Chiaruttini N.,ESPCI ParisTech |
Chiaruttini N.,University of Geneva |
Letellier L.,University Paris - Sud |
Viasnoff V.,ESPCI ParisTech |
And 3 more authors.
PLoS ONE | Year: 2013
We present an innovative method to couple electrophysiological measurements with fluorescence imaging of functionalized suspended bilayers. Our method combines several advantages: it is well suited to study transmembrane proteins that are difficult to incorporate in suspended bilayers, it allows single molecule resolution both in terms of electrophysiological measurements and fluorescence imaging, and it enables mechanical stimulations of the membrane. The approach comprises of two steps: first the reconstitution of membrane proteins in giant unilamellar vesicles; then the formation of a suspended bilayer spanning a 5 to 15 micron-wide aperture that can be visualized by high NA microscope objectives. We exemplified how the technique can be used to detect in real time the translocation of T5 DNA across the bilayer during its ejection from the bacteriophage capsid. © 2013 Chiaruttini et al.
Masters T.,MechanoBiology Institute of Singapore |
Engl W.,MechanoBiology Institute of Singapore |
Weng Z.L.,MechanoBiology Institute of Singapore |
Arasi B.,MechanoBiology Institute of Singapore |
And 3 more authors.
PLoS ONE | Year: 2012
Since protein patterning on 2D surfaces has emerged as an important tool in cell biology, the development of easy patterning methods has gained importance in biology labs. In this paper we present a simple, rapid and reliable technique to fabricate thin layers of UV curable polymer with through holes. These membranes are as easy to fabricate as microcontact printing stamps and can be readily used for stencil patterning. We show how this microfabrication scheme allows highly reproducible and highly homogeneous protein patterning with micron sized resolution on surfaces as large as 10 cm2. Using these stencils, fragile proteins were patterned without loss of function in a fully hydrated state. We further demonstrate how intricate patterns of multiple proteins can be achieved by stacking the stencil membranes. We termed this approach microserigraphy. © 2012 Masters et al.