PMC

(a) The temperature in the interrogation area of the device as a function of distance from the edge of the cold channel, with the cold and hot channels at 12 and 50 °C, respectively. Dots are experimental measurements; solid line is a FEMLAB numerical simulation with no fitting parameters. (b) Concentration of fluorescein (in relative units) in the interrogation channels as a function of channel number with 0, 25, and 50 ppm solution of fluorescein fed to the 0, 50, and 100% inlet, respectively. A concentration of 1 corresponds to a 50 ppm solution.

(a) Schematic drawings of various conformational states of α-synuclein with a micelle bound to it in the I_m state. Green and red spheres indicate the fluorescent dyes, Alexa 488 and 594, respectively. (b)–(c) Color-coded diagrams of apparent FRET efficiency, E_FRET, as a function of SDS concentration, [SDS], and temperature, with the protein conformations, U, I, F, and I_m, assigned to different domains. Black dots in (b) designate the 50% two-state transition midpoints between different states from temperature ramp experiments in Ref. . The dots are connected by lines to guide the eye. The temperature range in (c), 28 – 41°C, is half as broad as in (b), 21 – 47 °C.

Microfluidic device. (a) A schematic drawing of the microfluidic device. The flow channel network is shown in black. Three inlets of the device are labeled by the relative concentrations of the solutions fed to them, 0, 50, and 100%. The cyan, blue, and red channels (all 1 mm deep) are for circulation of room temperature, cold, and hot water, respectively. Arrows indicate flow directions. (b)–(d) Fluorescence micrographs of different parts of the gradient generator taken with 0, 25, and 50 ppm solutions of fluorescein in a pH 7.5 phosphate buffer fed to the 0, 50, and 100% inlets of the device, respectively. (b) Mixing channels of stages 0 – 2 and redistribution channels (horizontal strips) between them. (c) Mixing channels of stage 4 (25 µm wide straight channels with 25 µm partitions), a redistribution channel (horizontal strip), and entrance parts of the interrogation channels (8 µm wide with 8 µm partitions). (d) Interrogation area of the device with 100 parallel channels (vertical strips). Locations of the cold and hot circulation channel are marked by a blue and red horizontal strip, respectively. (e) Cross-sectional view of the microfluidic device along the dotted line in (a) with color-coded temperature profile as obtained from a two-dimensional FEMLAB thermal conductivity simulation with the boundaries of the cold, room temperature (RT), and hot channels, at 15, 20, and 55 °C, respectively (see for further details).