Recent advances in multiple emulsions and their application as templates

Bruno F.B. Silva, Carlos Rodrguez-Abreu, Neus Vilanova

PII: S1359-0294(16)30098-X
DOI: doi: 10.1016/j.cocis.2016.07.006
Reference: COCIS 1062

To appear in: Current Opinion in Colloid & Interface Science

Received date: 8 March 2016

Revised date: 7 July 2016
Accepted date: 8 July 2016

Please cite this article as: Silva Bruno F.B., Rodrguez-Abreu Carlos, Vilanova Neus,
Recent advances in multiple emulsions and their application as templates, Current Opinion
in Colloid & Interface Science (2016), doi: 10.1016/j.cocis.2016.07.006

This is a PDF le of an unedited manuscript that has been accepted for publication.
As a service to our customers we are providing this early version of the manuscript.
The manuscript will undergo copyediting, typesetting, and review of the resulting proof
before it is published in its nal form. Please note that during the production process
errors may be discovered which could aect the content, and all legal disclaimers that
apply to the journal pertain.
 ACCEPTED MANUSCRIPT

Recent advances in multiple emulsions and their application as templates

Bruno F. B. Silvaa, Carlos Rodrguez-Abreua* , Neus Vilanovab

a
International Iberian Nanotechnology Laboratory (INL). Av. Mestre Jos Veiga s/n. Braga 4715-

PT
330 Portugal

RI
b
Technische Universiteit Eindhoven, Institute for Complex Molecular Systems, Department of
Chemical Engineering and Chemistry, P.O. Box 513 5600 MB Eindhoven, the Netherlands.

SC
*Corresponding author. E-mail address: carlos.rodriguez@inl.int

NU
Keywords: multiple emulsions; microfluidics; formulation; encapsulation; porous particles
MA
Highlights
D

Microfluidic concepts and their use to prepare multiple emulsions are reviewed.
TE

Advantages and new prospects for microfluidics are discussed

Recent advances on bulk multiple emulsification are presented.
P

Methods to obtain porous particles from multiple emulsions are described.

CE
AC

1
 ACCEPTED MANUSCRIPT

Abstract

We review recent developments on the preparation of multiple emulsions and their

applications as templates for material fabrication. Emphasis is placed on microfluidic methods for

PT
accurate control of size and morphology and on new formulations that go beyond traditional
surfactant systems for increased complexity. The straightforward applicability of multiple

RI
emulsions in the fabrication of multihollow particles or capsules of various materials is illustrated
through representative examples.

SC
1. Introduction

NU
Multiple emulsions are soft materials formed by dispersed droplets (called outer droplets
herein) that contain in turn smaller droplets inside (called inner droplets herein). Their particular
MA
features as multicompartmental units with confined spaces in a continuum offer a wide range of
possibilities for different applications.
D

Following the conventional nomenclature for normal emulsions, multiple or double

TE

emulsions can be classified into water-in-oil-in-water (W/O/W) or oil-in-water-in-oil (O/W/O)

depending on the phase sequence at different scales; in such a nomenclature, water actually
P

means any aqueous or polar phase (even e.g. an ionic liquid [1]) whereas oil stands for any
CE

hydrophobic, i.e. water-insoluble phase. Such a phase sequence not only implies the necessity of
selective stabilization of droplets of the different phases but also finding a proper method to
AC

emulsify emulsions, namely, to produce small droplets inside bigger ones. The two traditional
and well known bulk methods to form multiple emulsions are the two-step method and the phase
inversion method. These methods usually render polydisperse droplets.

In the two-step method, briefly a simple W/O emulsion is produced first using a
hydrophobic surfactant. Then this emulsion is poured into an aqueous phase containing a
hydrophilic surfactant that helps to emulsify the initial emulsion so that a W/O/W multiple
emulsion is obtained. Alternatively, an O/W/O emulsion can be produced if one starts with an
O/W emulsion stabilized with a hydrophilic surfactant that is later emulsified in an oil phase
containing a hydrophobic surfactant.

2
 ACCEPTED MANUSCRIPT

The formation of multiple emulsions by phase inversion is driven by changes in the

interfacial curvature and in the hydrophile-lipophile balance of the surfactant. Phase inversion
can be transitional, as occurs at the phase inversion temperature (PIT), or catastrophic, as a result
of a change in the volume fraction of one of the phases. Readers are referred to the abundant

PT
literature on the subject for in-depth description of these phenomena.

RI
Multiple emulsions have potential applications in encapsulation, template-assisted
synthesis of materials or separation. In practice, however, their use in commercial applications

SC
remains challenging due to a lack of simple methods to create stable and reproducible multiple
emulsions. Hence, studies on new emulsifiers or other stabilization strategies as well as on

NU
methods that combine high throughput and size and morphology control are continuously
emerging
MA
Herewith, we review recent advances on the preparation of multiple emulsions,
particularly focusing on droplet microfluidics and new formulations, as well as the application of
D

these multiple emulsions as templates for the fabrication of porous particles and capsules.
TE

2. Double and multiple emulsion production by microfluidics

P

2.1 Microfluidics for droplet generation

CE

Microfluidics involve the flow and manipulation of fluids in channels with typical
dimensions of tens to hundreds of micrometers [2]. At such small length scales, the Reynolds
AC

number (Re) becomes small and inertial forces practically negligible. This leads to predictable,
reproducible and controllable flow regimes that can be used with great advantage to manipulate
materials for practical applications or studying natural phenomena. When two immiscible
flowing fluids merge in a microchannel, the balance between the viscous forces (induced by the
continuous phase) and the capillary forces (resulting from the interfacial tension between the two
liquids) leads to different flow regimes or breakup modes [3,4], with the most common being
dripping and jetting [5]. In the dripping regime, highly monodisperse droplets (coefficient of
variation ~1-5%) of tunable size form right at the mixing point, while in the jetting regime,
prevalence of viscous drag leads to the formation of a jet, which typically breaks into less
monodisperse droplets further downstream due to the Rayleigh-Plateau instability [2]. The
prevalence of each mode can be easily controlled by means of adjusting the flow rates.

3
 ACCEPTED MANUSCRIPT

For droplet production, many junction designs and combinations exist, but overall they
can be grouped into three main types [4,6]: T-junctions [7], co-flowing [8], and hydrodynamic
flow-focusing [3,9] Fig 1 (consult for instance Ref. [10] for a more detailed review on different
device designs). In a T-junction, fluids typically meet at an angle of 90, and droplet formation

PT
arises from the pressure drop in the continuous phase and squeezing of the dispersible phase [11].
In a co-flowing device the fluids meet in parallel streams, whereas in the flow-focusing design

RI
there is a geometric element that focuses/squeezes the inner fluid thread [4]. In both co-flowing

SC
and flow-focusing geometries, droplets of the dispersed phase typically form through the above-
mentioned dripping or jetting modes [5]. T-junctions are typically 2D devices, co-flow set-ups

NU
are made of 3D microcapillaries, while flow-focusing is available in both 2D and 3D geometries.
2D devices can be fabricated in glass by etching and in polymeric materials by soft lithography
(e.g. poly(dimethylsiloxane) PDMS, and poly(urethane)), thermal embossing and injection
MA
molding among other methods [10]. Simple 3D devices are typically assembled out of glass
capillaries but nowadays 3D devices can also be fabricated with polymeric materials, which
D

allow not only mimicking the coaxial structure of capillaries [12], but also the fabrication of
TE

complex 3D structures and networks [13,14].

Soft lithography fabrication methods, e.g. for making 2D or 3D PDMS devices, are
P

especially attractive due to the potential to include a high number of junction modules (e.g. T-
CE

junctions, flow-focusing, functions for sorting droplets, etc [15,16]), ideal to achieve more
complex assembly tasks such as a controlled production of multiple emulsions, or for
AC

parallelization of several identical processes for high-throughput. In addition, the possibility to

design true 3D devices allows for an enhanced control and potentiates the use of additional
functionalities on-chip. Nevertheless, the most typical co-flowing devices are still made of glass
and are ideal also for one-step emulsification of multiple emulsions [17]. Like PDMS devices,
they also allow cascading of multiple junctions to build ever-growing complex multi-emulsions,
but one drawback is the required expertise to assemble such devices out of glass capillaries, and
the difficulty in parallelization for a higher production rate of droplets. Recently, the possibility
to use 3D-printers to fabricate polymeric devices with 3D features has also gained much deserved
attention [14]. These fabrication methodologies are still somewhat limited in the resolution that
they offer, but they allow the facile design of very intricate 3D structures and networks that are
fabricated in a single step, without the need of cleanroom facilities and fabrication expertise. As

4
 ACCEPTED MANUSCRIPT

the technology is successfully reaching better and better resolution capabilities, 3D-printing
device fabrication will allow a facile, accessible, and affordable way to make complex
microfluidic devices.

PT
Another important parameter for droplet production in microfluidics is the wettability.
Typically, the dispersed phase (i.e. droplets) should not wet the device walls [18]. This is

RI
challenging in the case of multiple emulsions since some different sections of the same device
should wet the middle and outer phases which have different hydrophobicities. In glass

SC
microcapillary devices, the problem is avoided due to the 3D structure of the channels which help
to engulf the inner and middle phases. Fortunately, fabrication of PDMS on-chip devices with

NU
channels of different depths (so that they become closer to the 3D approach) is now
straightforward. Such channels help preventing the droplets from touching the walls, and under
MA
such conditions, wetting of the channels becomes less problematic [12]. Also, now,
functionalization of different sections of a polymeric device with different wetting properties is
easily achieved, which renders them extremely versatile for most multiple emulsion fabrication
D

applications [1922].
TE

Insert Figure 1
P
CE

Figure 1. Typical microfluidic geometries and droplet formation modes (dripping and jetting). (A) T-
junction [7]. (B) Co-flow [5]. (C) Flow-focusing through a constriction [9]. (D) Flow-focusing in a cross-
junction [3]. (E) Two T-junctions in series for double emulsion production in two steps [23]. (F) Flow-
AC

focusing geometry for double emulsion production in one step [24]. (G) Four flow-focusing cross-
junctions in series for the production of quadruple emulsions [20]. (H) Microcapillary coaxial flow-
focusing device combining co-flow with flow-focusing for the production of double emulsions in a single
step either in the dripping (I) or jetting (J) regimes [8]. (I) Double emulsion droplet production in the
dripping mode. (J) Coaxial jet before pinch off. Inset on the bottom: Double emulsion droplet pinch off
from the coaxial jet. Panels (H-J) from Ref. [8]. Reprinted with permission from AAAS
http://dx.doi.org/10.1126/science.1109164.

