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We propose a robust droplet fusion and sorting method for two parallel trains of droplets that is relatively insensitive to frequency and phase mismatch. Conventional methods of droplet fusion require an extremely precise control of aqueous/oil flows for perfect frequency matching between two trains of droplets. In this work, by combining our previous two methods (i.e., droplet synchronization using railroad-like channels and manipulation of shape-dependent droplets using guiding tracks), we realized an error-free droplet fusion/sorting device for the two parallel trains of droplets. If droplet pairs are synchronized through a railroad-like channel, they are electrically fused and the fused droplets transit to a middle guiding track to flow in a middle channel; otherwise non-synchronized non-fused droplets will be discarded into the side waste channels by flowing through their own guiding tracks. The simple droplet synchronization, fusion, and sorting technology will have widespread application in droplet-based chemical or biological experiments, where two trains of the chemically or biologically treated or pre-formed droplets yield a train of 100% one-to-one fused droplets at the desired outlet channel by sorting all the non-synchronized non-fused droplets into waste outlets.

We present a multiple-droplet clustering device that can perform sequential droplet trapping and storing. Shape-dependent droplet manipulation in forward and backward flows has been incorporated to achieve high trapping and storing efficiency in a 10 × 12 array of clustering structures (e.g., storing well, storing chamber, trapping well, and guiding track). In the forward flow, flattened droplets are trapped in each trapping well. In the backward flow, the trapped droplets are released from the trapping well and follow the guiding tracks to their corresponding storing wells. The guided droplets float up out of the confining channel to the super stratum of the storing chamber due to interfacial energy and buoyancy effects. This forward/backward flow-based trapping/storing process can be repeated several times to cluster droplets with different contents and samples in the storing chambers. We expect that the proposed platform will be a valuable tool to study complex droplet-based reactions in clustered droplets.

We present a simple method of water-in-oil droplet synchronization in a railroad-like channel network. The network consisted of a top channel, a bottom channel, and ladder-like channels interconnected between the two main channels. The presence of the pressure difference between the top and bottom channels resulted in the crossflow of carrier oil through the ladder network until the pressure in each channel was balanced automatically. The proposed model and method proved the feasibility of the parallel synchronization of two trains of droplets with up to 95% synchronization efficiency. Physical parameters that could improve the efficiency were investigated with the systematic variation of the droplet length and droplet generation frequency by controlling the flow rate in each channel. Under a subtle difference in the generation frequency, an unmatched droplet sandwiched between two matched droplets in the ladder network was switched and synchronized in turn. For perfect one-to-one droplet synchronization, the droplet length and the droplet generation frequency needed to be the same for both the top and bottom channels. In addition, one-to-multiple droplet synchronization was demonstrated by matching the product of the droplet length and the droplet generation frequency for both the top and bottom channels. The proposed method provides a simple unit operation for parallel synchronization of the trains of droplets that can be easily integrated with the conventional continuous-flow droplet-based microfluidic platform.

We present a simple method of guiding, distributing, and storing of a train of shape-dependent droplets by using side flows, cavity guiding tracks, and storage chambers. The squeezing flow makes a train of flattened droplets to align to one side of the wall and the pushing flow guides it to one of the designated guiding tracks. Then the guided droplets move along the guiding track due to the lowered surface energy when they flow along the track. In addition, simultaneous droplet guiding and storing process has been demonstrated. An array of storage chambers placed in each track could store each train containing differently concentrated droplets. The proposed method will be useful for distribution of droplets for further processes or storing for multiplex, large-scale, dynamic assays over time.

In this paper, we propose a generalized serial dilution module for universal microfluidic concentration gradient generators including N cascaded-mixing stages in a stepwise manner. Desired concentrations were generated by means of controlled volumetric mixing ratios of two merging solutions in each stage. The flow rates were adjusted by controlling channel length, which is proportional to fluidic resistance in each channel. A generalized mathematical model for generating any complex concentration and output flow rate gradients is presented based on the fact that there is an analogy between microfluidic circuits and electrical circuits. The pressure drop corresponds to a voltage drop, the flow rate to an electrical current, and the flow resistance to an electrical resistance. A simple equivalent electrical circuit model was generalized, and in the model each channel segment was represented by an electrical resistance. As a result of the mathematical modelling, the only variable parameter in the generalized serial dilution module was the channel length. By the use of the generalized serial dilution module with N = 4, three types of microfluidic gradient generators for linear, logarithmic and Gaussian gradients were successfully designed and tested. The proposed strategy is capable of generating universal monotonic gradients with a single module or arbitrary gradients with multiple modules ranging from linear to complex non-linear shapes of concentration gradients as well as arbitrary output flow rate gradients in a stepwise manner. The simple universal gradient generation technology using the generalized serial dilution module will find widespread use in the greater chemical and biological community, and address many challenges of gradient-dependent phenomena.