell electrophoresis: Automatic measurements by laser light scattering with Lazypher

The development of an automated cell electrophoresis analyzer

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