Microfluidic techniques for synthesizing particles

Adam R. Abate, Sebastian Seiffert, Andrew S. Utada, Anderson Shum, Rhutesh Shah, Julian Thiele, Wynter J. Duncanson, Alirezza Abbaspourad, Myung Han Lee, Ilke Akartuna, Daeyeon Lee, Assaf Rotem, David A. Weitz
a b

School of Engineering and Applied Sciences/Department of Physics, Harvard University, Cambridge, Massachusetts, USA. Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, Pennsylvania, USA.

Introduction Microfluidic devices are networks of micron scale channels that are integrated together to perform functions. The functions typically performed can be divided into two broad classes. In one class, devices perform chemical and biological assays. Cells, beads, and other reagents are introduced into the device, and the channels merge, mix, and split them, as needed for the reactions. This allows a variety of chemical and biological reactions to be executed with high precision, for combinatorial chemical screens, proteomics studies, genetic sequencing, and directed evolution [1–3]. In the second class of functions, drop formation devices are used to synthesize microparticles [4–7]. Solutions or melts of monomers or crosslinkable polymers are introduced into the device, along with an immiscible carrier phase; the devices disperse these solutions into equally sized micro-droplets, which can then be solidified by polymerization, crosslinking, or crystallization, thereby producing solid, monodisperse microparticles. The principal advantage of this microfluidic synthesis is that while the chemical composition of the particles is obtained by selecting which solutions to introduce into the device, the final particle structure is obtained by the fluidics. For example, a device that forms anisotropic particles can do so with a variety of fluids, yielding particles with a range of distinct chemical properties but identical structure. This ability to independently select structure and chemical composition is a significant advance over traditional bulk synthesis approaches, since in these cases the final structure is intrinsically linked to the chemistry in the reactor, for example, as it is particle synthesis by emulsion polymerization [8–10]. This greatly broadens not only the kinds of materials that can be formed into particles, but also the kinds of structures that can be formed, from simple monodisperse plastic spheres to anisotropic magnetic hydrogels, non-spherical Janus particles, and core-shell capsules of a variety of compositions. In this chapter, we provide an overview of the kinds of particles that can be synthesized with microfluidic techniques. We describe the two dominant types of microfluidic devices that are used to synthesize particles, beginning with glass capillary microfluidics and ending with lithographically fabricated poly(dimethylsiloxane) (PDMS) devices. Glass capillary devices afford several unique advantages for forming particles, including high chemical resistance and an ideal coaxial flow focusing geometry; this enables creation of particles of a wide range of compositions and structures. By contrast, lithographically fabricated PDMS devices do not match capillaries in these regards, but afford other advantages that make them superior for certain applications. This includes the ability to tailor channel networks to overcome specific challenges. As we will show, this

allows them to create new kinds of particles. The inherently parallel lithographic fabrication also allows these devices to be replicated in large numbers, making them attractive for largescale synthesis applications.

SECTION 1: Glass capillary microfluidics Single emulsion particle templating The first step to forming particles with microfluidic devices is to form monodisperse populations of micro-droplets. The principle of drop formation in microfluidic devices can be explained using a water faucet as an example. If we turn on a faucet at a low flow rate, water drips out one drop at a time. The drop size is a result of the balance between the surface forces of the hanging drop and its weight, and therefore depends on the surface tension of the fluid and the size of the faucet. Since both the surface tension and the faucet size are constant, all drops dripping from a faucet exhibit a narrow size distribution. However, if we gradually increase the flow rate through the faucet, a thin water stream, or a jet is formed. Although the jet eventually breaks up into drops as well, these have a larger size range [11–14]. The same principle can be employed in microfluidic channels that have sizes on the order of tens of micrometers. One significant difference between drop formation at a faucet and in microfluidic devices is that in the former case drops are formed in air, whereas in the latter case drops are formed in another immiscible liquid. Capillary microfluidics presents a way to controllably generate drops of one liquid in another immiscible liquid in devices that consist of coaxial assemblies of glass capillaries. One of the inherent advantages of these devices is that since they are made from glass, their wettability can be easily and precisely controlled by a surface reaction with an appropriate surface modifier. For example, a quick treatment of octadecyltrimethoxysilane will render the glass surface hydrophobic, whereas a treatment of 2-[methoxy(polyethyleneoxy)propyl]trimethoxysilane will make the surface more hydrophilic. An additional benefit of using glass is that the devices are both chemically resistant and rigid. Lastly and perhaps most importantly, these devices offer the distinct capability of creating truly three-dimensional flows, which is critical for some of the applications that we describe later. To build these devices, we begin with a circular glass capillary with an outer diameter of 1–2 mm. This capillary is heated and pulled using a pipette puller to create a tapered geometry that culminates in a fine orifice; this is our “faucet”. The precisely pulled circular capillary is carefully inserted into a square glass capillary to form a simple microfluidic device. We

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ensure coaxial alignment of the two capillaries by choosing the tubes such that the outer diameter of the circular capillary is the same as the inner dimension of the square capillary. Flowing one fluid inside the circular capillary while flowing a second fluid through the square capillary in the same direction results in a three-dimensional coaxial flow of the two fluids, as illustrated in Fig. 1a. This is known as coflow geometry [15, 16].

We observe two distinct dripping-to-jetting transitions when we vary the two flow rates, respectively, which is due to a different balance of forces on the jetting liquid. The first transition is driven by the flow rate of the outer fluid; as it is increased, drops formed at the tip decrease in size until the emerging fluid is finally stretched into a jet. At this point, drop breakup occurs downstream at the end of the thin jet, as shown in Fig. 1c. The second transition is driven by the flow rate of the inner fluid; as it is increased, the dripping drop is pushed downstream and ultimately pinches-off from the end of the resultant jet, shown in Fig. 1d.

Fig 2 Dependence of the transition between dripping and jetting on the capillary number of the outer flow and the Weber number of the inner flow of a coflow microcapillary device. Squares and diamonds: ηin/ηout = 0.01, with slightly different geometries; hexagons and circles: ηin/ηout = 0.1, with slightly different geometries; pentagons: ηin/ηout = 1; triangles, ηin/ηout = 10. Reprinted from [16]. Copyright 2007 American Physical Society.

Fig 1 Formation of single emulsions in a coflow microfluidic device. (a) Schematic of a coflow microcapillary device for making droplets. Arrows indicate the flow direction of fluids and drops. (b) Image of drop formation at low flow rates (dripping regime). (c) Image of a narrowing jet generated by increasing the flow rate of the continuous fluid above a threshold value, while keeping the flow rate of the dispersed phase constant. (d) Image of a widening jet generated by increasing the flow rate of the dispersed fluid above a threshold value, while keeping the flow rate of the continuous phase constant. (e) Monodisperse droplets formed using a microcapillary device. Parts (b), (c), and (d) are reprinted from [16]. Copyright 2007 American Physical Society.

When both fluids flow at low rates, individual monodisperse drops are formed periodically at the tip of the capillary orifice, in a process termed dripping, shown in Fig. 1b [15–17]. In the dripping regime, the drop at the end of the capillary tube experiences two competing forces: viscous drag pulling it downstream and forces due to surface tension holding it to the capillary. Initially, surface tension dominates but as the attached droplet grows, drag forces eventually become comparable; this is when the droplet breaks off and is carried away by the flow of the continuous phase [17]. If we increase the flow rate of either fluid beyond a certain critical limit, the result is a jet, which is a long stream of the inner fluid with drops forming downstream, as shown in Fig. 1c and 1d. Typically, these drops have a broader size distribution than drops formed from dripping because the point at which the drops separate from the jet changes with each drop.

To elucidate these processes, we plot them on a single phase diagram based on the relevant force balance that induces the transition. We find that the behavior of the system is determined by two non-dimensional numbers: the capillary number and the Weber number. The capillary number reflects the balance between the drag of the outer fluid pulling the drop downstream and surface tension forces that resist the flow in the jet as pinch-off occurs. The Weber number reflects a balance between inertial forces of the inner liquid pushing the drop downstream and, again, the surface tension forces resisting the flow. Since both numbers describe a balance between an applied force and surface tension forces, the boundary between dripping and jetting occurs when either number, or their sum, is approximately equal to 1, as shown in Fig. 2.

Fig 3 Schematic of a flow-focusing microcapillary device for making droplets.

An alternate geometry for drop formation in capillary devices is the flow-focusing geometry [18, 19]. In contrast to coflow capillary devices, the two fluids are introduced from the two ends of the same square capillary, from opposite directions. The inner fluid is hydrodynamically focused by the outer fluid through the narrow orifice of the tapered round capillary, as

