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  • Toward Realistic Large-Area Cell Membrane Mimics: Excluding Oil, Controlling Composition, and Including Ion Channels.

Toward Realistic Large-Area Cell Membrane Mimics: Excluding Oil, Controlling Composition, and Including Ion Channels.

Langmuir : the ACS journal of surfaces and colloids (2018-05-02)
Peter J Beltramo, Laura Scheidegger, Jan Vermant
ABSTRACT

Capacitance measurements provide unique insights into the thickness, compressibility, and composition of large-area membrane bilayers and are used here in addition to demonstrate the successful incorporation of model ion channels. The simultaneous ability to control the bilayer size, manipulate tension, and optically monitor and electrically stimulate freestanding membranes enables precise determination of their specific capacitance and thickness across a wide range of areas. We confirm that membranes formed by this recently developed technique have capacitive properties similar to those formed by existing protocols, including solvent-free approaches, and discuss the effect using either hexadecane or squalene as the oil solvent. The results obtained here are relevant for other methods where lipid membranes are reconstituted from a bulk oil solvent. Because biological membranes have a diverse phospholipid profile, we show that the technique can successfully reconstitute membranes with binary composition mixtures. As an outlook, we show the capability of model membrane proteins, specifically α-hemolysin and alamethicin, to be incorporated into the formed bilayers and measure ion transport.

MATERIALS
Product Number
Brand
Product Description

Avanti
4ME 16:0 PC, Avanti Research - A Croda Brand
Avanti
4ME 16:0 PC, Avanti Research - A Croda Brand
Avanti
18:1 (Δ9-Cis) PC (DOPC), Avanti Research - A Croda Brand
Avanti
18:1 (Δ9-Cis) PC (DOPC), Avanti Research - A Croda Brand
Avanti
18:1 (Δ9-Cis) PG, Avanti Research - A Croda Brand
Avanti
12:0 PC (DLPC), 1,2-dilauroyl-sn-glycero-3-phosphocholine, chloroform
Avanti
12:0 PC (DLPC), Avanti Research - A Croda Brand
Avanti
18:1 (Δ9-Cis) PG, Avanti Research - A Croda Brand