微液滴微流控芯片:微液滴的形成、操纵和应用

微流控芯片中形成的微液滴粒径均一、可控,与传统的连续流体系相比,具有能实现试剂的快速混合、通量更高等优点.本文介绍了微流控芯片中由微通道控制的微液滴的形成、分裂、合并、混合、分选和捕获等微液滴操纵技术,以及微液滴技术在纳米粒子、聚合物微粒的合成、纳米粒子自组装、蛋白质结晶研究和DNA、细胞分析等领域的研究进展.     

Fast on-demand droplet fusion using transient cavitation bubbles

A method for on-demand droplet fusion in a microfluidic channel is presented using the flow created from a single explosively expanding cavitation bubble. We test the technique for water-in-oil droplets, which are produced using a T-junction design in a microfluidic chip. The cavitation bubble is created with a pulsed laser beam focused into one droplet. High-speed photography of the dynamics reveals that the droplet fusion can be induced within a few tens of microseconds and is caused by the rapid thinning of the continuous phase film separating the droplets. The cavitation bubble collapses and re-condenses into the droplet. Droplet fusion is demonstrated for static and moving droplets, and for droplets of equal and unequal sizes. Furthermore, we reveal the diffusion dominated mixing flow and the transport of a single encapsulated cell into a fused droplet. This laser-based droplet fusion technique may find applications in micro-droplet based chemical synthesis and bioassays.     

Fusion and sorting of two parallel trains of droplets using a railroad-like channel network and guiding tracks

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.     

Hydrodynamic control of droplet division in bifurcating microchannel and its application to particle synthesis

We describe herein a microfluidic system for active and precise control of droplet division at a bifurcation point in a microchannel. Water-in-oil or oil-in-water droplets, which were initially formed at a T-junction, were introduced into the bifurcation point, and then divided into two daughter droplets. By continuously introducing 'tuning flow' into the downstream of one of the branch channels, and by controlling the flow rates distributed into the two branch channels, the sizes of the daughter droplets could be precisely tuned. The ratio of the volumetric flow rates into the branch channels was estimated by regarding the microchannel network as a resistive circuit. In addition, we performed synthesis of monodispersed polymer particles with controlled sizes utilizing the presented system. The ability to hydrodynamically control the droplet sizes will open new possibilities not only for producing useful emulsions, but also for conducting controlled chemical and biochemical reactions in a confined space.     

Microfluidic platform for the generation of organic-phase microreactors

Rapid prototyping photolithography of a thiolene-based resin was used to fabricate microfluidic devices stable to aliphatic and aromatic organic solvents. The swelling of the cross-linked polymer matrix in various organic solvents was quantified, and the solvent resistance properties of these microfluidic devices are described. Discrete droplets of hexanes and toluene of uniform size were generated in microfluidic devices inside a water matrix containing SDS surfactant (SDS = sodium dodecyl sulfate). Variation of water and organic flow rates in the fluidic channels was used to control droplet size and separation. Droplet composition could be controlled by varying flow rates of two joined organic streams. Organic-phase synthetic reactions within the droplets were demonstrated with the bromination of alkenes inside benzene droplets.     

Deformation and breakup of micro- and nanoparticle stabilized droplets in microfluidic extensional flows

Using a microfluidic flow-focusing device, monodisperse water droplets in oil were generated and their interface populated by either 1 μm or 500 nm amine modified silica particles suspended in the water phase. The deformation and breakup of these Pickering droplets were studied in both pure extensional flow and combined extensional and shear flow at various capillary numbers using a microfluidic hyperbolic contraction. The shear resulted from droplet confinement and increased with droplet size and position along the hyperbolic contraction. Droplet deformation was found to increase with increasing confinement and capillary number. At low confinements and low capillary numbers, the droplet deformation followed the predictions of theory. For fully confined droplets, where the interface was populated by 1 μm silica particles, the droplet deformation increased precipitously and two tails were observed to form at the rear of the droplet. These tails were similar to those seen for surfactant covered droplets. At a critical capillary number, daughter droplets were observed to stream from these tails. Due to the elasticity of the particle-laden interface, these drops did not return to a spherical shape, but were observed to buckle. Although increases in droplet deformation were observed, no tail streaming occurred for the 500 nm silica particle covered droplets over the range of capillary numbers studied.     