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drop formation occurs as soon as the inner fluid enters the circular orifice. 4a. An advantage of this method is that it allows us to make monodisperse drops with sizes smaller than that of the orifice.N’-tetramethyethylenediamine (TEMED). these microgels are being extensively evaluated for controlled delivery of water soluble drugs. As a result. it diffuses into the aqueous droplets triggering a redox reaction which causes simultaneous polymerization and gelation of monomers dissolved in the droplets. or magnetic nanoparticles. This feature is useful for making small droplets (~1 –5 μm in diameter). The scalebar in Panel b denotes 100 m. a crosslinker. and initiator. This is typically done by suspending such materials in the aqueous pre-gel mixture prior to emulsification. (b) Reproduced from [22]. Functional materials can be embedded inside such microgels to impart additional properties. Cross-sectional views at different points along the device length are shown in the second row. Hence. whereas under jetting conditions it occurs further downstream. shown in Figs. We begin with an aqueous pre-gel mixture which is emulsified in an oil phase using the device. we use a glass capillary single emulsion device [21. Such PNIPAm microgels swell and shrink reversibly in response to changes in temperature. N. We demonstrate this by fabricating microgels that are complexed with polymer particles. N’ methylene-bis-acrylamide (BIS). which is close to the human body temperature. N. Copyright 2007 Wiley-VCH Verlag GmbH & Co. To make these microgels. their diameter reduces to less than half the original diameter. essentially. (a) (b) Fig 4 (a) Schematic illustration of a capillary-based microfluidic device for fabricating monodisperse PNIPAm microgels. A reaction initiator.N’. as they are not chemically bonded to the polymer network but are only physically trapped within it. 3 . dissolved in kerosene. ammonium persulfate. and fluid C is the same oil as fluid B but contains a reaction accelerator that is both water. by adjusting flow rates. Copyright 2008 The Royal Society of Chemistry. TEMED is soluble in both oil as well as water. is introduced into the device through the left end of the right square capillary. KGaA. An aqueous phase containing the monomer. 22]. Low polydispersity of PNIPAm microgels is desirable for drug delivery applications as it could lead to narrow distribution of drug loading levels and uniform release kinetics. 22]. This size change occurs around 32 °C. The size change is reversible and reproducible over several cycles. when heated. hydrodynamically focuses the aqueous phase into the collection tube where the aqueous phase breaks into monodisperse drops. N-isopropylacrylamide.and oil-soluble. especially those from a particulate suspension. As an example of this “templating” process. shown in Fig 5d [21. and hence. pumped in from the right end of the left square capillary. kerosene. The use of a capillary with a larger orifice minimizes the probability of such tip clogging by the suspended particles or any entrapped debris. Fluid A is an aqueous suspension containing the monomer. Under dripping conditions. where the particles may clog the orifice in the coflow geometry [20]. is introduced from the left end of the left square capillary. Incorporation of voids improves the kinetics of size change of the microgels since the kinetics is a function of the rate of diffusion of water through the microgels. quantum dots. These spheres can then be solidified to produce particles of the same size and shape. we make monodisperse microgels that consist of a crosslinked network of poly(N-isopropylacrylamide). Templating thermoresponsive microgels from single emulsions The ability to form droplets with a microfluidic device allows one. 5(a–c) [23]. thereby creating uniform microgels. The resultant microgels exhibit excellent thermal response. Alternatively. to structure fluids – to disperse a continuous fluid into a series of equally-sized liquid spheres.shown in Fig. (a) Reproduced from [21]. it can be tuned by controlling drop size. microgels with embedded voids can also be fabricated by embedding polymeric particles inside microgels and subsequently dissolving the embedded particles. swollen with water.).N. which increases upon incorporation of voids. The accelerator diffuses into the drops and polymerizes the monomers to form monodisperse microgels. The monodisperse drops are then gelled just after formation. An oil. (b) Size change of PNIPAm microgels in water triggered by changing the temperature. The resulting monodisperse PNIPAm microgels are subsequently washed and redispersed in water. Since the size of the microgels follows the size of the emulsified drops. 3. A schematic of the device employed for fabricating PNIPAm microgels is shown in Fig. PNIPAm. as shown in Fig. fluid B is an oil. and a reaction initiator. 1b. The addition of such materials has no detrimental effect on the thermosensitive behavior of the microgels. crosslinker.

As an example. while the viscosity of the polymer solution is not too high. a small amount of glutaraldehyde is added to the aqueous mixture. we demonstrate the fabrication of monodisperse colloidosomes. Anionic magnetic beads are added to the aqueous mixture of the PNIPAm microgels and other monomers. as shown in Fig 7c. they can be of immense potential in applications that require targeted pulsed-release of active materials. (c) A bright-field microscope image of a microgel containing 10nm magnetic particles. Janus particles are biphasic particles with two sides of different composition and functionality. 7d. Once formed. Copyright 2007 Wiley-VCH Verlag GmbH & Co. (d) A bright-field microscope image of a microgel with embedded voids. alternatively known as supraparticles. (a) A fluorescence microscope image of a microgel containing fluorescently labeled 1. this allows each to be controlled independently. Glutaraldehyde molecules. We prepare these precursors by copolymerizing Nisopropylacrylamide and a DMMI-functionalized acrylamidederivative in a free-radical reaction in water. these experimental conditions guarantee that a space-filling polymer network can be formed inside each droplet. 7a. as shown in Fig. The overall schematic is presented in Fig. an intermediate range above the threshold for coil overlap. serve as connecting links between the amine-functionalized microgels through an amine-aldehyde condensation reaction. Reproduced from [21] and [23]. (b) A fluorescence microscope image of a microgel containing 19-nm quantum dots. An aqueous suspension of amine-functionalized submicrometer-sized PNIPAm microgels is emulsified in an oil using a single-emulsion microfluidic device. forming Janus supraparticles with a PNIPAm microgel-rich side and a PAAmrich side. Such colloidosomes exhibit thermosensitive behavior similar to that displayed by their constituent microgels: when the temperature is increased above the phase-transition temperature of PNIPAm. This aqueous mixture is emulsified in an oil and heated at 65 °C in an oven. ce*. c*. higher order particles. as shown in Fig. KGaA. 7b. Such magnetically anisotropic particles can be used to make magnetically actuated displays or other applications that require directional orientation or transportation of particles.(a) (b) (c) (d) 50μm Fig 5 Microgels with embedded materials.m diameter polystyrene particles. Upon heating. Since the microgel particles are cationic. and we control the molecular weight of the resultant copolymers by performing this polymerization in the presence of sodium formate [26]. Thus. as shown in Fig. Templating colloidosomes from single emulsions Besides just gelling a pre-particle droplet. Prior to emulsification. Prior to emulsification. We also dissolve 10 wt% acrylamide in the microgel suspension along with a crosslinker and a photoinitiator. microgel particles are obtained by droplet gelation. We fabricate monodisperse Janus supraparticles with a PNIPAm microgelrich side and a polyacrylamide (PAAm) rich side [28]. thus enabling microgel particles to be formed with well-controlled composition and functionality [26]. The colloidal PNIPAm microgels assemble at the oil- water interface within the emulsion droplets due to the presence of hydrophobic isopropyl groups and hydrophilic acrylamide groups. The advantage of this approach of particle fabrication is that it separates the polymer synthesis from the particle gelation. can be fabricated by controlling the assembly of nanoparticles/colloidal particles within microfluidically generated single emulsion droplets. which roughly translates to an 80% decrease in volume. a moiety that can be selectively transformed into dimers by UV irradiation [24. After forming pre-microgel droplets. yet below the onset of chain entanglement. Similar microgels can also be generated through controlled crosslinking of pre-fabricated PNIPAm polymer chains. To demonstrate this concept. the weakly associated PNIPAm microgel aggregate shrinks and becomes compacted on one side of the droplets by pushing the acrylamide containing water to the other side. as illustrated by the incorporation of magnetic nanoparticles in the microgel-rich side of the particles. Interlinking of the microgels at the oilwater interface results in the formation of colloidosomes. The acrylamide monomer is then polymerized and cross-linked by exposure to ultraviolet (UV) radiation. We emulsify aqueous precursor solutions with polymer concentrations in the semidilute unentangled regime. and are thus trapped only in the PNIPAm phase of the Janus particles. 6b. owing to their two reactive sites each. the magnetic beads electrostatically bind to the surface of the microgels. as shown in Fig. 6a. This is accomplished by emulsifying an aqueous suspension of aminefunctionalized microgels using a single-emulsion microfluidic device. microcapsules with a shell composed of tightly packed colloidal particles. 4 . the diameter of the colloidosomes decreases by 42%. achieved through photocrosslinking of the precursor polymers in the drops. we add a small amount of high molecular weight polyacrylic acid (PAAc) to the microgel suspension to induce clustering of microgels by electrostatic interactions between the ammonium ions of the microgels and the carboxyl groups of PAAc [29]. The functional dichotomy of such Janus particles can be further enhanced by embedding different materials selectively into either side of the particles. thus forming phase-separated Janus droplets. we use PNIPAm chains which are functionalized with dimethylmaleimide side groups. the resultant crosslinkable precursor polymers are used to create pre-microgel droplets in a microfluidic device. using colloidal PNIPAm microgels as building blocks [27]. 25]. Templating Janus particles from single emulsions Another class of supraparticles that can be made using microfluidically generated droplets as templates are Janus particles. thereby crosslinking the chains.

5 20 30 40 50 60 Temperature (oC) Fig 6 (a) Schematic representation of a technique used for fabricating colloidosomes using poly(N-isopropylacrylamide) microgel particles as building blocks and emulsion droplets as templates. (b) Aggregation and compaction of PNIPAm microgels on one side of premicrogel droplets upon heating (c) Fluorescent microscope image and SEM image of a Janus particle formed by photopolymerizing the monomers in the phase separated droplets. The thickness and properties of the shell layer directly affect the bubble lifetime and stability. However. Images were captured after allowing the sample to equilibrate for 30 min at each temperature. The scale bar denotes 100 µm. as we can precisely tune their properties. To enhance bubble lifetime or bubble stability. (d) Magnetically anisotropic Janus particles generated by embedding oppositely charged iron oxide particles selectively into the PNIPAm microgel rich phase. Reproduced from [28]. dispersed in an aqueous carrier phase. the contrast enhancement from free gas bubbles is limited as they are inherently unstable due to high surface tension at the gas-liquid interface [32]. high-shear batch processing methods produce bubbles with broad size distributions and a wide range of shell thicknesses [33]. chaotic.0 Relative diameter d/d20 0. but also allows gaseous fluids to be used. To form these bubbles. 32]. In one 5 . Thus. KGaA.8 PNIPAm colloidosomes T= 55 °C Cool 0. sugars. we use a microfluidic device to create an emulsion consisting of a gas bubble surrounded by an oil shell. as well as the acoustic response [34]. The sample was then cooled down to 20 °C using the same temperature steps. (a) H20 + + + + Photo initiator OIL 25 °C OIL 65 °C OIL 65 °C UV v NH2 COOH N H N H 2 COO- H2N 3N NH2 H2 N H2N H3N 2N H NH3 H2N -OOC 3N NH2 H2 N H2N COOH (b) (c) (d) bubbles are by far the most effective sound scatterers in liquids [30]. or polymers have been used to minimize the dissolution of gas [30. Free gas N H3 3 N H3 N H2 H2 N NH2 Fig 8 Images showing the textured morphology of AIM oil-shell stabilized bubbles (a) wet with a bright-field microscope and (b) dried with an SEM. The inset shows a magnified image of a single particle with the PNIPAm side attracted to a magnet. Traditionally. Size-change data of the constituent PNIPAm microgels over the same temperature range were collected using dynamic light scattering. we can form these same structures using a variety of gases and materials for the shells. As they oscillate under acoustic pressure waves. Copyright 2010 American Chemical Society. Due to the ability to independently control the structure of the bubbles and their chemical composition. Glass capillary microfluidics can form coated gas bubbles with controlled size and shell thickness.9 T= 20 °C Heat 0.7 0.6 Constituent PNIPAm microgels 0. (b) Equilibrium size change of PNIPAm colloidosomes and the constituent PNIPAm microgels.(a) (b) 1. Copyright 2009 Wiley-VCH Verlag GmbH & Co. H H Templating shell-stabilized bubbles from gas-filled single emulsions Microfluidic emulsification is not limited to the use of liquid fluids. Colloidosomes were dispersed in water and heated from 20 to 50 °C in fixed increments of 2 °C and then to 55 °C. shell coatings of lipids. control over shelled bubble fabrication allows us to design bubbles for specific applications. The PNIPAm microgels are tagged with Rhodamine B to enhance visual contrast between the two phases. in other words. they act as a point sound source to enhance contrast in ultrasound imaging [31]. This enables synthesis of microparticles that encapsulate gas-filled voids. Adapted and reproduced from [27]. N H NH2 H3N 3 Fig 7 (a) Schematic representation of a process for making Janus supraparticles. shell-stabilized bubbles. or.

