Pesquisar no resumo do documento
DOI: 10.1021/la903890h ALangmuir XXXX, XXX(XX), XXX–XXX
©XXXX American Chemical Society
Scalable, Shape-Specific, Top-Down Fabrication Methods for the Synthesis of Engineered Colloidal Particles
Timothy J. Merkel,† Kevin P. Herlihy,† Janine Nunes,† Ryan M. Orgel,† Jason P. Rolland,‡ and Joseph M. DeSimone*,†,‡,§
†Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, ‡Liquidia Technologies, Research Triangle Park, North Carolina 27709, and §Department of Chemical and
Biomolecular Engineering, North Carolina State University, Raleigh, North Carolina 27695
Received October 13, 2009. Revised Manuscript Received November 25, 2009
The search for amethod to fabricate nonspherical colloidal particles from a variety of materials is of growing interest. As the commercialization of nanotechnology continues to expand, the ability to translate particle-fabrication methods from a laboratory to an industrial scale is of increasing significance. In this feature article, we examine several of themost readily scalable top-down methods for the fabrication of such shape-specific particles and compare their capabilities with respect to particle composition, size, shape, and complexity as well as the scalability of the method. We offer an extensive examination of particle replication in nonwetting templates (PRINT) with regard to the versatility and scalability of this technique. We also detail the specific methods used in PRINT particle fabrication, including harvesting, purification, and surface-modification techniques, with an examination of both past and current methods.
The design and fabrication of particles with dimensions on the colloidal size scale (<10 μm) has become an area of great and continually growing interest formany applications. Particles have been researched and developed for such varied applications as catalysis, microelectronics, photovoltaics, coatings, cosmetics, smart fluids, and nanomedicine, among others. These small particles are of fundamental interest not only because of the changes in optical, electrical, and other material properties that occur when thematerial is reduced to the nanometer size scale but also because of other factors such as improved solubility proper- ties or the capability of encapsulating bioactive materials (e.g., therepeutics or imaging agents), which can be poorly soluble or toxic, lacking the protection of the particle. Anisotropic colloidal particles can assemble to form structures that are distinct from the hexagonal close-packed arrangements favored by spherical par- ticles and are of interest in the fields of memory storage, optical electronics, photonics, and sensors.1
Considerable effort has been devoted to the development of fabrication methods that can mass produce colloidal particles with fine control of size, shape, composition, cargo, and surface chemistry.We submit particle replication in nonwetting templates (PRINT) as a readily scalable method for the fabrication of such monodisperse colloids with precise control over size and shape and with broad capabilities with regard to material composition and chemical anisotropy. In addition to PRINT, we will present several other top-down particle-fabrication methods that have excellent potential for the scalable production of monodisperse, shape-specific particles on the colloidal size scale.
Two broad approaches are amenable to the fabrication of anisotropic particles: bottom-up and top-down techniques. Bottom-up approaches begin at the atomic ormolecular scale and build up to the desired particle size, whereas top-down methods
process a material on the desired size scale. The most commonly employed methods for the production of mass quantities of particles on the colloidal length scale are bottom-up synthesis approaches such as emulsion polymerization. In a typical process, a monomer is emulsified via rapid stirring in a mobile phase that contains an initiator and a surfactant. Upon heating to activate the initiator, spherical particles are nucleated in the surfactant micelles and grow to the desired size. Particles obtained by emulsion methods are typically spherical and can vary in size from tens of nanometers to as large as several micrometers in diameter. The particle size and the molecular weight of the polymers formed in emulsions are controlled with parameters such as the surfactant concentration and reaction temperature. Surfactant adsorbed onto the surface of these particles can be difficult to remove if undesired. Although this method is extre- mely scalable, the particles fabricated in this way are typically spherical in shape and are fairly polydisperse.
There is a growing need to generate particles with a diversity of nonspherical shapes. Complex particle shapes are desirable for a range of applications, including self-assembly,2 photonic materi- als,3 and microelectromechanical systems (MEMS).4 Particles with nonspherical shapes are of increasing interest in biomedical applications such as drug delivery, where the rodlike or corkscrew morphologies possessed by many viruses and bacteria are sus- pected to have derived an evolutionary advantage from their specific shapes.5 For many applications, the ability to control particle size and shape is of vital importance, and the ability to manufacture large quantities of such size- and shape-specific particles is a crucial factor in the utility of the fabrication method. For a particle-fabrication method to prove industrially useful, it must be scalable and capable of producing usefully large quan- tities of particles. For example, a particle tested for efficacy as a drug carrier will have very different production needs in a
*To whom correspondence should be addressed. E-mail: desimone@ unc.edu. (1) Glotzer, S. C.; Solomon, M. J. Nat. Mater. 2007, 6, 557.