2.2. Device geometry and experimental considerations for the production of double and multiple
emulsions
The production of double and multiple emulsions through microfluidic manipulation can
be achieved through two main approaches: (1) by cascading further droplet generator modules
downstream, or (2) by variations of the modules design so that multiple emulsions are produced
at a single step.

5
 ACCEPTED MANUSCRIPT

In the first approach, by placing two T-junctions in series in a 2D glass device, double
emulsions of W/O/W and O/W/O can be produced [23] (Fig 1E). Droplets of uniform size of the
inner phase form reproducibly in the middle phase at the first T-junction and this resulting stream
then breaks into droplets at the second junction, forming droplets of the middle phase (which in

PT
turn contain the inner phase) in the continuous phase. The internal and external droplet size and
number of encased droplets can be accurately controlled by manipulating the flow rates of the 3

RI
phases, whereas two droplets of the inner phase can be incorporated within a single middle phase

SC
droplet by replacing the first T-junction with a cross-shaped module. Later, similar devices were
also manufactured in PDMS with similar results [19].

NU
For the second approach, double emulsions were obtained using a 2D flow-focusing
device in poly(urethane), designed to form a coaxial jet of the inner and middle phases embedded
MA
in an outer fluid [24] (Fig 1F). Just by manipulation of the relative and absolute flow rates, the
double jet would break into droplets in such a way that it allowed control over the inner and outer
droplets size (hence, middle phase thickness). Importantly, by adjusting the flow rates of the
D

three fluids, jet breakup of the inner and middle fluids can be adjusted to incorporate a controlled
TE

number of inner droplets inside the outer one. Similar results were also achieved in coaxial flow-
focusing glass microcapillary devices, which combine the co-flow and the flow-focusing
P

geometries [8] (Fig 1H). The setup consists of two tapered cylindrical capillaries, aligned in the
CE

same axis and embedded within a square capillary. The inner fluid is injected through the
injection capillary, whereas the middle fluid is injected through the interstices between the
AC

injection (cylindrical) and squared capillaries. This forms a coaxial flow at the tip of the tapered
capillary. The outer fluid flows from the opposite side between the interstices of the collection
and squared capillary, and focuses the coaxial stream of the inner and middle fluid into the
collection capillary. Double emulsion droplets could be formed, with both inner and middle fluids
breaking into droplets either in the dripping or jetting regime, depending on the individual fluids
flow rates [8] (Fig 1I-J). This configuration allowed not only control over the outer and inner
droplets (hence the shell thickness), but also on the number of inner droplets. Interestingly, thin
shells (~3% of the total droplet volume) could be produced, which allowed the formation of
polymersomes, liposomes, polymer shells, and an array of other interesting materials [17,25].

6
 ACCEPTED MANUSCRIPT

Using combinations of these concepts, emulsions can grow in complexity either by

cascading more junctions downstream of the channel [20,26,27] (first approach), or varying the
design in order to allow the formation of more complex emulsions in a one-step fashion [28,29]
(second approach). Within the first strategy, by combining two or three coaxial co-flow junctions,

PT
monodisperse double or triple emulsions are formed in two or three steps respectively [26]. The
separation of the multiple emulsification steps allows enhanced control over each, by allowing

RI
manipulation of the individual inner, middle and outer flow rates. However, the fact that the outer

SC
fluid at one junction becomes the middle fluid at the following junction, still brings some
constraints into full control of the designer emulsion.

NU
An improvement for this strategy consists in incorporating two new modules to the
previous design: a connector and a liquid extractor [27] (Fig 2A). The connector allows the
MA
combination of two coaxial co-flow droplet generation devices in such a way that two different
droplets join in the same stream, flowing alongside and being co-encapsulated into an outer
droplet further downstream. The connector also allows addition of more fluid to increase the
D

spacing between the two droplet types, whilst the liquid extractor allows the removal of excess
TE

liquid to bring the droplets closer. These two new building blocks permit very different
combinations to produce highly hierarchical multiple emulsions with tailored designs such as
P

sextuple-component triple emulsions.

CE

The fabrication of these glass devices, including several aligned tapered microcapillaries,
AC

is difficult and labor intensive. In order to avoid the drawbacks associated with fabrication, the
same concept could also be translated to PDMS devices, whose wettability can be manipulated by
UV radiation through a patterned mask that allows only the regions of interest to be exposed [20].
The shape of the junctions is adjusted for a triggered drop formation (each nozzle is slightly
narrower than the incoming emulsion) in order to synchronize the formation of drops and allow
the production of triple, quadruple and quintuple emulsions. The control over the size and
monodispersity of the individual droplets was excellent, but with the geometry in question it was
not reported the incorporation of more than one droplet within the same hierarchical level as in
the preceding example.

As the continuous addition of further junctions downstream makes the device fabrication
very difficult in glass microcapillary-based devices and fluid operation increasingly more

7
 ACCEPTED MANUSCRIPT

complex, the introduction of variations on the droplet generation modules was also brought as an
alternative. Hence, a variation of the combined coaxial flow-focusing setup introduced before,
but now with a clever modification of the capillaries affinity for the different phases was
proposed. This brings the ability to produce triple and quadruple emulsions in a single step [28].

PT
Now, instead of flowing a single fluid through the interstices of the square and
collection/injection capillaries, two immiscible fluids are flowed together from each side (Fig

RI
2B). These fluids are stabilized against breakup by the confinement within the interstices, and by

SC
modification of the wettabilities of the square and circular capillaries. Hence, a flow scheme can
be produced to generate monodisperse quadruple W/O/W/O/W and O/W/O/W/O emulsions.

NU
Droplet formation is achieved in the dripping and jetting regime and determined by the flow rates
of the individual fluids. In the dripping regime, monodisperse highly regular onion-type
MA
structures are produced, whereas the jetting regime allows incorporation of several inner droplets
in the core.

Alternatively, by replacing the tapered inner fluid capillary of a coaxial flow-focusing

D

setup with a dual, triple and even quadruple bore injection capillary, one-step production of
TE

double emulsion with multiple different inner droplets can be achieved [29] (Fig 2C).
Importantly, the ability to independently change the flow rate and dripping to jetting regime of
P

both the outer droplet and each of the individual inner droplets, allows the production of a wide
CE

range of interesting double emulsions. An application was demonstrated for the triggered
coalescence of inner droplets, but applications for exquisite control of the structure of porous
AC

polymer particles could also be foreseen. In a slight modification of the device, by introducing
smaller capillaries inside a two- and four-bore injection capillary to produce segmented flows of
oil and water, and by hydrophobizing the inner surfaces of the injection capillary, very interesting
nonspherical double emulsions with multiple distinct water cores enveloped by very thin shells
could be produced [30].

One third approach to produce multiple emulsions using microfluidic methods is to revert
to a single junction where a complex mixture specifically tailored for the application is
emulsified in a suitable outer fluid [3136]. As the emulsion forms, phase separation within the
droplets occurs, leading to the formation of multiple emulsions (see more details in the following
section). The role of the microfluidic device here is more to control the size and monodispersity

8
 ACCEPTED MANUSCRIPT

of the outer droplets, while providing conditions for controlled mixing and diffusion (i.e. mass
transport) of the different components in and out of the emulsion droplet. Hence, the microfluidic
devices are here simplified, while the specific formulation (including the composition of the outer
phase) has to be specifically tailored to provide the out-of-equilibrium phase transformations

PT
within the droplets. Due to device simplicity these methods are potentially more attractive for
practical applications [6,31].

RI
2.3 Challenges for the microfluidic technology

SC
Each of the described approaches have advantages and disadvantages, but to a wide

NU
extent, the current technology seems to be sufficient for a wide range of applications. However,
two immediate limitations come presently to mind: achieving droplet dimensions of
submicrometer size (e.g. nanoemulsions and nanoparticles) and output scalability for industrial
MA
production.

Besides the flow conditions, droplet sizes typically also scale with the microchannel
D

dimensions according to Rdrop d -1, with d being the channel diameter [2]. Hence for production
TE

of submicrometer emulsions nano-sized channels are desired. However this is not an easy task.
The fabrication of channels of submicrometer dimensions is not straightforward and accessible,
P

and furthermore, pumping fluids through small nanometer sized channels becomes more
CE

challenging due to the huge hydrodynamic resistance that scales with Rhyd. ~ Ld-4 (with Rhyd.
being the hydrodynamic resistance, the fluid viscosity, and L the channel length). New ways for
AC

fluid transport may then need to be realized. In addition, in nanosized channels the cross-section
dimensions through which fluids flow become comparable to the screening lengths of double
layer and other interfacial forces [37], making the overall emulsification processes more difficult
to predict. One additional problem that cannot be overlooked is the difficulty for in-situ
visualization of the nanochannels with conventional laboratory instruments, making the
experiments much more complicated, and calls for the use of other in-situ characterization
methods such as X-ray scattering [38,39] and TEM [40]. For all of these reasons, the use of
nanofluidic devices for droplet production has been scarce. Recently one nanofluidic system was
successfully used for the preparation of very small multiple emulsions. Yet, the minimum size
reached was still around 1 m [41]. In that example, a clever use of a cross design, where the
liquid is injected into wide microchannels that branch out into nanochannels and an open end

9
 ACCEPTED MANUSCRIPT

kept at constant pressure, helps to control the pressure drop across the nanochannels and prevents
clogging due to dust particles. The fluids within the nanochannels form a jet in a flow-focusing
cross. The jet is stabilized by the nano-sized channels, but breaks into droplets when it reaches a
micron-sized reservoir, in a process called step-emulsification [42], where the height of the

PT
nanochannel is the most important parameter controlling the size of the droplets. By manipulation
of the fluid components, simple, multiple and Janus emulsions could be produced. With a related

RI
device, but by further decreasing the nanochannel dimensions, droplets of around 400 nm could

SC
be prepared, although in this case, no multiple emulsion preparation was reported [43].

