Combining patch-clamping and fluorescence microscopy for quantitative reconstitution of cellular membrane processes with Giant Suspended Bilayers

摘要: In vitro reconstitution and microscopic visualization of membrane processes is an indispensable source information about a cellular function. Here we describe conceptionally novel free-standing template that facilitates such quantitative remodelling at different scales. The Giant Suspended Bilayers (GSBs) spontaneously swell from lipid lamella reservoir deposited on microspheres. GSBs attached to the can be prepared virtually any composition following fast procedure. unilamellar vesicles further obtained by GSB detachment from stabilizes during deformations, mechanical micromanipulations, fluorescence microscopy observations, while GSB-reservoir boundary enables exchange small solutes with interior. These unique properties allow studying macro- nano-scale adding membrane-active compounds both sides applying patch-clamp based approaches, thus making versatile tool for quantification trafficking events.