(2) Glotzer, S. C. Science 2004, 306, 419. (3) Lu, Y.; Yin, Y.; et al. Adv. Mater. 2001, 13, 415. (4) Beebe, D. J.; Moore, J. S.; et al. Nature 2000, 404, 588. (5) Champion, J. A.; Katare, Y. K.; et al. J. Controlled Release 2007, 121, 3.
B DOI: 10.1021/la903890h Langmuir XXXX, XXX(XX), XXX–XXX
Invited Feature Article Merkel et al.
laboratory setting than on the industrial scale. The fabrication of milligram quantities of particles suffices for testing in the labora- tory, whereas the commercialization of this technology would require the production of gram and kilogram quantities to satisfy the needs of phase 1 or 2 clinical trials.
Although the focus of this feature article is on top-down methods, a brief review of bottom-up particle-fabrication meth- ods may be instructive by way of comparison. Bottom-up approaches are readily scalable but can lack fine control of particle size and dispersity and can be limited in the variety of shapes that can be produced. Crystals of many metals and metal oxides can be grown into both spherical and anisotropic shapes such as cubes, rods, discs, and faceted polyhedra using nucleation and arrested growth strategies.1 More complex branched struc- tures can be formed by the sequential growth of dots and rods of different materials.6 Selective crystallization and deposition methods have been developed to produce prisms, rods, arrows, and teardrop-shaped crystals of gold, silver, and cadmium sele- nide, respectively.7-9 Such nucleation and growth methods are limited to inorganics, and the shape selectivity is highly dependent on the material and its crystal structure. Furthermore, these inorganic materials often lack the capability to encapsulate cargo or undergo surface modification.
Bottom-up approaches to the fabrication of organic particles have received a great deal of attention for use in biomedical applications such as imaging, gene or drug delivery. These methods chiefly rely on self-assembly to create particles such as micelles, vesicles, liposomes, and polymersomes that are able to encapsulate cargo in their otherwise hollow interior. In a typical preparation, molecules are synthesized that possess both hydro- philic and hydrophobic domains. Upon exposure to water, the molecules orient themselves to present their hydrophilic portion to the water and the hydrophobic domains orient inward, result- ing in a sphere with a hydrophobic pocket. The hydrophobic interior is able to encapsulate hydrophobic cargo. Although spherical particles have been the standard, increasing attention has been directed toward higher-aspect-ratio particles that are achievable with control of the relative length of hydrophobic and hydrophilic domains.5,10 Recently, Discher et al. explored fila- mentous polymersomes called filomicelles for their circulation persistence and ability to encapsulate and deliver chemotherepeu- tics. The filomicelles were prepared from block copolymers with lipid-like amphiphilicity but a more symmetric ratio of hydro- philic to hydrophobic blocks than is found in lipids. Filomicelles were prepared with a hydrophilic block of poly(ethylene oxide) and a hydrophobic block of either poly(ethylethylene) or poly- (caprolactone). These filamentous particles persisted in circula- tion about 10 times longer than spheres with similar surface chemistry and were successful in encapsulating and delivering a hydrophobic chemotherapeutic to tumored mice.10 One criticism of these self-assembled systems is the dynamic nature of particles so assembled. With membrane components held together with attractive forces rather than with covalent bonds, the generated structures can add or lose components, making their size and shape more fluid than may be desired. The dynamic nature of these self-assembled systems has been addressed by Wooley
et al. with shell-cross-linked knedel-like particles (SCKs).11 These block copolymermicelles aremodifiedwith reactive groups on the surface that can be chemically cross linked after self-assembly, giving superior stability to their structure.
Top Down, Scalable Methods for Fabricating Shape-Specific Particles
Bottom-up fabrication methods tend to be readily scalable to large batches but typically offer limited control over size, size distribution, and particle shape. Whereas “crude” top-down methods, such as milling and grinding of bulk materials, have found utility in industry, they typically offer very little control over these same parameters of particle size, size distribution, and shape deviating greatly from spherical. Several top-down particle- fabrication methods are excellent targets for the mass production of shape-specific particles, having the potential for superior control over the above parameters with the potential for scal- ability. In addition to the PRINT method favored by us, hard- template methods, microfluidic reactors, and particle stretching all show promise in this regard (Table 1).
