Microfluidic encapsulation of DNAs in liquid beads for digital PCR application

Galogahi, Fariba Malekpour; Christie, Melodie; Yadav, Ajeet Singh; An, Hongjie; Stratton, Helen; Nguyen, Nam‐Trung

doi:10.1039/d3an00868a

Cited by 5 publications

(1 citation statement)

References 54 publications

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“…Following the collection, the formed core–shell droplets are exposed to a 24 W blue light (450–490 nm) source for 20 minutes to polymerize and form liquid beads, solid particles with a liquid core. 24 However, the throughput is lower than conventional ddPCR methods. 25 This is attributed to slower optimised flow rate and about 50% of the produced beads are core–shell.…”

Section: Methodsmentioning

confidence: 99%

Precise, wide field, and low-cost imaging and analysis of core–shell beads for digital polymerase chain reaction

et al. 2023

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Section: Methodsmentioning

confidence: 99%

Precise, wide field, and low-cost imaging and analysis of core–shell beads for digital polymerase chain reaction

et al. 2023

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Microfluidic Encapsulation of DNAs in Liquid Beads for Digital Loop‐Mediated Isothermal Amplification

Galogahi,

Strachan,

Yadav

et al. 2024

Advanced NanoBiomed Research

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Digital nucleic acid analysis has emerged as a prominent tool for the detection and absolute quantification of diverse pathogens. Digital loop‐mediated isothermal amplification (dLAMP) offers highly sensitive, specific, time‐efficient, and cost‐effective nucleic acid amplification. However, existing dLAMP techniques face challenges such as droplet merging, reliance on surfactants, restricted partition capacities, and the potential for sample loss during heating. Herein, these issues are addressed by introducing liquid beads for sample partitioning. Compared to microwells, our approach overcomes the limitations of chamber dimensions, enabling the analysis of an unlimited number of digitized targets. Furthermore, our novel approach effectively addresses sample loss and merging during thermal processing and eliminates the need for surfactants. Accurate and reproducible the quantitative detection of the gene cluster XALB1 of leaf scald disease is conducted using dLAMP based on liquid beads to verify its availability. The results demonstrate a high correlation between target concentration and positive signals, indicating the robust performance of our technique. A comparative analysis is then performed between dLAMP using liquid beads and using single droplets. Benchmarking these two techniques highlights the effectiveness of our innovative technique in overcoming existing challenges in dLAMP.

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Synthesis and active manipulation of magnetic liquid beads

Yadav,

Galogahi,

Vashi

et al. 2024

Biomed Microdevices

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We report the fabrication and characterisation of magnetic liquid beads with a solid magnetic shell and liquid core using microfluidic techniques. The liquid beads consist of a fluorinated oil core and a polymer shell with magnetite particles. The beads are generated in a flow-focusing polydimethylsiloxane (PDMS) device and cured by photo polymerisation. We investigated the response of the liquid beads to an external magnetic field by characterising their motion towards a permanent magnet. Magnetic sorting of liquid beads in a channel was achieved with 90% efficiency. The results show that the liquid beads can be controlled magnetically and have potential applications in digital microfluidics including nucleic acid amplification, drug delivery, cell culture, sensing, and tissue engineering. The present paper also discusses the magnetophoretic behaviour of the liquid bead by varying its mass and magnetite concentration in the shell. We also demonstrated the two-dimensional self-assembly of magnetic liquid beads for potential use in digital polymerase chain reaction and digital loop mediated isothermal amplification. Graphical abstract

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Microfluidic encapsulation of DNAs in liquid beads for digital PCR application

Abstract: Droplet-based microfluidics and digital polymerase chain reaction (PCR) hold significant promise for accurately detecting and quantifying pathogens. However, existing droplet-based digital PCR (ddPCR) applications have been relying exclusively on single...

Cited by 5 publications

References 54 publications

Precise, wide field, and low-cost imaging and analysis of core–shell beads for digital polymerase chain reaction

Precise, wide field, and low-cost imaging and analysis of core–shell beads for digital polymerase chain reaction

Microfluidic Encapsulation of DNAs in Liquid Beads for Digital Loop‐Mediated Isothermal Amplification

Synthesis and active manipulation of magnetic liquid beads

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