Microfluidic Single-cell Trapping and Cultivation for the Analysis of Host-viral Interactions

Ganguly, Reya; Lee, Byungjin; Kang, Solib; Kim, Yong Sic; Jeong, Seong-Geun; Kim, Jae Seong; Park, So Young; Yamauchi, Yohei; Lee, Chang‐Soo

doi:10.1007/s12257-020-0143-1

Cited by 8 publications

(4 citation statements)

References 37 publications

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“…In Figure , we outline typical competitive PCR-based DNA quantification in microdroplets. We designed a pyramidal microfluidic network with sequentially branched microchannels to achieve competitive PCR in microdroplets, allowing serial dilutions of the highest concentration in a linear gradient (Figure A). − Each dilution channel opens into a droplet-generating junction that produces microdroplets by flow-focusing. , The microdroplets with gradients are collected into a PCR tube for amplification, as shown in Figure B. In competitive PCR, target DNA, and competitor DNA levels maintain a ratio throughout the amplification process .…”

Section: Resultsmentioning

confidence: 99%

A Poisson-Independent Approach to Precision Nucleic Acid Quantification in Microdroplets

Ganguly,

Lee

2024

ACS Appl. Bio Mater.

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Digital PCR (dPCR) has become indispensable in nucleic acid (NA) detection across various fields, including viral diagnostics and mutant detection. However, misclassification of partitions in dPCR can significantly impact accuracy. Despite existing methods to minimize misclassification bias, accurate classification remains elusive, especially for nonamplified target partitions. To address these challenges, this study introduces an innovative microdroplet-based competitive PCR platform for nucleic acid quantification in microfluidic devices independent of Poisson statistics. In this approach, the target concentration (T) is determined from the concentration of competitor DNA (C) at the equivalence point (E.P.), where C/T is 1. Competitive PCR ensures that the ratio of target to competitor DNA remains constant during amplification, reflected in the resultant fluorescence intensity, allowing the quantification of target DNA concentration at the equivalence point. The unique amplification technique eliminates Poisson distribution, addressing misclassification challenges. Additionally, our approach reduces the need for post-PCR procedures and shortens analytical time. We envision this platform as versatile, reproducible, and easily adaptable for driving significant progress in molecular biology and diagnostics.

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Section: Resultsmentioning

confidence: 99%

A Poisson-Independent Approach to Precision Nucleic Acid Quantification in Microdroplets

Ganguly,

Lee

2024

ACS Appl. Bio Mater.

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“…Microelectrodes were used to detect the quantum secretion of dopamine from individual PC12 cell amperometricaly. The microfluidic device enabled manipulation of single-cell by injecting stimulants, and monitoring quantal secretion from the cell, therefore offering an appropriate platform to investigate physiological and pathological phenomena at an individual cellular level [ [51] , [52] , [53] ]. Similarly, the mercaptopropionic acid (MPA)-modified gold electrodes microchip were used to determine and monitor the release of dopamine from single PC12 cell [ 26 ].…”

Section: Single Neuron Dynamicsmentioning

confidence: 99%

Microfluidic platforms for single neuron analysis

Gupta

Shinde

Illath

et al. 2022

Materials Today Bio

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“…The concentration gradient of the drug doxorubicin hydrochloride model was successfully generated on a Christmas tree structure. Ganguly et al developed a microfluidic hydrodynamic single-cell trapping and culturing platform of host-viral interactions that reported the influenza A virus interactions analysis under the gradient concentration of bafilomycin A1 treatment [13]. This facilitated the biological analysis and experimentation of virus infection into host cells.…”

Section: Introductionmentioning

confidence: 99%

Complete experimental and theoretical characterization of nonlinear concentration gradient generator microfluidic device for analytical purposes

et al. 2021

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Microfluidic devices that generate stable concentration gradients are efficient instruments for automated calibration for analytical and bioanalytical systems. However, little attention has been paid to the development of reusable microfluidic concentration gradient generators, which can be useful for a range of species through mathematical characterization. In this work, we develop a microfluidic device based on three steps of serial dilution that were able to generate nonlinear concentration gradient for dyes and biomolecules. The microfluidic device was described mathematically, statistically and was suitable for reusable analytical and bioanalytical analysis. The device reproducibility was assessed by experimental tests, which have shown the same gradient concentration profile for different dyes and statistical reproducibility with 95% confidence interval for bovine serum albumin (BSA). Moreover, the experimental data converged well with those obtained by computational fluid dynamics simulation. Applicability was verified by coupling the microfluidic device to a surface plasmon resonance (SPR) biosensor, based on nanohole arrays with sensitivity of 358.7 nm RIU −1 determined by white-light SPR excitation exposed to different D-(+)-glucose aqueous solutions with 1.3361-1.4035 refractive index interval. The transmission light intensities obtained by the array of images allowed to quantify a pseudo-unknown BSA sample (160 µg mL −1 ) at 138 µg mL −1 . The SPR analysis has been validated in parallel by fluorescence emissions, which showed a concentration of 154.8 ± 16.6 µg mL −1 .

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Microfluidic Single-cell Trapping and Cultivation for the Analysis of Host-viral Interactions

Cited by 8 publications

References 37 publications

A Poisson-Independent Approach to Precision Nucleic Acid Quantification in Microdroplets

A Poisson-Independent Approach to Precision Nucleic Acid Quantification in Microdroplets

Microfluidic platforms for single neuron analysis

Complete experimental and theoretical characterization of nonlinear concentration gradient generator microfluidic device for analytical purposes

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