Under-oil open microfluidic systems for rapid phenotypic antimicrobial susceptibility testing

Li, Chao; McCrone, Sue; Warrick, Jay W.; Andes, David R.; Hite, Zachary; Volk, Cecilia; Rose, Warren E.; Beebe, David J.

doi:10.1039/d3lc00066d

Cited by 9 publications

(14 citation statements)

References 62 publications

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“…1c), high flow rate range (comparable to blood flow in circulatory systems), on-demand reversible open-fluid valves 30 , and new functions [e.g., autonomously regulated oxygen microenvironment (AROM) 32 ]. These advances in ELR-empowered UOMS together not only make open microfluidics comparable to the closed-chamber/channel microfluidics but also introduces new avenues to cellular 27,28,[30][31][32][33] and molecular 38 assays in biology and biomedicine.…”

Section: Resultsmentioning

confidence: 99%

Microliter whole blood neutrophil assay preserving physiological lifespan and functional heterogeneity

Li,

Hendrikse,

Mai

et al. 2023

Preprint

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For in vitro neutrophil functional assays, isolated neutrophils are typically used as the standard input. However, isolated neutrophils show a varying and limited lifespan as short as only a few hours ex vivo, significantly shorter than the lifespan of neutrophils in vivo up to 3-5 days. The limited assay time window leads to a significant loss of donor-specific information, which is attributed to non-specific activation (i.e., baseline cell death without operator-defined stimuli) of neutrophils after being pulled out of whole blood. In addition, due to neutrophils' inherently high sensitivity, removing neutrophils from whole blood generates operator-derived inconsistencies in the information extracted from the assays. Here we present a method - named Micro-Blood - that supports long-term (days) multiple phenotypic readouts of neutrophil function (including cell/nucleus morphology, motility, recruitment, and pathogen control) using a microliter of unprocessed whole blood. In autologous whole blood, neutrophils show sustained migration and limited non-specific activation kinetics [<0.1% non-specific activation >3 days]. By contrast, isolated neutrophils show altered activation kinetics [10-70% non-specific activation in 3 h and diminished (>90%) recruitment]. Using unprocessed whole blood, Micro-Blood captures a distinct neutrophil functional heterogeneity between healthy donors and cancer patients against microbial stimuli through days-long assays, showing the potential for in vitro immune assays with improved extraction of donor-specific information and assay consistency over time.

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Section: Resultsmentioning

confidence: 99%

Microliter whole blood neutrophil assay preserving physiological lifespan and functional heterogeneity

Li,

Hendrikse,

Mai

et al. 2023

Preprint

Self Cite

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“…These advances not only make ELR-empowered UOMS comparable to closed-chamber/channel microfluidics in functionality but also enables new applications in biology and biomedicine. [26,27] Enabled by Double-ELR, a selected pattern (e.g., two circular spots connected by a straight channel adopted in this study) can be readily prepared on a surface [e.g., glass, polystyrene (PS)] by under-oil sweep distribution (Figure 1a and 1b, Experimental Section). A hanging drop of the target media (e.g., collagen solution) at the end of a pipet tip is dragged across the Double-ELR chemically patterned surface (Figure 1c).…”

Section: Top Down -Double-elr Open Microfluidic Surface Patterningmentioning

confidence: 99%

Combining Top-Down and Bottom-Up: An Open Microfluidic Microtumor Model for Investigating Tumor Cell-ECM Interaction and Anti-Metastasis

Li,

Argall-Knapp

et al. 2024

Preprint

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Using a combined top-down (i.e., operator-directed) and bottom-up (i.e., cell-directed) strategy, we present an Under-oil Open Microfluidic System (UOMS)-based microtumor model for investigating tumor cell migration and anti-metastasis drug test. Compared to the mainstream closed microfluidics-based microtumor models, the UOMS microtumor model features: i) micrometer-scale lateral resolution of surface patterning with open microfluidic design for flexible spatiotemporal sample manipulation (i.e., top-down); ii) self-organized extracellular matrix (ECM) structures and tumor cell-ECM spontaneous remodeling (i.e., bottom-up); and iii) free physical access to the samples on a device with minimized system disturbance. The UOMS microtumor model is used to test an anti-metastasis drug (incyclinide, aka CMT-3) with a triple negative breast cancer cell line (MDA-MB-231). Thein vitroresults show a suppression of tumor cell migration and ECM remodeling echoing with thein vivomice metastasis results.

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“…Therefore, expanding the range of droplet size parameter generated by this method to include typical size ratios (0.5 < 𝜒 < 3) is necessary to enable its application in various field, such as digital microfluidics [44][45][46][47] and droplet cell culture. [48] Here, we performed systematic observation and expansion of the working regime of this hydrodynamic dispensing technology to fully exploit its applicability for droplet cell culture in digital microfluidics. We observed contact angle changes during aspiration and quantitatively explained the difference in the large (𝜒 > 0.5) and the small (𝜒 < 0.5) droplet pinch-off.…”

Section: Doi: 101002/smll202309062mentioning

confidence: 99%

“…Therefore, expanding the range of droplet size parameter generated by this method to include typical size ratios (0.5 < χ < 3) is necessary to enable its application in various field, such as digital microfluidics [ 44–47 ] and droplet cell culture. [ 48 ]…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Separating, Splitting, Collecting, and Dispensing by Droplet Pinch‐Off for Droplet Cell Culture

Bae,

Lee,

2023

Small

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Simultaneous separating, splitting, collecting, and dispensing a cell suspension droplet has been demonstrated by aspiration and subsequent droplet pinch‐off for use in microfluidic droplet cell culture systems. This method is applied to cell manipulations including aliquots and concentrations of microalgal and mammalian cell suspensions. Especially, medium exchange of spheroid droplets is successfully demonstrated by collecting more than 99% of all culture medium without damaging the spheroids, demonstrating its potential for a 3D cell culture system. Through dimensional analysis and systematic parametric studies, it is found that initial mother droplet size together with aspiration flow rate determines three droplet pinch‐off regimes. By observing contact angle changes during aspiration, the difference in the large and the small droplet pinch‐off can be quantitatively explained using force balance. It is found that the capillary number plays a significant role in droplet pinch‐off, but the Bond number and the Ohnesorge number have minor effects. Since the dispensed droplet size is mainly determined by the capillary number, the dispensed droplet size can be controlled simply by adjusting the aspiration flow rate. It is hoped that this method can contribute to various fields using droplets, such as droplet cell culture and digital microfluidics, beyond the generation of small droplets.

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Under-oil open microfluidic systems for rapid phenotypic antimicrobial susceptibility testing

Abstract: Antimicrobial susceptibility testing (AST) remains the cornerstone of effective antimicrobial selection and optimization in patients. Despite recent advances in rapid pathogen identification and resistance marker detection with molecular diagnostics (e.g.,...

Cited by 9 publications

References 62 publications

Microliter whole blood neutrophil assay preserving physiological lifespan and functional heterogeneity

Microliter whole blood neutrophil assay preserving physiological lifespan and functional heterogeneity

Combining Top-Down and Bottom-Up: An Open Microfluidic Microtumor Model for Investigating Tumor Cell-ECM Interaction and Anti-Metastasis

Simultaneous Separating, Splitting, Collecting, and Dispensing by Droplet Pinch‐Off for Droplet Cell Culture

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