Fluorescence correlation spectroscopy (FCS) of colloidal nanoparticles using a near-field fiber probe was numerically simulated.
The near-wall dynamics was simulated by accounting for the anisotropic mobility of nanoparticles owing to hydrodynamic interaction
with a wall (Stokes viscous force). By comparing the simulation results with theoretical model calculations, we found that
the influence of anisotropic diffusion is insignificant in near-field FCS autocorrelation analysis.
[Show abstract][Hide abstract] ABSTRACT: Fluorescence fluctuation spectroscopy is a versatile technique applied to in vitro and in vivo investigations of biochemical processes such as interactions, mobilities or densities with high specifity and sensitivity. The prerequisite of this dynamical fluorescence technique is to have, at a time, only few fluorescent molecules in the detection volume in order to generate significant fluorescence fluctuations. For usual confocal fluorescence microscopy this amounts to a useful concentration in the nanomolar range. The concentration of many biomolecules in living cell or on cell membranes is, however, often quite high, usually in the micro- to the millimolar range. To allow fluctuation spectroscopy and track intracellular interaction or localization of single fluorescently labeled biomolecules in such crowded environments, development of detection volumes with nanoscale resolution is necessary. As diffraction prevents this in the case of light microscopy, new (non-invasive) optical concepts have been developed. In this mini-review article we present recent advancements, implemented to decrease the detection volume below that of normal fluorescence microscopy. Especially, their combination with fluorescence fluctuation spectroscopy is emphasized.
Current Pharmaceutical Biotechnology 03/2006; 7(1):51-66. DOI:10.2174/138920106775789629 · 2.51 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Microfabricated integrated circuits revolutionized computation by vastly reducing the space, labor, and time required for calculations. Microfluidic systems hold similar promise for the large-scale automation of chemistry and biology, suggesting the possibility of numerous experiments performed rapidly and in parallel, while consuming little reagent. While it is too early to tell whether such a vision will be realized, significant progress has been achieved, and various applications of significant scientific and practical interest have been developed. Here a review of the physics of small volumes (nanoliters) of fluids is presented, as parametrized by a series of dimensionless numbers expressing the relative importance of various physical phenomena. Specifically, this review explores the Reynolds number Re, addressing inertial effects; the Péclet number Pe, which concerns convective and diffusive transport; the capillary number Ca expressing the importance of interfacial tension; the Deborah, Weissenberg, and elasticity numbers De, Wi, and El, describing elastic effects due to deformable microstructural elements like polymers; the Grashof and Rayleigh numbers Gr and Ra, describing density-driven flows; and the Knudsen number, describing the importance of noncontinuum molecular effects. Furthermore, the long-range nature of viscous flows and the small device dimensions inherent in microfluidics mean that the influence of boundaries is typically significant. A variety of strategies have been developed to manipulate fluids by exploiting boundary effects; among these are electrokinetic effects, acoustic streaming, and fluid-structure interactions. The goal is to describe the physics behind the rich variety of fluid phenomena occurring on the nanoliter scale using simple scaling arguments, with the hopes of developing an intuitive sense for this occasionally counterintuitive world.
Review of Modern Physics 10/2005; 77(3). DOI:10.1103/RevModPhys.77.977 · 29.60 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The microfabrication technologies of the semiconductor industry have made it possible to integrate increasingly complex electronic and mechanical functions, providing us with ever smaller, cheaper and smarter sensors and devices. These technologies have also spawned microfluidics systems for containing and controlling fluid at the micrometre scale, where the increasing importance of viscosity and surface tension profoundly affects fluid behaviour. It is this confluence of available microscale engineering and scale-dependence of fluid behaviour that has revolutionized our ability to precisely control fluid/fluid interfaces for use in fields ranging from materials processing and analytical chemistry to biology and medicine.
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