Microfluidic monitoring of Pseudomonas aeruginosa chemotaxis under the continuous chemical gradient.

This study presents a microfluidic approach for the rapid analysis of bacterial chemotaxis in response to chemical gradients. The diffusional mixing of laminar flow continuously generates a stable chemical gradient in a microfluidic device. For the proof of concept, we have investigated the effects of the attractant peptone and repellent trichloroethylene (TCE) on chemotactic responses of wild type Pseudomonas aeruginosa PAO1 and chemotactic mutant PC4. The microfluidic method clearly demonstrates that P. aeruginosa PAO1 is attracted to peptone and repelled from TCE, whereas PC4 shows non-chemotactic behavior. In addition, the analysis of PAO1 chemotaxis on 20 amino acids revealed the effective concentration range of each amino acid as a chemoeffector. Not only does the microfluidic approach facilitate the quantitative information of chemotaxis, which gives an insight into understanding the mechanism of P. aeruginosa motility, but it also provides a useful tool for the rapid monitoring of bacterial chemotaxis in a reproducible experimental manner.

Forces acting on a single particle in an evaporating sessile droplet on a hydrophilic surface.

The evaporating sessile droplet of a mono/didisperse colloid on a plate is a very useful and handy technique in micro/nano/bioapplications to separate, pattern, and control the particles. Although the fundamental nature of the evaporation phenomena and its applications have been extensively proposed, the crucial forces affecting a single particle motion in an evaporating droplet are not reported yet. To elucidate the impact of various forces including the drag, electrostatic, van der Waals, and surface tension forces on the particle motion in suspension, the magnitudes of them are compared using the scale analysis. In the early stage of the evaporation, in which the contact line is fixed, the motion of a single particle suspended in liquid are mostly affected by drag force. Later, with the incidence of the contact line recession, the surface tension force takes over the control of the single particle motion.

Behavior of particles in an evaporating didisperse colloid droplet on a hydrophilic surface.

It is well-known that the liquid and the nanoparticles in an evaporating colloid droplet on a hydrophilic surface move radially outward for the contact line to maintain its position. However, the motion of micro/nanoparticles in an evaporating didisperse colloid droplet has not been reported to date. In this study, an experiment on an evaporating didisperse colloid droplet on the hydrophilic surface is carried out. It is found that nanoparticles move radially outward and remain at the contact line while microparticles move inward toward the center of the droplet. Furthermore, the mechanism of the microparticles moving toward the center of the droplet is found to be due to the surface tension force of the liquid.

Quantitative analysis of single bacterial chemotaxis using a linear concentration gradient microchannel.

A microfluidic device to quantify bacterial chemotaxis has been proposed, which generates a linear concentration gradient of chemoattractant in the main channel only by convective and molecular diffusion, and which enables the bacteria to enter the main channel in a single file by hydrodynamic focusing technique. The trajectory of each bacterium in response to the concentration gradient of chemoattractant is photographed by a CCD camera and its velocity is acquired by a simple PTV (Particle Tracking Velocimetry) algorithm. An advantage of this assay is to measure the velocity of a single bacterium and to quantify the degree of chemotaxis by analyzing the frequency of velocities concurrently. Thus, the parameter characterizing the motility of wild-type Escherichia coli strain RP437 in response to various concentration gradients of L-aspartate is obtained in such a manner that the degree of bacterial chemotaxis is quantified on the basis of a newly proposed Migration Index.

Three-dimensional focusing of red blood cells in microchannel flows for bio-sensing applications.

Three-dimensional (3D) focusing of particles in microchannels has been a long-standing issue in the design of biochemical/biomedical microdevices. Current microdevices for 3D cell or bioparticle focusing involve complex channel geometries in view of their fabrication because they require multiple layers and/or sheath flows. This paper proposes a simple method for 3D focusing of red blood cells (RBCs) in a single circular microcapillary, without any sheath flows, which is inspired from the fluid dynamics phenomenon in that a spherical particle lagging behind a Poiseuille flow migrates toward and along the channel axis. More explicitly, electrophoresis of RBCs superimposed on the pressure-driven flow is utilized to generate an RBC migration mode analogous to this phenomenon. A particle-tracking scheme with a sub-pixel resolution is implemented to spatially position red blood cells flowing through the channel, so that a probability density function (PDF) is constructed to evaluate the tightness of the cell focusing. Above a specific strength of the electric field, approximately 90% of the sheep RBCs laden in the flow are tightly focused within a beam diameter that is three times the cell dimension. Particle shape effect on the focusing is discussed by making comparisons between the RBCs and the spherical particles. The lateral migration velocity, predicted by an existing theoretical model, is in good agreement with the present experimental data. It is noteworthy that 3D focusing of non-spherical particles, such as RBCs, has been achieved in a circular microchannel, which is a significant improvement over previous focusing methodologies.

Axisymmetric flow focusing of particles in a single microchannel.

Axisymmetric flow focusing of particles in a single microchannel has been proposed on the basis of the observation that a particle migrates toward the tube axis when it lags behind the fluid flow, which is supported by demonstrating that more than 90% of the particles are tightly focused within three times the particle diameter when negative electrophoretic mobility is imposed on the particles.

Quantification of the cell-substratum contact and cell lift-off under different intra/extracellular conditions.

Analyses of images of the cell-to-substratum region of contact have been carried out by the means of total internal reflection fluorescence (TIRF) microscopy during both the formation and the dissolution of cellular contacts. The evolutions of the cellular contacts are visualized during the adhesion process under normal and virus-infected intracellular conditions, and during the lift-off process under various toxicities of the extracellular medium fluid. Then, propositions are developed for quantifying the cell viabilities by estimating the increase in the area of contact for the adhesion process and by specifying the maximum intensity of the TIRF image for the lift-off process.