Cell membranes suspended across nanoaperture arrays.
We present a method for spreading large (>100 microm(2)) cell membrane fragments across nanoapertures in planar supports. Electron-beam and focused-ion-beam lithography were used to fabricate arrays of 50-600 nm diameter holes in free-standing silicon nitride (SiN) solid films 100-500 nm thick. By pressing adhering live cells onto the nanostructured SiN surface and then removing them, planar cell membrane sheets (CMSs) were transferred in a well-defined orientation onto the SiN support. We demonstrate the accessibility to both extracellular and intracellular surfaces of CMSs by targeting membrane constituents side-specifically with fluorescent markers. Our approach is of interest for studying ligand-receptor interactions using optical, electrical, and scanning probe techniques at the single-molecule level.
Danelon
Christophe
C
Laboratory of Physical Chemistry of Polymers and Membranes, Ecole Polytechnique Fédérale de Lausanne, 1015 Lausanne, Switzerland.
Perez
Jean-Baptiste
JB
Santschi
Christian
C
Brugger
Jürgen
J
Vogel
Horst
H
D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure*; D002478:Cells, Cultured; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D018613:Microscopy, Confocal; D036103:Nanotechnology; D018040:Receptors, Neurokinin-1 / Q000378:metabolism; D011985:Receptors, Serotonin / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017655:Silicon Compounds; D013499:Surface Properties
C485182:Cyan Fluorescent Protein; D018040:Receptors, Neurokinin-1; D011985:Receptors, Serotonin; D011993:Recombinant Fusion Proteins; D017655:Silicon Compounds; C078730:serotonin 5 receptor; D049452:Green Fluorescent Proteins; C032734:silicon nitride