Breakup dynamics for high‐viscosity droplet formation in a flow‐focusing device: Symmetrical and asymmetrical ruptures

The breakup mechanism of high‐viscosity thread for droplet formation in a flow‐focusing device is investigated using a high‐speed digital camera. Aqueous solution of 89.5%‐glycerol is used as the dispersed phase, while silicone oil as the continuous phase. The breakup process of the dispersed thread presents two categories: symmetrical rupture and asymmetrical rupture. Furthermore, the rupture behavior could be divided into two stages: the squeezing stage controlled by the squeezing pressure and the pinch‐off stage controlled by viscous stresses of both phases and surface tension. Specifically, it suggests that the differences in the shape of the liquid–liquid interface and the dynamics in the two breakup processes are caused by the disparity of the strain field at the point of detachment. Moreover, the thinning rate and the dynamics of the dispersed thread change with the viscosity of the continuous phase, but are less dependent of the flow rate of the continuous phase. © 2015 American Institute of Chemical Engineers AIChE J, 62: 325–337, 2016     

Self‐similar breakup of viscoelastic thread for droplet formation in flow‐focusing devices

The self‐similarity of the breakup of viscoelastic dispersed thread for droplet formation in flow‐focusing devices is investigated experimentally. A high‐speed camera is used to capture the evolution and angles of the cone‐shaped liquid‐liquid interface. The self‐similar profiles for the liquid‐liquid interface are obtained by normalizing the interface with the minimum width of the dispersed thread. The breakup of the dispersed thread transfers from a self‐similar power law scaling stage with an exponent of 0.36 to a self‐similar exponential scaling stage. The asymptotic cone angles prior to final breakup are consistent with the value of 125.5° and 151°, respectively. The viscoelasticity inhibits the development of finite‐time singularity for the breakup of the liquid‐liquid interface at microscale, similar to the capillary breakup at macroscale. The results demonstrate that the breakup of the viscoelastic dispersed thread for droplet formation exhibits self‐similarity at microscale. © 2017 American Institute of Chemical Engineers AIChE J, 2017     

Intra-Species Bacterial Quorum Sensing Studied at Single Cell Level in a Double Droplet Trapping System

In this paper, we investigated the intra-species bacterial quorum sensing at the single cell level using a double droplet trapping system. Escherichia coli transformed to express the quorum sensing receptor protein, LasR, were encapsulated in microdroplets that were positioned adjacent to microdroplets containing the autoinducer, N-(3-oxododecanoyl)- l-homoserine lactone (OdDHL). Functional activation of the LasR protein by diffusion of the OdDHL across the droplet interface was measured by monitoring the expression of green fluorescent protein (GFP) from a LasR-dependent promoter. A threshold concentration of OdDHL was found to induce production of quorum-sensing associated GFP by E. coli. Additionally, we demonstrated that LasR-dependent activation of GFP expression was also initiated when the adjacent droplets contained single E. coli transformed with the OdDHL synthase gene, LasI, representing a simple quorum sensing circuit between two droplets.     

微通道内卫星液滴生成机理与惯性分离机制

由于非线性动态特征,液液界面破裂过程常伴随卫星液滴的产生,对基于液滴的微流体技术生产液滴的均一性和精准化目标提出了挑战。阐释了微流体界面失稳的复杂动力学特征,剖析了界面失稳的影响因素,并分析了伴随界面失稳而产生的卫星液滴的现象与原理。结合惯性微流体新概念,总结了卫星液滴的惯性分离机制。展望了卫星液滴生成-惯性微流体分离一体化及其并行化数目放大的构想。相关工作的开展,有利于实现微流体技术生产单分散性液滴的精准化目标,为微流体与复杂流体相关的界面动力学行为与调控夯实基础。     

A high-throughput screen for porphyrin metal chelatases: application to the directed evolution of ferrochelatases for metalloporphyrin biosynthesis

Porphyrins are of particular interest in a variety of applications ranging from biocatalysis and chemical synthesis to biosensor and electronic technologies as well as cancer treatment. Recently, we have developed a versatile system for the high-level production of porphyrins in engineered E. coli cells with the aim of diversifying substitution patterns and accessing porphyrin systems not readily available through chemical synthesis. However, this approach failed to produce significant amounts of the metalloporphyrin in vivo from overproduced protoporphyrin due to insufficient metal insertion. Therefore, we systematically assessed the activity of the B. subtilis ferrochelatase in vivo and in vitro. A true high-throughput-screening approach based on catalytic in vivo ferrochelatase activity was developed by using fluorescence-activated cell sorting (FACS). This assay was used to screen a library of 2.4 x 10(6) ferrochelatase mutants expressed in protoporphyrin-overproducing recombinant E. coli cells. Several selected protein variants were purified, and their improved catalytic activity was confirmed in vitro. In addition to ferrochelatase activity, metal transport into E. coli was identified as another limitation for in vivo heme overproduction. Overexpression of the metal transporter zupT as part of the assembled pathway increased the overall metalloporphyrin production twofold. This report represents the most exhaustive in vitro evolution study of a ferrochelatase and demonstrates the effectiveness of our novel high-throughput-screening system for directed evolution of ferrochelatases based on their catalytic activity.     