Hard-Template Synthesis of Nanoparticles
For the last 20 years, hard-template methods have been used extensively to prepare nanowires, nanotubes, and nanorods.28 In this technology, porous templates are filled with one or more materials to fabricate monodisperse nanoparticles. The two most common templates used are anodized aluminum oxide (AAO) and track-etched polymer membranes. AAO, formed from the electrochemical oxidation of aluminum in an acid electrolyte, has a high density of uniform, parallel pores (as high as 1011 pores/ cm2) regularly spacedona hexagonal lattice. Templates have been fabricated with pore diameters as small as 5 nm.29 Track-etched polymermembranes are formed via the bombardment of polymer films, such as polycarbonate, with nuclear fission fragments to create damage tracks that are then chemically etched into pores. These polymer templates contain randomly distributed, uniform- diameter pores, with pore densities approaching 109 pores/cm2
and pore diameters as low as 10 nm. In this case, however, the pores are not parallel and so may intersect. Traditionally, the templates are filled via electrochemical means. This is accom- plished by first depositing a sacrificial metal layer, such as gold or silver, on one side of the template for electrical contact. Then, the material or materials of interest are electrochemically deposi- ted into the pores of the template; for example, metals can be synthesized using the appropriate electroplating solution, and conducting polymers can be synthesized via oxidative polymeri- zation. After deposition, the template can be dissolved; for example, AAO dissolves readily in acidic or basic solutions. This yields an ensemble of nanoparticles connected at the base to the
(6) Milliron, D. J.; Hughes, S. M.; et al. Nature 2004, 430, 190. (7) Jana, N. R.; Gearheart, L.; Murphy, C. J. J. Phys. Chem. B 2001, 105, 4065. (8) Manna, L.; Scher, E. C.; Alivisatos, A. P. J. Am. Chem. Soc. 2000, 122,
12700. (9) Jin, R.; Cao, Y.; et al. Science 2001, 294, 1901. (10) Geng, Y.; Dalhaimer, P.; et al. Nat. Nanotechnol. 2007, 2, 249. (11) Thurmond, K. B., 2nd; Remsen, E. E.; et al. Nucleic Acids Res. 1999, 27,
2966. (12) Hurst, S. J.; Payne, E. K.; et al. Angew. Chem., Int. Ed. 2006, 45, 2672.
(13) Hou, S.; Wang, J.; et al. J. Am. Chem. Soc. 2005, 127, 8586. (14) Hou, S.; Wang, J.; et al. Nano Lett. 2005, 5, 231. (15) Dendukuri, D.; Doyle, P. S. Adv. Mater. 2009, 21, 1. (16) Nisisako, T.; Torii, T. Lab Chip 2008, 8, 287. (17) Xu, S.; Nie, Z.; et al. Angew. Chem., Int. Ed. 2005, 44, 724. (18) Shepherd, R. F.; Conrad, J. C.; et al. Langmuir 2006, 22, 8618. (19) De Geest, B. G.; Urbanski, J. P.; et al. Langmuir 2005, 21, 10275. (20) Dendukuri, D.; Pregibon, D. C.; et al. Nat. Mater. 2006, 5, 365. (21) Shepherd, R. F.; Panda, P.; et al. Adv. Mater. 2008, 20, 4734. (22) Pregibon, D. C.; Toner, M.; et al. Science 2007, 315, 1393. (23) Hernandez, C. J.; Mason, T. G. J. Phys. Chem. C 2007, 111, 4477. (24) Badaire, S.; Cottin-Bizonne, C.; et al. J. Am. Chem. Soc. 2007, 129, 40. (25) www.liquidia.com, 2009. (26) Canelas, D. A.; Herlihy, K. P.; et al. Wiley Interdis Rev: Nanomed.
Nanobiotech. 2009, 1, 391. (27) Kelly, J. Y.; Desimone, J. M. J. Am. Chem. Soc. 2008, 130, 5438. (28) Cai, Z.; Martin, C. R. J. Am. Chem. Soc. 1989, 111, 4138. (29) Martin, C. R. Chem. Mater. 1996, 8, 1739.