The dashed line represents the predicted Rthread. To make these bubbles. for example. When the three fluids enter the collection tube. there are others in which a non-spherical shape is needed. as shown in Fig. the nanoparticles jam at the bubble surface. and different filling gases (N2. Accurate control of the size and structure of emulsions is often essential for these applications. (c-e) Double emulsion drops flowing through the collection tube. a double emulsion is formed. The inner fluid is pumped through the tapered circular capillary while the middle fluid. like a capsule. These structures have a core-shell architecture. The ratio of viscosities. which approach from the opposite end. we achieve good coaxial alignment. The outermost fluid flows through the outer capillary in the opposite direction and hydrodynamically flow focuses the coaxially flowing stream of the other two fluids. because the templating drops are themselves nearly always spheres. Nanoparticles can also be used to produce exceedingly stable “armored” bubbles [37. 40]. and poly(methyl methacrylate) (PMMA) for the shells. MF/ OF ≈ 0. The solid line represents the predicted Rdrop. While spheres are desirable for certain applications.system. The stability of the bubble shell against elastic instability-induced deformation strongly depends on the ratio of shell thickness to bubble radius. and compressed air) for the cores [35]. Rdrop values are represented with solid identical symbols. triangle). In this section. These solid-shelled bubbles exhibit superb stability. He. 8. smaller drops in its bulk. a polymer is dissolved in the oil phase. the stability of bubbles is improved by using polymers with higher stiffness and gas with lower solubility. providing a highly stable solid shell. Bubbles with smaller size or thicker shells exhibit enhanced stability against deformation. L-lactic-co-glycolic acid) (PLGA). By ensuring that the inner dimensions of the square capillary are the same as the outside diameters of the round capillaries. Double and triple emulsion particle templating As we have seen. single emulsion microfluidic devices afford unparalleled control over size and chemical composition of particles. and the volatile solvent is removed via evaporation. 38].08. The 6 . or in which the particles must have voids in them. we disperse hydrophobic silica nanoparticles in a toluene oil phase. which is immiscible with the inner and outer fluids. we describe a microfluidic technique that can create non-spherical particles and core-shell capsules. In addition. Fig 9 Fabrication of double emulsions in microfluidic devices. flows through the outer capillary in the same direction. Upon solvent evaporation. These observations provide guidance for the generation of stable bubbles with controlled physical properties. One design to do this combines both co-flow and flow focusing. A multiple emulsion is a drop containing additional. this approach almost always produces spherical drops. Schematic of a capillary microfluidic device that combines co-flow and flow focusing. This is achieved by forming double emulsions – drops with smaller drops contained inside – and then solidifying them to produce compound structures. and food applications. making them of potential use for ultrasound imaging and triggered or targeted imaging applications. This device consists of two circular capillaries arranged end-to-end within a square capillary. However. (b) The flat velocity profile of the flow as it enters the capillary tube. we use low-melting point AIM or paraffin oils for the shells and air as the inner phase. In this approach. The bubbles are collected in an ice water bath that instantly solidifies the shells. allowing them to be used for the encapsulation and release of materials in cosmetics. because these features directly affect the loading levels and the release kinetics of the encapsulated substances. and use this as the shell around a gas bubble [39. We can precisely control the shell thickness and bubble size by varying the flow rates during the microfluidic formation. drug delivery. for applications in ultrasound imaging and ultrasound-triggered release [36]. Another system that produces stable gas bubbles uses glassy polymers. CO2. as compartments into which to load drugs or other active materials. Fig 10 (a) Dependence of thread radius Rthread and drop radius Rdrop on the scaled flow rate QOF/Qsum. as shown in Fig 9 [18]. thereby generating a polymer shell at the air-water interface. polystyrene (PS). The high degree of control offered by glass capillary microfluidic devices enables fabrication of multiple emulsions. Half-filled triangles correspond to the radius of the internal droplets of the double emulsions. The open symbols represent the Rthread for different liquids and double emulsions consisting of a single silicon drop surrounded by a liquid shell (3QIF = QMF. such as poly(D.

Due to the resemblance of the structures to cell membranes. thereby yielding uniform PNIPAm microshells as shown in Fig. when PEG-b-PLA is dissolved in the shell phase of a W/O/W double emulsion. we form monodisperse polymersomes from a biodegradable. Subsequently. PEG is a hydrophilic block while PLA is a hydrophobic block. but these structures can have more elaborate shapes. These simple physical arguments highlight the versatility of this method in generating monodisperse double emulsions under different flow conditions (Fig. 41–44]. For a viscosity ratio of MF/ OF = 0. we can quantitatively predict the experimentally measured values. To illustrate this approach. such as permeability and selectivity. polymer vesicles. biocompatible diblock copolymer. Copyright 2007 Wiley-VCH Verlag GmbH & Co. Rthread is the radius of the fluid thread that breaks into drops. are tunable by varying the thickness and chemical composition of the bilayer membranes. All scale bars denote 200 m.1. Flow rates were controlled with stepper-motor-controlled syringe pumps (Harvard Apparatus). We use thermoresponsive pNIPAM as the matrix polymer to obtain environmentally sensitive microgel particles.82Rthread. The solvent drop is subsequently evaporated and a polymersome is formed. At the moment of their formation. The inner droplet is enclosed by a bilayer membrane that consists of the diblocks at the two interfaces. Upon evaporation of the oil phase.radii of drops shows that Rdrop = 1. (b) pNIPAAm microshells obtained from the experiment in Panel A. By multiplying the predicted thread diameter by a factor of two. have been shown to exhibit better mechanical stability than phospholipid vesicles. the PEG-b-PLA molecules adsorbs at the oil/water interfaces. where Qsum is the sum of the of inner and middle fluid flow rates. Double emulsion-templated vesicles Vesicles are compartments of fluid enclosed by bilayers of amphiphilic molecules. or polymersomes. these droplets are cured by UV exposure as they flow through a delay capillary a few centimeters downstream (not shown). while retaining many of their attractive properties such a tunable permeability and targeted release. thus they are widely used as encapsulating structures of active ingredients for applications ranging from pharmaceuticals to cosmetics. 11b. the drop diameter is approximately twice the diameter of the thread [13]. 11a. the size is controlled by the ratio of the flow rates of the combined inner and middle fluids to the outer fluid. At lower flow speeds. 10a. We use a glass capillary device to form monodisperse drops of a curable precursor solution that consists of a polymerizable or crosslinkable substance. as 7 . which is given by Eq. Fig 11 Microfluidic production of hollow microgel shells. as shown in Fig. respectively. Copyright 2010 The Royal Society of Chemistry. When drops form here. we again see very good agreement between the model and the data. To better control the size and composition of the double emulsions. 12c. and Rorifice is the radius of the collection tube where the drops are formed. which we either fabricate from monomeric NIPAM and BIS or which we gel from pre-fabricated. When a fluid thread breaks. as shown in the series of images in Fig. the shell is gelled by thermal monomer polymerization of by photochemical polymer-analogous gelation. we examine the physical mechanism of drop formation. poly(ethylene glycol)-block-poly(lactic acid) (PEG-bPLA) using a capillary microfluidic device [45]. in Fig. (c) Double-core microshells obtained upon slight variation of the flow rates in the experiment in Panel A (cf. and polypeptides. thereby creating a shell structure. we make monodisperse polymer microshells. Hence. (a) A glass microcapillary device is used to create an oil-water-oil double emulsion with a semidilute solution of crosslinkable pNIPAAm as aqueous phase. 11c. Their properties. 10a. We have developed an alternate approach for fabricating monodisperse vesicles from these polymers. To illustrate this templating process. these pre-microgel droplets are loaded with inner droplets of another oil. by employing monodisperse double emulsions as templates. materials encapsulated in them can potentially be released directly into cells through fusion of the vesicles with the target cells. By solving for Rthread/Rorifice in Eq. which is a reasonable assumption given the proximity of drop formation to the entrance of the collection tube. Operating the device with different flow rates produces shells with two cores. double emulsion can be used to template structures. (1) and plotting it as a function of Qsum/QOF. as shown by the dashed line in Fig. Qsum / Qof = Rthread / ( 2 shown in Fig. 10c–e). double emulsions typically form approximately one tube diameter downstream from the entrance. The resultant vesicles have great size uniformity and Templating microshells and capsules from double emulsions Just like single emulsions. Ref. QOF is the flow rate of the outer fluid. Reprinted from [16]. (1). and are thus a promising alternative. in agreement with what is expected theoretically [44]. Recently. We neglect the decrease in viscosity due to the contribution of the inner liquid because its volume fraction in the drop is small. 10b). A Oil B To UV Exposure Oil C Rorifice 2 Rthread ) [1] 2 This equation is valid for plug-flow (Fig. we emulsify an aqueous microgel precursor solution in a continuous oil phase. [26] for details). phospholipids. the double emulsion drops undergo a dewetting transition where the shell phase is collected into a single drop of solvent with the excess diblock copolymers. such as diblock copolymers. After droplet formation. Reproduced from [26]. within the collection tube [18. The experimentally measured diameters of different threads and the corresponding drops that pinch from these threads are shown as the open and closed symbols. To form the premicrogel droplets. the diameter of the resulting drop is proportional to the diameter of the thread as well as the viscosity ratio. KGaA. photocrosslinkable pNIPAM precursors.

