Microfluidic flow-flash: method for investigating protein dynamics.
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.
Toepke
Michael W
MW
Department of Chemical and Biomolecular Engineering and Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
Brewer
Scott H
SH
Vu
Dung M
DM
Rector
Kirk D
KD
Morgan
Joel E
JE
Gennis
Robert B
RB
Kenis
Paul J A
PJ
Dyer
R Brian
RB
D002248:Carbon Monoxide / Q000737:chemistry / Q000378:metabolism; D005453:Fluorescence; D007700:Kinetics; D007834:Lasers; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D018613:Microscopy, Confocal / Q000379:methods; D009211:Myoglobin / Q000737:chemistry* / Q000378:metabolism; D010100:Oxygen / Q000737:chemistry / Q000378:metabolism; D010782:Photolysis; D013050:Spectrometry, Fluorescence / Q000379:methods; D013056:Spectrophotometry, Ultraviolet / Q000379:methods
D009211:Myoglobin; D002248:Carbon Monoxide; D010100:Oxygen