RESUMO
We present a novel, to the best of knowledge, time-resolved, optical pump/NIR supercontinuum probe spectrometer suitable for oscillators. A NIR supercontinuum probe spectrum (850-1250 nm) is generated in a photonic crystal fiber, dispersed across a digital micromirror device (DMD), and then raster scanned into a single element detector at a 5 Hz rate. Dual modulation of pump and probe beams at disparate frequencies permits simultaneous measurement of both the bare reflectance R and its photoinduced change ΔR through lock-in detection, allowing for continuously self-normalized measurement of ΔR/R. Example data are presented on a germanium wafer sample that demonstrate for signals of order ΔR/R â¼ 10-3, a 2.87 nm spectral resolution and â²400 fs temporal resolution pre-recompression, and comparable sensitivity to standard time-resolved, amplifier-based pump-probe techniques.
RESUMO
Nanodiscs, which are disc-shaped entities that contain a central lipid bilayer encased by an annulus of amphipathic helices, have emerged as a leading native-like membrane mimic. The current approach for the formation of nanodiscs involves the creation of a mixed-micellar solution containing membrane scaffold protein, lipid, and detergent followed by a time consuming process (3-12 h) of dialysis and/or incubation with sorptive beads to remove the detergent molecules from the sample. In contrast, the methodology described herein provides a facile and rapid procedure for the preparation of nanodiscs in a matter of minutes (<15 min) using Sephadex® G-25 resin to remove the detergent from the sample. A panoply of biophysical techniques including analytical ultracentrifugation, dynamic light scattering, gel filtration chromatography, circular dichroism spectroscopy, and cryogenic electron microscopy were employed to unequivocally confirm that aggregates formed by this method are indeed nanodiscs. We believe that this method will be attractive for time-sensitive and high-throughput experiments.
