A microfluidic alternating-pull–push active digitization method for sample-loss-free digital PCR

Zhou, Xin; Ravichandran, Gopi Chandran; Zhang, Peng; Yang, Yang; Zeng, Yong

doi:10.1039/c9lc00932a

Cited by 32 publications

(12 citation statements)

References 52 publications

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“…Like with ELISA, microwell arrays for nucleic acid detection can be loaded using a range of techniques. Flow cell loading [ 91 , 92 ], vacuum-assisted loading and oil-driven digitization [ 93 , 94 , 95 , 96 ], pressure loading [ 97 ], dispensing robots [ 98 , 99 ], and self-induced partitioning via SlipChip [ 100 , 101 , 102 ] have all been used for loading microwell arrays for nucleic acid applications.…”

Section: Digital Biosensing For Nucleic Acidsmentioning

confidence: 99%

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Recent Advances in Digital Biosensing Technology

et al. 2022

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Digital biosensing assays demonstrate remarkable advantages over conventional biosensing systems because of their ability to achieve single-molecule detection and absolute quantification. Unlike traditional low-abundance biomarking screening, digital-based biosensing systems reduce sample volumes significantly to the fL-nL level, which vastly reduces overall reagent consumption, improves reaction time and throughput, and enables high sensitivity and single target detection. This review presents the current technology for compartmentalizing reactions and their applications in detecting proteins and nucleic acids. We also analyze existing challenges and future opportunities associated with digital biosensing and research opportunities for developing integrated digital biosensing systems.

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Section: Digital Biosensing For Nucleic Acidsmentioning

confidence: 99%

“…After the wells were fully loaded, the wells were sealed with an oil phase. The design offers ease of operation and high 99.5 ± 0.3% digitization efficiency for highly sensitive PCR [ 97 ].…”

Section: Digital Biosensing For Nucleic Acidsmentioning

confidence: 99%

Recent Advances in Digital Biosensing Technology

et al. 2022

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“…With the anti-evaporation layer, no discernible evaporation was observed after 16 hours (Supplementary Figure 1g). Compared to previously reported anti-evaporation methods [25][26][27][28] , our design is easy to fabricate, assemble, and can be applied to most standard double-layer microfluidic designs without the need of additional hardware.…”

Section: Batch Cell-free Expression Augmented With Continuous Dialysismentioning

confidence: 99%

Improved cell-free transcription-translation reactions in microfluidic chemostats augmented with hydrogel membranes for continuous small molecule dialysis

Lavickova

Grasemann

Maerkl

2022

Preprint

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Increasing protein production capacity of the PURE cell-free transcription-translation (TX-TL) system will be key to implementing complex synthetic biological circuits, and to establish a fully self-regenerating system as a basis for the development of a synthetic cell. Under steady-state conditions, the protein synthesis capacity of the PURE system is likely at least one order of magnitude too low to express sufficient quantities of all PURE protein components. This is in part due to the fact that protein synthesis can't be sustained during the entire dilution cycle, especially at low dilution rates. We developed a microfluidic chemostat augmented with semi-permeable membranes that combines steady-state reactions and continuous dialysis as a possible solution to enhance protein synthesis at steady-state. In batch operation, the continuous dialysis of low molecular weight components via the membranes extended protein synthesis by over an order of magnitude from 2 hours to over 30 hours, leading to a seven-fold increase in protein yield. In chemostat operation, continuous dialysis enabled sustained protein synthesis during the entire dilution cycle even for low dilution rates, leading to six-fold higher protein levels at steady state. The possibility to combine and independently manipulate continuous dialysis and chemostat operation renders our dialysis chemostat a promising technological basis for complex cell-free synthetic biology applications that require enhanced protein synthesis capacity.

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“…Figure 5 Related applications of microfluidic chips in medical field. (a )Schematic diagram of a microfluidic chip used in polymerase chain reaction (PCR) [47] . (b)Microfluidic intelligent devices for diagnosing infectious pathogens [50] .…”

Section: 化学发光(Cl)是物质在化学反应过程中伴随的⼀种光辐射现象，化学发光成像因其⾼灵敏度⼴泛应⽤mentioning

confidence: 99%