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Biomedical subjects

V Kachel

Publications and source records attributed to V Kachel.

At least 37 records · Page 2Linked to original sources

Clonal hybrid cell lines expressing cholinergic and adrenergic properties.

Different cholinergic cell lines were fused with an adrenergic neuroblastoma cell line (N115-BU-8). Its fusion with a cholinergic neuroblastoma-glioma hybrid produced a "hybrid-hybrid" line containing cholinergic and adrenergic enzyme activities. Both activities were also present in subclones of this line. The presence of catecholamines in single cells was confirmed by microspectrofluorimetry. These results are discussed with respect to the possibility of a simultaneous synthesis of noradrenaline and acetylcholine in single cells. The cholinergic and adrenergic enzyme activities are influenced by cell density, by dexamethasone, and by conditioned medium.

Acetylcholine↗

Fast imaging in flow: a means of combining flow-cytometry and image analysis.

The morphological identification of cells by flow cytometry is difficult. Usually cell sorting and microscopical analysis have to be used in addition. Morphological analysis is simplified by taking cell pictures from a range of particular interest immediately during flow cytometric analysis. Instruments using the video scanning technique for fluorescence imaging are slow and expensive (8, 10). Morphological information can also be obtained by transmission imaging of cells in flow, which requires shorter exposure times. Therefore a cell volume activated flow imaging device has been developed which operates at flow speeds up to 5 m/sec and which depicts transmission images of selected cells on a 16-mm film by a nsec flashlamp illumination. An electronic unit detects the particles in the optically accessible orifice, performs the pulse height analysis, triggers the flashlamp if particles are in the preselcted range of interest and feeds the film. The instrument is capable of delivering up to 150 pictures per second and works either as a flow microscope in which the cells in the preselected volume range are directly observed, or as a picture system in which the cell pictures are stored on the 16-mm film for documentation or for image analysis.

Cell Count↗

Simultaneous flow cytometric DNA and volume measurements of bone marrow cells as sensitive indicator of abnormal proliferation patterns in rat leukemias.

Simultaneous flow cytometric DNA and volume analysis of normal rat bone marrow cells shows three populations of nucleated cells with different mean volume. Each of these populations proliferates in a distinct cell cycle (alpha, beta, gamma). Normally the alpha-cell cycle has the highest amplitude, the beta-cell cycle is intermediate, and the gamma-cell cycle is low. The alpha-cell cycle was very significantly depressed and the beta + gamma-cell cycle was increased in three different rat leukemias (L5222, Shay, BNML), growing on three different rat strains (BDIX, Holtzmann, Brown Norway). The two parameter analysis further revealed that cells of the beta + gamma-cell cycle were slightly hyperdiploid and hypertetraploid in leukemic animals. The decrease of the alpha-cell cycle and the hyperploidies were more sensitive indicators for the abnormal proliferation pattern than the analysis of one parameter DNA distributions which remained within normal limits in all three leukemias.

Animals↗

Uniform lateral orientation, caused by flow forces, of flat particles in flow-through systems.

Recently, it was shown that the lateral orientation of sperm cells disturbs the deoxyribonucleic acid distribution measured by fluorescence in a laterally laser-illuminated flow system. The present results show how flat particles may be influenced to assume a uniform lateral orientation. This was achieved by choosing the geometrical dimensions of the hydrodynamic focusing flow path. High speed photographs of fixed chicken erythrocytes oriented in experimental chambers are presented.

Animals↗

Fluvo-metricell, a combined cell volume and cell fluorescence analyzer.

A new flow through instrument that simultaneously measures cell volume (resistance pulse technique) and cell fluorescence in the same orifice will be described. The fluorescence pulses of the hydrodynamically focussed cells are picked up by the optics via the axial direction (principle of Dittrich and Goehde, Z Naturforsch 24b:360, 1969). There is no coordination problem between the fluorescence and the resistance pulses to be observed because a new type of transducer is used. The electronic system provides gating of one or two parameter histograms. Function tests are performed with the incorporated two-parameter test spectrum generator. Different examples of using the instrument in practice are shown. The volume that may be measured with an orifice of 70 micron diameter ranges between 4 and 1400 micron3 (1:350). Coefficients of variation of the fluorescence below 2% are measured.

Cytological Techniques↗

Measurement of mammalian sperm deoxyribonucleic acid by flow cytometry. Problems and approaches.

Measurement of mammalian sperm deoxyribonucleic acid content is of importance in several areas of biomedical research. When measured in flow systems with orthogonal axes of illumination, flow and detection, an unexpected, distorted distribution consisting of a narrow peak with a lateral extension to the right is observed. Several lines of evidence lead to the conclusion that this effect is an optical-geometric artifact attributable to the flat shape and high index of refraction of mammalian sperm heads. This artifact disappears when an epiillumination flow system is used in which the optic axes for illumination and detection and the flow axis are all coincident. Other approaches also eliminate the artifact. The resulting coefficients of variation observed after acriflavine-Feulgen staining are 4-5%, short of the goal of 1.5% required to distinguish between human sperm bearing X and Y chromosomes and to develop a mutagen test system using mice.

Acriflavine↗

A volume-activated cell sorter.

A sorter that is activated by the resistance pulses of a Coulter orifice (volume detector) has been developed. The problem arising from the distortion of the volume signals produced by the electric droplet forming and charging voltages are avoided by special shielding and grounding of the volume detector. Experiments with beads and leukocytes are described. The maximal processing rate at this time is 2000 cells/sec, the purity of sorted fractions is better than 97%, and the viability of sorted cells is better than 80%.

Cell Separation↗

Basic principles of electrical sizing of cells and particles and their realization in the new instrument "Metricell".

To understand the basic events during the passage of particles through the Coulter orifice, three experiments were performed. (1) The denpendence of the volume results on the particle path has been shown by ink-colored particle beams. (2) The deformation and alignment of cells during their passage through the orifice have been photographed by a nano-second photographing technique. (3) The absolute volume evaluation of particles has been studied with model particles in enlarged model orifices of different lengths. A new compact sizing instrument, "Metricell," equipped with a particle-independent electrical calibrating system, is described.

Animals↗