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A Facile and Rapid Route to Self-Digitization of Samples into a High Density Microwell Array for Digital Bioassays

By Xu Cui, Tianbao Hu, Qiang Chen, Qiang Zhao, Yin Wu, Tengbao Xie, Pengyong Liu, Xi Su, Gang Li

Posted 01 Feb 2021
bioRxiv DOI: 10.1101/2021.01.31.428989

Digital bioassays are powerful methods to detect rare analytes from complex mixtures and study the temporal processes of indi-vidual entities within biological systems. In digital bioassays, a crucial first step is the discretization of samples into a large num-ber of identical independent partitions. Here, we developed a rapid and facile sample partitioning method for versatile digital bio-assays. This method is based on a detachable self-digitization (DSD) chip which couples a reversible assembly configuration and a predegassing-based self-pumping mechanism to achieve an easy, fast and large-scale sample partitioning. The DSD chip consists of a channel layer used for loading sample and a microwell layer used for holding the sample partitions. Benefitting from its de-tachability, the chip avoids a lengthy oil flushing process used to remove the excess sample in loading channels and can compart-mentalize a sample into more than 100,000 wells of picoliter volume with densities up to 14,000 wells/cm2 in less than 30 s. We also demonstrated the utility of the proposed method by applying it to digital PCR and digital microbial assays.

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