However, small organic molecules can exchange between adjacent droplets 4, 12, limiting their utility for high-throughput screening applications. These tiny reaction chambers are created, analyzed, and sorted at kHz rates, typically using polydimethylsiloxane (PDMS) microfluidic devices 4. Additionally, emulsion droplets are easily generated with highly monodisperse and reproducible droplet size, making them suitable to study different aspects of physics and chemistry 10, 11, 12, 13, 14. Accordingly, these emulsions have been exploited for numerous biological applications, including rapid parallel transcriptome profiling of thousands of cells with single-cell resolution 1, 2, high-throughput drug screening 3, 4, 5, analysis of products secreted by individual cells 6, directed evolution of desired enzymes 7, 8, and construction of synthetic cells 9. Emulsions made with fluorosurfactant-containing fluorinated oil have been shown to be biologically inert and, due to fluorinated oil’s capacity for dissolved oxygen, are suitable for cell culture. We used a panel of dendritic oligo-glycerol-based surfactants to demonstrate that a high degree of inter- and intramolecular hydrogen bonding, as well as the dendritic architecture, contribute to high droplet stability in PCR thermal cycling and minimize inter-droplet transfer of the water-soluble fluorescent dye sodium fluorescein salt and the drug doxycycline.įluorosurfactant-stabilized, water-in-fluorinated-oil (w/o) droplets, with volumes of pico- to nanoliters, have facilitated a variety of powerful research techniques. Here, we provide insights into designing surfactants that form robust microdroplets with improved stability and resistance to inter-droplet transfer. Therefore, the use of droplet-based platforms for ultrahigh-throughput combination drug screening and polymerase chain reaction (PCR)-based rare mutation detection has been limited. In addition, the microdroplets typically coalesce at temperatures higher than 80 ☌. However, current surfactants cannot completely prevent inter-droplet transfer of small organic molecules encapsulated or produced inside the droplets. Fluorosurfactant-stabilized microfluidic droplets are widely used as pico- to nanoliter volume reactors in chemistry and biology.
0 kommentar(er)