High-throughput sorting of nanoliter droplets enabled by a sequentially addressable dielectrophoretic array

Droplet microfluidics has emerged as a powerful tool for a diverse range of biomedical and industrial applications such as single-cell analysis, directed evolution, and metabolic engineering. In these applications, droplet sorting has been effective for isolating small droplets encapsulating molecules, cells, or crystals of interest. Recently, there is an increased interest in extending the applicability of droplet sorting to larger droplets to utilize their size advantage. However, sorting throughputs of large droplets have been limited, hampering their wide adoption. Here, we report our demonstration of high-throughput fluorescence-activated droplet sorting of 1 nL droplets using an upgraded version of the sequentially addressable dielectrophoretic array (SADA), which we reported previously. The SADA is an array of electrodes that are individually and sequentially activated/deactivated according to the speed and position of a droplet passing nearby the array. We upgraded the SADA by increasing the number of driving electrodes constituting the SADA and incorporating a slanted microchannel. By using a ten-electrode SADA with the slanted microchannel, we achieved fluorescence-activated droplet sorting of 1 nL droplets at a record high throughput of 1752 droplets/s, twice as high as the previously reported maximum sorting throughput of 1 nL droplets.     

A single-cell encapsulation method based on a microfluidic multi-step droplet splitting system

Single cell analysis is of great significance to understand the physiological activity of organisms.Microfluidic droplet is an ideal analytical platform for single-cell analysis. We developed a microfluidic droplet splitting system integrated with a flow-focusing structure and multi-step splitting structures to form 8-line droplets and encapsulate single cells in the droplets. Droplet generation frequency reached1021 Hz with the aqueous phase flow rate of 1 m L/min and the oil phase flow rate of 15 mL /min. Relative standard deviation of the droplet size was less than 5% in a single channel, while less than 6% in all the8 channels. The system was used for encapsulating human whole blood cells. A single-cell encapsulation efficiency of 31% was obtained with the blood cell concentration of 2.5× 10~4cells/mL, and the multicellular droplet percentage was only 1.3%. The multi-step droplet splitting system for single cell encapsulation featured simple structure and high throughput.     

Plateau Rayleigh instability simulation

The well-known phenomena of Plateau-Rayleigh instability has been simulated using computational fluid dynamics (CFD). The breakup of a liquid film into an array of droplets on a cylindrical element was simulated using a volume-of-fluid (VOF) solver and compared to experimental observations and existing theory. It is demonstrated that the VOF method can correctly predict the breakup of thins films into an array of either axisymmetric droplets or clam-shell droplets, depending on the surface energy. The existence of unrealistically large films is precluded. Droplet spacing was found to show reasonable agreement with theory. Droplet motion and displacement under fluid flow was also examined and compared to that in previous studies. It was found that the presence of air flow around the droplet does not influence the stable film thickness; however, it reduces the time required for droplet formation. Novel relationships for droplet displacement were derived from the results.     

Sn-Pb合金颗粒异质成核及其冷却凝固行为预测

The Sn-5%Pb droplets with sizes of 150 and 185 μm were generated by Uniform Droplet Spray（UDS） under N2 atmosphere with 1. 36 μmol / L oxygen. The appearance of Sn-5%Pb droplets under the optical microscope showed that the droplets are uniform and spherical. The method employed non-adiabatic calorimetry to determine the nucleation point and undercooling of droplets. The fraction covered by oxide was calculated as a function of time and temperature. The model for heterogeneous nucleation catalyzed by oxidation on the droplet surface was developed,which was a reasonable expression of the heterogeneous nucleation and solidification behavior of Sn-Pb droplets. The CCT curves were computed using above experimental results under the heterogeneous surface nucleation of droplet. The same model can be applied to predict the heterogeneous nucleation behavior of the droplets for any type of cooling schedule. The calculation results are very reliable based on the experimental data.     

