Digital polymerase chain reaction (dPCR) is emerging as a powerful method for nucleic acid detection due to its unprecedented sensitivity and precision. However, most current dPCR platforms are inherently limited by their low multiplexing ability due to primer-pair cross interactions and spectral overlap of available fluorophores. Here, we present a novel and robust method for multiplexing dPCR that is free from primer dimerization and fluorescence channel number limitation, enabling highly precise and multiplexed detection of nucleic acid targets.
View Article and Find Full Text PDFACS Appl Mater Interfaces
May 2022
This paper proposes a simple and robust method for spontaneously digitizing aqueous samples into a high-density microwell array. The method is based on an oil-triggered template-confined dewetting phenomenon. To realize the dewetting-induced sample digitization, an aqueous sample is first infused into a networked microwell array (NMA) through a pre-degassing-based self-pumping mechanism, and an immiscible oil phase is then applied over the surface of NMA chip to induce the templated dewetting.
View Article and Find Full Text PDFIn this work, we present a simple and equipment-free system for discretizing samples into tens of thousands of discrete volumes in tens of seconds. Unlike conventional sample discretization systems that require bulky syringe pumps, pressure controllers, or vacuum equipment, our system requires only a sheet of water-soluble film, a hand-operated syringe, and a microfluidic device containing a high-density microchamber array. In this system, the water-soluble film seals the device inlet to form a closed channel-chamber system, while the syringe is used to create a vacuum in the closed system.
View Article and Find Full Text PDFDigital bioassays are powerful methods to detect rare analytes from complex mixtures and study the temporal processes of individual entities within biological systems. In digital bioassays, a crucial first step is the discretization of samples into a large number of identical independent partitions. Here, we developed a rapid and facile sample partitioning method for versatile digital bioassays.
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