DOI: 10.1021/la903890h CLangmuir XXXX, XXX(XX), XXX–XXX
Merkel et al. Invited Feature Article
substrate metal. It is occasionally useful to leave the nanoparticles connected for electronic applications; however, the connecting sacrificial metal layer can also be dissolved to generate a mono- disperse suspension of high-aspect-ratio nanoparticles. Both tubes and solid wires or rods can be prepared depending on the surface treatment of the template. For example, gold nanowires can be synthesized galvanostatically in AAO; however, if the template is first treated with (2-cyanoethyl)triethoxysilane, then the gold preferentially deposits on the walls of the template pores and thus forms nanotubes instead of solid nanowires.30 Additionally, whereas the nanotubes are generally preparedwith uncapped ends, they can be synthesizedwith one end capped.31 This has interesting implications with respect to the ability of nanotubes to act as effective biological delivery vectors.31 Nonelectrochemical meth- ods have also been developed to fill the templates, such as sol-gel32
and layer-by-layer33 approaches to the fabrication of semiconduc- tor (cadmiumselenide, zinc oxide, and graphitic) nanoparticles and nonconducting polymer particles.34
Remarkably, Martin and co-workers13,14 have extended the template method to the fabrication of biological nanoparticles such as DNA and protein nanotubes. For the case of DNA nanotubes, this was made possible by pretreating the template pores with alternating R,ω-diorganophosphate (R,ω-DOP) Zr- (IV) chemistry. The first DNA segment binds to Zr(IV) through the phosphonate end of the DNA strand. Subsequent DNA segments are incorporated into the nanotube through hybridiza- tion with the preceding DNA segment. Thus, the resulting DNA nanotube has an outer skin composed of the (R,ω-DOP) Zr(IV) layer.13 A similar alternating chemistry approach was used in the fabrication of protein nanotubes;the AAO pores were first treated with 3-amino propylphosphonic acid, followed by protein immobilization agent glutaraldehyde. Protein then reacted with glutaraldehyde, and additional alternating layers of glutaralde- hyde and protein were deposited to thicken the nanotube walls.14
One crucial factor in these biological systems is the release of nanotubes from the template because the AAO template is dissolved under acidic or basic conditions. For the release of these nanotubes, the template was dissolved under very mild acidic conditions (1.5-5% phosphoric acid)13,14 at 0 C for 24 h. The biological activity of the nanotubes was confirmed after release. However, in the case of the protein nanotubes, it was noted that many of the tubes were broken during the dissolution and filtration steps.14
Onemajor advantage of the templatemethod is that segmented or multicomponent nanoparticles can be straightforwardly fab- ricated (Figure 1).12,35 This has led to an impressive breadth of research into the area of postfabrication modifications of seg- mented nanoparticles. Multicomponent nanorods can be selec- tively functionalized for different multiplexing, sensing, and self- assembly applications.36-38 Mirkin et al. developed on-wire lithography (OWL) to create nanowires with nanoscale gaps along the length of the wire. For example, a segmented nano- wire with alternating gold and nickel segments was synthesized electrochemically in an AAO template.37 After the nanowires were released and dispersed onto a substrate, gold or silicon dioxide was coated onto one side. The rodswere then redispersed,
T a b le 1 . C o m p a ri so n o f S ev er a l H ig h ly
S ca la b le , T o p -D
o w n , S h a p e- S p ec if ic P a rt ic le -F a b ri ca ti o n M et h o d s
li m it s
sh a p e sp ec if ic it y
p a rt ic le co m p o si ti o n /m
a te ri a l
li m it a ti o n s
fa b ri ca ti o n ca p a ci ty