Droplet size based separation by deterministic lateral displacement-separating droplets by cell--induced shrinking

We present a novel method for passive separation of microfluidic droplets by size at high throughput using deterministic lateral displacement (DLD). We also show that droplets containing Saccharomyces cerevisiae shrink significantly during incubation while droplets containing only yeast media retain or slightly increase their size. We demonstrate the DLD device by sorting out shrunken yeast-cell containing droplets from 31% larger diameter droplets which were generated at the same time containing only media, present at a >40-fold excess. This demonstrates the resolving power of droplet separation by DLD and establishes that droplets can be separated for a biological property of the droplet contents discriminated by a change of the physical properties of the droplet. Thus suggesting that this technique may be used for e.g. clonal selection. The same device also separates 11 μm from 30 μm droplets at a rate of 12,000 droplets per second, more than twofold faster than previously demonstrated passive hydrodynamic separation devices.     

Alternating droplet generation and controlled dynamic droplet fusion in microfluidic device for CdS nanoparticle synthesis

A multifunctional and high-efficiency microfluidic device for droplet generation and fusion is presented. Through unique design of the micro-channels, the device is able to alternately generate droplets, generating droplet ratios ranging from 1 ratio 5 to 5 ratio 1, and fuse droplets, enabling precise chemical reactions in several picoliters on a single chip. The controlled fusion is managed by passive control based on the channel geometry and liquid phase flow. The synthesis of CdS nanoparticles utilizing each fused droplet as a microreactor for rapid and efficient mixing of reagents is demonstrated in this paper. Following alternating droplet generation, the channel geometry allows the exclusive fusion of alternate droplets with concomitant rapid mixing and produces supersaturated solution of Cd2+ and S2- ions to form CdS nanoparticles in each fused droplet. The spectroscopic properties of the CdS nanoparticles produced by this method are compared with CdS prepared by bulk mixing.     

Physical/chemical separations in the break-up of highly charged droplets from electrosprays

Highly-charged droplets, as formed by an electrospray process, are known to undergo asymmetric fission to form smaller droplets. We have observed a chemical and physical separation phenomenon that occurs in the droplet break-up process and is related to a compound's surface activity in solution. Two experimental approaches demonstrated that the smaller satellite droplets and the progeny droplets generated by the spray formation and asymmetric fission processes to be surfactant-enriched. These smaller droplets were also effectively separated from the larger primary and residual droplets because of their smaller inertia and high surface charge density, and a region attributed to the initially formed smaller satellite droplets was found to be strikingly confined in a narrow periphery region of the electrospray. The phenomenon may have utility for chemical separations and have significant implications for the sensitivity and selectivity of electrospray ionization-mass spectrometry.     

Microfluidic separation of satellite droplets as the basis of a monodispersed micron and submicron emulsification system

Emulsions are widely used to produce sol-gel, drugs, synthetic materials, and food products. Recent advancements in microfluidic droplet emulsion technology has enabled the precise sampling and processing of small volumes of fluids (picoliter to femtoliter) by the controlled viscous shearing in microchannels. However the generation of monodispersed droplets smaller than 1 microm without surfactants has been difficult to achieve. Normally, the generation of satellite droplets along with parent droplets is undesirable and makes it difficult to control volume and purity of samples in droplets. In this paper, however, several methods are presented to passively filter out satellite droplets from the generation of parent droplets and use these satellite droplets as the source for monodispersed production of submicron emulsions. A passive satellite droplet filtration system and a dynamic satellite droplet separation system are demonstrated. Satellite droplets are filtered from parent droplets with a two-layer channel geometry. This design allows the creation and collection of droplets that are less than 100 nm in diameter. In the dynamic separation system, satellite droplets of defined sizes can be selectively separated into different collecting zones. The separation of the satellite droplets into different collecting zones correlates with the cross channel position of the satellite droplets during the breakup of the liquid thread. The delay time for droplets to switch between the different alternating collecting zones is nominally 1 min and is proportional to the ratio of the oil shear flows. With our droplet generation system, monodispersed satellite droplets with an average radius of 2.23 +/- 0.11 microm, and bidispersed secondary and tertiary satellite droplets with radii of 1.55 +/- 0.07 microm and 372 +/- 46 nm respectively, have been dynamically separated and collected.     

