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Biomedical subjects

B Thorell

Publications and source records attributed to B Thorell.

At least 37 records · Page 2Linked to original sources
PubMed1979

Quantitative reflection contrast microscopy of living cells.

Mammalian cells in culture (BHK-21, PtK2, Friend, human flia, and glioma cells) have been observed by reflection contrast microscopy. Images of cells photographed at two different wavelengths (546 and 436 nm) or at two different angles of incidence allowed discrimination between reflected light and light that was both reflected and modulated by interference. Interference is involved when a change in reflected intensity (relative to glass/medium background reflected intensity) occurs on changing either the illumination wavelength or the reflection incidence angle. In cases where interference occurs, refractive indices can be determined at points where the optical path difference is known, by solving the given interference equation. Where cells are at least 50 nm distant from the glass substrate, intensities are also influenced by that distance as well as by the light's angle of incidence and wavelength. The reflected intensity at the glass/medium interface is used as a standard in calculating the refractive index of the cortical cytoplasm. Refractive indices were found to be higher (1.38--1.40) at points of focal contact, where stress fibers terminate, than in areas of close contact (1.354--1.368). In areas of the cortical cytoplasm, between focal contacts, not adherent to the glass substrate, refractive indices between 1.353 and 1.368 were found. This was thought to result from a microfilamentous network within the cortical cytoplasm. Intimate attachment of cells to their substrate is assumed to be characterized by a lack of an intermediate layer of culture medium.

Animals↗
PubMed1978

The effect of atebrine and an acridine analog (BCMA) on the coenzyme fluorescence spectra of cultured melanoma and Ehrlich ascites (EL2) cells.

Coenzyme fluorescence spectra of single living cells are due to free pyridine nucleotides (folded configuration), bound pyridine nucleotides (unfolded configuration) and a third component, possibly a mixture or flavins. Such spectra can be used to recognize possible differences in coenzyme composition between cell lines or changes of metabolic pathways due to chemicals acting at levels below or above cytotoxicity, by high resolution spectrofluorometry. A study of spectra recorded from cultured Ehrlich ascites (EL2), and Harding Passey melanoma cells (HPM-67 and HPM-73 line) grown under comparable conditions, shows that free NAD(P)H predominates in HPM-67 and EL2, while this coenzyme is bound in HPM-73. The free/bound ratio may be profoundly modifed by chemicals, e.g. in the HPM-73 increase of free and decrease of bound NAD(P)H occurred upon treatment with 10(-6) oligomycin. When atebrine at levels (10(-6) M) below cytotoxicity was added, there was a decrease of the free NAD(P)H spectrum possibly through energy transfer from NAD(P)H to atebrine. Consideration of long range energy transfer i.e., excitation of atebrine by fluorescence of NAD(P)H vs. short range transfer of excitation energy from free NAD(P)H to atebrine, favors the latter mechanism. A transient (reversible) increase in atebrine fluorescence is seen following intracellular microinjection of substrate (e.g. glucose-6-P) leading to an increase in free NAD(P)H. At cytotoxic levels of atebrine (e.g 2 x 10(-5) M) an irreversible increase of atebrine fluorescence is seen. The microspectrofluorometric technique appears therefore well suited to study physiological processes at the level of intracellular coenzymes, as well as possible processes of intermolecular energy transfer in the microenvironment.

Acridines↗
PubMed1978

Characterization of the normoblast population in beta-thalassaemic blood by rapid-flow cytofluorometry.

Samples of peripheral blood from patients with beta-thalassaemia major which contained significant numbers of nucleated normoblasts were stained with acridine orange and analyzed with rapid-flow cytofluorometry. The pyknotic normoblast-nuclei gave less green 'DNA' fluorescence than the (diploid) leucocytes and constituted a separate, distinct subpopulation. Mean values of the fluorescence intensities and standard deviations as displayed by multichannel analyses gave a numerical value for normoblasts with regard to their maturation stages. These mean values correlated with the differential counts of 'early and late' normoblasts in the light microscope under rigidly standardized conditions. Rapid-flow cytofluorometry thus provides an objective and quantitative way to monitor and define peripheral blood normoblast populations as a measure of the severity of 'erythropoietic stress'.

Adolescent↗
PubMed1978

Metabolic rates and intercellular transfer of molecules in cultures of human glia and glioma cells.

Metabolic rates and intercellular transfer of metabolites were studied in human glia and glioma culture cells via topographic scan of NAD(P)H fluorescence by multichannel microfluorometry in conjunction with microinjection of glucose-6-P + allosteric activators. Metabolic rates evaluated from NAD(P) in equilibrium NAD(P)H transients and the required substrate levels were 3--4 times lower in glioma cells as compared to glia cells. Both glia and glioma cells showed variability in the occurrence of intercellular metabolite transfer, detectable via observation of a transient in a neighbour of the cell injected with substrate. On this basis "multicellular integrated states" can be defined in clusters of glioma and glia cells interconnected by cell-to-cell contact and a mesh-like network of intercellular processes. Such multicellular steady states and the associated metabolic rates or their impairment can be used in turn to classify different culture lines in reference to cell physiology and pathology.

Adenosine Diphosphate↗
PubMed1977

Mithramycin fluorescence for quantitative determination of deoxyribonucleic acid in single cells.

The use of the antibiotic drug mithramycin for cytoflurometric assessment of deoxyribonucleic acid in single cells has been studied in smears of a standard cell population of rat thymocytes. The optimal staining conditions have been determined including the influence of fixation (freeze-drying, formalin and ethanol). The drug equilibration time has been estimated in relation to the concentration of the mithramycin and to Mg++ was found to enhance the fluorescence intensity produced by the mithramycin-deoxyribonucleic acid interaction. The stability and the reproducibility of the fluorescence reaction are discussed.

Animals↗
PubMed1975

Multichannel microspectrofluorometry for topographic and spectral analysis of NAD(P)H fluorescence in single living cells.

Starting from a previously described prototype microspectrofluorometer a more versatile apparatus has been developed with rapid optional operation on a topographic mode for the simultaneous multisite evaluation of NAD(P) reduction-reoxidation transients or on a spectral mode for the analysis of natural and exogenous fluorochromes, in single living cells. On the topographic mode, adetailed kinetic analysis of NAD or NAD P-linked dehydrogenases can be made from 50-100 cell points imultaneously via automatic recording of topographic scans upt to 16 times a second, in correlation with microelectrophoretic intracellular inuection of metabolites (e.g. nearly immediate response to glucose 6-phosphate, 20-25 s delay for 6-phosphogluconate). Rapid shifts from topographic to spectral operation make possible the detection of a change in fluorescence intensity at a specific intracellular site and the immediate verification of its nature (NAD(P)H or exogenous fluorochrome) by spectral observations.

Evaluation Studies as Topic↗