Single molecule correlation spectroscopy in continuous flow mixers with zero-mode waveguides.
Zero-Mode Waveguides were first introduced for Fluorescence Correlation Spectroscopy at micromolar dye concentrations. We show that combining zero-mode waveguides with fluorescence correlation spectroscopy in a continuous flow mixer avoids the compression of the FCS signal due to fluid transport at channel velocities up to approximately 17 mm/s. We derive an analytic scaling relationship [equation: see text] converting this flow velocity insensitivity to improved kinetic rate certainty in time-resolved mixing experiments. Thus zero-mode waveguides make FCS suitable for direct kinetics measurements in rapid continuous flow.
Liao
David
D
Department of Physics, Princeton University, Princeton, NJ 08544, USA. dliao@princeton.edu
Galajda
Peter
P
Riehn
Robert
R
Ilic
Rob
R
Puchalla
Jason L
JL
Yu
Howard G
HG
Craighead
Harold G
HG
Austin
Robert H
RH
D004867:Equipment Design; D005456:Fluorescent Dyes / Q000494:pharmacology; D007700:Kinetics; D008027:Light; D015233:Models, Statistical; D008962:Models, Theoretical; D036103:Nanotechnology / Q000379:methods; D055095:Optics and Photonics; D017785:Photons; D013050:Spectrometry, Fluorescence / Q000295:instrumentation* / Q000379:methods*; D013997:Time Factors
D005456:Fluorescent Dyes