Microfluidic Cell Analysis Research Articles

High-efficiency extraction of target particles in viscoelastic contraction-expansion microchannels.

The efficient and precise extraction of target particles is a crucial prerequisite for achieving accurate detection and analysis in microfluidic cell analysis. In this study, a symmetrical contraction-expansion microchannel with sheath flow was designed, aiming to extract target larger particles from particles of different sizes within the channel. This paper conducted numerical simulations to investigate the three-dimensional migration mechanisms of particles and performed experimental studies to examine the separation performance of particles with different sizes under varying flow rate ratios and different numbers of contraction-expansion structures. The experimental results indicate that at moderate sample flow rates and higher flow rate ratios, microchannels with fewer contraction-expansion structures are likely to achieve better performance in extracting target particles compared to microchannels with a greater number of these structures. Our work advances the application of viscoelastic contraction-expansion microchannels in particle separation. This device is easy to set up in parallel and significantly enhances throughput, providing an accurate and efficient solution for future particle separation applications.

Bridging the academic–industrial gap: application of an oxygen and pH sensor-integrated lab-on-a-chip in nanotoxicology

Translation of advanced cell-based assays exhibiting a higher degree of automation, miniaturization, and integration of complementary sensing functions is mainly limited by the development of industrial-relevant prototypes that can be readily produced in larger volumes. Despite the increasing number of academic publications in recent years, the manufacturability of these microfluidic cell cultures systems is largely ignored, thus severely restricting their implementation in routine toxicological applications. We have developed a dual-sensor integrated microfluidic cell analysis platform using industrial specifications, materials, and fabrication methods to conduct risk assessment studies of engineered nanoparticles to overcome this academic-industrial gap. Non-invasive and time-resolved monitoring of cellular oxygen uptake and metabolic activity (pH) in the absence and presence of nanoparticle exposure is accomplished by integrating optical sensor spots into a cyclic olefin copolymer (COC)-based microfluidic platform. Results of our nanotoxicological study, including two physiological cell barriers that are essential in the protection from exogenous factors, the intestine (Caco-2) and the vasculature (HUVECs) showed that the assessment of the cells' total energy metabolism is ideally suited to rapidly detect cytotoxicities. Additional viability assay verification using state-of-the-art dye exclusion assays for nanotoxicology demonstrated the similarity and comparability of our results, thus highlighting the benefits of employing a compact and cost-efficient microfluidic dual-sensor platform as a pre-screening tool in nanomaterial risk assessment and as a rapid quality control measure in medium to high-throughput settings.

Advances in numerical approaches for microfluidic cell analysis platforms

Microfluidics has demonstrated enormous potential through its role in recent advances in biological sciences. However, designing a new and customized microfluidic platform, gaining a better understanding of its function and the underlying physics still pose significant technical challenges. On the one hand, experimental approaches have been commonly used for the development of microfluidic devices since they are accurate and evidence-based methods. However, these approaches are expensive and laborious. Numerical approaches, on the other hand, are now recognized as a reliable complementary method to reducing cost, time, and effort and being relatively accurate. This paper systematically reviews the capability of numerical approaches in developing efficient microfluidic technologies for cell analysis. Moreover, this paper provides an initial insight for researchers who are interested in establishing numerical approaches for microfluidic cell analysis platforms.

