4.8 Article

Microfluidic flow-flash: Method for investigating protein dynamics

期刊

ANALYTICAL CHEMISTRY
卷 79, 期 1, 页码 122-128

出版社

AMER CHEMICAL SOC
DOI: 10.1021/ac061193x

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资金

  1. NHLBI NIH HHS [R01 HL016101, HL 16101] Funding Source: Medline
  2. NIGMS NIH HHS [GM 068036, P01 GM068036] Funding Source: Medline
  3. NATIONAL HEART, LUNG, AND BLOOD INSTITUTE [R01HL016101, R37HL016101] Funding Source: NIH RePORTER
  4. NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [P01GM068036] Funding Source: NIH RePORTER

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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 O-2 binding to myoglobin after the laser-induced photolysis of CO from myoglobin by UV/visible absorbance spectral imaging.

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