[Show abstract][Hide abstract] ABSTRACT: We report the application of a microfluidic device for semi-contact temperature measurement in picoliter volumes of aqueous media. Our device, a freely positionable multifunctional pipette, operates by a hydrodynamic confinement principle, i.e., by creating a virtual flow cell of micrometer dimensions within a greater aqueous volume. We utilized two fluorescent rhodamines, which exhibit different fluorescent responses with temperature, and made ratiometric intensity measurements. The temperature dependence of the intensity ratio was calibrated and used in a model study of the thermal activation of TRPV1 ion channels expressed in Chinese hamster ovary cells. Our approach represents a practical and robust solution to the specific problem of measuring temperature in biological experiments in vitro, involving highly localized heat generation, for example with an IR-B laser.
[Show abstract][Hide abstract] ABSTRACT: Hydrophobic side groups on a stimuli-responsive polymer, encapsulated within a single giant unilamellar vesicle, enable membrane attachment during compartment formation at elevated temperatures. We thermally modulated the vesicle through implementation of an IR laser via an optical fiber, enabling localized directed heating. Polymer-membrane interactions were monitored using confocal imaging techniques as subsequent membrane protrusions occurred and lipid nanotubes formed in response to the polymer hydrogel contraction. These nanotubes, bridging the vesicle membrane to the contracting hydrogel, were retained on the surface of the polymer compartment, where they were transformed into smaller vesicles in a process reminiscent of cellular endocytosis. This development of a synthetic vesicle system containing a stimuli-responsive polymer could lead to a new platform for studying inter/intramembrane transport through lipid nanotubes.
Full-text · Article · Nov 2011 · Journal of the American Chemical Society