Streamlined digital bioassays with a 3D printed sample changer

Menezes, Roberta; Dramé-Maigné, Adèle; Taly, Valérie; Rondelez, Yannick; Gines, Guillaume

doi:10.1039/c9an01744e

Cited by 13 publications

(9 citation statements)

References 69 publications

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“…Although a moving interface and syringe pump are widely used to convey samples and reagents, ,, application for rare cells is hampered by cell loss due to sedimentation and mechanical damage during flow. To address this issue, we designed a TFI chamber (Figure B-⑤) to directly connect to the chip inlet.…”

Section: Resultsmentioning

confidence: 99%

“…External forces, including sound waves, laser irradiation, and magnetic fields, have been employed to facilitate mixing, − but the accompanying instrumentation requirements increase the difficulty of integration . Pump-activated micromixers have also been used, but they rely on accurate coordination of pneumatic valves and complicated chip fabrication. − In addition, extensive investigation of the heterogeneity of CTC population requires isolation of a single CTC distribution with low background interferences, minimal cell damage, and accurate manipulation. − In this circumstance, positive pressurization is preferable for driving the on-chip immunoassay because of its precise control, minimal tubing retention, and low cross-contamination while providing a stable force to protect cells from shear forces and avoid over-pressuring. , However, pressurization in microfluidic devices leads to difficulties in precise flow regulation and bubble removal. Longwell and Fordyce creatively allowed bubble escape through a bypass in their microIO system and obtained minimal cross-contamination .…”

Section: Introductionmentioning

confidence: 99%

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A Fully Automated and Integrated Microfluidic System for Efficient CTC Detection and Its Application in Hepatocellular Carcinoma Screening and Prognosis

Wang

et al. 2021

ACS Appl. Mater. Interfaces

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Analysis of circulating tumor cells (CTCs) is regarded as a useful diagnostic index to monitor tumor development and guide precision medicine. Although the immunoassay is a common strategy for CTC identification and heterogeneity characterization, it is challenged by poor reaction efficiency and laborious manipulations in microdevices, which hinder the sensitivity, throughput, simplification, and applicability. To meet the need for rapid, sensitive, and simple CTC analysis, we developed an efficient CTC detection system by integrating a 3D printed off-chip multisource reagent platform, a bubble retainer, and a single CTC capture microchip, which can achieve CTC capture and identification within 90 min. Compared with traditional CTC identification methods, this system decreases immunostaining time and antibody consumption by 90% and performs the on-chip immunoassay in a fully automated manner. Using this system, CTCs from the peripheral blood of 19 patients with various cancers were captured, detected, and compared with clinical data. The system shows great potential for early screening, real-time monitoring, and precision medicine for hepatocellular carcinoma (HCC). With the advantages of automation, stability, economy, and user-friendly operation, the proposed system is promising for clinical scenarios.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A Fully Automated and Integrated Microfluidic System for Efficient CTC Detection and Its Application in Hepatocellular Carcinoma Screening and Prognosis

Wang

et al. 2021

ACS Appl. Mater. Interfaces

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“…As a proof of principle, we prepared four samples spiked with 0 or 3 pM of microRNA mir39 (α switch) and let7a (β switch). Each sample is barcoded with a combination of two fluorescent dextrans and serially emulsified using a homemade sample changer . After incubation, the droplets are imaged by fluorescence microscopy (see the Material and Methods section, Figure b–d).…”

Section: Resultsmentioning

confidence: 99%

“…Each sample is barcoded with a combination of two fluorescent dextrans and serially emulsified using a homemade sample changer. 35 After incubation, the droplets are imaged by fluorescence microscopy (see the Material and Methods section, Figure 6b−d). While we recorded a few false-positive events, we achieved accurate quantification of the two microRNAs within 12 ± 6% errors (which could be partially explained by concentration uncertainties from the serial dilution of the targets).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Multiplex Digital MicroRNA Detection Using Cross-Inhibitory DNA Circuits

Rondelez

Gines

2020

ACS Sens.

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Ubiquitous post-transcriptional regulators in eukaryotes, microRNAs are currently emerging as promising biomarkers of physiological and pathological processes. Multiplex and digital detection of microRNAs represents a major challenge toward the use of microRNA signatures in clinical settings. The classical reverse transcription polymerase chain reaction quantification approach has important limitations because of the need for thermocycling and a reverse transcription step. Simpler, isothermal alternatives have been proposed, yet none could be adapted in both a digital and multiplex format. This is either because of a lack of sensitivity that forbids single molecule detection or molecular cross-talk reactions that are responsible for nonspecific amplification. Building on an ultrasensitive isothermal amplification mechanism, we present a strategy to suppress cross-talk reactions, allowing for robust isothermal and multiplex detection of microRNA targets. Our approach relies on target-specific DNA circuits interconnected with DNA-encoded inhibitors that repress nonspecific signal amplification. We demonstrate the one-step, isothermal, digital, and simultaneous quantification of various pairs of important microRNA targets.

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“…To this end, autosamplers are emerging solutions with promises of scalability and automation 28 – 36 . Autosamplers typically store multiple samples and leverage various sample switching mechanisms (e.g., motorized stage 28 , 35 , 36 , rotational stage 29 , high-performance liquid chromatography 30 , injection loops 31 ) to load multiple samples through a single tubing to the droplet microfluidic device. The second approach replaces tubings with more convenient reservoirs and tanks.…”

Section: Introductionmentioning

confidence: 99%

Facile and scalable tubing-free sample loading for droplet microfluidics

Shao

Hsieh

Zhang

et al. 2022

Sci Rep

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Droplet microfluidics has in recent years found a wide range of analytical and bioanalytical applications. In droplet microfluidics, the samples that are discretized into droplets within the devices are predominantly loaded through tubings, but such tubing-based sample loading has drawbacks such as limited scalability for processing many samples, difficulty for automation, and sample wastage. While advances in autosamplers have alleviated some of these drawbacks, sample loading that can instead obviate tubings offers a potentially promising alternative but has been underexplored. To fill the gap, we introduce herein a droplet device that features a new Tubing Eliminated Sample Loading Interface (TESLI). TESLI integrates a network of programmable pneumatic microvalves that regulate vacuum and pressure sources so that successive sub-microliter samples can be directly spotted onto the open-to-atmosphere TESLI inlet, vacuumed into the device, and pressurized into nanoliter droplets within the device with minimal wastage. The same vacuum and pressure regulation also endows TESLI with cleaning and sample switching capabilities, thus enabling scalable processing of many samples in succession. Moreover, we implement a pair of TESLIs in our device to parallelize and alternate their operation as means to minimizing idle time. For demonstration, we use our device to successively process 44 samples into droplets—a number that can further scale. Our results demonstrate the feasibility of tubing-free sample loading and a promising approach for advancing droplet microfluidics.

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Streamlined digital bioassays with a 3D printed sample changer

Abstract: Off-chip sample changer device increase the sample throughput of droplet digital bioassays.

Cited by 13 publications

References 69 publications

A Fully Automated and Integrated Microfluidic System for Efficient CTC Detection and Its Application in Hepatocellular Carcinoma Screening and Prognosis

A Fully Automated and Integrated Microfluidic System for Efficient CTC Detection and Its Application in Hepatocellular Carcinoma Screening and Prognosis

Multiplex Digital MicroRNA Detection Using Cross-Inhibitory DNA Circuits

Facile and scalable tubing-free sample loading for droplet microfluidics

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