A facile and rapid route to self-digitization of samples into a high density microwell array for digital bioassays

[摘要] Digital 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. 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 the sample and a microwell layer used for holding the sample partitions. Benefitting from its detachability, the chip avoids a lengthy oil flushing process used to remove the excess sample in loading channels and can compartmentalize a sample into more than 100,000 wells of picoliter volume with densities up to 14,000 wells/cm(2) in less than 30 s. We also demonstrated the utility of the proposed method by applying it to digital PCR and digital microbial assays.

[发布日期] 2021-10-01

[关键词] Digital bioassay;Self-digitization;High-density microwell array;Absolute quantification;Single bacteria growth kinetics