Abstract

Centrifugal Step Emulsification Allows Miniaturized Digital Droplet-RPA,-LAMP and -PCR on the Centrifugal Microfluidic Platform

Felix von Stetten, Deputy Head, Hahn-Schickard-Gesellschaft für angewandte Forschung e.V.

The talk begins with a brief introduction to the centrifugal microfluidic LabDisk platform. The platform consists of a processing device, a disposable test carrier with prestored reagents and a validated set of microfluidic unit operations, as well as a scalable production process for test carriers.

The major focus of the talk is on a novel unit operation, designated as centrifugal step emulsification. This unit operation enables centrifugal droplet generation, DNA amplification and fluorescence detection within one single cavity of a disposable chip. This enables significant miniaturization of future devices for digital assays. Two different implementation scenarios are outlined: First, the processing of the microfluidic chip on off the shelf laboratory devices, namely a mini centrifuge for droplet generation, a thermocycler for amplification and a microarray scanner for readout will be brought to the fore. Second, the processing of the chip by one single miniaturized and integrated device comprising a centrifuge, incubator and fluorescence CCD camera for readout will be explained.

Results for absolute quantification of DNA by digital droplet-RPA,-LAMP and -PCR amplification using the novel unit operation are in good agreement with those obtained by a commercial state of the art dPCR system.