(PS-b-PEO) [47]. making these capsules ideal for applications that require simultaneous release of incompatible actives.encapsulation efficiency as demonstrated by the encapsulation of a model fluorescent dye shown in Fig. (b) Encapsulation of a fluorescent dye in polymersomes. and then in separate channels before forming two separate inner droplets without mixing. Using this double emulsion-templated approach. if the two inner compartments are close to each other. Copyright 2010 American Chemical Society. Reprinted from [45]. Using such a device. To make the encapsulating structures more robust. and the scale bar is 10 m. we can also incorporate homopolymers in the membrane and tune the properties of the polymersomes [45]. as shown in Fig. thus the droplets can be cooled below the melting point of the oil to yield solid capsules. the actives in the Fig 13 (a) Schematic of a capillary microfluidic device for generating doublecore double emulsions. this technique can be extended 8 . the desired reactions can be triggered by coalescence of the inner droplets and subsequent mixing of the reactants. If the two inner compartments are far enough apart. Release of the actives from these capsules can be easily triggered by melting the shells. The scale bar denotes 400 m.8 s. Reprinted from [49]. We demonstrate this using a modified glass capillary device. 12b. Thus. By dissolving functional homopolymers in the shell phase of the double emulsion templates. illustrated in Fig. To complete this picture. we now introduce another method to form multiple emulsions: sequential coflow emulsification. This approach for forming polymersomes is general and has also been applied to form polymersomes of poly(normal-butyl acrylate)-block-poly(acrylic acid) (PBA-b-PAA) [46] and polystyrene-block-poly(ethylene oxide). However. by increasing the osmolality of the environment and bursting the vesicles by sucking water out. the inner droplets remain separate as long as the capsule is solid. Scale bar is 100 m. By freezing the double emulsion drops quickly to speed up their solidification. The ability to fabricate multicore double emulsions and capsules creates new opportunities for encapsulation-related applications in personal care products. Making triple emulsions with glass capillary microfluidics So far we have described the formation of double emulsions by concurrent coflow and flow-focusing emulsification. As we will show. If multiple reactants can be stored separately within the same double emulsion droplets. these capsules can act as micro-reactors in which mixing of reactants is triggered by heating. double emulsion drops with multiple separate inner droplets for the different actives are needed. (d) Shrinkage and breakage of a polymersome after an osmotic shock. (c) Wax capsules with two compartments formed by freezing double emulsion drops with a crystallizable shell phase. Microfluidic devices are apt for fabricating such structures due to their easily customized geometries. The scale bar is 10 m. Copyright 2008 American Chemical Society. (b) Double emulsion drops with two inner drops encapsulating a red and a blue dye separately. (a ) solven t Diblock copolymer (b ) Solvent Removal Double Emulsion Drop (c) Vesicle (d ) t=0 min t=8 min t=11 min t=12 min t=13 min separate droplets are not pre-mixed before triggered release. we use a molten crystallizable oil as the shell phase of the double emulsion. By carefully tuning the morphology of such capsules. The actives can be released with a simple osmotic shock. we have fabricated double emulsion droplets with two cores. The scale bar is 400 m. food and beverages as well as cosmetics. as shown in Fig. as shown in Fig. 13b. for instance. By designing devices with more complicated geometries that enable separate injection of multiple inner phases. the two actives will be released to the outer phase separately. they will coalesce before release to the surroundings. To achieve this. (a ) Fig 12 (a) Schematic illustrating a double-emulsion-templated approach for fabricating polymersomes. 13c. The model encapsulants. (b ) (c) Templating multi-core capsules Double emulsions with a single-phase inner droplet can be used directly for encapsulating actives or indirectly as templates for more sophisticated capsules. we have also fabricated monodisperse phospholipid vesicles with high encapsulation efficiency [48]. An important technical challenge in this area is the co-encapsulation of multiple incompatible actives without cross-contamination. it is possible to manipulate the release profile of actives encapsulated in this fashion. it is possible to encapsulate more than two incompatible actives or reactants. 12d. Thus. Successive images are taken at intervals of 1. wright stain (blue) and rhodamine B (red) dyes flow separately into the device. (c) Dewetting transition of a double emulsion drop. 13a [49]. which has an injection tube with two separate internal channels.

KGaA. and Q3 = 5000 L/hr. middle (II). the variation ranges for the flow rates of the inner. Again. to generate additional hierarchical levels of multiple emulsions. 14d and 14e. for example. and those of the inner fluid are Q1 = 20. and outer fluids are Q1 ≈ 20–600. as shown in Fig. Q3 = 2000. Fig 14 Capillary microfluidic device and the formation of precisely controlled monodisperse double emulsions. The variation ranges for the flow rates are Q1 ≈ 5–100. Furthermore. The flow rates of the inner. Q2 = 2000. as illustrated by the series of triple emulsions with one to seven innermost drops. respectively. and the transition tube (a second cylindrical capillary with an inner diameter D2). and one to three middle drops in each outer drop (Fig. The devices consist of a coflow junction followed by additional coflow junctions. as shown in Fig. (b) and (c) High-speed optical micrographs of the first (b) and second (c) emulsification stages. and injecting the outermost fluid to flow coaxially around this tube to form the third level of emulsification at its outlet. thus enabling us Fig 15 Generation of highly controlled monodisperse triple emulsions. the variance of the diameters of triple emulsions is less than 1. (a) Schematic diagram of the device geometry. 85. 55. to produce even higher-order emulsion droplets. leading to flow-focusing of the first middle fluid into the 9 . This method for fabricating multiple emulsions has the advantage that it can be extended very easily. of the inner droplets of the double emulsions. Q2 ≈ 200–1000. 150. 14b and 14c. the technique can clearly be sequentially scaled to even higher levels of emulsification if desired. This preparation is accomplished by adding a second transition tube at the outlet of the first. 225. and outer fluids in (b) and (c) are Q1 = 350. Q3 is larger than Q2). Q3 ≈ 2000–3500. as shown in Fig. second (c). 15a. middle. quadruple emulsions could be made by adding an additional stage. 200. Q2 ≈ 1600–5000. We illustrate this concept by fabricating monodisperse triple emulsions.graphs displaying the first (b). Images of each stage of emulsification are shown in Fig. 15(b–d). Although there are large deformations of the droplets during their formation as they flow through the tapered regions of the capillaries. All scale bars are 200 m. 70. In (d). (d) Optical micrographs of monodisperse double emulsions containing a controlled number of monodisperse single emulsions. Reproduced from [50]. (e) Optical micrographs of triple emulsions that contain a controlled number of inner and middle droplets. The outer fluid must be immiscible with the middle fluid and the middle fluid must be immiscible with the inner fluid. All double emulsions were made in the same device and with the same fluids.indefinitely. (e) Optical micrographs of monodisperse double emulsions showing controlled increase of the diameter of the inner droplets while the number is constant. which consist of water-in-oil-in-water-in-oil (W/O/W/O) drops.5%. and third (d) emulsification stages. 14a. Q2 = 500. In all experiments. respectively. Using the exquisite control over the number and size of the internal droplets of the double emulsions. and 240 L/hr. the flow rates of middle and outer fluids are fixed at Q2 = 2000 and Q3 = 5000 L/hr. The individual steps of drop formation leading to the triple emulsions are shown in Fig. Copyright 2007 Wiley-VCH Verlag GmbH & Co. In (e). They are composed of the injection tube (a cylindrical capillary with a tapered end). (a) Schematic diagram of the extended capillary microfluidic device for generating triple emulsions. middle (I). we are able to vary the number and size. and Q4 = 5000 L/hr. from the left top to the right bottom. (b)–(d) High-speed optical micro. 15e). middle. and outer fluids in (b)–(d) are Q1 = 50. and Q3 ≈ 2000–8000 L/hr (in each case. both the diameter and the number of the individual drops at every level can be precisely controlled. (f) Schematic diagram detailing an alternate method for generating triple emulsions where the middle fluid (II) is injected from the entry side of the first square tube. and Q4 = 5000 L/hr. The flow rates of the inner. the emulsification process remains stable.

Reproduced from [50]. The light passes through the transparent regions of the photomask but is blocked by the black parts. The extra layer surrounding the microcapsule in (b)–(e) is water that is squeezed out from the hydrogel shell as it shrinks. are all black except for the regions that are to become the channels. KGaA. The coalescence of the expelled inner oil with the outer oil cannot be resolved. (b)–(e) Optical micrograph time series showing the forced expulsion of the oil and water droplets contained within the micro-capsule when the temperature is rapidly increased from 25 to 50°C. and initiator in the outer aqueous layer. the inner middle fluid is also oil. The variation ranges for the flow rates in (g)–(j) are Q1 = 50–200. This is important because this thickness determines the height of the final microfluidic channels. The photons that penetrate create radicals in the photoresist that crosslink these regions. Q2 = 1600–2500. Upon heating from 25 °C to 50 °C. The result is a microcapsule consisting of a shell of thermosensitive hydrogel that encapsulates an oil drop containing several water drops. SECTION 2: PDMS microfluidics Introduction The microfluidic devices described to this point are built by aligning and gluing glass microcapillary tubes together. providing spontaneous. and Q4 = 5000–12000 L/hr. Since each fluid layer is sandwiched between two immiscible fluids. The scale bar in all images is 200 m. particular for forming double emulsions. pulsed release of the innermost water droplets into the continuous oil phase. we form a W/O/W/O triple emulsion and add monomer. where the continuous liquid is oil. Fig 16 Temperature-sensitive hydrogel microcapsule for pulsed release. Copyright 2007 Wiley-VCH Verlag GmbH & Co. (a) Optical micrograph of a microcapsule with a shell comprised of a thermosensitive hydrogel containing aqueous droplets dispersed in oil. Further refinements could adjust the thickness of the layers and the number of droplets. refraction. to tailor the channels to overcome specific challenges. the nature of the fabrication. As we show. which would facilitate their highly controlled release. The wafer is then heated for a few minutes and placed in a solvent wash to removes the unexposed. since both liquids have the same index of 10 . For the process we describe. This structure has a Trojan-horse like behavior. Fabricating PDMS devices using photolithography The general concept in photolithography is to fabricate three-dimensional microfluidic devices from drawings of the channels. the coating is solid but fragile and can easily be removed with a solvent. normally SU8. At this point. the hydrogel shell is aqueous. usually a few inches in diameter. (g) and (h) High-speed optical micrographs showing the formation of double emulsions in a one-step process in the transition capillary (g) and the subsequent formation of triple emulsions in the collection capillary (h). the hydrogel shell shrinks by expelling water. The ability to precisely control the formation of multiple emulsions offers new opportunities to engineer novel materials. these regions are left transparent. hardening them. 16a. the thermosensitive hydrogel rapidly shrinks by expelling water. This works by first printing a to-scale picture of the device on transparency plastic. In this section. crosslinker. Nevertheless. This capsule was generated from a triple emulsion. called photomasks. 16b-e. thus enabling fine control over diffusion of compounds contained within the innermost droplets. to allow light to pass through. the shaping of the capillary tips by heating and stretching them. the thickness of the coating can be controlled to micron precision. protecting the inner most water droplets in the hydrogel shell until their temperature-induced release. we use the triple emulsion structure to fabricate monodisperse microcapsules made with a thermo-sensitive. the device drawings. KGaA. We also add an accelerator to the inner oil phase. Moreover. Q3 = 4000–8000. and then transferring this image to a substrate using the photolithographic process. and cooled to solidify the coating. all in a continuous oil phase. the hydrogel shell breaks. is coated with photoresist. as shown in Fig. This is achieved by pouring a glob of the resist on the wafer and spinning it at a high speed. a drawback to these devices is that the manual fabrication makes it difficult to construct more than a few devices at a time. The scale bar is 200 m. we are able to perform a polymerization reaction in a specific layer. This allows very small channels to be made. It also affords greater flexibility when designing devices. as shown in Fig. it also highlights the potential of this microfluidic device to create highly engineered structures for controlled release of active substances. (i) and (j) Optical micrographs of triple emulsions that contain a different number of double emulsions. poly(N-isopropylacrylamide) hydrogel that can be used for controlled release of active substances [50]. Upon increase of the temperature. and the innermost droplets are aqueous. This experiment demonstrates the utility of our technique to generate highly controlled capsules with multiple internal volumes that remain separate from each other. The photomask is placed on top of the coated wafer and the two are exposed to ultra-violet (UV) light.transition capillary. To create a device from the photomask. In this case. where it diffuses into the outer aqueous shell and speeds polymerization of the hydrogel. for the synthesis of small particles. Depending on the viscosity of the photoresist and the spin rate and duration. because of the incompressibility of the inner oil. allowing fabrication of devices down to a micron in width. a silicon wafer. this allows creation of new kinds of structures. Copyright 2007 Wiley-VCH Verlag GmbH & Co. The time series begins once the temperature reaches 50°C. Although quite simple. This process utilizes photolithography in poly(dimethylsiloxane) (PDMS) [51] and is thus inherently parallel and precise. The process therefore remains challenging for producing large number of identical devices. we describe an alternative process that sacrifices the idyllic flow conditions in microcapillaries to achieve greater control and reproducibility. Reproduced from [50]. this process enables the creation of devices with superb flow properties. however. is limited in precision and reproducibility. The coated wafer is then heated to evaporate solvent. To illustrate this potential.