Delayed coalescence behavior of droplets with completely miscible liquids

Because of capillary forces, sessile droplets usually fuse instantaneously after contact. We find however a delay of the droplet fusion by many seconds if the droplets consist of different but completely miscible liquids. After the initial contact, the main bodies of the droplets remain separated, connected only through a shallow conduit with a flow from the low to the high surface tension liquid. Sporadically, this connecting film can thicken with turbulent or pulsating flows. The droplets will finally fuse when the flow has sufficiently reduced the difference in composition and surface tension. We present calculations which explain this delayed droplet fusion with the compensation of the fusion-promoting capillary pressure by a droplet-separating dynamic pressure caused by the flow between the droplets. Droplets with high contact angles fuse instantaneously. In this case, no separation-stabilizing dynamic pressure can build up because the interdroplet flow becomes turbulent.     

Surface modification of droplet polymeric microfluidic devices for the stable and continuous generation of aqueous droplets

Droplet microfluidics performed in poly(methyl methacrylate) (PMMA) microfluidic devices resulted in significant wall wetting by water droplets formed in a liquid-liquid segmented flow when using a hydrophobic carrier fluid such as perfluorotripropylamine (FC-3283). This wall wetting led to water droplets with nonuniform sizes that were often trapped on the wall surfaces, leading to unstable and poorly controlled liquid-liquid segmented flow. To circumvent this problem, we developed a two-step procedure to hydrophobically modify the surfaces of PMMA and other thermoplastic materials commonly used to make microfluidic devices. The surface-modification route involved the introduction of hydroxyl groups by oxygen plasma treatment of the polymer surface followed by a solution-phase reaction with heptadecafluoro-1,1,2,2-tetrahydrodecyl trichlorosilane dissolved in fluorocarbon solvent FC-3283. This procedure was found to be useful for the modification of PMMA and other thermoplastic surfaces, including polycyclic olefin copolymer (COC) and polycarbonate (PC). Angle-resolved X-ray photoelectron spectroscopy indicated that the fluorination of these polymers took place with high surface selectivity. This procedure was used to modify the surface of a PMMA droplet microfluidic device (DMFD) and was shown to be useful in reducing the wetting problem during the generation of aqueous droplets in a perfluorotripropylamine (FC-3283) carrier fluid and could generate stable segmented flows for hours of operation. In the case of PMMA DMFD, oxygen plasma treatment was carried out after the PMMA cover plate was thermally fusion bonded to the PMMA microfluidic chip. Because the appended chemistry to the channel wall created a hydrophobic surface, it will accommodate the use of other carrier fluids that are hydrophobic as well, such as hexadecane or mineral oils.     

Impact of diblock copolymers on droplet coalescence, emulsification, and aggregation in immiscible homopolymer blends