Regarding practical applications, one of the most concerning drawbacks of microfluidic methods

NU
for emulsion preparation and particle synthesis is its low output, which typically does not exceed
more than 1 mL/h per device, completely impractical for the industry. One way of partially
MA
overcoming this limitation is to scale-up by parallelizing several copies of the device within the
same chip, so that a single set of pumps can be used to drive the fluids through the device array
[4446]. Here, soft lithography methods provide an obvious advantage compared to
D

microcapillary methods, since scaling-up many copies of the same device on a wafer is
TE

straightforward. Still, a few problems have to be overcome in order to make the procedure
feasible and entirely scalable. One of these problems is that very minor differences in the several
P

copies will increase the polydispersity of the droplets (e.g. even just a small tilt in the silicon
CE

wafer during the spin-coating process can provoke minor variations in channel height from
device to device [45]). Another problem is that since the different copies are fed by the same
AC

liquid source, droplet formation in one copy creates pressure oscillations that transmit to the
source and influence the other copies. This cross-talk can be dampened by increasing the
hydrodynamic resistance of the channels between the droplet makers and the common feeding
channels [45]. By increasing the hydrodynamic resistance of the droplet maker feeding channels
and lowering the resistance of the common supply channels, it is also ensured that the pressure
drop is the same in each channel, and therefore the flow rates and droplet sizes are also identical
across the different copies [46]. Another issue to overcome is the wettability of the channels.
Typically the dispersed phase should not wet the walls of the device, which is challenging when
producing double and multiple emulsions. As mentioned earlier, devices with sections with
different wettabilities are easy to make nowadays, but in a scaled-up parallelized device the
challenge is still considerable. One way to avoid this is to simply ignore the different

10
 ACCEPTED MANUSCRIPT

wettabilities, and tune the size of the channels at each junction so that the droplets formed do not
touch the microchannel walls [12,46]. Having these criteria in mind, a device containing 15
parallelized double emulsion drop makers was able to produce drops at rates of 1 Kg/day with a
polydispersity inferior to 6%, whereas continuous scaling to a device fulfilling 1 L of volume

PT
could produce around 30 tons of double emulsion droplets per year [46]. Even so, this figure is
relatively small for industrial standards, which makes material synthesis with microfluidics

RI
economically viable only for materials with very high added value, such as pharmaceuticals and

SC
cosmetics [17].

Insert Figure 2

NU
Figure 2. Multiple emulsion production with glass microcapillary devices. (A) Five-component triple
emulsion formation using a multi-step microfluidic system. (a1) schematic of the microfluidic assembly
MA
made out of three different modules: connector, liquid extractor and droplet generator. (a2-a11) different
multiple emulsion droplet combinations obtained with different scheme arrangements of the modules
shown in a1. The scale bar represents 200 m. Reproduced and adapted from Ref. [27] with permission
from The Royal Society of Chemistry http://dx.doi.org/10.1039/c1lc20065h. (B) Microfluidic one-step
D

production of quadruple emulsions. (b1) Scheme of the surface-modified microcapillary coaxial flow-
focusing device. Orange walls are hydrophobic and blue walls are hydrophilic. (b2) Snapshot of quadruple
TE

emulsion formation in the microfluidic device. (b3) Collected monodisperse quadruple emulsions, along
with a very small amount of single and double emulsions. Reproduced and adapted with permission from
P

[28]. Copyright 2011 John Wiley and Sons. (C) Microfluidic one-step production of multi-component
double emulsions. (c1) Schematic of a coaxial flow-focusing device with a two-bore injection capillary for
CE

the production of double emulsions with a controlled number of multi-component inner drops. (c2)
Snapshot of double emulsion droplet generation containing two different inner drops. (c3) Color optical
micrograph of double monodispersed double emulsions produced in a2, each containing one red and one
AC

blue inner droplets. The scale bars represent 100 m. Reproduced and adapted from Ref. [29] with
permission from The Royal Society of Chemistry http://dx.doi.org/10.1039/c2sm25953b.

3.Innovative formulations for multiple emulsions

3.1 New copolymer stabilizers for one-step preparation of multiple emulsions

The preparation of multiple emulsions with bulk methods generally involves a time
consuming two-step procedure and the combination of hydrophilic and hydrophobic stabilizers,
as indicated at the beginning of this paper. In this context, amphiphilic block copolymers or
biopolymers are increasingly being used as emulsifiers. They do not only decrease the interfacial

11
 ACCEPTED MANUSCRIPT

tension, but also increase emulsion stability by multiple anchoring which translates into a higher
energy of adsorption at the interface. Moreover, the high efficiency of polymers means that they
perform at relatively low concentrations which can have positive impacts on industrial costs.
With this in mind, major efforts are dedicated on exploring new copolymers that allow preparing

PT
stable multiple emulsions in one step. Nowadays, studies are focused on understanding the
correlation between the copolymer structure and its emulsification performance, as the

RI
interactions with the other emulsion components were shown to be decisive [47]. For instance,

SC
Poly(ethylene glycol)/Polypropylene (PEG/PPG)-glycol-dimethicone copolymers with different
hydrophilic-hydrophobic length segments have been used to prepare silicone multiple emulsions

NU
[48,49], because of their dual compatibility with aqueous and PDMS phases. There are also few
examples where a unique copolymer is used to stabilize inner and outer droplets, thus ruling out
MA
possible destabilization pathways that might be present in surfactant mixtures. Promising
formulations were found using pH-responsive polystyrene-2-(dimethylamino)ethyl methacrylate
(PS-DMAEMA) based copolymers with different asymmetric architectures [50,51]. Upon
D

increasing the pH, the system reaches a sufficient low interfacial tension to form transient
TE

bicontinuous structures. The asymmetric fragmentation of this structure, caused by the

asymmetric partition of the copolymer, results in the formation of a discontinuous oil phase with
P

nested water droplets, subsequently evolving to a multiple emulsion. Similarly, asymmetric block
CE

PEG-lysine copolymers mixed with various electrolytes at different concentrations were also
investigated. In this particular case, the interactions between the electrolyte anion and the cation
AC

residues of the lysine aminoacid seemed to be the main responsible for the long-term stability
[52]. Additionally, the hydrogen-bonds between the peptide segments provided an extra
mechanical stabilization. Contrary to these examples, a symmetric PEG-b-PS based copolymer
showed to be a superior stabilizer compared to its asymmetric counterparts [53]. Here, the
stability was ascribed to the high surfactant coverage in both interfaces, providing at the same
time steric repulsion. Despite the advantages that the one-step processes offer in terms of
processing, they still do not allow a control over the emulsion morphology as accurate as the two-
step processes.

Insert Figure 3

12
 ACCEPTED MANUSCRIPT

Figure 3. Formation of multiple emulsions via different pathways. A) Schematic representation

of the in-situ functionalization of silica@PEI nanoparticles, placed at the interfaces of an
emulsion, with an aldehyde dissolved in the organic phase (left) and a microscopy image of the
resulting Pickering multiple emulsion (right). Adapted with permission from [54]. Copyright
2014 American Chemical Society. B) Evolution in time (in seconds) of a diethyl
phthalate/ethanol/water (0.41/0.42/0.17 vol%) mixture from a singlet droplet to a quintuple

PT
droplet. Scale bar is 100 m. Adapted with permission from [35]. Copyright 2014 Wiley-VCH.
C) Single droplets of a hexaneperfluorohexane mixture in water. Above 22.65 oC (Tc), both

RI
organic solvents are miscible, below the Tc a phase separation is induced, which yields a double
emulsion. Scale bar is 200 m. Reprinted with permission from [36]. Copyright 2015 Macmillan
Publishers Ltd: Nature.

SC
3.2 Multiple Pickering emulsions

NU
The use of particles as emulsifiers is a well-established strategy to produce highly stable
emulsions, coined as Pickering emulsions. Inspired by results on simple emulsions, research on
MA
multiple Pickering emulsions has developed. Actual formulations include both emulsions co-
stabilized by surfactants and particles or solely by particles. In the latter case, solid particles of
opposite wettability are required, analogous to surfactants with different hydrophile-lipophile
D

balance (HLB) values for conventional multiple emulsions. As expected, the morphology,
TE

rheological properties and hence stability, of multiple Pickering emulsions can be regulated by
adjusting the particles/surfactant ratio or the ratio between different types of particles. Particles
P

used as stabilizers include inorganic particles (SiO2 [55], Fe3O4 [56,57]) or hybrid [58], food-
CE

grade particles such as cellulose derivatives [59] or quinoa [60], or even soft particles as
PNIPAM nanogels [61]. Given that the use of SiO2 particles is the most extended strategy,
AC

several more sophisticated silica-based particles have emerged. For instance, the wettability of
hydrophilic poly(ethylene imine)-functionalized silica particles (silica@PEI) can be modified in-
situ (within the emulsion), by coupling an hydrophobic aldehyde dissolved in the oil phase.
Through such in-situ interfacial reaction, the hydrophobicity of the particles increases enough to
stabilize the W/O interface and yield multiple emulsions that are stable for months [54] (Fig 3A).
Similar to surfactant-stabilized multiple emulsions, Pickering multiple emulsions via one-step
processes have also been produced by using anisotropic cellulose particles [62] or stimuli-
responsive styrene-acrylic acid particles [63]. However, as for surfactant-stabilized emulsions,
the two-step method is still the most adequate to produce stable multiple Pickering emulsions
with a certain control over their structure (droplet and globule sizes).