” another polymer.uncrosslinked photoresist. The drops form in the nozzle channel. as shown in Fig.01). 17b. However. drops are formed. including sol-gel and polymer coatings. 17b. Another difference is that T-junctions tend to yield more monodisperse drops at low flow rates. The devices can also be treated with Aquapel. to make drops.e. forming covalent Si–O–Si linkages that create an irreversible seal. Whichever drop formation mechanism used. the dispersed phase is injected into the central inlet and the continuous phase into the two side inlets. drop size can be controlled by adjusting flow rates. as shown in Fig. which are an inverse of the photomask. stretched. At high flow rates when viscous forces become comparable to surface tension (Ca > 0. This leaves behind the crosslinked parts. but they all share the same basic structure. At this point fluids can be pumped through the channels to use the device. specifically. which is the diameter 11 . 17a. due to the enhanced flow stability of having only a single continuous phase inlet [54]. eventually pinching off a drop. where these two fluids meet. even at high Ca plugging effects still likely play a role in these devices. becoming a clear. it is sheared by the drag of the continuous phase. protruding) channels where the light exposed the resist. monodisperse drops can be formed with a size that can be adjusted by changing flow rates. constricting the path of the continuous phase. and in contrast to the unbounded flows of capillary co-flow drop makers. For example. again. Flow-focus junctions yield better emulsions at higher flow rates. This creates “positive” (i. (a) In a T-junction geometry the dispersed phase is injected from one channel (left) and the continuous phase from another channel (top). shearing forces start to play a role. In this geometry. As will be shown in later sections. however. due to the highly confined nature of the flows. especially those templated from multiple emulsions.01). As with T-junction drop formation the physics of the process depends on flow rates. T-junction and flow-focus drop makers behave quite similarly. in these devices the physics by which the drops form changes as a function of flow rate. in most cases either geometry will suffice. Single emulsion drop formation The first drop formation geometry developed for PDMS microfluidics was a so-called “T-junction” drop maker [52]. drops form where these two channels intersect. to make the devices more chemically resilient. Fig 17 Schematics of two most common drop formation geometries for PDMS microfluidic devices. The edges of the solid PDMS are sliced with a scalpel and the slab is peeled from the master. monodisperse drops can be formed with a size that can be controlled by adjusting flow rates. the interface is Another way to produce drops is to use a so-called “flow-focus” geometry [53]. creating the PDMS microchannels. and the choice is a matter of personal preference. so that drops form as a consequence of plugging and squeezing [52]: the inflating tip of the dispersed phase blocks the downstream nozzle. When this shear force is equal to the tensile force adhering the drop to the inlet. especially with respect to the size and production rate of the drops for a given set of flow conditions. is poured over it. The bonding is achieved by oxidizing the surfaces of the blocks using oxygen plasma etching. which continues to be pumped in. squeezes on the dispersed phase. in which two channels intersect to form a four-way cross. to make them even more hydrophobic. the dispersed phase is injected from a side channel and the continuous phase from a vertical channel. In this device. (b) In a flow-focus geometry the dispersed phase is injected from a central channel (top) and the continuous phase from the two side channels (left and right). In both cases. Again. This geometry has many variants. The PDMS hardens when baked in an oven. rubbery material. In fact. shearing off a drop. dominated by plugging and squeezing at low flow rates and shearing at high flow rates. PDMS is hydrophobic by default. Valve-based drop formation As described in previous sections. The resulting block is imprinted with negative features of the SU8 channels. for the emulsification of other kinds of solvents. At low flow rates viscous forces are small compared to surface tension (Ca < 0. As the dispersed phase bulges into the nozzle. a commercial glass treatment that fluorinates the surfaces. there is another parameter on which drop size depends. as shown in Fig. poly(dimethylsiloxane) (PDMS). though there are subtle differences between them. These fluids meet in the nozzle where. to create emulsions. this ability to functionalized PDMS channels is essential when forming structures with the devices. on the ratio of the dispersedto-continuous phase flow rates. in turn. Nevertheless. the wettability of the device must be controlled using a chemical treatment. To mold a microfluidic device from this “master. drop size depends on flow conditions. often additional processing must occur to control the surface properties of the channels. Interestingly. Silanol groups generated by the plasma undergo condensation reactions with silanols on the other block. Other coatings can also be deposited. two channels intersect to form a T shape. this causes a pressure rise in the continuous phase that. since the centered position of the dispersed phase tends to enable dripping at higher flow speeds. In practice. enabling the production of water-in-oil emulsions with move solvents. Holes are punched in the block where it is to be interfaced with tubing and the imprinted side is bonded to a flat substrate – either another block of PDMS or a glass slide – enclosing and sealing the channels.

Nevertheless. this parameter is rarely used as a means to adjust drop size. the wettability properties of the channels are of critical importance in determining the types of drops formed – for example. In these regions. The drop size depends on the nozzle dimensions. drops must be encapsulated in drops. since doing so requires fabrication of different devices. they combine the best attributes of glass capillaries with the reproducibility and scalability of lithographically fabricated devices. like additional drop makers. For example. This allows drop size to be adjusted in real time. drops of the same type are formed. the particles are flowed at high velocities down a long channel such that the Reynolds number approaches one. In this technique. By contrast. the in-flow of the objects is normally random. there are strategies that can be used to relax these constraints. The scale bars denote 100 m. making it larger or smaller as needed. when creating double or higher order multiple emulsions. The reason for this difference has to do with the way in which the fluids are injected into PDMS drop makers. There is a technique. When the dispersed phase enters such a device. as though the device were two opposing T-junctions. the larger the drops. the 12 . for example. which is called valve-based flow focusing [55]. water will be lifted off the walls by the oil. it must be lifted off these surfaces and surrounded by the continuous phase. Wetting is of such importance in these devices that even if the inlets for the oil and water are switched. relatively independent of channel wettability. lowering overall encapsulation efficiency. However. if water is injected into the oil inlet and oil into the water inlet of a hydrophobic flow-focus device. If. can form drops of either type. Copyright 2009 American Institute of Physics The dominance of wetting in PDMS devices Whichever drop formation geometry used. Using a combination of geometrical control and shear. To be formed into a drop. instead normally there are stretches of ordered particles separated by empty gaps of continuous phase. resulting in water drops. inertial effects become important. (a) Schematic of valve-based flow focusing device. This can be accomplished using inertial particle ordering [57. and especially so when forming multiple emulsions. to vary the size of the drops. however. as shown in Fig 18b-c. but they will drip from the two side channels. and are the subject of the final section of this chapter. These PDMS devices mimic the superior flow properties of glass capillary devices. cells and beads must be encapsulated in drops. single-layer membrane valves [56] are used to constrict the size of the drop formation nozzle. empty drops are formed. due to their circular coaxial geometry. by exploiting threedimensional channel graduations. injection channels. For these reasons. These fluids are injected into the nozzle. This is in contrast to glass capillary devices which. water drops will still be formed. a unique challenge when encapsulating objects in drops is that while the drop formation is controlled and periodic. this allows the drop size to be adjusted without changing flow rates. a challenge with this technique is that the high velocities required can preclude stable drop formation. generally. Despite the usual dominance of wetting in these devices. or sorters. when performing biological assays.of the drop formation orifice: the larger the orifice. lifting off the oil. For example. One strategy to beat Poisson statistics is to order the objects so that they enter the drop maker at more regular intervals. it is possible to form either type of emulsion in a device with fixed wettability. This is useful when using drop makers in combination with other components. instead leading to jetting and polydisperse drops. This circumstance makes it difficult to synchronize drop formation with the objects. whether oil drops in water or water drops in oil are formed. this is not attempted and the objects are encapsulated at random. (d) Sequence of images for different actuation states of the valves. producing oil drops. where drops are formed. thereby loading the objects according to Poisson statistics and leading to a large number of improperly filled drops and only a few containing single-objects. generally. However. Essentially. water is injected downward into the central inlet and oil from the two side inlets. which can be adjusted using the single layer membrane valves. creating forces between the particles that cause them to order. in which the behavior of the devices can be very sensitive to changes in flow rate. the wetting of PDMS devices must be controlled carefully when forming emulsions. it is initially in contact with the upper and lower surfaces of the channels. Reproduced from [55]. Applications of PDMS single emulsification Close packed encapsulation There are occasions when using microfluidics in which it is necessary to encapsulate objects inside of drops. without changing flow rates. if the channels are hydrophilic then the water clings to the walls. In this technique. 58]. that can adjust orifice size without having to fabricate additional devices. (b) Formation of large drops with the valves un-actuated and (c) formation of smaller drops by constricting the nozzle with the valves. At these velocities. Whether this happens depends on the wettability properties of the channels. Fig 18 Valve-based flow focusing drop formation. as shown in Fig 18a. channels are hydrophobic. Another challenge is that it can be difficult to order all the particles. delay lines.