Using rheo-optical techniques, we investigated the impact of interfacial wetting of symmetric diblock copolymers (BCPs) on the coalescence and aggregation of polydimethylsiloxane (PDMS) droplets in immiscible polyethylene-propylene (PEP) homopolymers. Anionic polymerization was used to synthesize well-defined matrix homopolymers and symmetric 16 kg/mol-to-16 kg/mol PDMS-b-PEP diblock copolymers with low polydispersity (PDI ≈ 1.02) as characterized with size exclusion chromatography and nuclear magnetic resonance spectroscopy. Blends were formulated to match the viscosities between the droplets and the matrix. Moreover, molecular weights of these components were varied to ensure that the inner block of the copolymer inside the droplet was collapsed and dry, whereas the outer block of the copolymer outside of the droplet was stretched and wet. Droplet breakup and coalescence as well as interfacial tensions were measured using rheo-optical experiments with Linkam shearing stage and an optical microscope. Subsequent to droplet breakup at high shear rates, we found that the BCPs mitigated shear-induced coalescence at lower shear rates. Based on surface tension measurements, the stretching of the BCP increased in lower molecular weight matrices, causing the droplet surface to saturate at lower coverage in line with theoretical predictions. Droplet aggregation was detected with further reductions in shear rate, which was attributed to the dewetting or the expulsion of the matrix from a saturated brush. Ultimately, the regions of droplet coalescence and aggregation were scaled by balancing the forces of shear with those due to the attraction between BCP-coated droplets.     

Concentration and binary separation of micro particles for droplet-based digital microfluidics

This paper describes a concept of concentration and binary separation of particles and its experimental confirmations for digital microfluidics where droplets are driven by the mechanism of electrowetting-on-dielectric (EWOD). As a fundamental separation unit, a binary separation scheme is developed, separating two different types of particles in one droplet into two droplets, one type each. The separation scheme consists of three distinctive steps, each with their own challenges: (1) isolate two different types of particles by electrophoresis into two regions inside a mother droplet, (2) physically split the mother droplet into two daughter droplets by EWOD actuation so that each type of particle is concentrated in each daughter droplet, and (3) free the daughter droplets from the separation site by EWOD to ready them for follow-up microfluidic operations. By applying a similar procedure to a droplet containing only one type of particle, two daughter droplets of different particle concentrations can be created. Using negatively charged carboxylate modified latex (CML) particles, 83% of the total particles are concentrated in a daughter droplet. Successful binary separation is also demonstrated using negatively charged CML particles and no-charge-treated polystyrene particles. Despite the undesired vortex developed inside the mother droplet, about 70% of the total CML particles are concentrated in one daughter droplet while about 70% of the total polystyrene particles are concentrated in the other daughter droplet.     

Characterization of microdroplets using optofluidic signals

We develop a simple method to determine the microdroplet features in a microfluidic chip fabricated by conventional soft lithography. Different sizes of microdroplets are generated through a typical microfluidic T-junction by adjusting the flow rates of the two immiscible liquids. Droplet size and content can be determined by monitoring the optofluidic signals reflected at the fluid-polydimethylsiloxane (PDMS) interface. The demonstrated droplet characterization system can be readily integrated with other microfluidic networks, making it promising for biochemical and biosensing applications.     

Efficient Generation of Microdroplets Using Tail Breakup Induced with Multi-Branch Channels

In recent years, research on the application of microdroplets in the fields of biotechnology and chemistry has made remarkable progress, but the technology for the stable generation of single-micrometer-scale microdroplets has not yet been established. In this paper, we developed an efficient and stable single-micrometer-scale droplet generation device based on the fragmentation of droplet tails, called “tail thread mode”, that appears under moderate flow conditions. This method can efficiently encapsulate microbeads that mimic cells and chemical products in passively generated single-micrometer-scale microdroplets. The device has a simple 2D structure; a T-junction is used for droplet generation; and in the downstream, multi-branch channels are designed for droplet deformation into the tail. Several 1–2 µm droplets were successfully produced by the tail’s fragmentation; this continuous splitting was induced by the branch channels. We examined a wide range of experimental conditions and found the optimal flow rate condition can be reduced to one-tenth compared to the conventional tip-streaming method. A mold was fabricated by simple soft lithography, and a polydimethylsiloxane (PDMS) device was fabricated using the mold. Based on the 15 patterns of experimental conditions and the results, the key factors for the generation of microdroplets in this device were examined. In the most efficient condition, 61.1% of the total droplets generated were smaller than 2 μm.