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Measurement of cell volume changes by fluorescence self-quenching

dc.creatorHamann, Steffen
dc.creatorKiilgaard, Jens Folke
dc.creatorLitman, Thomas
dc.creatorAlvarez-Leefmans, Francisco J.
dc.creatorWinther, Benny R.
dc.creatorZeuthen, Thomas
dc.date.accessioned2017-06-29T04:27:50Z
dc.date.available2017-06-29T04:27:50Z
dc.date.issued2002es_ES
dc.identifier364es_ES
dc.identifier.issn1053-0509es_ES
dc.identifier.urihttp://repositorio.inprf.gob.mx/handle/123456789/5055
dc.identifier.urihttps://doi.org/10.1023/A:1016832027325es_ES
dc.language.isoenges_ES
dc.relation12 (2) 139-145 p.es_ES
dc.relationversión del editores_ES
dc.rightsacceso cerradoes_ES
dc.subject.meshAnalytical Chemistryes_ES
dc.subject.meshBiochemistry, generales_ES
dc.subject.meshBiophysicses_ES
dc.subject.meshBiomedical Physicses_ES
dc.titleMeasurement of cell volume changes by fluorescence self-quenchinges_ES
dc.typearticlees_ES
dc.contributor.affiliationDepartment of Medical Physiology, The Panum Institute, University of Copenhagen, Blegdamsvej 3, DK-2200, Copenhagen N, Denmarkes_ES
dc.contributor.emailshamann@mfi.ku.dkes_ES
dc.relation.jnabreviadoJ FLUORESCes_ES
dc.relation.journalJournal of Fluorescencees_ES
dc.identifier.placePaíses Bajoses_ES
dc.date.published2002es_ES
dc.identifier.organizacionInstituto Nacional de Psiquiatría Ramón de la Fuente Muñizes_ES
dc.identifier.eissn1573-4994es_ES
dc.identifier.doi10.1023/A:1016832027325es_ES
dc.description.monthJunes_ES
dc.description.abstractotrodiomaAt high concentrations, certain fluorophores undergo self-quenching, i.e., fluorescence intensity decreases with increasing fluorophore concentration. Accordingly, the self-quenching properties can be used for measuring water volume changes in lipid vesicles. In cells, quantitative determination of water transport using fluorescence self-quenching has been complicated by the requirement of relatively high (mM) and often toxic loading concentrations. Here we report a simple method that uses low (_M) loading concentrations of calcein-acetoxymethyl ester (calcein-AM) to obtain intracellular concentrations of the fluorophore calcein suitable for measurement of changes in cell water volume by self-quenching. The relationship between calcein fluorescence intensity, when excited at 490 nm (its excitation maximum), and calcein concentration was investigated in vitro and in various cultured cell types. The relationship was bell-shaped, with the negative slope in the concentration range where the fluorophore undergoes fluorescence self-quenching. In cultured retinal pigment epithelial cells, calcein fluorescence and extracellular osmolarity were linearly related. A 25-mOsm hypertonic challenge corresponded to a decrease in calcein fluorescence with high signal-to-noise ratio (>15). Similar results were obtained with the fluorophore BCECF when excited at its isosbestic wavelength (436 nm). The present results demonstrate the usefulness of fluorescence self-quenching to measure rapid changes in cell water volumees_ES
dc.subject.koWater transportes_ES
dc.subject.koCell volumees_ES
dc.subject.koCalceines_ES
dc.subject.koSelf-quenchinges_ES
dc.subject.koEpithelial cellses_ES


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