Microfluidic flow-flash: method for investigating protein dynamics |
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Authors: | Toepke Michael W Brewer Scott H Vu Dung M Rector Kirk D Morgan Joel E Gennis Robert B Kenis Paul J A Dyer R Brian |
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Affiliation: | Department of Chemical and Biomolecular Engineering and Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA. |
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Abstract: | We report a new method, microfluidic flow-flash, for measuring protein reaction kinetics. The method couples a microscope imaging detection system with a microfluidic flow cell to reduce data acquisition times and sample consumption. This combination allows for the simultaneous collection of spectral and temporal information. The microfluidic flow cell design utilizes three-dimensional sheath flow to reduce sample dispersion and minimize sample consumption. The ability to alter the flow rates in the microfluidic flow cells allows a variety of time scales to be studied with submillisecond time resolution. The imaging detection system can be coupled with several spectroscopic probes including fluorescence and UV/visible absorbance spectroscopy. Here, we utilize the microfluidic flow-flash method to probe the kinetics of CO recombination or O2 binding to myoglobin after the laser-induced photolysis of CO from myoglobin by UV/visible absorbance spectral imaging. |
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