13
 ACCEPTED MANUSCRIPT

3.3 Interfacial complexation

In the food industry, multiple emulsions are thoroughly investigated as promising

reduced-fat formulations. Unfortunately, the allowed surfactant concentrations in this field are

PT
relatively low and there is only a limited number of food emulsifiers approved. To solve this
problem, current studies are focused on expanding the range of edible stabilizers in the market

RI
including, besides the edible particles, protein-polysaccharide mixtures [64,65]. A better degree
of food structuring is obtained when the biopolymers are added individually, whilst the addition

SC
of the preformed complex yields a much more effective release system. Differences are attributed
to a different arrangement of the biopolymers at the interface.

NU
3.4 Gelation routes MA
Emulsion stabilization has also been achieved by adding a thickener or a gelling agent
such as whey protein [66], sodium caseinate [67], starch [68], xanthan gum [69], alginate [70] or
D

carrageenan [71] into one or more of the phases. By reducing the mobility of the droplets, its
TE

coalescence rate is decreased. This also allows to reduce the concentration of surfactant or even
to avoid its use. In addition, when gelling the inner phase, the resulting multiple emulsion appears
P

to be less sensitive towards an imbalance in the osmotic pressure. As a function of the thickener
CE

or gelling agent used, multiple emulsions with a wide variety of rheological properties can be
prepared. Such capacity to form different gel structures is attractive for the cosmetic and food
AC

industries, where textures are particularly important.

3.5 Multiple emulsion formation assisted by an induced-phase separation

Even though in most of the cases a phase separation seems to be a detrimental process, an
induced-phase separation of a simple droplet can also be a powerful tool to form multiple
emulsions. Normally, the dispersed phase contains a cosolvent that is more soluble in the
continuous phase than in the dispersed phase. Hence when the dispersed phase-cosolvent mixture
gets in contact with the outer fluid, a mass transport from the cosolvent into the outer fluid and
vice versa occurs, inducing a phase separation within the droplet mixture [31,34,35]. That is why
current multiple emulsion advances also comprise the study of the underlying mechanisms of
phase separation processes. By understanding and mapping these processes, the formation of

14
 ACCEPTED MANUSCRIPT

complex systems such as onion-like multiple emulsions is simplified and can even be predicted
[35] (Fig 3B). For instance, hexane-perfluorohexane multiple emulsion showed to have
transitions from a multiple emulsion to a Janus droplet or even a phase inversion that could be
triggered by changing the temperature (Fig 3C), irradiating the system or adding a surfactant

PT
[36]. The key of this approach was to play with the solubility of the components and adjust the
interfacial tension by using stimuli-responsive surfactants. Janus emulsions have been also

RI
prepared with other oil mixtures [72][73]. Along the same rational, through the use of different

SC
immiscible polymers, formulations were prepared for the production of W/W/W emulsions
[32,33]. Applying similar dewetting-wetting concepts but following a different strategy, Deng

NU
and coworkers [74,75] were able to produce multiple emulsions from lower-order emulsion by a
drop-engulfing approach within a simple microfluidic device.
MA
3.6 Aerated multiple emulsions

Another appealing multiple emulsion-based system is that in which either the inner phase
D

[76,77] or the intermediate phase (antibubble) [78] is a gas. Such structures present many
TE

potential and varied applications, from aerated food products to ultrasound contrast agents.
Unfortunately, the traditionally used ultrasonic methods lack control over the size of the air
P

bubbles and the resulting systems present short lifetimes, as gas is prone to be dissolved in one of
CE

the phases. Recently, researchers began to use microfluidic technology to control the flow
patterns to form highly monodisperse encapsulated air bubbles. For these complex architectures,
AC

a noticeably enhanced stability has also been observed when colloids are used as stabilizers.

3.7 Multiple nano-emulsions

Multiple emulsions with outer droplet sizes below one micrometer, i.e. within the nanometer
range, are rare, due to the intrinsic difficulties in producing outer droplets of such small size
either by microfluidics or by bulk methods. However, there are a few examples of multiple nano-
emulsions in the literature. One of the first reports concerned the stabilization of nanoscale
double W/O/W emulsions by single-component block copolypeptides [79]. The double emulsions
were prepared by ultrasonication followed by high pressure homogenization; outer droplets with
size below 200 nm were obtained, as evidenced by cryogenic transmission electron microscopy.
Sub-micrometer multiple W/O/W emulsions have also been prepared by membrane

15
 ACCEPTED MANUSCRIPT

emulsification using a two-step process [80]; these emulsions proved to enhance intestinal
absorption of biopharmaceutics. Multiple nano-emulsions have been tested as drug delivery
systems [96,97] and as vehicles for Magnetic Resonance Imaging (MRI) contrast agents.[83]

PT
4. Multiple emulsions as templates for porous particles and capsules

4.1 Hardening multiple emulsions and their evolution during the process

RI
Among the different ways to produce multi-compartmentalized particles the multiple

SC
emulsion templating route is one of the most widely used. As multiple emulsions already have a
hierarchical architecture, the additional use of porogen agents is not needed. The ability to

NU
generate multiple emulsions with different compositions and morphologies paved the way to
form a wide range of different porous particles by simply solidifying the middle phase.
MA
Moreover, the fact that the inner phase is separated by a physical barrier from the external
medium makes this type of particles potentially useful for the encapsulation and isolation of
D

active agents. Encapsulation can be accomplished either within the inner phase only, or within
TE

the inner and the middle phase, which allows encapsulating incompatible active agents within the
same vehicle without cross-contamination. The most established routes to solidify the middle
P

phase of multiple emulsions are polymerization, solvent-evaporation and temperature-induced

CE

solidification. In the polymerization route, the middle phase contains a monomer solution, whose
chemical or physical crosslinking can be triggered at a specific time by UV irradiation [84,85],
AC

temperature or adding an initiator [57,8689]. Interestingly, the rigidity of the material can be
easily tuned by regulating the crosslinking degree [90]. Although most of the research performed
is focused onto polymer particles, the synthesis of inorganic particles is also feasible by using
silica [91,92] or zirconium dioxide [93] precursors. In the solvent evaporation-based method, the
middle phase is a solution of normally a polymer or a lipid in a volatile solvent. Upon removing
the solvent, the material precipitates creating the solid matrix. The most used polymers are the
polyesters (poly(lactic acid) [94], poly(lactic-co-glycolic acid) [95,96]) and polyacrylates, which
in some cases might even provide some stability without the need of surfactants [97]. When using
templates produced with a microfluidic device, challenging structures with ultrathin shells such
as vesicles [98] or polymersomes [99] can also be fabricated. In the third approach, the emulsion
is prepared using as an intermediate phase a material above its melting temperature. When the
system is cooled down, the material crystalizes yielding the porous particle. As this is a solvent-
16
 ACCEPTED MANUSCRIPT

free route, it has been widely used to prepare lipid vehicles for pharmaceutical drug delivery
purposes [100,101].

It is obvious that in all cases the morphology of the final particle closely resembles the

PT
emulsion template, especially when the polymerization or the temperature-induced solidification
routes are used. In the case of the solvent-evaporation route, the resulting particles are

RI
significantly smaller than the template, given that a fraction of the middle phase is evaporated.
Such shrinkage could in turn modify the distribution and size of the pores, so that the final

SC
morphology is less predictable. Nevertheless, when the template is produced by bulk
emulsification methods, the particle and pore size as well as the pore structure (isolated or

NU
interconnected) can be adjusted by playing with several formulation parameters, such as the
stabilizer concentration or the ratio between the phases (Fig 4A) [57,102,103]. However, their
MA
morphologies can only be controlled up to some extent. Difficulties might arise when a much
more well-defined morphology is required. Fortunately, the fine control over the liquid flow rates
afforded by the microfluidic technology can be exploited to tackle this problem. For instance,
D

capsules can be synthesized from multiple emulsions with a single inner droplet. The mechanical
TE

properties and permeability of these capsules can be tuned by varying the wall thickness and the
chemical composition of the middle phase (Fig 4B-C) [104107]. However, sometimes the
P

structure of the final particle not only depends on the morphology of the template but also on its
CE

evolution during the solidification process. When a single droplet is generated, it tends to sink
because of a density mismatch with the middle phase. Polymerization of this eccentric emulsion
AC

results in an open structure. Several strategies have been used to avoid this phenomenon, such us
the use of highly viscous middle phases or generating the emulsion in the jetting regime [84].

Solidification processes could also be accompanied by a phase separation, but as pointed

out above, this can be used to synthesize exotic particles with open structures. Different solid
particles with shape- and/or composition-anisotropy have been prepared by controlling the degree
of phase separation within the droplet (from a complete separation to partial separation) during
the solidification process (Fig 4D) [34,100,108,109]. Control over dewetting can be achieved by
using surfactants or changing the composition of the middle phase, which governs the interfacial
tension or adhesion force between the phases. Recently, a more sophisticated route based on the
use of a poly(4-vinylpyridine)-oleic acid supramolecular complex, whose amphiphilicity can be

17
 ACCEPTED MANUSCRIPT

modulated upon changing the pyridine-acid ratio, has been described [110]. Phase separation in
double emulsion can also be used to prepare vesicles, circumventing the problem of the presence
of non-desired organic solvent remnants [111].

PT
Interconnecting porous polymer microspheres were prepared using high internal water-
phase double emulsions (W/O/W) generated via catastrophic phase inversion [112]. Interestingly,

RI
these emulsions were stabilized by a single surfactant (12-acryloxy-9-octadecenoic acid). These
particles are designated as poly(HIPE) microspheres in analogy to bulk porous polymers derived

SC
from high-internal-phase ratio (HIPE) emulsions.

