Scanning Probe Lithography on Fluid Lipid Membranes

Scanning probe lithography (SPL) is applied to pattern fluid lipid membranes on a solid borosilicate substrate. Grids of metal lines, prepatterned onto the substrate by electron beam lithography, serve to partition the supported membrane into an array of isolated fluid pixels. By toggling the pH of the surrounding solution, the effect of the probe tip on the membrane can be regulated. Alkaline conditions favor membrane removal, while neutral pH favors membrane deposition. Arbitrary membrane patterns with spatial dimensions limited by the underlying grid size can be constructed by sequential SPL membrane removal followed by refill with a different membrane type. In the present study, bilayers of unique composition fill 1 × 1 μm corrals and were positioned 100 nm apart

Topographical Imaging of an Intermembrane Junction by Combined Fluorescence Interference and Energy Transfer Microscopies

Topographical Imaging of an Intermembrane Junction by Combined Fluorescence Interference and Energy Transfer Microscopie

Scanning Probe Lithography on Fluid Lipid Membranes

Scanning probe lithography (SPL) is applied to pattern fluid lipid membranes on a solid borosilicate substrate. Grids of metal lines, prepatterned onto the substrate by electron beam lithography, serve to partition the supported membrane into an array of isolated fluid pixels. By toggling the pH of the surrounding solution, the effect of the probe tip on the membrane can be regulated. Alkaline conditions favor membrane removal, while neutral pH favors membrane deposition. Arbitrary membrane patterns with spatial dimensions limited by the underlying grid size can be constructed by sequential SPL membrane removal followed by refill with a different membrane type. In the present study, bilayers of unique composition fill 1 × 1 μm corrals and were positioned 100 nm apart

Surface Binding Affinity Measurements from Order Transitions of Lipid Membrane-Coated Colloidal Particles

Lipid bilayers can be assembled onto the surface of colloidal silica particles to form a continuous and fluid supported membrane coating. In this configuration, the collective behavior of the colloidal dispersion is governed by interactions between particles and exhibits a sensitive dependency on chemical features of the membrane surface. Protein binding to membrane surface receptors can trigger macroscopic changes in the colloidal order, which provides a label-free readout of such binding events. Here, the relationship between order in the colloidal dispersion and the surface concentration of bound protein is characterized quantitatively in terms of the radial pair distribution function. Using parallel fluorescence measurements for comparison, we construct a scalar measure of the distribution function that exhibits linear proportionality with surface protein binding. This is used to determine binding affinity based only on observations of the colloidal distribution

Scanning Probe Lithography on Fluid Lipid Membranes

Scanning probe lithography (SPL) is applied to pattern fluid lipid membranes on a solid borosilicate substrate. Grids of metal lines, prepatterned onto the substrate by electron beam lithography, serve to partition the supported membrane into an array of isolated fluid pixels. By toggling the pH of the surrounding solution, the effect of the probe tip on the membrane can be regulated. Alkaline conditions favor membrane removal, while neutral pH favors membrane deposition. Arbitrary membrane patterns with spatial dimensions limited by the underlying grid size can be constructed by sequential SPL membrane removal followed by refill with a different membrane type. In the present study, bilayers of unique composition fill 1 × 1 μm corrals and were positioned 100 nm apart

Formation and Spatio-Temporal Evolution of Periodic Structures in Lipid Bilayers

Miscibility phase separation in lipid bilayers is widely implicated as an organizing principle in living cell membranes. However, the chemical and physical aspects of how membrane phase separation modulates protein activity remain obscure. Herein, we describe formation of ordered superstructures of coexisting liquid phases in bilayer membranes. Metastable stripe and hexagonal domain lattices are observed, as well as transitions between them. The high degree of order achieved by these methods facilitates statistical analysis of domain spatial distributions and enables measurement of domain interactions. Such long-range ordering principles may exist in more complicated membrane systems

Formation and Spatio-Temporal Evolution of Periodic Structures in Lipid Bilayers

Miscibility phase separation in lipid bilayers is widely implicated as an organizing principle in living cell membranes. However, the chemical and physical aspects of how membrane phase separation modulates protein activity remain obscure. Herein, we describe formation of ordered superstructures of coexisting liquid phases in bilayer membranes. Metastable stripe and hexagonal domain lattices are observed, as well as transitions between them. The high degree of order achieved by these methods facilitates statistical analysis of domain spatial distributions and enables measurement of domain interactions. Such long-range ordering principles may exist in more complicated membrane systems

Formation and Spatio-Temporal Evolution of Periodic Structures in Lipid Bilayers

Miscibility phase separation in lipid bilayers is widely implicated as an organizing principle in living cell membranes. However, the chemical and physical aspects of how membrane phase separation modulates protein activity remain obscure. Herein, we describe formation of ordered superstructures of coexisting liquid phases in bilayer membranes. Metastable stripe and hexagonal domain lattices are observed, as well as transitions between them. The high degree of order achieved by these methods facilitates statistical analysis of domain spatial distributions and enables measurement of domain interactions. Such long-range ordering principles may exist in more complicated membrane systems

Formation and Spatio-Temporal Evolution of Periodic Structures in Lipid Bilayers

Miscibility phase separation in lipid bilayers is widely implicated as an organizing principle in living cell membranes. However, the chemical and physical aspects of how membrane phase separation modulates protein activity remain obscure. Herein, we describe formation of ordered superstructures of coexisting liquid phases in bilayer membranes. Metastable stripe and hexagonal domain lattices are observed, as well as transitions between them. The high degree of order achieved by these methods facilitates statistical analysis of domain spatial distributions and enables measurement of domain interactions. Such long-range ordering principles may exist in more complicated membrane systems

Curvature-Modulated Phase Separation in Lipid Bilayer Membranes

Cellular membranes exhibit a variety of controlled curvatures, with filopodia, microvilli, and mitotic cleavage furrows being only a few of many examples. Coupling between local curvature and chemical composition in membranes could provide a means of mechanically controlling the spatial organization of membrane components. Although this concept has surfaced repeatedly over the years, controlled experimental investigations have proven elusive. Here, we introduce an experimental platform, in which microfabricated surfaces impose specific curvature patterns onto lipid bilayers, that allows quantification of mechanochemical couplings in membranes. We find that, beyond a critical curvature value, membrane geometry governs the spatial ordering of phase-separated domain structures in membranes composed of cholesterol and phospholipids. The curvature-controlled ordering, a consequence of the distinct mechanical properties of the lipid phases, makes possible a determination of the bending rigidity difference between cholesterol-rich and cholesterol-poor lipid domains. These observations point to a strong coupling between mechanical bending and chemical organization that should have wide-reaching consequences for biological membranes. Curvature-mediated patterning may also be useful in controlling complex fluids other than biomembranes