Centrifugally automated Solid-Phase Extraction of DNA by immiscible liquid valving and chemically powered centripetal pumping of peripherally stored reagents

Kinahan, David J.; Burger, Robert; Lawlor, Daryl; Early, Philip L.; Vembadi, Abhishek; McArdle, Niamh A.; Kilcawley, Niamh A.; Glynn, Macdara; Ducrée, Jens

doi:10.1016/j.biosx.2021.100085

Cited by 3 publications

(10 citation statements)

References 55 publications

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“…The tests were run manually, with the disc performance monitored using the stroboscopically coupled camera. The disc is rotated at 30 Hz. The FC-40 is pumped into the routing structure to block a DF film with an immiscible plug of liquid . The primers and mineral oil are pumped into the amplification chambers.…”

Section: Methodsmentioning

confidence: 99%

“…A key benefit of this technology is the ability to accept samples by direct pipetting rather than needing complex loading and priming steps. , These features make the LoaD platform particularly appealing for a wide range of point-of-care and point-of-use applications. − ,, . Nucleic acid (NA) methods have been implemented on the LoaD by a number of research groups. ,− ,,− LoaD has also been used for the detection of foodborne pathogens using NA methods. − …”

Section: Introductionmentioning

confidence: 99%

“…Modeling this using Boyle’s Law is facile, , and through careful design, automation of 8 LUOs has been demonstrated using these valves . Both rotationally actuated “burst” DF valves and “event-triggered” dissolvable film valves have been applied to a wide range of applications ,,− with good success. However, a limitation of DF valves remains that the actuation of these valves is dependent on either tuning the valve opening spin-speed, which necessitates limiting the number of LUOs on the disc, or determining the dissolve time of the DF (usually ∼40 s), limiting the ability to perform sometimes assay critical incubations on the LoaD.…”

Section: Introductionmentioning

confidence: 99%

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Low–High–Low Rotationally Pulse-Actuated Serial Dissolvable Film Valves Applied to Solid Phase Extraction and LAMP Isothermal Amplification for Plant Pathogen Detection on a Lab-on-a-Disc

Julius,

Saeed,

Kuijpers

et al. 2024

ACS Omega

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The ability of the centrifugal Lab-on-a-Disc (LoaD) platform to closely mimic the "on bench" liquid handling steps (laboratory unit operations (LUOs)) such as metering, mixing, and aliquoting supports on-disc automation of bioassay without the need for extensive biological optimization. Thus, well-established bioassays, normally conducted manually using pipettes or using liquid handling robots, can be relatively easily automated in self-contained microfluidic chips suitable for use in point-of-care or point-of-use settings. The LoaD's ease of automation is largely dependent on valves that can control liquid movement on the rotating disc. The optimum valving strategy for a true low-cost and portable device is rotationally actuated valves, which are actuated by changes in the disc spin-speed. However, due to tolerances in disc manufacturing and variations in reagent properties, most of these valving technologies have inherent variation in their actuation spin-speed. Most valves are actuated through stepped increases in disc spin-speed until the motor reaches its maximum speed (rarely more than 6000 rpm). These manufacturing tolerances combined with this "analogue" mechanism of valve actuation limits the number of LUOs that can be placed on-disc. In this work, we present a novel valving mechanism called low−high−low serial dissolvable film (DF) valves. In these valves, a DF membrane is placed in a dead-end pneumatic chamber. Below an actuation spinspeed, the trapped air prevents liquid wetting and dissolving the membrane. Above this spin-speed, the liquid will enter and wet the DF and open the valve. However, as DFs take ∼40 s to dissolve, the membrane can be wetted, and the disc spin-speed reduced before the film opens. Thus, by placing valves in a series, we can govern on which "digital pulse" in spin-speeding a reagent is released; a reservoir with one serial valve will open on the first pulse, a reservoir with two serial valves on the second, and so on. This "digital" flow control mechanism allows the automation of complex assays with high reliability. In this work, we first describe the operation of the valves, outline the theoretical basis for their operation, and support this analysis with an experiment. Next, we demonstrate how these valves can be used to automate the solid-phase extraction of DNA on on-disc LAMP amplification for applications in plant pathogen detection. The disc was successfully used to extract and detect, from a sample lysed off-disc, DNA indicating the presence of thermally inactivated Clavibacter michiganensis ssp. michiganensis (Cmm), a bacterial pathogen on tomato leaf samples.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Low–High–Low Rotationally Pulse-Actuated Serial Dissolvable Film Valves Applied to Solid Phase Extraction and LAMP Isothermal Amplification for Plant Pathogen Detection on a Lab-on-a-Disc

