Stable water-in-oil emulsion is essential to digital PCR and many other bioanalytical reactions that employ droplets as microreactors. We developed a novel technology to produce monodisperse emulsion droplets with high efficiency and high throughput using a bench-top centrifuge. Upon centrifugal spinning, the continuous aqueous phase is dispersed into monodisperse droplet jets in air through a micro-channel array (MiCA) and then submerged into oil as a stable emulsion. We performed dPCR reactions with a high dynamic range through the MiCA approach, and demonstrated that this cost-effective method not only eliminates the usage of complex microfluidic devices and control systems, but also greatly suppresses the loss of materials and cross-contamination. MiCA-enabled highly parallel emulsion generation combines both easiness and robustness of picoliter droplet production, and breaks the technical challenges by using conventional lab equipment and supplies.

The Tamsui River basin is located in Northern Taiwan and encompasses the most metropolitan city in Taiwan, Taipei City. The Taiwan Environmental Protection Administration (EPA) has established 38 water quality monitoring stations in the Tamsui River basin and performed regular river water quality monitoring for the past two decades. Because of the limited budget of the Taiwan EPA, adjusting the monitoring program while maintaining water quality data is critical. Multivariate analysis methods, such as cluster analysis (CA), factor analysis (FA), and discriminate analysis (DA), are useful tools for the statistically spatial assessment of surface water quality. This study integrated CA, FA, and DA to evaluate the spatial variance of water quality in the metropolitan city of Taipei. Performing CA involved categorizing monitoring stations into three groups: high-, moderate-, and low-pollution areas. In addition, this categorization of monitoring stations was in agreement with that of the assessment that involved using the simple river pollution index. Four latent factors that predominantly influence the river water quality of the Tamsui River basin are assessed using FA: anthropogenic pollution, the nitrification process, seawater intrusion, and geological and weathering processes. We plotted a spatial pattern using the four latent factor scores and identified ten redundant monitoring stations near each upstream station with the same score pattern. We extracted five significant parameters by using DA: total organic carbon, total phosphorus, As, Cu, and nitrate, with spatial variance to differentiate them from the polluted condition of the group obtained by using CA. Finally, this study suggests that the Taiwan EPA can adjust the surface water-monitoring program of the Tamsui River by reducing the monitoring stations to 28 and the measured chemical parameters to five to lower monitoring costs.

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The realization of the vast potential of digital PCR (dPCR) to provide extremely accurate and sensitive measurements in the clinical setting has thus far been hindered by challenges such as assay robustness and high costs. Here we introduce a lossless and contamination-free dPCR technology, termed CLEAR-dPCR, which addresses these challenges by completing the dPCR sample preparation, PCR, and readout all in one tube. Optical clearing of the droplet dPCR emulsion was combined with emerging light-sheet fluorescence microscopy, to acquire a three-dimensional (3D) image of a half million droplets sealed in a tube in seconds. CLEAR-dPCR provides ultrahigh-throughput readout results in situ and fundamentally eliminates the possibility of either sample loss or contamination. This approach exhibits improved accuracy over existing dPCR platforms and enables a greatly increased dynamic range to be comparable to that of real-time quantitative PCR.