变截面通道内流动聚焦微液滴生成数值研究

化学分析研究的不断深入,对液滴微流控精度提出了更高的要求。设计了不同孔径尺寸的流动聚焦通道模型,模拟油-水两相流动剪切微液滴生成过程。采用Level Set方法处理两相流动界面,研究了流动聚焦模型下通道尺寸和油-水两相流量比对生成液滴尺寸的影响。研究结果对实现微液滴生成过程的精准操控具有一定的参考意义。     

Ultrahigh‐Throughput FACS‐Based Screening for Directed Enzyme Evolution

Directed enzyme evolution has proven to be a powerful tool for improving a range of properties of enzymes through consecutive rounds of diversification and selection. However, its success depends heavily on the efficiency of the screening strategy employed. Fluorescence‐activated cell sorting (FACS) has recently emerged as a powerful tool for screening enzyme libraries due to its high sensitivity and its ability to analyze as many as 10⁸ mutants per day. Applications of FACS screening have allowed the isolation of enzyme variants with significantly improved activities, altered substrate specificities, or even novel functions. This review discusses FACS‐based screening for enzymatic activity and its potential application for the directed evolution of enzymes, ribozymes, and catalytic antibodies.     

Droplets formation and merging in two-phase flow microfluidics

Two-phase flow microfluidics is emerging as a popular technology for a wide range of applications involving high throughput such as encapsulation, chemical synthesis and biochemical assays. Within this platform, the formation and merging of droplets inside an immiscible carrier fluid are two key procedures: (i) the emulsification step should lead to a very well controlled drop size (distribution); and (ii) the use of droplet as micro-reactors requires a reliable merging. A novel trend within this field is the use of additional active means of control besides the commonly used hydrodynamic manipulation. Electric fields are especially suitable for this, due to quantitative control over the amplitude and time dependence of the signals, and the flexibility in designing micro-electrode geometries. With this, the formation and merging of droplets can be achieved on-demand and with high precision. In this review on two-phase flow microfluidics, particular emphasis is given on these aspects. Also recent innovations in microfabrication technologies used for this purpose will be discussed.     

Dynamic formation and scaling law of hollow droplet with gas/oil/water system in dual‐coaxial microfluidic devices

Based on the one‐step microfluidic method of producing hollow droplet with thin film, this article studies the effect of water and oil flow rate, gas pressure, and viscosity of aqueous phase on the dynamic formation and size of hollow droplet by analyzing large amounts of data acquired automatically. The results show that the filling stage of hollow droplet is similar to that of microbubble formation, while the necking stage is similar to that of droplet formation process. Furthermore, based on the data and mathematical model describing droplet formation mechanism, a filling stage model including Capillary number of continuous phase is developed. Considering the dynamic interface breakup and displacement of droplet in necking stage, a necking stage model is developed. The results show that the model results considering filling and necking stage fit well with the experimental data, and the relative error is less than 5%. Finally, the same model with parameters is used to predict the size of hollow droplet with other systems and devices, and the model is proved to be relative precise in our experimental conditions. The results presented in this work provide a more in‐depth understanding of the dynamic formation and scaling law of hollow droplet with G/L/L systems in microfluidic devices. © 2017 American Institute of Chemical Engineers AIChE J, 64: 730–739, 2018     

Electrohydrostatics of capillary switches

A capillary switch is a system of two liquid drops, one sessile and the other pendant, obtained by overfilling a hole of radius R in a plate. When surface tension dominates gravity, the equilibrium shapes of the drops are spherical sections of equal radii. If the combined volume of the top V_T and bottom V_B drops exceeds , the system has three equilibrium states of which two are stable. This bistability is exploited in applications by toggling the system between its two stable states. Here, we examine the effectiveness of using an electric field for toggling. Bifurcation diagrams are obtained that depict how the system's response varies with applied field strength E, and show loss of stability at turning points and the possibility of hysteresis. A phase diagram in space is presented to readily infer when an electric field is an effective means for toggling. © 2014 American Institute of Chemical Engineers AIChE J, 60: 1451–1459, 2014     

Activity‐Fed Translation (AFT) Assay: A New High‐Throughput Screening Strategy for Enzymes in Droplets

There is an increasing demand for the development of sensitive enzymatic assays compatible with droplet‐based microfluidics. Here we describe an original strategy, activity‐fed translation (AFT), based on the coupling of enzymatic activity to in vitro translation of a fluorescent protein. We show that methionine release upon the hydrolysis of phenylacetylmethionine by penicillin acylase enabled in vitro expression of green fluorescent protein. An autocatalytic setup where both proteins are expressed makes the assay highly sensitive, as fluorescence was detected in droplets containing single PAC genes. Adding a PCR step in the droplets prior to the assay increased the sensitivity further. The strategy is potentially applicable for any activity that can be coupled to the production of an amino acid, and as the microdroplet volume is small the use of costly reagents such as in vitro expression mixtures is not limiting for high‐throughput screening projects.     

微通道内液滴/气泡破裂动力学分析

微化学工程与技术是现代化学工程学科的前沿领域。微通道内液滴及气泡破裂动力学是决定多相过程并行微通道数目放大的基础与难点。破裂流型转换条件、界面动力学和尺寸调控等三方面是微通道内液滴与气泡破裂动力学的主要研究对象。讨论了对称微通道、非对称微通道、多级微通道、旁路微通道、含有障碍物的微通道内气泡和液滴破裂行为及影响因素,指出了目前微尺度下气泡与液滴破裂行为相关研究工作存在的不足,并对该领域未来的发展进行了展望。     

高密度差体系中单液滴运动的数值模拟

采用VOSET方法捕捉液液两相运动界面,对高密度差体系中单液滴的运动进行数值模拟,分析不同时刻的表面图和流线图及瞬态速度变化图,研究了单液滴运动过程的速度变化及形变。研究结果表明：液滴形变随密度差和液滴粒径以及界面张力减小而变得剧烈;液滴上升速度随密度差和界面张力的增大而增大;液滴速度振荡幅度随粒径减小而减小;液滴形变的频率随界面张力增大或粒径减小而变小。