Exploring operational boundaries for acoustic concentration of cell suspensions.

The development of a standardized, generic method for concentrating suspensions in continuous flow is challenging. In this study, we developed and tested a device capable of concentrating suspensions with an already high cell concentration to meet diverse industrial requirements. To address typical multitasking needs, we concentrated suspensions with high solid content under a variety of conditions. Cells from Saccharomyces cerevisiae, Escherichia coli, and Chinese hamster ovary cells were effectively focused in the center of the main channel of a microfluidic device using acoustophoresis. The main channel bifurcates into three outlets, allowing cells to exit through the central outlet, while the liquid evenly exits through all outlets. Consequently, the treatment separates cells from two-thirds of the surrounding liquid. We investigated the complex interactions between parameters. Increasing the channel depth results in a decrease in process efficiency, attributed to a decline in acoustic energy density. The study also revealed that different cell strains exhibit distinct acoustic contrast factors, originating from differences in dimensions, compressibility, and density values. Finally, a combination of high solid content and flow rate leads to an increase in diffusion through a phenomenon known as shear-induced diffusion. KEY POINTS: • Acoustic focusing in a microchannel was used to concentrate cell suspensions • The parameters influencing focusing at high concentrations were studied • Three different cell strains were successfully concentrated.

Scanning defocusing particle tracking for the experimental characterization of flows in demanding microfluidic systems.

A novel scanning particle image velocimetry technique, to the best of our knowledge, is proposed to characterize flows in microfluidic applications. Three-dimensional information is acquired by oscillating the target sample over a fixed focal plane, allowing the reconstruction of particle trajectories with micrometer accuracy over an extended depth. This technology is suited for investigating acoustic flows with unprecedented precision in microfluidic applications. In this contribution, we describe the experimental setup and the data processing pipeline in detail; we study the technique's performance by reconstructing pressure-driven flow; and we report the three-dimensional trajectory of a 2 µm particle in an acoustic flow in a 525µm×375µm microchannel with micrometric accuracy.

C-Term Reduction in 3 µm Open-Tubular High-Aspect-Ratio Channels in AC-EOF Vortex Chromatography Operation.

The performance of liquid chromatography operation in open-tubular channels, the ideal chromatographic column format, is limited by slow mass transport between the mobile and stationary phase. We recently introduced a lateral mixing methodology ("vortex chromatography") to reduce Taylor-Aris dispersion by employing (small) AC-EOF (alternating current electroosmotic flow) fields oriented perpendicular to the conventionally applied, axially oriented pressure gradient, resulting in the reduction of the C-term by a factor of 3, studied in 40 × 20 µm2 (aspect ratio (AR) = 2) channels under unretained conditions. In the present contribution, a further increased performance gain for channel dimensions relevant for chromatographic applications is demonstrated. The impact of the applied voltage and salt concentration is studied for 3 × 20 and 5 × 20 µm2 channels in ARs of up to 6.7, revealing a C-term reduction potential of a factor of up to 5 for large molecules (dextran) under unretained conditions. The decrease in κaris in a 5 µm channel (reduction of 80%) was larger than the decrease in a 3 µm channel (reduction of 44%).

Concepts and recent advances in microchip electrophoresis coupled to mass spectrometry: Technologies and applications.

The online coupling of microchip electrophoresis (ME) as a fast, highly efficient, and low-cost miniaturized separation technique to mass spectrometry (MS) as an information-rich and sensitive characterization technique results in ME-MS an attractive tool for various applications. In this paper, we review the basic concepts and latest advances in technology for ME coupled to MS during the period of 2016-2021, covering microchip materials, structures, fabrication techniques, and interfacing to electrospray ionization (ESI)-MS and matrix-assisted laser desorption/ionization-MS. Two critical issues in coupling ME and ESI-MS include the electrical connection used to define the electrophoretic field strength along the separation channel and the generation of the electrospray for MS detection, as well as, a miniaturized ESI-tip. The recent commercialization of ME-MS in zone electrophoresis and isoelectric focusing modes has led to the widespread application of these techniques in academia and industry. Here we summarize recent applications of ME-MS for the separation and detection of antibodies, proteins, peptides, carbohydrates, metabolites, and so on. Throughout the paper these applications are discussed in the context of benefits and limitations of ME-MS in comparison to alternative techniques.

Rail configuration for lateral particle transport across intact co-flows: effect of wall and rail geometry on liquid and particle transport.

Layer-by-Layer coating technology is of great importance for many applications of microparticles whereby exposure of the particles to various reagents is needed. Mutual contamination of the reagents during this process is a key challenge, and this undesired effect should be avoided. Here we introduce a device that provides subsequent exposure of particles to various liquids and minimizes mixing of the liquids at the same time. The key element of the device is a rail (groove) at the bottom of a microfluidic channel. The rail forms an angle (between 0 and 90 degrees) and thus enables passive transport of particles through the intact co-flows of the different fluids. To avoid the undesirable effect of reagent stream mixing, internal walls are introduced to separate the different flows. Various designs of the proposed device are considered, and their performance is experimentally analyzed.

