Low-cost wide-adaptation centrifugal digital droplet generation method and device

Abstract

The invention discloses a low-cost wide-adaptation centrifugal digital droplet generation method and device, an upper-layer water phase centrifugal unit is embedded in a lower-layer oil phase centrifugal unit, a micron tube is arranged at the lower part of the upper-layer water phase centrifugal unit, and under the action of centrifugal force, the upper water phase overcomes the surface tension effect, liquid drops generated through the micro-tube channel enter the lower oil phase to form water-in-oil liquid drops with uniform sizes, and the sizes of the water-in-oil liquid drops are adjustedby adjusting the diameters of the micro-tubes and / or the centrifugal force. A mutually soluble laser-induced curing agent can be added into the oil phase, and the oil phase is quickly crosslinked andcured under the action of an induction source; the invention is applied to nucleic acid isothermal amplification to realize digital nucleic acid amplification. According to the invention, water-in-oildroplets with uniform size and high quality can be generated. The micro-liter aqueous phase solution can quickly generate high-flux water-in-oil droplets within a few minutes, the high-flux water-in-oil droplets are combined with isothermal amplification of nucleic acid, and meanwhile, ultrafast solidification of the droplets can be realized by virtue of a solidification platform.

Description

technical field [0001] The invention belongs to the field of analysis and sensing, and mainly relates to a low-cost and widely applicable centrifugal digital droplet generation method and device, which can be used for single-molecule nucleic acid detection and the like. Background technique [0002] In recent years, digital nucleic acid detection technology has played a huge role in the fields of biological detection, chemical sensing, and medical diagnosis. As a third-generation nucleic acid amplification method, the sample is diluted to the single-molecule level by distributing the sample into tens of millions of units using the technical concept of micro-droplets. Each droplet contains nucleic acid or does not contain nucleic acid. After amplification, the droplet containing nucleic acid has a fluorescent signal, and the number of fluorescent droplets conforms to the Poisson distribution. Absolute quantitative detection of samples such as target nucleic acids can be achi...

AI Technical Summary

Problems solved by technology

Among the droplet generating devices studied in recent years, CN109536590A proposes a single-cell gene detection method based on a microporous chip, but the production of this multi-layer chip is cumbersome and the processing procedure is complicated

CN110841734A proposes a single-pump droplet generation system, but the syringe pump introduces the risk of mixed contamination

In 2018 and 2020, Mou Ying's research group proposed a thermal curing and UV curing solution respectively, which effectively alleviated the problem of droplet expansion, but there is also the problem of too long curing time

[0006] 1) The structure of the droplet generating device is generally complicated, and it is not easy to promote on a large scale;

[0007] 2) The pump in the microfluidic chip introduces the risk of external contamination;

[0008] 3) Droplets may flow, agglomerate, or cure for too long during heating or UV light

Images