Cellular point-of-care diagnostics using an inexpensive layer-stack microfluidic device

Lucas, Kilean; Oh, Juhyun; Hoelzl, Jan; Weissleder, Ralph

doi:10.1039/d2lc00162d

Cited by 8 publications

(10 citation statements)

References 28 publications

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“…We developed a specially constructed flow chamber to facilitate rapid cycling and preservation of reagents, with no need for harsh conditions (e.g., paraformaldehyde to fix and adhere EVs or hydrogen peroxide to bleach fluorochromes), thus allowing unequivocal analyses of a single EV. This chamber uses an acrylic pressure-sensitive adhesive to bond a treated coverglass to a microscope slide 28 . The shape of the pressure-sensitive adhesive (width 4 mm, length 12 mm, height 50 µm) allows pump-free flushing with flow rates of ∼ 1 µL/s within the ∼2 µL channel.…”

Section: Resultsmentioning

confidence: 99%

“…The coverglass surface was maximally activated for subsequent attachment of EV. Microfluidic devices were prepared following a recent description of an inexpensive device for cell staining 28 . Channels were cut from 50 µm thick double-sided 3M VHB adhesive tape (F9460PC) using a Silhouette Cameo 4 craft cutting tool (Silhouette USA).…”

Section: Device Fabrication and Assemblymentioning

confidence: 99%

“…The footprint of the adhesive ( ∼22 mm x 24 mm ) accommodates multiple channels per single base slide. The shape of the pressure-sensitive adhesive cut-out (width 4 mm length 12 mm height 50 µm) allows pump-free flushing driven via Laplacian pressure 28 . Using this design, flow rates of ∼1 µL/s could be achieved within the 2 µL channel.…”

Section: Supplemental Informationmentioning

confidence: 99%

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Multiplexed analysis of a single EV (MASEV) reveals unique biomarker composition with diagnostic impact

Spitzberg

Ferguson

Yang

et al. 2022

Preprint

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Exosomes and extracellular vesicles (EV) are increasingly being explored as circulating biomarkers, but their heterogenous composition will likely mandate the development of single EV technologies. Highly multiplexed analyses of single EVs have been challenging to implement beyond a few colors during spectral sensing. We use a multiplexed analysis of the single EV technique (MASEV) to interrogate thousands of individual EVs during 5 cycles of multi-channel fluorescence staining for 15 EV biomarkers. Contrary to the common belief, we show that i) several “ubiquitous” markers are much less common than believed; ii) that multiple biomarkers concur in single vesicles but only in small fractions, iii) that affinity purification can lead to loss of rare EV subtypes, and iv) that deep profiling allows detailed analysis of EV potentially improving the diagnostic content. These findings establish the potential of MASEV for uncovering fundamental EV biology and heterogeneity and increasing diagnostic specificity. Abstract Figure Graphical abstract Multiplexed analysis of single EV (MASEV) allows robust protein profiling at the single EV level, a prerequisite for early cancer detection or organ of origin determination.

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Section: Resultsmentioning

confidence: 99%

Section: Device Fabrication and Assemblymentioning

confidence: 99%

Section: Supplemental Informationmentioning

confidence: 99%

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Multiplexed analysis of a single EV (MASEV) reveals unique biomarker composition with diagnostic impact

Spitzberg

Ferguson

Yang

et al. 2022

Preprint

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“…Lately, double-sided adhesives (DSA) based micro uidic fabrication strategies are being developed for multilayered microphysiological systems and cellular diagnostics (Hosic et al 2021;Lucas et al 2022).…”

Section: Introductionmentioning

confidence: 99%

High-throughput microbead assay system with a portable, cost-effective Wi-Fi imaging module, and disposable multi-layered microfluidic cartridges for virus and microparticle detection, and tracking

Castro

Sommerhage²,

Khanna

et al. 2022

Preprint

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In recent years biomedical scientific community has been working towards the development of high-throughput devices that allow a reliable, rapid and parallel detection of several strains of virus or microparticles simultaneously. One of the complexities of this problem lies on the rapid prototyping of new devices and wireless rapid detection of small particles and virus alike. By reducing the complexity of microfluidics microfabrication and using economic materials along with makerspace tools (Avra Kundu, Ausaf, and Rajaraman 2018) it is possible to provide an affordable solution to both the problems of high-throughput devices and detection technologies. We present the development of a wireless, standalone device and disposable microfluidics chips that rapidly generate parallel readouts for selected, possible virus variants from a nasal or saliva sample, based on motorized and non-motorized microbeads detection, and imaging processing of the motion tracks of these beads in micrometers. Microbeads and SARS-CoV-2 COVID-19 Delta variant were tested as proof-of-concept for testing the microfluidic cartridges and wireless imaging module. The Microbead Assay (MA) system kit consists of a WiFi readout module, a microfluidic chip, and a sample collection/processing sub-system. Here, we focus on the fabrication and characterization of the microfluidic chip to multiplex various micrometer-sized beads for economic, disposable, and simultaneous detection of up to six different viruses, microparticles or variants in a single test, and data collection using a commercially available, WiFi-capable, and camera integrated device (Fig. 1).

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“…There is minimal bleed through of signal into consecutive cycles, providing an effective method for multiplexed imaging and cancer diagnosis. Reproduced with permission [7]. Copyright 2022, Royal Society of Chemistry.…”

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confidence: 99%

Cycling, Fasting, Fishing, and Other Things: The Emerging World of Biocompatible Chemistries

Raman

2022

Advanced Biology

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Cellular point-of-care diagnostics using an inexpensive layer-stack microfluidic device

Abstract: Passively pumped layer stack microfluidics allow for simple, inexpensive cyclic imaging with reduced reagent usage.

Cited by 8 publications

References 28 publications

Multiplexed analysis of a single EV (MASEV) reveals unique biomarker composition with diagnostic impact

Multiplexed analysis of a single EV (MASEV) reveals unique biomarker composition with diagnostic impact

High-throughput microbead assay system with a portable, cost-effective Wi-Fi imaging module, and disposable multi-layered microfluidic cartridges for virus and microparticle detection, and tracking

Cycling, Fasting, Fishing, and Other Things: The Emerging World of Biocompatible Chemistries

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