NU
4.2 Pickering-based porous particles

The growing research on Pickering multiple emulsions has also evolved towards their use
MA
as scaffolds for particle synthesis. The reaction (usually a polymerization) can occur within the
particle-stabilized droplet, leading to formation of a porous particle decorated with the stabilizing
D

particles. Highly porous particles without throat pores can be obtained [55], which is difficult to
TE

achieve using surfactant-stabilized emulsions. One of the main advantages of using Pickering
emulsions as templates is the possibility to endow a specific performance to the system by simply
P

using functional particles as stabilizers. For instance, some studies have demonstrated the
CE

possibility of adding magnetic properties to the system by using Fe3O2 nanoparticles as stabilizers
[5658]. Alternatively, capsules can be obtained by crosslinking the particles adsorbed at the
AC

interface. This requires the use of chemically modified colloids. An example of this is the use of
amine-functionalized silica colloids that can subsequently react with an epoxy-derivate molecule
dissolved in one of the phases [113]. The resulting particle film showed to be strong enough to
withstand the drying process, hence, yielding colloidosomes-in-colloidosomes, which might offer
extra encapsulating advantages compared to the conventional colloidosomes.

Insert Figure 4

Figure 4. Internal-structured particles formed from multiple emulsions. A) Scanning Electron

Microscopy (SEM) images of silicone particles with different inner structures obtained from
W/O/W multiple emulsions formulated with 5, 18 and 30% of inner aqueous phase. Adapted with
the permission from [102]. Copyright 2013 American Chemical Society. B) The permeability of
methacrylate capsules can be tuned by adding different amounts of polyacrylic acid-methyl
methacrylate (PAA-b-PMMA, a pH-responsive material), what results in a different hole density

18
 ACCEPTED MANUSCRIPT

and sizeAdapted with permission from [104]. Copyright 2013 American Chemical Society. C)
Fluorescent microscopy pictures of lipid capsules with different shell thicknesses (C1-2). The red
and the green colors denote the encapsulation of a hydrophilic and a hydrophobic drug,
respectively. A SEM image of a broken capsule is also shown (C3). Adapted with permission
from [100]. Copyright 2013 American Chemical Society. D) Optical microscopy pictures (inset)
of diverse emulsion templates with different ethoxylated trimethylolpropane triacrylate (ETPTA)

PT
concentrations in the middle phase and SEM images of the resulting particles. By varying the
amount of ETPTA, the adhesion of the inner droplet with the outer phase changes, hence

RI
resulting in particles with fishbowl-, bowl- or truncated-sphere-shapes. Scale bar is 200 m.
Adapted with permission from [61]. Copyright 2013 Wiley-VCH.

SC
5. Summary and outlook

The broad spectrum of potential applications of multiple emulsions is constantly catalyzing the

NU
development of better preparation methods and stabilization strategies. Concerning preparation,
microfluidic devices have attracted much interest due to their capability to produce droplets with
MA
extremely fine control over size and structure. However, droplet production throughput to meet
the industrial scale is still a significant challenge, making the synthesis of materials viable only
D

for high value products, such as pharmaceuticals and cosmetics. On a different note, the advent of
new fabrication methods such as 3D printing, will become accessible to a much wider range of
TE

scientists due to affordability and automated single-step fabrication. This promises to bring even
P

more innovative ideas to the field of multiple emulsion production and microfluidics in general.
CE

Advances on the formulation of multiple emulsions, particularly those related with the use of
block copolymers and particles for emulsion stabilization, have expanded the field beyond
AC

traditional surfactant mixtures, with uses that range from edible multiple emulsions to systems for
environmental purposes.

The availability of multiple emulsions with controlled size and structure have helped to
consolidate them as robust and versatile soft templates for the fabrication of porous particles and
capsules with complex morphologies through several strategies for the hardening of the
intermediate phases. New matrix and wall materials, with their associated functionalities, are
expected to come.

Acknowledgments

N.V. acknowledges The Netherlands Organization for Scientific Research (NWO-ECHO-STIP

Grant 717.013.005) for financial support.

19
 ACCEPTED MANUSCRIPT

C.R-A is grateful to the European Regional Development Fund (ON.2 O Novo Norte Program).

PT
References

[1] Li J, Zhang J, Han B, Zhao Y, Yang G. Formation of multiple water-in-ionic liquid-in-

RI
water emulsions. J Colloid Interface Sci 2012;368:3959. doi:10.1016/j.jcis.2011.10.083.

SC
*[2] Squires TM, Quake SR. Microfluidics: Fluid physics at the nanoliter scale. Rev Mod Phys
2005;77:9771026. doi:10.1103/RevModPhys.77.977. Essential comprehensive review

NU
focusing on the multiple physical phenomena that emerges when fluids flow under micro
and nanoscale confinement. The review goes beyond the flow of immiscible fluids and
MA
emulsion formation that we focus on in this work but is highly recommended to someone
new in the field
D

**[3] Cubaud T, Mason TG. Capillary threads and viscous droplets in square microchannels.
TE

Phys Fluids 2008;20:053302. doi:10.1063/1.2911716. Comprehensive study about the

physics governing the different flow regimes of two immiscible fluids
P

*[4] Anna SL. Droplets and Bubbles in Microfluidic Devices. Annu Rev Fluid Mech
CE

2016;48:285309. doi:10.1146/annurev-fluid-122414-034425. Current up-to-date review

focusing on the physics of multiphase flow and droplet formation in microfluidic devices
AC

[5] Utada AS, Fernandez-Nieves A, Stone HA, Weitz DA. Dripping to Jetting Transitions in
Coflowing Liquid Streams. Phys Rev Lett 2007;99:094502.
doi:10.1103/PhysRevLett.99.094502.

[6] Clegg PS, Tavacoli JW, Wilde PJ. One-step production of multiple emulsions:
microfluidic, polymer-stabilized and particle-stabilized approaches. Soft Matter
2016;12:9981008. doi:10.1039/C5SM01663K.

[7] Thorsen T, Roberts RW, Arnold FH, Quake SR. Dynamic pattern formation in a vesicle-
generating microfluidic device. Phys Rev Lett 2001;86:41636.
doi:10.1103/PhysRevLett.86.4163.

20
 ACCEPTED MANUSCRIPT

**[8] Utada AS, Lorenceau E, Link DR, Kaplan PD, Stone HA, Weitz DA. Monodisperse
Double Emulsions Generated from a Microcapillary Device. Science 2005;308:53741.
doi:10.1126/science.1109164. Original contribution for one-step double emulsion
formation in a glass microcapillary device

PT
[9] Anna SL, Bontoux N, Stone HA. Formation of dispersions using flow focusing in

RI
microchannels. Appl Phys Lett 2003;82:3646. doi:10.1063/1.1537519.

SC
*[10] Vladisavljevi GT, Khalid N, Neves MA, Kuroiwa T, Nakajima M, Uemura K, Ichikawa
S, Kobayashi I. Industrial lab-on-a-chip: Design, applications and scale-up for drug

NU
discovery and delivery. Adv Drug Deliv Rev 2013;65:162663.
doi:10.1016/j.addr.2013.07.017. Very comprehensive review focusing on many different
principles in microfluidics, including many different chip designs and fabrication methods
MA
[11] Garstecki P, Fuerstman MJ, Stone HA, Whitesides GM. Formation of droplets and bubbles
in a microfluidic T-junction-scaling and mechanism of break-up. Lab Chip 2006;6:43746.
D

doi:10.1039/b510841a.
TE

*[12] Rotem A, Abate AR, Utada AS, Van Steijn V, Weitz DA. Drop formation in non-planar
P

microfluidic devices. Lab Chip 2012;12:4263-4268. doi:10.1039/c2lc40546f. Example of

CE

how to use PDMS microfluidic devices with a 3D structure mimicking glass

microcapillaries, and avoiding many wettability issues in the production of multiple
AC

emulsions

[13] Zhang M, Wu J, Wang L, Xiao K, Wen W. A simple method for fabricating multi-layer
PDMS structures for 3D microfluidic chips. Lab Chip 2010;10:1199.
doi:10.1039/b923101c.

*[14] Ho CMB, Ng SH, Li KHH, Yoon Y-J. 3D printed microfluidics for biological
applications. Lab Chip 2015;15:362737. doi:10.1039/C5LC00685F. Review highlighting
current 3D printing methods, and how they are predicted to revolutionize microfluidic
technology, either by bringing new device design and complexity possibilities, but also
ease in fabrication

21
 ACCEPTED MANUSCRIPT

[15] Abate AR, Agresti JJ, Weitz DA. Microfluidic sorting with high-speed single-layer
membrane valves. Appl Phys Lett 2010;96:13. doi:10.1063/1.3431281.

[16] Leman M, Abouakil F, Griffiths AD, Tabeling P. Droplet-based microfluidics at the

PT
femtolitre scale. Lab Chip 2015;15:75365. doi:10.1039/c4lc01122h.

[17] Shah RK, Shum HC, Rowat AC, Lee D, Agresti JJ, Utada AS, Chu L-Y, Kim J-W,

RI
Fernandez-Nieves A, Martinez CJ, Weitz DA. Designer emulsions using microfluidics.

SC
Mater Today 2008;11:1827.

[18] Li W, Nie Z, Zhang H, Paquet C, Seo M, Garstecki P, Kumacheva E. Screening of the

NU
effect of surface energy of microchannels on microfluidic emulsification. Langmuir
2007;23:80104. doi:10.1021/la7005875.
MA
[19] Barbier V, Tatoulian M, Li H, Arefi-Khonsari F, Ajdari A, Tabeling P. Stable modification
of PDMS surface properties by plasma polymerization: Application to the formation of
D

double emulsions in microfluidic systems. Langmuir 2006;22:52302.

TE

doi:10.1021/la053289c.