Selecting active matter according to motility in an acoustofluidic setup: self-propelled particles and sperm cells.

Active systems - including sperm cells, living organisms like bacteria, fish, birds, or active soft matter systems like synthetic "microswimmers" - are characterized by motility, i.e., the ability to propel using their own "engine". Motility is the key feature that distinguishes active systems from passive or externally driven systems. In a large ensemble, motility of individual species can vary in a wide range. Selecting active species according to their motility represents an exciting and challenging problem. We propose a new method for selecting active species based on their motility using an acoustofluidic setup where highly motile species escape from the acoustic trap. This is demonstrated in simulations and in experiments with self-propelled Janus particles and human sperm. The immediate application of this method is selecting highly motile sperm for medically assisted reproduction (MAR). Due to the tunable acoustic trap, the proposed method is more flexible than the existing passive microfluidic methods. The proposed selection method based on motility can also be applied to other active systems that require selecting highly motile species or removing immotile species.

Performance of laterally elongated pillar array columns in capillary electrochromatography mode.

In the present study, cylindrical and laterally elongated pillar array columns were investigated for use in capillary electrochromatography. Minimal theoretical plate heights of H = 1.90 and 1.46 µm (in absence of sidewall effect) were obtained for coumarin C440 under unretained conditions for cylindrical and rectangular (laterally elongated, aspect ratio 4) pillar array columns, respectively. By comparing dispersion at the entire channel width to that at the central zone only, it appears that sidewall related dispersion significantly contributes to overall dispersion. A 40% reduction of the plate height was observed by taking into account only the central channel zone. A kinetic plot analysis was performed to evaluate the potential of the studied geometries by considering a maximum operating voltage of 20 kV as limiting parameter. It was demonstrated that rectangular radially elongated pillars produce a higher efficiency than cylindrical pillars and other microfabricated column structures for microchip capillary electrochromatography previously studied.

Anisotropic Exclusion Effect between Photocatalytic Ag/AgCl Janus Particles and Passive Beads in a Dense Colloidal Matrix.

Synthetic nano- and micromotors interact with each other and their surroundings in a complex manner. Here, we report on the anisotropy of active-passive particle interaction in a soft matter system containing an immobile yet photochemical Ag/AgCl-based Janus particle embedded in a dense matrix of passive beads in pure water. The asymmetry in the chemical gradient around the Janus particle, triggered upon visible light illumination, distorts the isotropy of the surrounding electric potential and results in the repulsion of adjacent passive beads to a certain distance away from the Janus particle. This exclusion effect is found to be anisotropic with larger distances to passive beads in front of the Ag/AgCl cap of the Janus particle. We provide insight into this phenomenon by performing the angular analysis of the radii of exclusion and tracking their time evolution at the level of a single bead. Our study provides a novel fundamental insight into the collective behavior of a complex mixture of active and passive particles and is relevant for various application scenarios, e.g., particle transport at micro- and nanoscale and local chemical sensing.

Achieving a Peak Capacity of 1800 Using an 8 m Long Pillar Array Column.

In the present study, the peak capacity potential of ultralong porous cylindrical pillar array columns is investigated. Coupling 4 columns of 2 m long allows for working near the minimal separation impedance of small molecules under retained conditions at a maximal pressure load of 250 bar. Minimal plate heights of H = 5.0 µm, H = 6.3 µm, and H = 7.7 µm were obtained for uracil (unretained), butyrophenone (k = 0.85), and valerophenone (k = 1.94), respectively, corresponding to a number of theoretical plates of N = 1.6 × 106, N = 1.2 × 106, and N = 1.0 × 106. The optimal linear velocities were 0.60 mm/s for a retained compound and 0.74 mm/s for an unretained compound. Based on a mixture of 9 compounds, the peak capacity nc was determined as a function of gradient time (tG). Peak capacities (tG-based) of 1103 and 1815 were obtained when applying 650 min and 2050 min gradients (tG/t0 = 4.5 and 14, respectively, with tG as the gradient time and t0 as the void time). These values are much higher than earlier reported peak capacity values for small molecules.

Study of peak capacities generated by a porous layered radially elongated pillar array column coupled to a nano-LC system.

The performance of a porous-layered radially elongated pillar (PLREP) array column in a commercial nano-LC system was examined by performing separation of alkylphenones and peptides. The mesoporous silica layer was prepared by sol-gel processing of a mixture of tetramethoxysilane and methyltrimethoxysilane on REPs filling a 16.5 cm long, 1 mm wide channel (three lanes of 5.5 cm long channels connected by turns). The minimum plate height of 1.4 µm for octanophenone (k = 2.21) observed in isocratic mode is 5 times smaller than the smallest off-column plate height previously reported for porous pillar array columns for a retained component. This advantage is related to the earlier introduced shape of the radially elongated pillar bed that outperforms the cylindrically shaped pillar bed in terms of the plate height. In gradient mode, maximum conditional peak capacities of 220 (for a mixture of thiourea and 7 alkylphenones, tG = 180 min) and 160 (for a cytochrome c digest, tG = 150 min) were obtained. These results indicate excellent potential for implementation of this sol-gel layer in pillar array column formats.