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K Zeiller

Publications and source records attributed to K Zeiller.

At least 19 recordsLinked to original sources
PubMed1979

Cytofluorometric analysis of R-Thy-1. antigens in various rat lymphocytes with different electrophoretic mobility and organ distribution.

Small bone marrow lymphocytes, which had been previously enriched by velocity sedimentation, thymocytes, lymph node cells and spleen cells were electrophoretically separated, stained with fluorescein conjugated rabbit a-rat-Thy-1. globulin and their fluorescence intensities analyzed with a flow cytophotometer. Thy-1. antigens were found in 80% of the bone marrow small lymphocytes showing low electrophoretic mobility (EPM), in all thymocytes, about 80% of which show low and the rest medium to high EPM, and in a few lymph node cells of high EPM. Thy-1. positive cells were not observed in the spleen. All fluorescence intensity histograms obtained were modal and could be properly fitted with normal curves showing coefficients of variation (C.V.) in the range of 20% to 30%. It was observed that the thymocytes of low EPM had an antibody binding affinity significantly different from that of the other stained lymphocytes. Moreover the surface antigen density decreased in the sequence: thymocytes of low EPM, bone marrow lymphocytes of low EPM and thymocytes of high EPM. The fluorescence intensity of stained lymph node cells of high EPM appeared similar to that of thymocytes of high EPM but was not evaluated precisely. Thus the two dimensional cell analysis provided by a combination of EPM and surface fluorescence of Thy-1.+ cells, allows the characterization of different lymphocyte populations which cannot be clearly identified with normal one dimensional techniques. The biological significance of the results is discussed briefly.

Animals↗
PubMed1978

Characterization of rat bone marrow lymphoid cells. I. A study of the distribution parameters of sedimentation velocity, volume and electrophoretic mobility.

Various cell populations in rat bone marrow were characterized by means of a two dimensional separation using velocity sedimentation and free flow electrophoresis and by electrical sizing of the separated cells. Up to 4.5 mm/hr five different populations with discrete distributions in volume (coefficient of variation 10% to 13%) and sedimentation velocity (coefficient of variation 6% to 10%) were observed. Three of the small sized populations represented lymphocytes and small normoblasts and two of the larger sized populations represented myeloid cells. Almost all of these cells were in the G0/G1 cycle phase. In the faster sedimenting fractions which contained immature myeloid, erythroid and undefined blast cells and two S phase populations, discrete volume distributions were not evaluated. The cell populations with homogeneous volume (particularly the small lymphocytes) showed high density variations which condiserably impair the separation resolution. The cells sedimenting slower than 3.5 mm/hr were further separated by means of free flow electrophoresis into three peaks differing in electrophoretic mobility (EPM). The peaks of low and high EPM contained two populations and the peak of medium EPM contained three populations all characterized by normal volume distributions of uniform coefficient of variation between 11% and 14%. The small cells in the peaks of high and medium EPM were normolblasts and the other cells were lymphocytes. The biological significance of these results is discussed.

Animals↗
PubMed1976

Analysis of velocity sedimentation techniques in cell separation. Influence of apparative and sample properties on separative power, resolution and sensitivity.

In the present investigation the velocity sedimentation technique was analysed with respect to separation resolution, power and sensitivity. It was found that apparative modifications do not influence the resolution, which is a function of the contribution of apparative errors to the dispersion. A surprisingly small parameter of 0.15 was determined and it seems unlikely that this value can be improved. On the other hand an apparative modification is presented which improves the separation power and makes sample loading independent of the gradient filling. If cells (from rat bone marrow) were separated, a several times higher dispersion for a given cell volume was observed than was due to the apparative error. It was concluded that density variations were the major source of this dispersion. Since cell volume and density apparently show independent variations within a biological cell population the cell density cannot be disregarded if velocity sedimentation profiles are discussed in physical terms as is often done.

Cell Separation↗
PubMed1976

B lymphocyte subpopulations in the mouse spleen. A study of the differentiation pathway using free flow electrophoretically separated subpopulations of direct PFC progenitor cells.

