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dc.contributor.authorGarner, Aen
dc.contributor.authorFox, Brian Wen
dc.date.accessioned2011-06-22T09:09:30Z
dc.date.available2011-06-22T09:09:30Z
dc.date.issued1981-08
dc.identifier.citationMeasurements of daunorubicin uptake of Yoshida sarcoma cells in culture using flow cytofluorimetry. 1981, 36 (2):189-210 Chem Biol Interacten
dc.identifier.issn0009-2797
dc.identifier.issn10.1016/0009-2797(81)90020-X
dc.identifier.pmid7273243
dc.identifier.urihttp://hdl.handle.net/10541/134136
dc.description.abstractA comparative measurement of the transport and localisation of daunorubicin into Yoshida sarcoma cells, was undertaken by a biochemical extraction process and a flow cytometric method. An advantage of this latter procedure would be to identify subpopulation of cells which have enhanced or impaired daunorubicin incorporation as well as the ability to exclude any non-specific incorporation into cell debris, which would otherwise interfere with the overall estimation. It has been possible to use the Biophysics argon ion laser at a wavelength of 488 nm which coincides with the visible absorption bands of daunorubicin and doxorubicin (adriamycin) and the cytofluorimetric estimations of daunorubicin incorporation have now been compared with biochemically determined uptake in Yoshida cells. A high lethal dose of 10 microM was required to achieve the direct measurement by cytofluorimetry procedures on the Biophysics instrument. From cell fractionation and CHCl3/amyl alcohol extraction, it was possible to show that during a 5-h exposure period to daunorubicin (10 microM), the uptake into the nucleus was at first rapid and that into the cytoplasm was much slower. After about 3-h incubation, the level in the cytoplasm decreased, followed by a decrease from the nucleus 1 h later. This could be equated when observed microscopically to the gain in fluorescent cell debris. If all nuclear binding is to DNA, then at the level of (10 microM) concentration in the medium, the number of base pairs to daunorubicin would be 9 : 1, respectively. Cytofluorimetry showed a broad spread of intracellular daunorubicin fluorescence which increases with cell size. Increasing external concentration caused a more rapid incorporation as well as a quicker release from the cells.
dc.language.isoenen
dc.subject.meshAnimals
dc.subject.meshBiological Transport
dc.subject.meshCell Line
dc.subject.meshCell Nucleus
dc.subject.meshDaunorubicin
dc.subject.meshFlow Cytometry
dc.subject.meshKinetics
dc.subject.meshModels, Structural
dc.subject.meshNeoplasm Proteins
dc.subject.meshSarcoma, Yoshida
dc.titleMeasurements of daunorubicin uptake of Yoshida sarcoma cells in culture using flow cytofluorimetry.en
dc.typeArticleen
dc.contributor.departmentPaterson Laboratories, Christie Hospital & Holt Radium Institute, Manchester, M20 9BX, (United Kingdom)en
dc.identifier.journalChemico-Biological Interactionsen
html.description.abstractA comparative measurement of the transport and localisation of daunorubicin into Yoshida sarcoma cells, was undertaken by a biochemical extraction process and a flow cytometric method. An advantage of this latter procedure would be to identify subpopulation of cells which have enhanced or impaired daunorubicin incorporation as well as the ability to exclude any non-specific incorporation into cell debris, which would otherwise interfere with the overall estimation. It has been possible to use the Biophysics argon ion laser at a wavelength of 488 nm which coincides with the visible absorption bands of daunorubicin and doxorubicin (adriamycin) and the cytofluorimetric estimations of daunorubicin incorporation have now been compared with biochemically determined uptake in Yoshida cells. A high lethal dose of 10 microM was required to achieve the direct measurement by cytofluorimetry procedures on the Biophysics instrument. From cell fractionation and CHCl3/amyl alcohol extraction, it was possible to show that during a 5-h exposure period to daunorubicin (10 microM), the uptake into the nucleus was at first rapid and that into the cytoplasm was much slower. After about 3-h incubation, the level in the cytoplasm decreased, followed by a decrease from the nucleus 1 h later. This could be equated when observed microscopically to the gain in fluorescent cell debris. If all nuclear binding is to DNA, then at the level of (10 microM) concentration in the medium, the number of base pairs to daunorubicin would be 9 : 1, respectively. Cytofluorimetry showed a broad spread of intracellular daunorubicin fluorescence which increases with cell size. Increasing external concentration caused a more rapid incorporation as well as a quicker release from the cells.


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