*[20] Abate AR, Weitz DA. High-order multiple emulsions formed in poly(dimethylsiloxane)
P

microfluidics. Small 2009;5:20302. doi:10.1002/smll.200900569. Work describing the

CE

production of multiple emulsions by cascading several droplet generator junctions in a

PDMS device. Simple onion structures of up to five-layer emulsion droplets were obtained
AC

[21] Bauer W-AC, Fischlechner M, Abell C, Huck WTS. Hydrophilic PDMS microchannels for
high-throughput formation of oil-in-water microdroplets and water-in-oil-in-water double
emulsions. Lab Chip 2010;10:18149. doi:10.1039/c004046k.

[22] Arriaga LR, Amstad E, Weitz DA. Scalable single-step microfluidic production of single-
core double emulsions with ultra-thin shells. Lab Chip 2015;15:333540.
doi:10.1039/C5LC00631G.

**[23]Okushima S, Nisisako T, Torii T, Higuchi T. Controlled production of monodisperse

double emulsions by two-step droplet breakup in microfluidic devices. Langmuir
2004;20:99058. doi:10.1021/la0480336. Original contribution for double emulsion

22
 ACCEPTED MANUSCRIPT

formation in a 2D microfluidic device with two T-junctions in series

**[24] Nie Z, Xu S, Seo M, Lewis PC, Kumacheva E. Polymer particles with various shapes and
morphologies produced in continuous microfluidic reactors. J Am Chem Soc

PT
2005;127:805863. doi:10.1021/ja042494w. Original contribution for one-step double
emulsion formation in a 2D flow-focusing device

RI
SC
[25] Park J Il, Saffari A, Kumar S, Gnther A, Kumacheva E. Microfluidic Synthesis of
Polymer and Inorganic Particulate Materials. Annu Rev Mater Res 2010;40:41543.

NU
doi:10.1146/annurev-matsci-070909-104514.

[26] Chu L-Y, Utada AS, Shah RK, Kim J-W, Weitz DA. Controllable monodisperse multiple
MA
emulsions. Angew Chemie Int Ed 2007;46:89704. doi:10.1002/anie.200701358.

*[27] Wang W, Xie R, Ju X-J, Luo T, Liu L, Weitz DA, Chu L-Y. Controllable microfluidic
D

production of multicomponent multiple emulsions. Lab Chip 2011;11:158792.

TE

doi:10.1039/c1lc20065h. Work describing the production of multiple emulsions by

cascading several glass-microcapillary droplet generator devices. Multiple droplet
P

configurations of controlled size were obtained

CE

*[28] Kim S-H, Weitz DA. One-step emulsification of multiple concentric shells with capillary
AC

microfluidic devices. Angew Chem Int Ed 2011;50:87314. doi:10.1002/anie.201102946.

Work describing the production of multiple emulsions in a single step through the use of a
hydrophobically-modified glass microcapillary device. Onion-like quadruple emulsions
were obtained

*[29] Adams LLA, Kodger TE, Kim S-H, Shum HC, Franke T, Weitz DA. Single step
emulsification for the generation of multi-component double emulsions. Soft Matter
2012;8:1071924. doi:10.1039/c2sm25953b. Work describing the production of multicore
double-emulsions in a single step using a glass microcapillary device with a two, three and
four-bore injection capillary

[30] Lee SS, Abbaspourrad A, Kim SH. Nonspherical double emulsions with multiple distinct

23
 ACCEPTED MANUSCRIPT

cores enveloped by ultrathin shells. ACS Appl Mater Interfaces 2014;6:1294300.

doi:10.1021/am405283j.

*[31] Zhao CX, Middelberg APJ. Microfluidic mass-transfer control for the simple formation of

PT
complex multiple emulsions. Angew Chemie - Int Ed 2009;48:720811.
doi:10.1002/anie.200902485. Original contribution to the combination of microfluidics

RI
with phase-separation phenomena for the production of multiple emulsion droplets. The
role of the microfluidic device is to provide controlled outer droplet sizes and controlled

SC
mixing, whereas the internal structure is controlled by the fluids compositions

NU
[32] Ziemecka I, van Steijn V, Koper GJM, Kreutzer MT, van Esch JH. All-aqueous core-shell
droplets produced in a microfluidic device. Soft Matter 2011;7:987880.
doi:10.1039/c1sm06517c.
MA
*[33] Song Y, Shum HC. Monodisperse w/w/w double emulsion induced by phase separation.
Langmuir 2012;28:120549. doi:10.1021/la3026599. Work reporting on the use of phase-
D

separation phenomena for the production of all-aqueous (W/W/W) monodisperse multiple

TE

emulsion droplets. The low interfacial tension between the aqueous phases required the
use of an external mechanical vibrator to produce droplets instead of jets
P
CE

[34] Choi CH, Weitz DA, Lee CS. One step formation of controllable complex emulsions:
From functional particles to simultaneous encapsulation of hydrophilic and hydrophobic
AC

agents into desired position. Adv Mater 2013;25:253641. doi:10.1002/adma.201204657.

**[35]Haase MF, Brujic J. Tailoring of high-order multiple emulsions by the liquid-liquid phase
separation of ternary mixtures. Angew Chemie - Int Ed 2014;53:117937.
doi:10.1002/anie.201406040. Work focused on a deeper understanding of the phase
separation events occurring inside emulsion droplets formed in a microfluidic device. A
relationship between the composition in the initial equilibrium phase diagram and the
number of layers in the resulting multiple emulsion is proposed

**[36]Zarzar LD, Sresht V, Sletten EM, Kalow J a, Blankschtein D, Swager TM. Dynamically
reconfigurable complex emulsions via tunable interfacial tensions. Nature 2015;518:520
4. doi:10.1038/nature14168. This paper introduces reconfigurable multiple emulsion,

24
 ACCEPTED MANUSCRIPT

whose structural changes can be triggered through different stimuli. The clever concepts
applied to induce such phase separation processes provide a new means for the design of
multiple emulsions

PT
[37] Sparreboom W, van den Berg a, Eijkel JCT. Principles and applications of nanofluidic
transport. Nat Nanotechnol 2009;4:71320. doi:10.1038/nnano.2009.332.

RI
[38] Polte J, Erler R, Thnemann AF, Sokolov S, Ahner TT, Rademann K, Emmerling F,

SC
Kraehnert R. Nucleation and Growth of Gold. ACS Nano 2010;4:107682.
doi:10.1021/nn901499c.

NU
[39] Silva BFB, Zepeda-Rosales M, Venkateswaran N, Fletcher B, Carter L, Matsui M, Weiss
T, Han J, Li Y, Olsson U, Safinya CR. Nematic director reorientation at solid and liquid
MA
interfaces under flow: SAXS studies in a microfluidic device. Langmuir 2015;31:436171.
doi:10.1021/la5034614.
D

[40] Hoppe SM, Sasaki DY, Kinghorn AN, Hattar K. In-situ transmission electron microscopy
TE

of liposomes in an aqueous environment. Langmuir 2013;29:995861.

doi:10.1021/la401288g.
P

**[41]Malloggi F, Pannacci N, Attia R, Monti F, Mary P, Willaime H, Tabeling P, Cabane B,

CE

Poncet P. Monodisperse Colloids Synthesized with Nanofluidic Technology. Langmuir

2010;26:236973. doi:10.1021/la9028047. A step-emulsification strategy using nano-sized
AC

channels is used for the production of single and double emulsion droplets. The minimum
sizes achieved were about 1 m for single, and 2-4 m for Janus and multiple emulsions

[42] Li Z, Leshansky AM, Pismen LM, Tabeling P. Step-emulsification in a microfluidic

device. Lab Chip 2015;15:102331. doi:10.1039/C4LC01289E.

[43] Shui L, Van Den Berg A, Eijkel JCT. Scalable attoliter monodisperse droplet formation
using multiphase nano-microfluidics. Microfluid Nanofluidics 2011;11:8792.
doi:10.1007/s10404-011-0776-7.

[44] Nisisako T, Torii T. Microfluidic large-scale integration on a chip for mass production of
monodisperse droplets and particles. Lab Chip 2008;8:28793. doi:10.1039/b713141k.

25
 ACCEPTED MANUSCRIPT

*[45] Li W, Greener J, Voicu D, Kumacheva E. Multiple modular microfluidic (M3) reactors for
the synthesis of polymer particles. Lab Chip 2009;9:271521. doi:10.1039/b906626h.
Comprehensive study on many issues associated with the on-chip parallelization of
multiple drop generator modules for the production of polymer particles. A particle

PT
production rate of about 1 Kg/day was achieved

RI
**[46]Romanowsky MB, Abate AR, Rotem A, Holtze C, Weitz DA. High throughput production
of single core double emulsions in a parallelized microfluidic device. Lab Chip

SC
2012;12:8027. doi:10.1039/c2lc21033a. Parallelization of an on-chip microfluidic device
capable of producing double emulsions at a rate of 1 Kg/day, which could be further

NU
scaled up for small industry applications

[47] Huang X, Fang R, Wang D, Wang J, Xu H, Wang Y, Zhang X. Tuning Polymeric

MA
Amphiphilicity via Se-N Interactions: Towards One-Step Double Emulsion for Highly
Selective Enzyme Mimics. Small 2015;11:153741. doi:10.1002/smll.201402271.
D

[48] Zhang W, Li F. Preparation and characterization of multiple emulsions (W/Si/W) by

TE

single-step emulsification. Colloids Surfaces A Physicochem Eng Asp 2013;423:98103.

doi:10.1016/j.colsurfa.2013.02.004.
P
CE

[49] Li F, Zhang W. Stability and rheology of W/Si/W multiple emulsions with

polydimethylsiloxane. Colloids Surfaces A Physicochem Eng Asp 2015;470:2906.
AC

doi:10.1016/j.colsurfa.2015.02.001.

[50] Besnard L, Marchal F, Paredes JF, Daillant J, Pantoustier N, Perrin P, Guenoun P.

Multiple emulsions controlled by stimuli-responsive polymers. Adv Mater 2013;25:2844
8. doi:10.1002/adma.201204496.

