A numbering-up strategy of hydrodynamic microfluidic filters for continuous-flow high-throughput cell sorting

Ozawa, Ryoken; Iwadate, Hideki; Toyoda, Hajime; Yamada, Masao; Seki, Minoru

doi:10.1039/c9lc00053d

Cited by 25 publications

(18 citation statements)

References 51 publications

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“…In recent years microfluidic technologies have emerged as practical platforms to precisely manipulate micrometer-sized objects, mostly cells. For instance, various types of unique cell-sorting mechanisms have been devised utilizing specific flow profiles formed in microchannels. − In microfluidic cell sorters, cells/particles with different physical characteristics (e.g., size, shape, stiffness, and deformability) flow along different streamlines, because of the application of external forces, the generation of particle inertia, or the hydrodynamic effect caused by channel geometries. Such sorting mechanisms have been adopted to the medium exchange for cells in a continuous process.…”

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confidence: 99%

Laborless, Automated Microfluidic Tandem Cell Processor for Visualizing Intracellular Molecules of Mammalian Cells

Toyoda

Kimura

et al. 2019

Anal. Chem.

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Visualization and quantification of intracellular molecules of mammalian cells are crucial steps in clinical diagnosis, drug development, and basic biological research. However, conventional methods rely mostly on labor-intensive, centrifugation-based manual operations for exchanging the cell carrier medium and have limited reproducibility and recovery efficiency.Here we present a microfluidic cell processor that can perform four-step exchange of carrier medium, simply by introducing a cell suspension and fluid reagents into the device. The reaction time period for each reaction step, including fixation, membrane permeabilization, and staining, was tunable in the range of 2 to 15 min by adjusting the volume of the reaction tube connecting the neighboring exchanger modules. We double-stained the cell nucleus and cytoskeleton (F-actin) using the presented device with an overall reaction period of ∼30 min, achieving a high recovery ratio and high staining efficiency. Additionally, intracellular cytokine (IL-2) was visualized for T cells to demonstrate the feasibility of the device as a pretreatment system for downstream flow-cytometric analysis. The presented approach would facilitate the development of laborless, automated microfluidic systems that integrate cell processing and analysis operations and would pave a new path to high-throughput biological experiments.

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confidence: 99%

Laborless, Automated Microfluidic Tandem Cell Processor for Visualizing Intracellular Molecules of Mammalian Cells

Toyoda

Kimura

et al. 2019

Anal. Chem.

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“…This result clearly indicated that the particle inertia did not dominate the separation mechanism of the presented method. Because a relatively high separation throughput was demonstrated, we expect that a high-throughput processing would be possible using the presented concept by further optimizing the operating conditions and/or microchannel geometries, including microchannel parallelization [34].…”

Section: Resultsmentioning

confidence: 99%

Hydrodynamic Microparticle Separation Mechanism Using Three-Dimensional Flow Profiles in Dual-Depth and Asymmetric Lattice-Shaped Microchannel Networks

et al. 2019

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We herein propose a new hydrodynamic mechanism of particle separation using dual-depth, lattice-patterned asymmetric microchannel networks. This mechanism utilizes three-dimensional (3D) laminar flow profiles formed at intersections of lattice channels. Large particles, primarily flowing near the bottom surface, frequently enter the shallower channels (separation channels), whereas smaller particles flowing near the microchannel ceiling primarily flow along the deeper channels (main channels). Consequently, size-based continuous particle separation was achieved in the lateral direction in the lattice area. We confirmed that the depth of the main channel was a critical factor dominating the particle separation efficiencies, and the combination of 15-μm-deep separation channels and 40-μm-deep main channels demonstrated the good separation ability for 3–10-μm particles. We prepared several types of microchannels and successfully tuned the particle separation size. Furthermore, the input position of the particle suspension was controlled by adjusting the input flow rates and/or using a Y-shaped inlet connector that resulted in a significant improvement in the separation precision. The presented concept is a good example of a new type of microfluidic particle separation mechanism using 3D flows and may potentially be applicable to the sorting of various types of micrometer-sized objects, including living cells and synthetic microparticles.

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“…It is well known that cell manipulation and sorting are essential steps in cell biology research and in many diagnostics and therapeutic methods and have been accomplished in microfluidic systems using mostly biochemical labels. [ 254,267 ] Further, there have been also interest in taking advantages of other physicochemical properties of cells such as size, electrical polarity, and hydrodynamic properties. An early application of cell‐based microfluidics included combining 2D dielectrophoretic forces with field‐flow‐fractionation in a microfluidic device for separating cells according to their dielectric properties.…”

Section: Physically Active Bioreactors—main Typesmentioning

confidence: 99%

Physically Active Bioreactors for Tissue Engineering Applications

et al. 2020

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Tissue engineering (TE) is a dynamic and growing scientific field that merges the knowledge of different areas such as biology, physics, medicine, and engineering. [1] The TE discipline was first coined at a National Science Foundation sponsored meeting in 1987, thus originating a field that employs life sciences and engineering with the purpose of developing biologic or synthetic supports to restore, keep, or enhance tissue functions or damaged organs. [2] The classical TE approach has been mainly focusing on associating cells with a supporting matrix, also called biomaterials or scaffolds, that essentially acts as a passive template for tissue formation in vitro by allowing cells to adhere, migrate, differentiate, and produce tissue. Usually, the cells are seeded onto the scaffolds and, occasionally, growth factors are also added. The combination of cells, growth factors, and scaffold is often referred as the TE triad. [3] Nevertheless, novel approaches in TE that include the application of a biophysical stimuli to the scaffolds through the use of a bioreactor are emerging. [4] Tissue engineering (TE) is a strongly expanding research area. TE approaches require biocompatible scaffolds, cells, and different applied stimuli, which altogether mimic the natural tissue microenvironment. Also, the extracellular matrix serves as a structural base for cells and as a source of growth factors and biophysical cues. The 3D characteristics of the microenvironment is one of the most recognized key factors for obtaining specific cell responses in vivo, being the physical cues increasingly investigated. Supporting those advances is the progress of smart and multifunctional materials design, whose properties improve the cell behavior control through the possibility of providing specific chemical and physical stimuli to the cellular environment. In this sense, a varying set of bioreactors that properly stimulate those materials and cells in vitro, creating an appropriate biomimetic microenvironment, is developed to obtain active bioreactors. This review provides a comprehensive overview on the important microenvironments of different cells and tissues, the smart materials type used for providing such microenvironments and the specific bioreactor technologies that allow subjecting the cells/ tissues to the required biomimetic biochemical and biophysical cues. Further, it is shown that microfluidic bioreactors represent a growing and interesting field that hold great promise for achieving suitable TE strategies.

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A numbering-up strategy of hydrodynamic microfluidic filters for continuous-flow high-throughput cell sorting

Abstract: A numbering-up strategy of hydrodynamic filters was presented to dramatically increase the throughput of cell/particle sorting up to ∼15 mL min−1.

Cited by 25 publications

References 51 publications

Laborless, Automated Microfluidic Tandem Cell Processor for Visualizing Intracellular Molecules of Mammalian Cells

Laborless, Automated Microfluidic Tandem Cell Processor for Visualizing Intracellular Molecules of Mammalian Cells

Hydrodynamic Microparticle Separation Mechanism Using Three-Dimensional Flow Profiles in Dual-Depth and Asymmetric Lattice-Shaped Microchannel Networks

Physically Active Bioreactors for Tissue Engineering Applications

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