but both formed from different macromolecular precursors. 20c. they spontaneously order into a crystalline array. in this example. This allows every drop to be encapsulated with exactly one particle. ordered particles is shown in Fig 19a. as evidenced by the middle and lower row of micrographs in Fig. the core remains unaffected. an image of packed. as shown in Fig 20a – b. 21b. This is achieved by injecting the particles into a device at very high volume fraction. Because the particles are monodisperse and packed together. these particles are applicable for encapsulation and controlled release purposes: when the pNIPAAm shell is swollen. 21a [60]. Reproduced from [59]. 22a and detailed in Fig. to encapsulate exactly one particle per drop. in this example. a random number of particles is encapsulated per drop. three or even four particles per drop. and since they are monodisperse. Fig 19 (a) Polyacrylamide gel particles injected at high volume fraction into the inlet of a microfluidic drop maker. (b) Encapsulation of single particles in drops with a wide nozzle. their rate with respect to the drop rate can be increased. (b) Intensity time traces or particles moving through the boxed region shown in the image. The resultant particles consist of a hydrophilic polymer core nested in a hydrophilic polymer shell. 22b: while the shell collapses due to the volume phase transition of pNIPAAm. they naturally order into a regular lattice. In the first junction we add a semidilute. (c) Encapsulation of particles in which the particle flow is not ordered. In this example. we use a shell phase that is tagged with a green fluorescent tracer polymer. as shown in Fig. To substantiate this finding. slight differences in the frequency of the drop formation and particle flow can lead to occasional empty drops.Our approach for beating Poisson statistics is to use mechanical forces to order the particles [59]. These profiles show that the red-tagged core material and the green-tagged shell material are well separated. and then we wrap these particles in an aqueous polymer shells using a microfluidic device. The device consists of two cross-junctions in series. it is porous and permeable. Since the ordered array moves at constant velocity. layered pre-microgel drops. Copyright 2009 The Royal Society of Chemistry. Due to this selective sensitivity. since this reduces the overall compressive force on each particle by its neighbors. The particle frequency can be synchronized with the drop formation by adjusting particle velocity. 21b are plotted in Fig. along with red-tagged core microgel particles. In the second junction we add oil to form bi- In an implementation of this approach. the particles are nearly the same size as the drop formation nozzle. as shown in Fig. To demonstrate the utility of this technique. this allows us to visualize the formation of core-shell structures which exhibit a well-controlled number of core particles in each shell and a well-controlled shell-thickness. Due to the high volume fraction. we create monodisperse hydrogel particles. in which drop formation is not triggered. The encapsulation efficiency with this technique is far better than can be achieved with random encapsulation. Fig 20 (a) Encapsulation of single particles using close-packed ordering. as shown in Fig 19b and inset into Fig 19c. 21b-c. To realize this concept. the particles enter the drop maker at regular intervals. we incorporate nonthermo-responsive polyacrylamide (pAAm) particles (not labeled) into thermo-responsive pNIPAAm shells (green fluorescently labeled) [60]. Reproduced from [59]. the particles pack together. By increasing the velocity of the particles. allowing them to perturb the flows. aqueous solution of crosslinkable pNIPAAm chains as shell phase. 21d. There is also no interpenetration of the shell material into the core. spatially resolved profiles of the fluorescence intensity across the micrographs in the left column of Fig. 13 . to encapsulate two. to trigger drop formation. due to the regular flow of the particles. there is a tall peak at the frequency of the particle flow. which show separate visualizations of the green-labeled shells and the red-labeled cores of the microgels in the upper row. this introduces them into the lower junction at a regular frequency. The particles are about 25 m in diameter. (c) Power spectrum of the intensity time trace. Copyright 2009 The Royal Society of Chemistry. The behavior of these core-shell particles upon increase of the temperature to 35 °C is visualized in Fig. Close-packed particle encapsulation for the formation of coreshell microgels The ability to encapsulate controlled numbers of microgels into drops offers a powerful means to template multi-layered microgel particles. We then lock in these structures by crosslinking the pNIPAAm chains in the shell. both crosslinked and swollen in water. as sketched in Fig.

dubbed “two-step” formation. this can be achieved by using two drop makers in series [61–63]. Since the UV beam can be shaped using holes. which is called flow patterning [64]. operate on different principles. the thermo-responsive shell collapses and encapsulates these actives in the pAAm core. pNIPAAm or polyacrylamide. slits. the actives are rapidly released. 22c. In addition to geometrical considerations like this. when the shells are swollen. 14 . Pictures in the upper row of Panel B show an overlay of the micrographs in the middle and lower row. In PDMS devices. This application is substantiated in Fig. to form an oil-in-water-in-oil double emulsion. can be achieved using either cascading T-junctions or flow-focus drop makers. (d) Spatially resolved intensity profiles of the red and green fluorescence in the single-core particle shown in Panel B. For the simple patterns needed for these devices. (a) Schematic of a microfluidic device forming aqueous pNIPAAm droplets that are loaded with a well-defined number of pre-fabricated microgel particles of a similar material. in double and higher order multiple emulsion formation spatially patterned wettability is required. Thus. However. c) The flow rates of the inner particle phase (red-tagged pNIPAAm). Intensity 0 -120 -60 0 r ( m) 60 120 Fig 21 Microfluidic fabrication of microgel capsules that consist of two miscible yet distinct layers. which produces the outer drops. However. Then. in the case of double emulsion devices only a very simple wettability pattern is needed. One technique is to use a photo-patterning approach. The scale bar denotes 100 m and applies to all micrographs in Panel B and C. in contrast to single emulsion formation in which only uniform wetting is required. and the outer oil phase control the number of core particles in each shell (b) as well as the shell thickness (c). the pAAm core remains unaffected by temperature. the particles can be loaded with hydrophilic low molecular weight or mesoscopic additives. which depict separate visualizations of the green-tagged pNIPAAm shell and the red-tagged pNIPAAm core. Upon increase of the temperature. (b. a different approach is available that trades versatility in the pattern shape for simplicity in the treatment process. providing stability of shape. This technique. which are fed into the inlet of the second drop maker. Copyright 2010 American Chemical Society. coupled with the fact double emulsions consist of drops of at least two immiscible phases. Generally. D B Microgel Particles pNIPAAm Oil 1 To UV Exposure C Norm. Reprinted from [60]. A double emulsion consists of at least one liquid droplet encapsulated in another droplet of an immiscible phase. it is also necessary to control the wetting properties of the devices. the oil drops must be formed in a hydrophilic part of the device and the water drops in a hydrophobic part. This fact. which shows a sequence of images from an experiment where RITC–1 tagged dextran (M = 10. because the symmetric injection of the middle and continuous phases helps prevent wetting of the drops on the channels. This type of double emulsification. Spatially-controlling wettability in PDMS devices In PDMS devices channel wettability determines the type of emulsion formed: oil-in-water emulsions are formed in hydrophilic channels. evidencing only very little interpenetration of its two phases. double emulsions can be formed in PDMS devices. serial flow-focus devices are used. means forming them in PDMS devices requires channels with at least two distinct wettabilities. By contrast. Subsequent droplet gelation leads to microgels with a distinct core-shell architecture. are formed in hydrophobic channels. To create such a structure with a microfluidic device requires emulsification of the inner phase followed by emulsification of the outer phase. and lenses. and the added complication of having to align the photo-pattern with the microchannels can make this approach unattractive in these instances. Double emulsification with PDMS devices As with glass capillary devices. For example. this approach allows for the creation of complex wettability patterns. The need to spatially control the wettability in PDMS devices has stimulated development techniques to spatially modify device surface properties. in which the devices are filled with a solution that only reacts with the channels under exposure to intense UV light [63]. in fact. all A surrounding feed material can be removed and the loaded particles can be stored at elevated temperatures. however. though the devices look quite different and. The first drop maker produces the inner drops. making the formation more robust.000 g mol ) is released from pAAm– pNIPAAm microgels. as soon as the temperature is decreased. water-in-oil emulsions.whereas it becomes non-porous and impermeable when it collapses. while the inverse. the middle polymer phase (green-tagged pNIPAAm).

These two phases are then surrounded by the continuous phase. The dotted lines are guides to the eye. yielding a coaxial nested jet in the second junction. pAAm core plus pNIPAAm shell. whereas light blue squares represent only the pAAm core. To create this wettability pattern. d. one-step formation can exceed these flow rates. the flow rates are set such that a jet of the inner phase is surrounded by the middle phase. (a). the first drop maker junction must be hydrophobic and the second hydrophilic. operating far into the jetting regime. as shown in Fig 24b. Dark blue circles represent the diameter of the entire particle. These are ideal for encapsulation because they require minimal volume of shell material per unit of material encapsulated. one-step formation can only produce single-core double emulsions. whereas it collapses at elevated temperatures (lower row). this process is dubbed “two step” formation. Copyright 2010 American Chemical Society. The reactive solution changes the wettability only in the treated A regions. Two-step versus one-step double emulsification Double emulsions can be formed in PDMS devices in two different processes that depend on flow conditions. wettability patterns for W/O/W or O/W/O double emulsions can be created. the temperature remains above 33°C. an inert solution is injected into the inner phase inlet and a reactive solution into the outlet of the device. as shown to the right in Fig 24b and 25a. A specific advantage of this is that it allows formation of double emulsions with exceedingly thin shells. These two processes have different limitations and advantages. and is illustrated in Fig 24a. T.. confining it to the lower portion 15 . Whereas two-step formation can produce multicore double emulsions containing several inner drops. To make a water-in-oil-in-water double emulsion. inner drops form in the first drop maker and are encapsulated in the outer drops in the second drop maker. At ambient temperatures (upper row). “one step” formation. pinching into double emulsions in a single step. whereas two-step formation is limited to flow rates that enable dripping in the first junction. as shown in Fig 25b.e. the double jet becomes unstable due to the widening channel. no drops are formed in the first junction. while the untreated areas retain their default wettability. the shell is swollen. i. as depicted in Fig 23. As the temperature decreases. In the second process. All scalebars denote 100 m. This allows creation of double emulsions with a greater range of shell thicknesses. In the course of the first ten seconds of this experiment. respectively. this process is dubbed.000 g mol–1) from the particles in Panel A. By contrast. (b) Detailed plot of the particle-diameter. Because the inner and outer drops are formed in two locations in two subsequent steps. However. instead.patterns wettability by introducing a reactive solution into select portions of a device and preventing it from going using the flow of an inert blocker solution. There. B d ( m) 30 C 120 Shell 60 Core 0 25 27 29 31 T ( C) 33 35 35 C C 0 5 10 15 20 t (s) Fig 22 Thermo-responsive behavior and controlled release application of pAAm–pNIPAAm core-shell microgels. as a function of temperature. Fig 23 Schematic of flow patterning used to spatially control the wetting properties of the microfluidic devices. Reprinted from [60]. a spontaneous release of the active incorporated in the particles is triggered by the swelling of the pNIPAAm shells. (c) Controlled release of RITC-dextran (M = 10. as shown in Fig 1a–b and 1c–d. The inert solution blocks the reactive solution. the core dimension remains unaffected by the same changes of temperature. Because all jets pinch at the same location and time. In the first process. By controlling how the solutions are injected. (a) Fluorescence images (left column) and bright field micrographs (right column) of microgels consisting of a 60 m untagged pAAm core encapsulated in a green-tagged pNIPAAm shell. and the particles remain sealed (left three pictures).

