Label-Free Measurement of T-Cell Activation by Microfluidic Acoustophoresis

Dabbi, Jayanth M.; Shi, Yulei; Mërtiri, Alket; Christianson, Rebecca J.; Fiering, Jason

doi:10.1109/transducers50396.2021.9495614

Cited by 3 publications

(1 citation statement)

References 5 publications

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“…As label-free systems allow cells to remain in their natural state and are better compatible with downstream separation or analysis methods, negative magnetophoresis has also become a topic of interest for use in integrated microfluidic systems. [58,59] One example demonstrated by Liu et al separated circulating tumor cells (CTCs), the cells of interest, from white blood cells (WBCs) via negative magnetophoresis independent of their physical size and surface antigens. [60] In this system, the cell mixture was suspended in a water-based ferrofluid whose magnetization could be tuned by changing the volume fraction of magnetic materials in the ferrofluid.…”

Section: Inertial and Hydrodynamic Focusingmentioning

confidence: 99%

Integrated Microfluidics for Single‐Cell Separation and On‐Chip Analysis: Novel Applications and Recent Advances

Kutluk,

Viefhues,

Constantinou

2024

Small Science

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From deciphering infection and disease mechanisms to identifying novel biomarkers and personalizing treatments, the characteristics of individual cells can provide significant insights into a variety of biological processes and facilitate decision‐making in biomedical environments. Conventional single‐cell analysis methods are limited in terms of cost, contamination risks, sample volumes, analysis times, throughput, sensitivity, and selectivity. Although microfluidic approaches have been suggested as a low‐cost, information‐rich, and high‐throughput alternative to conventional single‐cell isolation and analysis methods, limitations such as necessary off‐chip sample pre‐ and post‐processing as well as systems designed for individual workflows have restricted their applications. In this review, a comprehensive overview of recent advances in integrated microfluidics for single‐cell isolation and on‐chip analysis in three prominent application domains are provided: investigation of somatic cells (particularly cancer and immune cells), stem cells, and microorganisms. Also, the use of conventional cell separation methods (e.g., dielectrophoresis) in unconventional or novel ways, which can advance the integration of multiple workflows in microfluidic systems, is discussed. Finally, a critical discussion related to current limitations of integrated microfluidic single‐cell workflows and how they could be overcome is provided.

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Section: Inertial and Hydrodynamic Focusingmentioning

confidence: 99%

Integrated Microfluidics for Single‐Cell Separation and On‐Chip Analysis: Novel Applications and Recent Advances

Kutluk,

Viefhues,

Constantinou

2024

Small Science

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Microparticles with tunable, cell-like properties for quantitative acoustic mechanophenotyping

2023

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Mechanical properties of biological cells have been shown to correlate with their biomolecular state and function, and therefore methods to measure these properties at scale are of interest. Emerging microfluidic technologies can measure the mechanical properties of cells at rates over 20,000 cells/s, which is more than four orders of magnitude faster than conventional instrumentation. However, precise and repeatable means to calibrate and test these new tools remain lacking, since cells themselves are by nature variable. Commonly, microfluidic tools use rigid polymer microspheres for calibration because they are widely available in cell-similar sizes, but conventional microspheres do not fully capture the physiological range of other mechanical properties that are equally important to device function (e.g., elastic modulus and density). Here, we present for the first time development of monodisperse polyacrylamide microparticles with both tunable elasticity and tunable density. Using these size, elasticity, and density tunable particles, we characterized a custom acoustic microfluidic device that makes single-cell measurements of mechanical properties. We then applied the approach to measure the distribution of the acoustic properties within samples of human leukocytes and showed that the system successfully discriminates lymphocytes from other leukocytes. This initial demonstration shows how the tunable microparticles with properties within the physiologically relevant range can be used in conjunction with microfluidic devices for efficient high-throughput measurements of mechanical properties at single-cell resolution.

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Rosette-induced separation of T cells by acoustophoresis

et al. 2022

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Breakthrough cell therapies for the treatment of cancers require the separation of specific cells, such as T cells, from the patient's blood. Current cell therapy processes rely on magnetic separation, which adds clinical risk and requires elevated manufacturing controls due to the added foreign material that constitutes the magnetic beads. Acoustophoresis, a method that uses ultrasound for cell separation, has demonstrated label-free enrichment of T cells from blood, but residual other lymphocytes limit the ultimate purity of the output T cell product. Here, to increase the specificity of acoustophoresis, we use affinity reagents to conjugate red blood cells with undesired white blood cells, resulting in a cell–cell complex (rosette) of increased acoustic mobility. We achieve up to 99% purity of T cells from blood products, comparable to current standards of magnetic separation, yet without the addition of separation particles.

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Label-Free Measurement of T-Cell Activation by Microfluidic Acoustophoresis

Cited by 3 publications

References 5 publications

Integrated Microfluidics for Single‐Cell Separation and On‐Chip Analysis: Novel Applications and Recent Advances

Integrated Microfluidics for Single‐Cell Separation and On‐Chip Analysis: Novel Applications and Recent Advances

Microparticles with tunable, cell-like properties for quantitative acoustic mechanophenotyping

Rosette-induced separation of T cells by acoustophoresis

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