Shape design for stabilizing microparticles in inertial microfluidic flows

Design of microparticles which stabilize at the centerline of a channel flow when part of a dilute suspension is examined numerically for moderate Reynolds numbers ($10 \le Re \le 80$). Stability metrics for particles with arbitrary shapes are formulated based on linear-stability theory. Particle shape is parametrized by a compact, Non-Uniform Rational B-Spline (NURBS)-based representation. Shape-design is posed as an optimization problem and solved using adaptive Bayesian optimization. We focus on designing particles for maximal stability at the channel-centerline robust to perturbations. Our results indicate that centerline-focusing particles are families of characteristic "fish"/"bottle"/"dumbbell"-like shapes, exhibiting fore-aft asymmetry. A parametric exploration is then performed to identify stable particle-designs at different k's (particle chord-to-channel width ratio) and Re's ($0.1 \le k \le 0.4, 10 \le Re \le 80$). Particles at high-k's and Re's are highly stabilized when compared to those at low-k's and Re's. A comparison of the modified dumbbell designs from the current framework also shows better performance to perturbations in Fluid-Structure Interaction (FSI) when compared to the rod-disk model reported previously (Uspal & Doyle 2014) for low-Re Hele-Shaw flow. We identify basins of attraction around the centerline, which span larger release-angle-ranges and lateral locations (tending to the channel width) for narrower channels, which effectively standardizes the notion of global focusing in such configurations using the current stability-paradigm. The present framework is illustrated for 2D cases and is potentially generalizable to stability in 3D flow-fields. The current formulation is also agnostic to Re and particle/channel geometry which indicates substantial potential for integration with imaging flow-cytometry tools and microfluidic biosensing-assays.

Development of a Multifunctional Nanobiointerface Based on Self-Assembled Fusion-Protein rSbpA/ZZ for Blood Cell Enrichment and Phenotyping.

We present a multifunctional nanobiointerface for blood cell capture and phenotyping applications that features both excellent antifouling properties and high antibody activity. Multifunctionality is accomplished by modifying polymeric materials using self-assembled S-layer fusion-protein rSbpA/ZZ to immobilize high density antibodies at the two protein A binding sites of the rSbpA/ZZ nanolattice structure. Controlled orientation and alignment of the antibodies reduced antibody consumption 100-fold and increased cell capture efficiency 4-fold over standard methodologies. Cell analysis in complex samples was made possible by the remarkable antifouling properties of the rSbpA domain, while at the same time reducing unspecific binding and forgoing tedious blocking procedures. An automated microfluidic in situ cell analysis platform for isolation and phenotyping of primary peripheral blood mononuclear cells was developed as practical application. Results obtained using our automated microfluidic cell analysis platform showed that the multifunctional nanobiointerface can discriminate among T helper and cytotoxic T cells, and thymocytes. Additionally, on-chip cell capture under flow conditions using a high affinity CD 3 selective nanobiointerface preferentially isolated cells with strong surface marker expression. This means that our dynamic microfluidic cell purification method allows the enrichment of 773 CD 8 positive cytotoxic T cells out of a total blood cell population of 7728 PBMCs, which is an increase in cell enrichment of 8-fold with a purity of 85%.

Three-dimensional (3D) hydrodynamic focusing for continuous sampling and analysis of adherent cells

A simple three-dimensional (3D) hydrodynamic focusing microfluidic device integrated with continuous sampling, rapid dynamic lysis, capillary electrophoretic (CE) separation and detection of intracellular content is presented. One of the major difficulties in microfluidic cell analysis for adherent cells is that the cells are prone to attaching to the channel surface. To solve this problem, a cross microfluidic chip with three sheath-flow channels located on both sides of and below the sampling channel was developed. With the three sheath flows around the sample solution-containing cells, the formed soft fluid wall prevents the cells from adhering to the channel surface. Labeled cells were 3D hydrodynamically focused by the sheath-flow streams and smoothly introduced into the cross-section one by one. The introduction of sheath-flow streams not only ensured single-cell sampling but avoided blockage of the sampling channel by adherent cells as well. The maximum rate for introduction of individual cells into the separation channel was about 151 cells min(-1). With electric field applied on the separation channel, the aligned cells were driven into the separation channel and rapidly lysed within 400 ms at the entry of the channel by sodium dodecylsulfate (SDS) added in the sheath-flow solution. The microfluidic system was evaluated by analysis of reduced glutathione (GSH) and reactive oxygen species (ROS) in single HepG2 cells. The average analysis throughput of ROS and GSH in single cells was 16-18 cells min(-1).