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Continuous Pore-Spanning Lipid Bilayers on Silicon Oxide-Coated Porous Substrates
journal contribution
posted on 2017-11-17, 00:00 authored by Nelli Teske, Jeremias Sibold, Johannes Schumacher, Nikolas K. Teiwes, Martin Gleisner, Ingo Mey, Claudia SteinemA number
of techniques has been developed and analyzed in recent
years to generate pore-spanning membranes (PSMs). While quite a number
of methods rely on nanoporous substrates, only a few use micrometer-sized
pores to be able to individually resolve suspending membranes by means
of fluorescence microscopy. To be able to produce PSMs on pores that
are micrometer in size, an orthogonal functionalization strategy resulting
in a hydrophilic surface is highly desirable. Here, we report on a
method to prepare PSMs based on the evaporation of a thin layer of
silicon monoxide on top of the porous substrate. PM-IRRAS experiments
demonstrate that the final surface is composed of SiOx with 1 < x < 2. The hydrophilic
surface turned out to be well suited to spread giant unilamellar vesicles
forming PSMs. As the method does not rely on a gold coating as frequently
used for orthogonal functionalization, fluorescence micrographs provide
information not only from the freestanding membrane areas but also
from the supported ones. The observation of the entire PSM area enabled
us to observe phase-separation in these membranes on the freestanding
and supported parts as well as protein binding and possible lipid
reorganization of the membranes induced by binding of the protein
Shiga toxin.
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orthogonal functionalization strategyuse micrometer-sized poresorthogonal functionalizationspread giant unilamellar vesiclesfreestanding membrane areasPM-IRRAS experimentspore-spanning membranesgold coatingSiO xnanoporous substratessilicon monoxidePSM areamethodContinuous Pore-Spanning Lipid Bilayerssurfacelipid reorganizationprotein Shiga toxinfluorescence microscopyprotein bindingSilicon Oxide-Coated Porous Substratesfluorescence micrographs
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