A low-cost high throughput microfluidics system for multi-thousand droplets per second sorting based on time resolved fluorescence measurement

Abstract

Time-resolved fluorescence measurement screening and sorting has been proven to be interesting for biological applications. Fluorescence lifetime (FLT) measurement provides additional information to the measurement of the intensity or polarization of the fluorescence emission. Furthermore, droplet microfluidics enables higher throughput than well plate readers. Few researches have been recently done about the microfluidic droplet sorting based on the fluorescence lifetime measurement. Some of the proposed solutions support high throughput but they do not extract the FLT directly from the fluorescence decay. In this paper, we present an alternative low-cost system for the microfluidic droplets sorting. We implemented a FLT measurement system based on the time-correlated single photon counting (TCSPC) technique on a cyclone V SoC-FPGA. For the excitation light source, we use a simple pulsed laser diode and a single photon avalanche diode (SPAD) as a photodetector. The optical system is an ad hoc microscope. The droplet generation is done with flow focusing technique in a PDMS-based microfluidics chip. This system was successfully tested in real-time at a droplet rate of more than 3000 droplets per second.