and stabilizers for emulsions. it is not possible to create single emulsions from immiscible fluids. by solidifying the monomer solutions using photo-polymerization. (b). As described in the section on capillary microfluidics. Fig 25 (a) Pinch off location of the inner and outer drops for double emulsions. spatially configured chemical properties of these particles. (a) Oil-in-water-in-oil double emulsions are first formed from two monomer solutions using a double flow-focus junction device. they have many uses in pharmaceutics and in industrial processes. because of their miscibility. which is achieved by switching the inlets into which the reactive and inert solutions. By flowing these double emulsions through a confining channel. which pinches into double emulsions in a one-step process. Applications of PDMS double emulsification Templating Janus particles from double emulsions In this section. Copyright 2010 The Royal Society of Chemistry. requires miscible fluids that. As the flow rate ratio is increased. a Janus particle can be composed of an organophilic material on one side and a hydrophilic material on the other. (b). Reproduced from [66]. preventing synthesis of Janus particles with very dissimilar halves. By exploiting the inability of flows to mix quickly at these scales. Copyright 2010 The Royal Society of Chemistry. Copyright 2010 The Royal Society of Chemistry. The drops can then be solidified. However. because it is necessary to first form drops. This. we describe a microfluidic approach to circumvent this constraint. Reproduced from [65]. the inner phase jets into the second junction. we adopt a double emulsion strategy. drops can be formed of two distinct fluids localized in the two halves. often with distinct chemical properties. Rather than using a single emulsion strategy. to lock in this biphasic structure. Janus particles can be formed with controlled properties using microfluidic single emulsification. Reproduced from [64]. tend to have similar chemical composition. enabling synthesis of Janus particles with two very dissimilar halves [66]. Janus particles are biphasic structures that have two halves. as carriers for drugs and other actives. As the flow rate ratio is increased the shell thickness decreases. (c). a limitation of single emulsion formation is that. causing drops to form in the first junction as well as in the second junction. Copyright 2009 American Chemical Society. allowing them to be solidified upon UV irradiation. in general. only fluid combinations that can both be emulsified by the same continuous phase can be used. Reproduced from [65]. Fig 24 Formation of double emulsions using a two-step process. (a). At low ratios the inner phase is in the dripping regime. Fig 26 Creation of organophilic-hydrophilic Janus particles using double emulsions as templates. (b) Shell thickness as a function of flow rate ratio. because a larger fraction of the double emulsion consists of the inner droplet. we induce circulation in their interior that 16 . Both drops contain polymerizable monomers. We encapsulate an organophilic (oily) drop in a hydrophilic (aqueous) drop. a coaxial jet of the inner inside the middle phase is created in the second junction.of the device (b). The drops are solidified while in the device to lock in the Janus structure. Due to the mixed. as shown in Fig 26a. For example. By increasing the flow rate of the inner phase to cause it to jet. To create oil-in-water-in-oil double emulsion the inverse pattern is needed. in which the inner drops are formed in the first junction and then encapsulated in the outer drops in the second junction. (d). in this case the inner and middle phases pinch off at nearly the same place. as a function of the inner-to-middle phase flow rate ratio.

Fig 27 Electron micrograph (upper right) of Janus particles formed from double emulsion templates. because their interaction with magnetic fields makes it possible to detect them through tissue. The number of beads can be controlled by adjusting flow rates. as shown in Fig. shown for different rotation angles in the smaller images. 27. To lock in this structure. Due to the immiscibility of these ingredients. By rotating the 17 . producing double emulsion droplets with two distinct halves. Copyright 2009 Wiley-VCH Verlag GmbH & Co. as shown in the small images in Fig. as illustrated in Fig 26b. Templating magnetic hydrogel particles with anisotropic structure Magnetic microparticles are useful for contrast imaging in the body. in addition to being magnetically responsive. the dye does not go into the organophilic half. because their ability to be manipulated by magnetic fields allows them to be guided to target sites in the body. In this section. To demonstrate the magnetic responsiveness. However. this provides a high contrast image of the internal Janus structure. The inner droplet of the double emulsion is composed of an organophilic monomer/ferrofluid mixture. the rotation is eccentric. we image them with a confocal microscope at several different focal depths. This biphasic structure is maintained provided the double emulsion flows through the constricting channel. providing a high-resolution image of the particle interior. Due to the anisotropic structure of the particle. Because of the small size of the particles. Fig 29 (a) Velocity field of fluid stirred up by rotating a magnetic hydrogel using a magnetic field. To make these structures. This creates the desired Janus particles in which the two halves have very dissimilar chemical properties. to encapsulate one or several cores per double emulsion. 28. Reproduced from [67]. we place a particle in an aqueous fluid containing 2 m polystyrene beads. producing hydrogels with magnetic beads inside. Moreover. We encapsulate a small magnetic bead in a biocompatible hydrogel shell. because of its low solubility in oil. as shown in Fig. causing them to appear bright in the confocal images while the organophilic parts appear dark. Reproduced from [67]. we infuse the hydrophilic halves of the particles with fluorescein dye. The resulting slices are reconstructed into a three-dimensional image of the Janus particle. we create a double emulsion consisting of a ferrofluidic/organopholic monomer for the inner droplet surrounded by an aqueous monomer outer droplet. which freely diffuses into these parts because of the high solubility. The velocity field is averaged over a full rotation cycle. Reproduced from [66]. To see this structure with greater clarity. we image a Janus particle with a scanning electron microscope. (b) Fluid speed averaged over all rotation angles. Because the bead is surrounded by the shell. Copyright 2009 Wiley-VCH Verlag GmbH & Co. it is difficult to spatially resolve the details of the Janus structure.causes the inner drop to migrate towards the back interface. also useful for targeted drug delivery applications. we image them with a confocal fluorescence microscope. the toxicity of the iron oxide nanoparticles that make it magnetic is reduced. This dyes only the hydrophilic portions of the particles. The double emulsions are solidified by polymerizing the monomer solution. KGaA. The flow disturbance peaks at a distance equal to about one particle diameter. Fig 28 Schematic of a microfluidic double emulsion device used to form magnetic hydrogels with anisotropic structure. and have the ability to take up drugs and other active reagents. To visualize the internal Janus structure of the particles. To enable fluorescence imaging. Copyright 2009 American Chemical Society. we describe the synthesis of such biocompatible magnetic microparticles using PDMS double emulsion templating [67]. the hydrogel material can be chosen to maximize the absorption of drug compounds. 27. to be useful. while the outer droplet is composed of a hydrophilic monomer. To confirm the Janus structure of the particles. They are The resulting particles can be infused with compounds and manipulated with a magnetic field. the magnetic bead is entirely encapsulated within the shell of the hydrogel. KGaA. we solidify both drops by exposing them to an intense UV beam. the particles must be biocompatible. The resulting images show a peanut-shaped particle with two distinct halves. as a function of the distance from the center of the rotation.

