Droplet microfluidics has revolutionized single-cell analysis, allowing for ultra-high-throughput sorting of droplets to meet the demands of single-cell sorting. However, accurately and efficiently dispensing target droplets into specific containers for downstream analysis has remained a challenge. Here, we present an integrated microfluidic droplet dispenser based on the electrohydrodynamic (EHD) principle, capable of on-demand single droplet dispensing with high accuracy. Our system uses optical signal activation and EHD actuation to isolate and dispense droplets. The microfluidic droplet auto dispenser consists of four main modules: a pump-drive chip module, an optical automatic signal recognition module, a high voltage control module, and an automated X-Y translation stage module. By optimizing the chip structure and voltage parameter, this platform successfully isolates single Escherichia coli (E. coli) cells onto a Petri dish for downstream cultivation and heterogeneity analysis. In addition, the dispenser microfluidic system dispenses the sorted droplets in a "one droplet, one well" manner, providing an automated method for droplet interface from the micro- to macro- platform. This dispenser system provides a platform for phenotypic screening of single colony pick and single cell analysis
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