[51] Besnard L, Protat M, Malloggi F, Daillant J, Cousin F, Pantoustier N, Guenoun P, Perrin

P. Breaking of the Bancroft rule for multiple emulsions stabilized by a single stimulable
polymer. Soft Matter 2014;10:707387. doi:10.1039/c4sm00596a.

[52] Zhang Y, Gou J, Sun F, Geng S, Hu X, Zhang K, Lin X, Xiao W, Tang X. Impact of
electrolytes on double emulsion systems (W/O/W) stabilized by an amphiphilic block
copolymer. Colloids Surfaces B Biointerfaces 2014;122:36874.

26
 ACCEPTED MANUSCRIPT

doi:10.1016/j.colsurfb.2014.07.008.

[53] Sun G, Liu M, Zhou X, Hong L, Ngai T. Influence of asymmetric ratio of amphiphilic
diblock copolymers on one-step formation and stability of multiple emulsions. Colloids

PT
Surfaces A Physicochem Eng Asp 2014;454:1622. doi:10.1016/j.colsurfa.2014.04.015.

*[54] Williams M, Warren NJ, Fielding LA, Armes SP, Verstraete P, Smets J. Preparation of

RI
double emulsions using hybrid polymer/silica particles: New pickering emulsifiers with

SC
adjustable surface wettability. ACS Appl Mater Interfaces 2014;6:2091927.
doi:10.1021/am505581r. A report on the wettability modification of silica particles for

NU
emulsion stabilization, a principle that could be applied to other types of nanoparticles as
well MA
[55] Yin D, Li B, Liu J, Zhang Q. Structural diversity of multi-hollow microspheres via
multiple Pickering emulsion co-stabilized by surfactant. Colloid Polym Sci 2014;293:341
7. doi:10.1007/s00396-014-3401-y.
D
TE

[56] Zhu W, Ma W, Li C, Pan J, Dai X. Well-designed multihollow magnetic imprinted

microspheres based on cellulose nanocrystals (CNCs) stabilized Pickering double
P

emulsion polymerization for selective adsorption of bifenthrin. Chem Eng J

CE

2015;276:24960. doi:10.1016/j.cej.2015.04.084.

[57] Zou S, Liu H, Yang Y, Wei Z, Wang C. Multihollow nanocomposite microspheres with
AC

tunable pore structures by templating Pickering double emulsions. React Funct Polym
2013;73:123141. doi:10.1016/j.reactfunctpolym.2013.06.006.

*[58] Wu R, Pan J, Dai X, Qiu D, Zhu H, Ma Y, Shi W, Yan Y. A hierarchical rippled and
crumpled PLA microstructure generated through double emulsion: the interesting roles of
Pickering nanoparticles. Chem Commun 2015;51:162514. doi:10.1039/c5cc06516j. A
protocol to create rippled and crumpled soft polymeric particles by means of gas
generation within the emulsion droplets

[59] Cunha AG, Mougel JB, Cathala B, Berglund LA, Capron I. Preparation of double
pickering emulsions stabilized by chemically tailored nanocelluloses. Langmuir
2014;30:932735. doi:10.1021/la5017577.

27
 ACCEPTED MANUSCRIPT

[60] Matos M, Timgren A, Sj M, Dejmek P, Rayner M. Preparation and encapsulation

properties of double Pickering emulsions stabilized by quinoa starch granules. Colloids
Surfaces A Physicochem Eng Asp 2013;423:14753. doi:10.1016/j.colsurfa.2013.01.060.

PT
**[61]Wang W, Zhang MJ, Xie R, Ju XJ, Yang C, Mou CL, Weitz DA, Chu LY. Hole-shell
microparticles from controllably evolved double emulsions. Angew Chemie - Int Ed

RI
2013;52:80847. doi:10.1002/anie.201301590. Control over the final capsule morphology
is achieved by varying the adhesion force between the inner droplets and the outer phase

SC
by means of changing the composition of the middle phase

NU
[62] Carrillo CA, Nypel TE, Rojas OJ. Cellulose nanofibrils for one-step stabilization of
multiple emulsions (W/O/W) based on soybean oil. J Colloid Interface Sci 2015;445:166
73. doi:10.1016/j.jcis.2014.12.028.
MA
[63] Tu F, Lee D. One-step encapsulation and triggered release based on Janus particle-
stabilized multiple emulsions. Chem Commun 2014;50:1554952.
D

doi:10.1039/c4cc07854c.
TE

[64] Hernndez-Marn NY, Lobato-Calleros C, Vernon-Carter EJ. Stability and rheology of

P

water-in-oil-in-water multiple emulsions made with protein-polysaccharide soluble

CE

complexes. J Food Eng 2013;119:1817. doi:10.1016/j.jfoodeng.2013.05.039.

[65] Mohammadi A, Jafari SM, Esfanjani AF, Akhavan S. Application of nano-encapsulated

AC

olive leaf extract in controlling the oxidative stability of soybean oil. Food Chem
2016;190:5139. doi:10.1016/j.foodchem.2015.05.115.

[66] Oppermann AKL, Renssen M, Schuch A, Stieger M, Scholten E. Effect of gelation of

inner dispersed phase on stability of (w1/o/w2) multiple emulsions. Food Hydrocoll
2015;48:1726. doi:10.1016/j.foodhyd.2015.01.027.

[67] Matos M, Gutirrez G, Coca J, Pazos C. Preparation of water-in-oil-in-water (W1/O/W2)

double emulsions containing trans-resveratrol. Colloids Surfaces A Physicochem Eng Asp
2014;442:6979. doi:10.1016/j.colsurfa.2013.05.065.

[68] Mun S, Choi Y, Park KH, Shim JY, Kim YR. Influence of environmental stresses on the

28
 ACCEPTED MANUSCRIPT

stability of W/O/W emulsions containing enzymatically modified starch. Carbohydr Polym

2013;92:150311. doi:10.1016/j.carbpol.2012.10.050.

[69] Seddari S, Moulai-Mostefa N. Double Emulsions Stabilized by Xanthan in the Absence of

PT
Hydrophilic Surfactant. J Dispers Sci Technol 2016;37:5305.
doi:10.1080/01932691.2015.1048807.

RI
[70] Perez-Moral N, Watt S, Wilde P. Comparative study of the stability of multiple emulsions

SC
containing a gelled or aqueous internal phase. Food Hydrocoll 2014;42:215-222.
doi:10.1016/j.foodhyd.2014.05.023.

NU
[71] Patel AR, Dumlu P, Vermeir L, Lewille B, Lesaffer A, Dewettinck K. Rheological
characterization of gel-in-oil-in-gel type structured emulsions. Food Hydrocoll
MA
2015;46:8492. doi:10.1016/j.foodhyd.2014.12.029.

**[72] Guo R. Droplet topology control of Janus emulsion prepared in one-step high energy
D

mixing. Soft Matter 2014;10:4498505. doi:10.1039/c4sm00456f. Interesting article on

TE

how to use multiple emulsion to create Janus emulsions. The topology of the droplets was
found to be dependend on the contact angle of the three liquids
P

[73] Kovach I, Koetz J, Friberg SE. Janus emulsions stabilized by phospholipids. Colloids
CE

Surfaces A Physicochem Eng Asp 2014;441:6671. doi:10.1016/j.colsurfa.2013.08.065.

AC

*[74] Deng NN, Wang W, Ju XJ, Xie R, Weitz DA, Chu LY. Wetting-induced formation of
controllable monodisperse multiple emulsions in microfluidics. Lab Chip 2013;13:4047
52. doi:10.1039/c3lc50638j. The article presents a novel method to form highly ordered
multiple emulsions from lower-order emulsions. By adjusting the wetting properties
between the phases, droplets can engolf droplets of immiscible liquids

[75] Deng NN, Mou CL, Wang W, Ju XJ, Xie R, Chu LY. Multiple emulsion formation from
controllable drop pairs in microfluidics. Microfluid Nanofluidics 2014;17:96772.
doi:10.1007/s10404-014-1381-3.

[76] Wang WT, Chen R, Xu JH, Wang YD, Luo GS. One-step microfluidic approach for
controllable production of gas-in-water-in-oil (G/W/O) double emulsions and hollow

29
 ACCEPTED MANUSCRIPT

hydrogel microspheres. RSC Adv 2014;4:164448. doi:10.1039/c4ra01526f.

[77] Wang WT, Chen R, Xu JH, Wang YD, Luo GS. One-step microfluidic production of gas-
in-water-in-oil multi-cores double emulsions. Chem Eng J 2015;263:4128.

PT
doi:10.1016/j.cej.2014.11.030.

[78] Silpe JE, Nunes JK, Poortinga AT, Stone HA. Generation of antibubbles from core-shell

RI
double emulsion templates produced by microfluidics. Langmuir 2013;29:87827.

SC
doi:10.1021/la4009015.

*[79] Hanson J a, Chang CB, Graves SM, Li Z, Mason TG, Deming TJ. Nanoscale double

NU
emulsions stabilized by single-component block copolypeptides. Nature 2008;455:858.
doi:10.1038/nature07197. To our knowledge, this is the first work on the formation of
MA
nanoscale double emulsions providing robust experimental evidence

[80] Koga K, Takarada N, Takada K. Nano-sized water-in-oil-in-water emulsion enhances

D

intestinal absorption of calcein, a high solubility and low permeability compound. Eur J
TE

Pharm Biopharm 2010;74:22332. doi:10.1016/j.ejpb.2009.09.004.

[81] Schwarz JC, Klang V, Karall S, Mahrhauser D, Resch GP, Valenta C. Optimisation of
P

multiple W/O/W nanoemulsions for dermal delivery of aciclovir. Int J Pharm

CE

2012;435:6975. doi:10.1016/j.ijpharm.2011.11.038.
AC

[82] Sigward E, Mignet N, Rat P, Dutot M, Muhamed S, Guigner JM, Scherman D, Brossard
D, Crauste-Manciet S. Formulation and cytotoxicity evaluation of new self-emulsifying
multiple W/O/W nanoemulsions. Int J Nanomedicine 2013;8:61125.
doi:10.2147/IJN.S35661.