quadruple. we rotate the bead. Due to their extreme mechanical stability and ability to be ruptured by osmotic shock or changes in temperature. Thus the double-emulsion droplets burst downstream. To maintain the stability of the polymersomes. the diblock-containing solvent must be close to the point of precipitation for the diblock copolymers. the two interfaces of the shell inside the double emulsions are not sufficiently stabilized. by contrast. The microfluidic devices are solgel-coated to increase the resistance of the channel walls against organic solvents. by injecting them into different inlets.magnetic field. (d). 29. The precipitates adhere to the surface of the channels. The disturbance is small at the center of the rotation and grows to a maximum at a fixed distance radially outward. (b) Some of the precipitates are observed in the shell phase of the doubleemulsion drops formed. Fig 31 Serial drop maker arrays used to form multiple emulsions. The geometry also allows us to control the flow rate of each solvent independently. triple. Reprinted from [68]. an example of which is the subject of this section. four. like PDMS devices. KGaA. This demonstrates that the stirring of the fluid is localized close to the particle. A single emulsion droplet takes one drop maker to create (a). and (e). building up a thick layer. they are attractive for applications that require efficient encapsulation and controlled release. we compute the velocity as a function of the distance away from the center. as expected due to the laminar flow properties at this scale. the diblocks would immediately precipitate. b) Formation of PEG-b-PLA stabilized W/O/W double emulsions using premixed mixtures of toluene and chloroform. the motion of which we record by tracking the polystyrene tracer particles. we exploit the customizability of PDMS devices [68]. averaged over all angles. agricultural products. while the coating in the lower part is rendered hydrophilic due to functionalization by grafted poly(acrylic acid). The sol-gel coating in the upper half of the device is hydrophobic. and quintuple emulsions take two. and hence. the jet breaks up into stable double emulsion droplets approximately 1 mm downstream in the outlet channel. fouling them. (b). three. as shown in Fig. which initially connects the double emulsions. because to do so would require large numbers of identical devices – a feat that is not possible with the existing fabrication techniques. However. The non-Newtonian nature of the PVA solution causes the middle phase to develop a tail. However. KGaA. As described in a previous section. changing their wetting. (c) Novel microfluidic device forming PEG-b-PLA stabilized W/O/W double emulsions. (a) Most of the diblock copolymer forms a precipitate after the more volatile chloroform starts to evaporate in the microfluidic device. especially in PDMS devices. Fig 30 Formation of copolymer-stabilized W/O/W double emulsions using a conventional microfluidic device with two cross junctions for injecting premixed organic solvents (left). are much easier to fabricate with identical properties and in large numbers. the osmolarity of the inner and outer phase of the double emulsion templates is balanced by adding glucose to the inner phase and polyvinyl alcohol (PVA) to the outer phase. Templating polymersomes from double emulsions One of the principal advantages of PDMS-based microfluidics is that the lithographic fabrication allows construction of sophisticated channel networks and channels with very fine details. This allows us to obtain 18 . This can be exploited to tailor the devices Lithographically fabricated devices. Copyright 2010 Wiley-VCH Verlag GmbH & Co. The rotation of the particle disturbs the fluid around it. Reprinted from [69]. due to the amphiphilic properties of the diblock copolymers and their interaction with organic solvents needed to emulsify them. glass capillary devices can form monodisperse polymersomes: vesicles composed of a diblock copolymer membrane that encapsulates an active material. as shown in Fig. The wettability of the multiple emulsion devices has been patterned so that it inverts between hydrophilic and hydrophobic from one junction to the next. Copyright 2009 Verlag GmbH & Co. (c). as shown in a failing device to the left in Fig. right half. and five drop makers in series to create. We separate the inlets into which the good solvent saturated with diblock copolymers and the unsaturated poor solvent are injected. forming double emulsions with these solutions is extremely difficult. When such a solution is injected into a PDMS device the diblocks immediately precipitate on the channel walls. The disturbance peaks at a distance equal to the particle diameter and falls off steadily for greater distances. 29b. The challenge is that for the double emulsions to dewet to birth the polymersomes. and preventing the formation of double emulsions. glass capillary devices cannot form them in large quantities. If these two solvents were pre-mixed before reaching the drop-making junctions. including of drugs. as shown by the velocity field summed over a full rotation cycle in Fig. to tune the ratio of the two solvents. as shown in Fig. 29a. (a. Scale bar for all panels denotes 100 µm. they do not mix until after encapsulation in a double emulsion. To quantify the size of the disturbance. 30. However. 29b. and cosmetics. 30. to overcome specific challenges. and using a novel device design allowing separate injection of organic solvents (right). However. Fig. respectively. whereas double. To overcome this problem. Since the organic solvent phase is depleted of the copolymer before the formation of double emulsions occurs.

To form quintuple emulsions. Fig 33 The formation of a double emulsion (water-in-oil-in-water (W/O/W)) in PDMS devices with one and two thicknesses (a). The outer diameters of the quintuple emulsions are 120 m. as shown in Fig. The goal of this section is to describe strategies to relax the constraints of wetting in PDMS devices [70] by mimicking the three-dimensional sheath flow of capillaries. the final quintuple emulsions are identical. glass capillary microfluidic devices can form emulsions of almost any We describe three realizations of geometrically controlled double emulsification in PDMS devices. such that the middle phase never contacts the channel surfaces. 32. encapsulating quadruple emulsions in yet another drop. Because the drops in each stage are monodisperse and the formation is synchronized. which is sheared 19 . where there is a single height step. 31b. This allows formation of emulsions of either type. we use a two-step. multi-thickness device. To form double emulsions. This is the best geometry for forming double emulsions of any composition without having to control wetting. For each junction. (b) Single layered PDMS device that cannot form double emulsions. independent of channel wetting. A high-order multiple emulsion is the logical continuation of a double emulsion to larger numbers of nested drops: A triple emulsion is a double emulsion encapsulated within a drop. Reprinted from [69]. respectively. In this approach. making oil drops energetically favorable. demonstrating how the continuous aqueous phase shields the middle phase from wetting the channels. To make triple or quadruple emulsions. To create these structures requires a microfluidic device having multiple drop makers aligned in series [69]. as shown in Fig. Forming high-order multiple emulsions serial drop maker arrays One of the clearest advantages of PDMS-based microfluidics is the ability to customize the devices. Copyright Assaf Rotem. 31c–d. an ability that is important for forming vesicles form other copolymers. we form emulsions of different orders with serial drop formation devices. we describe an application that exploits this ability. This produces a coaxial jet of the inner and middle phases. KGaA. and centered vertically and horizontally with respect to the nozzle. Reproduced from [70]. For example. to construct sophisticated channel networks. shown in Fig. the device could be uniformly hydrophobic so that it easily forms water-in-oil emulsions. as shown in Fig 33. the inlet channel leading into the nozzle is smaller in crosssectional size than the nozzle. Relaxing the constraints of wetting: double emulsification through geometrical control In contrast to PDMS devices in which wettability properties are crucial when forming emulsions. A simpler version of the device that works nearly as well but is easier to fabricate is to use a two-step.solvent ratios that normally could not be used in polymersome synthesis because of precipitation. To illustrate this. we also pattern wettability. the wetting of the device is of a uniform composition such that one type of emulsion is always preferred. Fig 32 Optical microscope image of quintuple emulsions formed with serial drop formation device. We begin by forming single emulsions using a single drop maker. two-thickness approach. we add another drop maker. while a quadruple emulsion is merely a triple emulsion inside yet another drop. (c) Two-thickness device with the same hydrophobic wetting properties that can easily form double emulsions. 31e. two thickness device. scaling up device complexity to form high-order multiple emulsions. (e) Top and Side views of a similar device with two thicknesses. immiscible fluids. This is used in the first junction to form the inner drops and then a step is used in the second junction to form the outer drops. because the middle oil phase wets the hydrophobic walls better than the continuous phase. Water-inoil drops form at the first junction and oil-in-water form at the geometrically controlled junction. Copyright 2009 Verlag GmbH & Co. 31a. as shown in Fig. independent of how they wet the device surfaces. (d). we simply add third and fourth junctions. A third implementation is to use a one-step. In this section. all fluids can be injected into a single junction. for example. because the threedimensional channel graduations reduce the importance of wetting. we add a fifth junction. in capillary devices the axially symmetric injection into the orifice imposes a topology on the phases. This difference stems from the distinctive geometrical properties of these devices: Whereas in PDMS devices all fluids are initially in contact with the channels and must be lifted off the surface by preferential wetting. as shown in Fig. but the multiple height steps needed make fabrication somewhat labor intensive. so that the inner drops are encapsulated within larger drops of an immiscible phase. In the first.

including polymer spheres. and core-shell capsules. stepping both up and down where the channels widen. illustrated in Fig. which occurs after the water diffuses away into the blocks while in an oven set to 65°C. we use water as a lubricating fluid. mechanically aligning the two blocks so that the channel features are aligned. chemical composition is determined by selecting which fluids to introduce into the device. To do this. a challenge in commercializing these techniques is to increase production yields: current systems typically produce only microliters-tomilliliters of product per hour. Available methods utilizing bifurcating channel networks lack the uniformity needed. during this process the water evaporates. An especially useful property of using mechanical alignment is that the device remains well aligned even during the heating step.S. because they can be replicated in large numbers with precision. development of such a system is critical for these techniques to achieve their potential in particle synthesis applications. (b) and (c) Two replicates of the PDMS device are cured and then bonded facing each other using plasma bonding. allowing the silanol groups in both blocks to react. The friction of the locks and keys maintains the alignment through the heating process. which are then aligned and bonded to other multi-height channels in a second block of PDMS. To lubricate the blocks and allow the key to explore the lock. and will likely comprise the next stages of microfluidic research. a droplet of water is placed on the freshly plasma oxidized surfaces. thousands or hundreds-of-thousands could be used in parallel. friction holds the blocks in place. Reproduced from [70]. Copyright Assaf Rotem. Nevertheless. (d) The protruding “key” on one PDMS block slides into the sunken “lock” on the other half. including therapeutics. resulting in polydisperse particles. This allows construction of devices in which the channel steps are symmetric in the vertical direction. compressibility of the solutions. is a research fellow of the German Academy Fig 35 Fabrication process for two thickness emulsification devices. Copyright Assaf Rotem. After aligning the blocks. and the Massachusetts Life Sciences Center. while a known and challenging problem. and cosmetics. Conclusions The principal advantage of microfluidic synthesis is the ability to independently choose the chemical composition and structure of the particles formed. we use multi-layer lithography to create multi-height channels. we use a simple approach in which matching physical locks and keys mechanically align the PDMS blocks. and Christian Holtze for their help. One of the PDMS blocks has a hole (the lock) of a particular shape and at a specific location. While the structure of the particles is determined by the flow properties in the devices. 35. we use standard fabrication techniques in which a silicon wafer is coated with photoresist and exposed to UV light through different photomasks in several iterations. This enables synthesis of a range of particles with distinct composition and structure. To fabricate these multi-height devices. Rather than a single particle synthesis device operating at a time. as shown in Fig. S. W/O/W double emulsions are formed with different volume fractions. (e) An image of a sample device including a channel and a lock and key. irreversible seal. 34. the other block has a protrusion (the key) of the same shape and at the same location. To allow the keys to explore the locks. from 1:1 (inner:shell) volume fraction in the left image to 25:1 (inner:shell) volume fraction on the right. Acknowledgments We thank Jinwoong Kim. with volume fractions of 1:25 shell/inner phase. the key slides into the lock. One strategy to increase yield is to parallelize the devices.into double emulsions by a sheath flow from the continuous phase. crop protection. as shown in Fig. and large hydrodynamic resistance of the dispensing channels lead to long-lived pressure oscillations and uneven flow. medical imaging. 35 d. Fig 34 Microfluidic devices with two different hights for the formation of double emulsions in one step. since the compliance of the devices. This should increase production rates to the levels needed for commercial applications. an open challenge to achieving controlled parallel production is the development of a uniform fluid distribution system. However. Reproduced from [70]. maintaining the alignments through the bonding process. When the two faces of the PDMS are brought into contact. the Harvard MRSEC (DMR-0820484). However. Bingjie Sun. The water retains the silanol bonds critical for covalent bonding of the blocks. creating an 20 . a more challenging aspect of the fabrication is aligning and bonding the two PDMS channel slabs together. In this respect PDMS devices hold the greatest potential. the device is placed in an oven set to 65 °C for 1 hr. To make the multilayer channels. This work was supported by the NSF (DMR-0602684). when thermal expansion of the PDMS could break the alignment. Many of these particles have uses in commercial applications. (a) A twoheight master is prepared using photolithography. Janus particles. nested multiple emulsions. This produces double emulsion with very thin shells. However.

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