Please use this identifier to cite or link to this item: http://doi.org/10.25358/openscience-1623
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dc.contributor.authorFine, Tamir
dc.date.accessioned2010-01-22T15:07:32Z
dc.date.available2010-01-22T16:07:32Z
dc.date.issued2010
dc.identifier.urihttps://openscience.ub.uni-mainz.de/handle/20.500.12030/1625-
dc.description.abstractAs the elastic response of cell membranes to mechanical stimuli plays a key role in various cellular processes, novel biophysical strategies to quantify the elasticity of native membranes under physiological conditions at a nanometer scale are gaining interest. In order to investigate the elastic response of apical membranes, elasticity maps of native membrane sheets, isolated from MDCK II (Madine Darby Canine kidney strain II) epithelial cells, were recorded by local indentation with an Atomic Force Microscope (AFM). To exclude the underlying substrate effect on membrane indentation, a highly ordered gold coated porous array with a pore diameter of 1.2 μm was used to support apical membranes. Overlays of fluorescence and AFM images show that intact apical membrane sheets are attached to poly-D-lysine coated porous substrate. Force indentation measurements reveal an extremely soft elastic membrane response if it is indented at the center of the pore in comparison to a hard repulsion on the adjacent rim used to define the exact contact point. A linear dependency of force versus indentation (-dF/dh) up to 100 nm penetration depth enabled us to define an apparent membrane spring constant (kapp) as the slope of a linear fit with a stiffness value of for native apical membrane in PBS. A correlation between fluorescence intensity and kapp is also reported. Time dependent hysteresis observed with native membranes is explained by a viscoelastic solid model of a spring connected to a Kelvin-Voight solid with a time constant of 0.04 s. No hysteresis was reported with chemically fixated membranes. A combined linear and non linear elastic response is suggested to relate the experimental data of force indentation curves to the elastic modulus and the membrane thickness. Membrane bending is the dominant contributor to linear elastic indentation at low loads, whereas stretching is the dominant contributor for non linear elastic response at higher loads. The membrane elastic response was controlled either by stiffening with chemical fixatives or by softening with F-actin disrupters. Overall, the presented setup is ideally suitable to study the interactions of the apical membrane with the underlying cytoskeleton by means of force indentation elasticity maps combined with fluorescence imaging.en_GB
dc.language.isoeng
dc.rightsInCopyrightde_DE
dc.rights.urihttps://rightsstatements.org/vocab/InC/1.0/
dc.subject.ddc540 Chemiede_DE
dc.subject.ddc540 Chemistry and allied sciencesen_GB
dc.titleMapping the elastic response of epithelial apical cell membranes suspended across porous arrayen_GB
dc.typeDissertationde_DE
dc.identifier.urnurn:nbn:de:hebis:77-21740
dc.identifier.doihttp://doi.org/10.25358/openscience-1623-
jgu.type.dinitypedoctoralThesis
jgu.type.versionOriginal worken_GB
jgu.type.resourceText
jgu.description.extent108 S.
jgu.organisation.departmentFB 09 Chemie, Pharmazie u. Geowissensch.-
jgu.organisation.year2009
jgu.organisation.number7950-
jgu.organisation.nameJohannes Gutenberg-Universität Mainz-
jgu.rights.accessrightsopenAccess-
jgu.organisation.placeMainz-
jgu.subject.ddccode540
opus.date.accessioned2010-01-22T15:07:32Z
opus.date.modified2010-01-27T12:24:27Z
opus.date.available2010-01-22T16:07:32
opus.subject.dfgcode00-000
opus.subject.otherAFM, Elasticity, Apical Membrane, Indentationen_GB
opus.organisation.stringFB 09: Chemie, Pharmazie und Geowissenschaften: Institut für Physikalische Chemiede_DE
opus.identifier.opusid2174
opus.institute.number0906
opus.metadataonlyfalse
opus.type.contenttypeDissertationde_DE
opus.type.contenttypeDissertationen_GB
jgu.organisation.rorhttps://ror.org/023b0x485
Appears in collections:JGU-Publikationen

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