Abstract
Combinatorial droplet microfluidic devices with programmable microfluidic valves have recently emerged as a viable approach for performing multiplexed experiments in microfluidic droplets. However, the serial operation in these devices restricts their throughput. To address this limitation, we present a parallelized combinatorial droplet device that enhances device throughput via droplet bifurcation, parallelized droplet fusion, and parallelized droplet detection. In this device, sample droplets split evenly at bifurcating Y-junctions before multiple independent reagent droplets are injected directly into the split sample droplets for robust droplet fusion. Finally, the fused sample and reagent droplets can be imaged in parallel via microscopy. The combination of these approaches enabled us to improve the throughput over traditional, serially-operated combinatorial droplet devices by 16-fold—with ready potential for further enhancement. Given its current performance and prospect for future improvements, we believe the parallelized combinatorial droplet device has the potential to meet the demand as a flexible and cost-effective tool that can perform high throughput screening applications.
Highlights
Recent years have seen the emergence of microfluidic droplet technology in a variety of chemical and biochemical applications [1,2], ranging from single-cell analysis [3,4,5,6] to enzyme kinetics measurements [7,8] and even nanoparticle synthesis [9,10,11]
Droplet microfluidic devices that utilize programmable microfluidic valves to mix combinations of samples and reagents on-demand [23,24,25,26,27,28,29] present a viable approach to performing multiplexed experiments in microfluidic droplets
We demonstrate that microfluidic droplets can split evenly at all Y-junctions across all four bifurcation stages in our device
Summary
Recent years have seen the emergence of microfluidic droplet technology in a variety of chemical and biochemical applications [1,2], ranging from single-cell analysis [3,4,5,6] to enzyme kinetics measurements [7,8] and even nanoparticle synthesis [9,10,11]. Microfluidic droplets have proven pivotal in digital analysis of individual biological samples at the single-molecule level [12,13,14,15] While this technology offers the capacity to divide a homogeneous reaction (i.e., one sample against one set of reagents) into a large number of reactions in droplets, it faces challenges in generating a large combination of sample-reagent mixtures in a multiplexed manner. Droplet microfluidic devices that utilize programmable microfluidic valves to mix combinations of samples and reagents on-demand [23,24,25,26,27,28,29] present a viable approach to performing multiplexed experiments in microfluidic droplets. In some of these devices, the conseDrvNaAtio[2n6o,2f8p] oansidticohnaarlaoctredriezraotifoenaocfhmdartorpixlemt eatfatellropcoromtebaisneast[o2r5i,a2l9]a.sIsmempobrltyanetnlya,binlesosmpaetoiaflthinedseexing as andeevffiececst,ivtheemcoenasnesrvtaotiotnraocfkpdorsoitpiolneatsl o[r2d7e,2r 9o]f. eaUchnfdorrotpulnetataefltye,r caomkebyinalitmoriitaaltaiossnemtoblytheisnapbllaestform is its stphartoiaulginhdpeuxti.ngBaescaaunseefftehcetivceommebainnsattoortiraalckdrdorpolpeltestsa[r2e7,g29e]n.eUrantfeodrtusneaqtueleyn, taiaklleyy alinmditasteiorinaltloy, the rate otahfnidsanspealraliytafsollirysm, itshiseliimrtastiettehodrfoubagnyhalptyhusetis. rBiasetceliamuasitteewdthhbeiycchtohmdebrroianptaelteoatrstiawcl ahdnircohbpedlertaosspsaleertmes bgcleaenndebraaetneaddssesdemeqtbuelecendtetidaanllidyn the orderdoefteacstesdeminbtlhye(Forigduerreof1Aas)s.eAmbdlyes(iFgingutrhea1tAca).nAindcerseiganseththate ctahnroinucgrheapsuettohef sthurcohugmhipcurot voaflsvuec-hbased combminiacrtoovriaalvl ed-rboapseldetcdomevbiicneastoisritahl edrreofpolreet dneeveidceesdi.s needed
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