High-throughput microfluidic platform for high-density immobilization and parallelized quantitative real-time measurements of droplets

Unmet Need:
Droplet platforms, which compartmentalize samples into encapsulated droplets for high-throughput analysis, have been used to detect and analyze diverse molecules, including nucleic acids, proteins, and single cells, at a higher sensitivity than traditional methods. Despite advantages in sensitivity and throughput, most existing droplet analysis platforms are only capable of endpoint analysis, limiting detection and analysis of the droplet to a single time point. To enhance the capabilities of single-molecule analyses, there is a need for platforms that allow for real-time analysis for applications such as melting curve analysis, cell growth monitoring, and enzyme kinetics studies. Additionally, current droplet platforms suffer from inefficiencies in droplet capture, which decreases the resolution at which rare molecules and biomarkers can be identified.

Technology Overview:
Researchers at Johns Hopkins have developed a real-time droplet platform that enhances the overall throughput of real-time droplet analyses by maximizing imaging footprint, and increasing operational flexibility and droplet capture efficiency. The platform consists of a microfluidic droplet trapping device that has 100% efficiency in the loading of droplets ranging from 100 pL to 1 nL and a thermal-optical platform, which allows for the real-time analyses of up to 30,000 droplets in parallel across a broad range of temperatures. The inventors have demonstrated that the platform is capable of detecting heterogeneous DNA methylation of the tumor suppressor gene CDO1 through digital droplet PCR followed by digital High Resolution Melt (HRM) analysis.

Stage of Development:
The inventors have developed the platform and validated its time-lapse analyses of up to 30,000 droplets in parallel for use in single-molecule genetic and epigenetic profiling applications.
 

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