Free-flow electrophoretic separation of mouse spleen cells provides three distinct progenitor cells of direct PFC, showing high, medium and low electrophoretic mobility. All progenitor cells possess surface immunoglobulin and mouse B-lymphocyte specific antigen. The progenitor cells of high electrophoretic mobility show high cycling turnover, a spleen seeking capacity of 16%, provide PFC with a maximum 8 days after transfer and reveal an isometrical increase of the PFC dose response line as a function of the graft size. The progenitor cells of medium electrophoretic mobility are low cycling, 16% home to the spleen, a maximum of PFC is developed eight days after transfer and the PFC dose response line increases allometrically. The progenitor cells of low EPM show low cycling activity, 20% home to the spleen, a maximum of PFC is attained six days after transfer and the PFC dose response line rises isometrically. These results suggest that the electrokinetically different PFC progenitors represent biologically distinct subsets. In double transfer experiments, some evidence was obtained that progenitor cells of low electrophoretic mobility are derived from progenitors of higher electrophoretic mobility. The same observation accounts also for the formation of B lymphocytes of low EPM. Since it seemed likely that the PFC progenitor cells represent virgin cells of a single lineage, the results were discussed in the terms of differentiation pathways of B lymphocytes. A model is considered in which a progenitor of medium electrophoretic mobility provides those of high electrophoretic mobility which after passing a transient cycling stage finally produce mature resting B lymphocytes of low electrophoretic mobility.

Animals↗
PubMed1975

[Free-flow electrophoresis. I. Theoretical and experimental investigations of the influence of mechanical and electrokinetic variables on the efficiency of the method].

In the present paper, various types of band-broadening effects in free-flow electrophoresis were investigated. They resulted from the velocity profiles of the liquid curtain and electroosmosis, temperature gradient, thermal diffusion and sample inlet geometry. An analytical free-flow electrophoresis apparatus permitted easy observation of these parameters. The experiments showed that the influence of the temperature gradient was negligible, whereas the effect of the velocity profiles on band broadening was higher than theoretically expected. In preparative work this is of utmost importance, since overlap of bands is not desirable. In this case, the zeta potential of the walls can be adjusted to that of the material to be separated, resulting in a considerable reduction of band broadening. Various approaches are indicated. Further attention was given to the influence of the relaxation effect on the separation. Conditions are shown where particles are separated either according to their surface charge density or to their size. The practical relevance of the results is discussed.

Electrophoresis↗
PubMed1975

Free-flow electrophoresis. II. Analysis of the method with respect to preparative cell separation.

Electrophoretic cell separation by means of free-flow electrophoresis in an FF5 apparatus was investigated with respect to band resolution, separation capacity, reproducibility and influence on cell viability. Very sharp bands and a large separation capacity were achieved using triethanolamine/acetate buffered glycine media as liquid curtain. Acid buffer ions such as N-2-hydroxyethylpiperazine-N'-ethanesulfonic acid (HEPES) or phosphate produced broader bands. Osmotic expanders such as saccharides, though preserving cell viability excellently, decrease electrophoretic velocity and thus separation capacity. The decrease in cell viability observed in glycine media could be compensated for by addition of Ca2. Band broadening caused by methodologically specific velocity flow profiles could be reduced to a negligible level by coating the chamber walls with albumin and by appropriate adjustment of sample flow rate and liquid curtain velocity. Under the optimum conditions described, selective cell loss and artificial change in electrophoretic mobility of the cells during operation can be disregarded. The main reason for cell loss was cell aggregation at low ionic strength, which can be prevented or reversed by treatment of the cells with deoxyribonuclease.

Animals↗
PubMed1974

T and B lymphocytes: striking differences in surface membranes.

Lymphocytes can be separated by electrophoresis into cells of high and of low electrophoretic mobility. Cells of high mobility are T cells and those of low mobility are B cells, with distinct immunological functions. Biophysical and biochemical studies on physically isolated and immunologically defined T and B cells in CBA mice showed striking differences in the chemical composition of the surface membranes.

Animals↗