2022
DOI: 10.1002/smll.202205567
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Native Planar Asymmetric Suspended Membrane for Single‐Molecule Investigations: Plasma Membrane on a Chip

Abstract: of the plasma membrane are essential for numerous cellular activities, such as protein complex formation and receptor-mediated signaling. [1] However, obtaining structural and dynamic information on membrane protein complexes is one of the most significant challenges in cell and molecular biology. Most of the in vitro platforms to study membrane protein interactions can be grouped into two broad categories: curved or planar bilayers. Curved bilayers can be achieved with liposomes in the format of small unilame… Show more

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Cited by 10 publications
(15 citation statements)
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“…Together with single-vesicle fusion assays, [49][50][51][52][53][54][55] we can dissect the roles of individual molecular components in the insulin biphasic secretion process. This includes our patented suspended lipid membrane platform (SLIM), 56 which uses synthetic reconstitutions or native lipid membranes to perform highthroughput fusion assays while also providing the precision to observe single molecule localization at the same time.…”
Section: Discussionmentioning
confidence: 99%
“…Together with single-vesicle fusion assays, [49][50][51][52][53][54][55] we can dissect the roles of individual molecular components in the insulin biphasic secretion process. This includes our patented suspended lipid membrane platform (SLIM), 56 which uses synthetic reconstitutions or native lipid membranes to perform highthroughput fusion assays while also providing the precision to observe single molecule localization at the same time.…”
Section: Discussionmentioning
confidence: 99%
“…However, it is possible to directly count the number of SNAREpins associated with each ready-release vesicle in the reconstituted vesicle-bilayer fusion system using well-established single-molecule methods involving TIRF. We employed a variation of the suspended bilayer chip [12] specially adapted to enable TIRF imaging [30] thereby allowing singlemolecule counting by step-photobleaching of SNAREpins labeled with fluorescent Complexin [16].…”
Section: Refining the Model With Synaptophysin To Template Snarepinsmentioning
confidence: 99%
“…For TIRF experiments, we created the suspended bilayer using a silicon chip, with buffer supplemented with 45% OptiPrep gradient media for index matching 69 . Optiprep on the top was washed out with 1X buffer after bilayer formation.…”
Section: Suspended Bilayer and Vesicle Preparationmentioning
confidence: 99%