[83] Sigward E, Corvis Y, Doan BT, Kindsiko K, Seguin J, Scherman D, Brossard D, Mignet
N, Espeau P, Crauste-Manciet S. Preparation and Evaluation of Multiple Nanoemulsions
Containing Gadolinium (III) Chelate as a Potential Magnetic Resonance Imaging (MRI)
Contrast Agent. Pharm Res 2015;32:298394. doi:10.1007/s11095-015-1680-8.

[84] Wu B, Gong HQ. Formation of fully closed microcapsules as microsensors by microfluidic

double emulsion. Microfluid Nanofluidics 2013;14:63744. doi:10.1007/s10404-012-

30
 ACCEPTED MANUSCRIPT

1083-7.

[85] Wang J, Cheng Y, Yu Y, Fu F, Chen Z, Zhao Y, Gu Z. Microfluidic Generation of Porous

Microcarriers for Three-Dimensional Cell Culture. ACS Appl Mater Interfaces

PT
2015;7:270359. doi:10.1021/acsami.5b10442.

[86] Zieringer MA, Carroll NJ, Abbaspourrad A, Koehler SA, Weitz DA. Microcapsules for

RI
Enhanced Cargo Retention and Diversity. Small 2015;11:29039.

SC
doi:10.1002/smll.201403175.

[87] Dias , Fidalgo J, Machado J, Moniz J, Mendes AM, Magalhes FD. Study of

NU
multivesiculated polyester particles synthesis by double emulsion process. Eur Polym J
2013;49:66474. doi:10.1016/j.eurpolymj.2013.01.004.
MA
[88] Menzel C, Cambn A, Yeates SG. Double emulsion template suspension polymerisation:
Towards the synthesis of polyelectrolyte core porous hydrophobic shell particles for
D

environmental applications. J Mater Chem A 2013;1:125539. doi:10.1039/c3ta12556d.

TE

[89] Liu L, Wu F, Ju XJ, Xie R, Wang W, Niu CH, Chu LY. Preparation of monodisperse
calcium alginate microcapsules via internal gelation in microfluidic-generated double
P

emulsions. J Colloid Interface Sci 2013;404:8590. doi:10.1016/j.jcis.2013.04.044.

CE

[90] Vilanova N, Kolenko Y V, Solans C, Rodrguez-Abreu C. Multiple emulsions as soft

AC

templates for the synthesis of multifunctional silicone porous particles. J Colloid Interface
Sci 2015;437:23543. doi:10.1016/j.jcis.2014.09.006.

[91] Yan H, Kim C. Formation of monodisperse silica microparticles with various shapes and
surface morphologies using double emulsion templates. Colloids Surfaces A Physicochem
Eng Asp 2014;443:8895. doi:10.1016/j.colsurfa.2013.10.049.

[92] Sousa FL, Almeida A, Giro A V, Fateixa S, Trindade T. Multiple emulsion templating of
hybrid Ag/SiO<inf>2</inf> capsules for antibacterial applications. Part Part Syst Charact
2015;32:5616. doi:10.1002/ppsc.201400168.

*[93] Zhang L, Hao S, Liu B, Shum HC, Li J, Chen H. Fabrication of ceramic microspheres by
diffusion-induced sol-gel reaction in double emulsions. ACS Appl Mater Interfaces

31
 ACCEPTED MANUSCRIPT

2013;5:1148993. doi:10.1021/am402028w. Protocol to synthesise monodisperse ceramic

capsules combining the microfluidic technology and a sol-gel reaction

[94] Cavallaro G, Craparo EF, Sardo C, Lamberti G, Barba AA, Dalmoro A. PHEA-PLA

PT
biocompatible nanoparticles by technique of solvent evaporation from multiple emulsions.
Int J Pharm 2015;495:71927. doi:10.1016/j.ijpharm.2015.09.050.

RI
[95] Gong A, Ma X, Xiang L, Ren W, Shen Z, Wu A. Improved double emulsion technology

SC
for fabricating autofluorescent microcapsules as novel ultrasonic/fluorescent dual-modality
contrast agents. Colloids Surfaces B Biointerfaces 2014;116:5617.

NU
doi:10.1016/j.colsurfb.2014.01.038.

[96] Qian L, Zhang H. One-step synthesis of protein-encapsulated microspheres in a porous

MA
scaffold by freeze-drying double emulsions and tuneable protein release. Chem Commun
2013;49:88335. doi:10.1039/c3cc45012k.
D

[97] Hu Y, Zou S, Yang Y, Tong Z, Wang C. Facile fabrication of macroporous PLGA

TE

microspheres via double-pickering emulsion templates. Macromol Chem Phys

2015;216:71420. doi:10.1002/macp.201400574.
P

*[98] Arriaga LR, Datta SS, Kim SH, Amstad E, Kodger TE, Monroy F, Weitz DA. Ultrathin
CE

shell double emulsion templated giant unilamellar lipid vesicles with controlled
microdomain formation. Small 2014;10:9506. doi:10.1002/smll.201301904. Challenging
AC

production of giant unilamellar vesicles with a controlled phase separated lipid

microdomains (rafts)

[99] Kim SH, Nam J, Kim JW, Kim DH, Han SH, Weitz DA. Formation of polymersomes with
double bilayers templated by quadruple emulsions. Lab a Chip - Miniaturisation Chem
Biol 2013;13:13516. doi:10.1039/c3lc41112e.

[100] Windbergs M, Zhao Y, Heyman J, Weitz DA. Biodegradable core-shell carriers for
simultaneous encapsulation of synergistic actives. J Am Chem Soc 2013;135:79337.
doi:10.1021/ja401422r.

[101] Zhao G, Hu C, Sun R, Ni S, Li Q, Xia Q. Development of novel composite antioxidant

32
 ACCEPTED MANUSCRIPT

multiple lipid particles from combination of W/O/W multiple emulsions and solid lipid
nanoparticles. Eur J Lipid Sci Technol 2015;117:105665. doi:10.1002/ejlt.201400398.

[102] Vilanova N, Solans C, Rodrguez-Abreu C. Preparation of novel silicone

PT
multicompartment particles by multiple emulsion templating and their use as encapsulating
systems. Langmuir 2013;29:1541422. doi:10.1021/la403134c.

RI
[103] Baimark Y, Srisuwan Y. Hollow chitosan microspheres prepared by an oil1-in-water-in-

SC
oil2 double emulsion method. Powder Technol 2013;249:43642.
doi:10.1016/j.powtec.2013.09.011.

NU
[104] Abbaspourrad A, Datta SS, Weitz DA. Controlling release from pH-responsive
microcapsules. Langmuir 2013;29:12697702. doi:10.1021/la403064f.
MA
[105] Abbaspourrad A, Duncanson WJ, Lebedeva N, Kim SH, Zhushma AP, Datta SS, Dayton
PA, Sheiko SS, Rubinstein M, Weitz DA. Microfluidic fabrication of stable gas-filled
D

microcapsules for acoustic contrast enhancement. Langmuir 2013;29:123527.

TE

doi:10.1021/la402598p.

[106] Abbaspourrad A, Carroll NJ, Kim SH, Weitz DA. Polymer microcapsules with
P

programmable active release. J Am Chem Soc 2013;135:774450. doi:10.1021/ja401960f.

CE

[107] Vilanova N, Rodrguez-Abreu C, Fernndez-Nieves A, Solans C. Fabrication of novel

AC

silicone capsules with tunable mechanical properties by microfluidic techniques. ACS

Appl Mater Interfaces 2013;5:524752. doi:10.1021/am4010896.

[108] Huang X, Qian Q, Wang Y. Anisotropic particles from a one-pot double emulsion induced
by partial wetting and their triggered release. Small 2014;10:141220.
doi:10.1002/smll.201302743.

[109] Guo S, Yao T, Ji X, Zeng C, Wang C, Zhang L. Versatile preparation of nonspherical

multiple hydrogel core PAM/PEG emulsions and hierarchical hydrogel microarchitectures.
Angew Chemie - Int Ed 2014;53:75049. doi:10.1002/anie.201403256.

[110] Wang D, Huang X, Wang Y. Managing the phase separation in double emulsion by tuning
amphiphilicity via a supramolecular route. Langmuir 2014;30:144608.

33
 ACCEPTED MANUSCRIPT

doi:10.1021/la5043525.

[111] Deshpande S, Caspi Y, Meijering AEC, Dekker C. Octanol-assisted liposome assembly on

chip. Nat Commun 2016;7:10447. doi:10.1038/ncomms10447.

PT
[112] Li Z, Liu H, Zeng L, Liu H, Yang S, Wang Y. Preparation of high internal water-phase
double emulsions stabilized by a single anionic surfactant for fabricating interconnecting

RI
porous polymer microspheres. Langmuir 2014;30:1215463. doi:10.1021/la502564r.

SC
*[113]Williams M, Armes SP, Verstraete P, Smets J. Double emulsions and colloidosomes-in-
colloidosomes using silica-based pickering emulsifiers. Langmuir 2014;30:270311.

NU
doi:10.1021/la500219m. A procedure on how to make solvent-free liposomes
MA
D
P TE
CE
AC

34
 ACCEPTED MANUSCRIPT

PT
RI
SC
NU
MA
Figure 1
D
TE
P
CE
AC

35
 ACCEPTED MANUSCRIPT

PT
RI
SC
NU
MA
D
TE
P
CE
AC

Figure 2

36
 ACCEPTED MANUSCRIPT

PT
RI
SC
NU
MA
D
TE
P

Figure 3
CE
AC

37
 ACCEPTED MANUSCRIPT

PT
RI
SC
NU
Figure 4 MA
D
TE
P
CE
AC

38
 ACCEPTED MANUSCRIPT

PT
RI
SC
NU
MA
D
P TE
CE

Graphical abstract
AC

39