A more efficient device for preparing model-membrane liposomes by the rapid solvent exchange method.

We modified the original design for a rapid solvent exchange (RSE) device with the intent of making the RSE method (i) more efficient and (ii) easier to adopt and implement. Our modifications improved solvent-removal kinetics by a factor of 2, while reducing sample-prep time by a factor of 3. In this paper, we develop the kinetic model that informed the device revision and we address several RSE parameters that have not yet been discussed in the literature.

Stern-Volmer modeling of steady-state Forster energy transfer between dilute, freely diffusing membrane-bound fluorophores.

Two different metrics are used to assess Forster resonance energy transfer (FRET) between fluorophores in the steady state: (i) acceptor-quenching of donor fluorescence E (also known as transfer efficiency) and (ii) donor-excited acceptor fluorescence F(A) (Dex). While E is still more widely used, F(A) (Dex) has been gaining in popularity for practical reasons among experimentalists who study biomembranes. Here, for the special case of membrane-bound fluorophores, we present a substantial body of experimental evidence that justifies the use of simple Stern-Volmer expressions when modeling either FRET metric under dilute-probe conditions.

High-resolution mapping of phase behavior in a ternary lipid mixture: do lipid-raft phase boundaries depend on the sample preparation procedure?

For some time now, we have been using a fluorescence resonance energy transfer (FRET)-based strategy to conduct high-resolution studies of phase behavior in ternary lipid-raft membrane mixtures. Our FRET experiments can be carried out on ordinary, polydisperse multilamellar vesicle suspensions, so we are able to prepare our samples according to a procedure that was designed specifically to guard against artifactual phase separation. In some respects (i.

Steady-state probe-partitioning fluorescence resonance energy transfer: a simple and robust tool for the study of membrane phase behavior.

An experimental strategy has been developed specifically for the study of composition-dependent phase behavior in multicomponent artificial membranes. The strategy is based on steady-state measurements of fluorescence resonance energy transfer between freely diffusing membrane probe populations, and it is well suited for the rapid generation of large data sets. Presented in this paper are the basic principles that guide the experiment's design, the derivation of an underlying mathematical model that serves to interpret the data, and experimental results that confirm the model's predictive power.

Fluorescence methods to detect phase boundaries in lipid bilayer mixtures.

Phase diagrams of lipid mixtures can show several different regions of phase coexistence, which include liquid-disordered, liquid-ordered, and gel phases. Some phase regions are small, and some have sharp boundaries. The identity of the phases, their location in composition space, and the nature of the transitions between the phases are important for understanding the behavior of lipid mixtures.

Phospholipid solubility determined by equilibrium distribution between surface and bulk phases.

A general strategy is proposed for determining the very low aqueous solubility limits of bilayer-forming phospholipids. The strategy exploits the inherent surface activity of phospholipids and has been termed EDSB, which stands for Equilibrium Distribution between Surface and Bulk phases. In this report, EDSB has been used to determine the critical bilayer concentration of dilauroylphosphatidylycholine (DLPC), a short-chain bilayer-forming phospholipid.