Julius,

Saeed,

Kuijpers

et al. 2024

ACS Omega

Self Cite

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“…However, the efficiency of this technique usually relies on the ratio of the flow resistances of the inlet and outlet channels and the use of a powerful motor to vigorously change the rate of rotation. In a notable exception, using immiscible liquid check valves with pumping structures has permitted efficient pumping at more sedate acceleration/deceleration profiles [24]. Chemical pumping, where gas is released from a chemical reaction and displaces a liquid inwards, has also been demonstrated to have a good effect [24][25][26].…”

Section: Introductionmentioning

confidence: 99%

“…In a notable exception, using immiscible liquid check valves with pumping structures has permitted efficient pumping at more sedate acceleration/deceleration profiles [24]. Chemical pumping, where gas is released from a chemical reaction and displaces a liquid inwards, has also been demonstrated to have a good effect [24][25][26].…”

Section: Introductionmentioning

confidence: 99%

Dissolvable Film-Controlled Buoyancy Pumping and Aliquoting on a Lab-On-A-Disc

et al. 2023

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Lab-on-a-Disc (LoaD) has great potential for applications in decentralised bioanalytical testing where speed and robustness are critical. Here, a disc-shaped microfluidic chip is rotated to pump liquid radially outwards; thus, all microfluidic structures must be fitted into the available radial length. To overcome this limitation, several centripetal pumping technologies have been developed. In this work, we combine buoyancy pumping, enabled by displacing aqueous samples and reagents centripetally inwards by a dense liquid (fluorocarbon FC-40), with dissolvable film (DF) to automate a multi-step assay. The DF dissolves in the presence of water but is not in contact with the FC-40. Therefore, the FC-40 can be stored behind the DF membranes and is autonomously released by contact with the arriving aqueous sample. Using this technology, tasks such as blood centrifugation can be located on the disc periphery where ‘disc real estate’ is less valuable and centrifugal forces are higher. To demonstrate this, we use the combination of the buoyancy-driven centripetal pumping with DF barriers to implement a fully automated multi-parameter diagnostic assay on the LoaD platform. The implemented steps include plasma extraction from a structure, automatically triggered metering/aliquoting, and the management of five onboard stored liquid reagents. Critically, we also demonstrate highly accurate aliquoting of reagents using centripetal pumping. We also provide a mathematical model to describe the pumping mechanism and apply lumped-element modelling and Monte Carlo simulation to estimate errors in the aliquoting volumes caused by manufacturing deviations.

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Design and Performance Analysis of a Double-Acting Type External Centripetal Plunger Fluid-powered Pump

2023

ijfans

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This research focuses on the development and analysis of a centripetal plunger fluid-powered pump of a double-acting type external impeller. The pump utilizes an innovative external impeller inner stator structure, incorporating oil absorption and oil pressing centripetal cams to drive the plungers in a straight reciprocating motion along the centripetal direction. The pump achieves efficient pressure oil generation and employs a unique oil distribution system using a sliding valve. The design ensures reliable sealing, minimal leakage, and symmetrical stress distribution, while maintaining a compact structure, high volume efficiency, and low abrasion.

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Centrifugally automated Solid-Phase Extraction of DNA by immiscible liquid valving and chemically powered centripetal pumping of peripherally stored reagents

Cited by 3 publications

References 55 publications

Low–High–Low Rotationally Pulse-Actuated Serial Dissolvable Film Valves Applied to Solid Phase Extraction and LAMP Isothermal Amplification for Plant Pathogen Detection on a Lab-on-a-Disc

Low–High–Low Rotationally Pulse-Actuated Serial Dissolvable Film Valves Applied to Solid Phase Extraction and LAMP Isothermal Amplification for Plant Pathogen Detection on a Lab-on-a-Disc

Dissolvable Film-Controlled Buoyancy Pumping and Aliquoting on a Lab-On-A-Disc

Design and Performance Analysis of a Double-Acting Type External Centripetal Plunger Fluid-powered Pump

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