ca rg o
h a rd
te m p la te
m et h o d s
1 0 n m - 2 μ m
fo r tr a ck -
et ch ed
b ra n es ,
5 n m - 2 6 7 n m
fo r A A O
li m it ed
to ro d /
cy li n d er
m o rp h o lo g ie s:
h ig h a sp ec t ra ti o s p o ss ib le
m et a ls , p o ly m er s, in o rg a n ic
co m p o u n d s,
se m ic o n d u ct o rs ,1 2 a ci d /
b a se -c o m p a ti b le m a te ri a ls
te m p la te s w it h u p to
1 0 1 1
p o re s/ cm
D N A
a n d p ro te in
1 3 ,1 4
m ic ro fl u id ic s -
d ro p le t- b a se d
5 - 2 0 0 μ m
sp h er ic a l d er iv a ti v es , d ef in ed
b y ch a n n el a rc h it ec tu re
p h o to p o ly m er iz a b le m a te ri a ls ,
lo w -m
el ti n g -p o in t o il s,
so lu b le p o ly m er s
3 2 0 m L /h
f 5 g /m
in o f
9 6 μ m
a cr y la te
p a rt ic le s1
6 q u a n tu m
d o ts ,1 7 co ll o id s, 1 8
p ro te in s1
m ic o fl u id ic s -
fl o w m et h o d s
1 - 2 0 0 μ m
d ef in ed
b y ch a n n el a rc h it ec tu re
a n d li th o g ra p h ic m a sk
p h o to p o ly m er iz a b le m a te ri a ls
o n ly , P D M S -c o m p a ti b le
u p to
6 0 0 0 p a rt ic le s/ m in
fo r g 1 0 μ m
p a rt ic le s2
0 co ll o id s, 2 1 D N A
p a rt ic le st re tc h in g
6 0 n m - 1 0 0 μ m
5 sp h er ic a l d er iv a ti v es
w it h
co m p le x g eo m et ri es
p o ly st y re n e, P L G A
1 0 8 - 1 0 1 0 p a rt ic le s p er
st re tc h in g a p p a ra tu s5
a p ro te in s5
p h o to li th o g ra p h y
th ic k n es s g 1 0 0 n m ,2 3 la te ra l
en si o n s g 8 0 0 n m
d ef in ed
b y m a sk
- a n y a rb it ra ry
sh a p e d ra w n w it h co m p u te r-
a id ed
d es ig n (C
A D )
p h o to p o ly m er iz a b le m a te ri a ls
b u t o p ti m a l fo r p h o to re si st s
1 0 8 p a rt ic le s/ m in
2 3 b
d y es , in o rg a n ic
n a n o p a rt ic le s2
P R IN
T 1 0 n m - 2 0 0 μ m
sh a p e d ef in ed
b y a m a st er
te m p la te
w it h n a n o m et er
re so lu ti o n
p ro te in s, a ct iv e th er ep eu ti cs ,
so lu b le o r m el t p ro ce ss a b le
p o ly m er s, p o ly m er iz a b le
m a te ri a ls
4 0 0 fe et
o f m o ld /h
f 2 0 m g /
o f su b -2 0 0 -n m
p a rt ic le s, 5 0 0 m g /m
in o f
5 μ m
p a rt ic le s2
co ll o id s a n d im
a g in g
a g en ts ,2 6 fl u o ro p h o re s,
p ro te in s, 2 7 d ru g s2
a T h e sc a la b il it y o f th es e m et h o d s h a s n o t b ee n d em
o n st ra te d ,a n d p a rt ic le y ie ld s a re d if fi cu lt to
es ti m a te o n a g /m
in b a si s.
b T h is re p re se n ts a n es ti m a te ,w
h ic h a ss u m es th e in co rp o ra ti o n o f a n a u to m a te d ro b o ti c
tr a ck
sy st em
fo r th e ex p o su re
o f w a fe rs o f p a rt ic le s, a n d d o es
n o t in cl u d e th e ti m e a ss o ci a te d w it h p o st ex p o su re
b a k e o r d ev el o p in g st ep s.
c T h es e v a lu es
re p re se n t th e o p ti m iz ed
o p er a ti o n o f a n a u to m a te d ,r ee l- to -
re el P R IN
T p a rt ic le fa b ri ca ti o n sy st em
em p lo y ed
b y L iq u id ia
T ec h n o lo g ie s. 2 5
(30) Brumlik, C. J.; Martin, C. R. J. Am. Chem. Soc. 1991, 113, 3174. (31) Gasparac, R.; Kohli, P.; et al. Nano Lett. 2004, 4, 513. (32) Hulteen, J. C.; Martin, C. R. J. Mater. Chem. 1997, 7, 1075. (33) Hou, S.; Harrell, C. C.; et al. J. Am. Chem. Soc. 2004, 126, 5674. (34) Liang, Z.; Susha, A. S.; et al. Adv. Mater. 2003, 15, 1849. (35) Park, S.; Lim, J.-H.; et al. Science 2004, 303, 348. (36) Martin, B. R.; Dermody, D. J.; et al. Adv. Mater. 1999, 11, 1021. (37) Banholzer, M. J.; Qin, L.; et al. Nat. Protoc. 2009, 4, 838. (38) Wei, W.; Li, S.; et al. Angew. Chem., Int. Ed. 2009, 48, 4210.
D DOI: 10.1021/la903890h Langmuir XXXX, XXX(XX), XXX–XXX
Invited Feature Article Merkel et al.
and the nickel was etched, resulting in nanowires with predefined gaps and disks. These architecturally complex structures have been found to have very interesting properties that are useful in nanoelectronics12 and surface-enhanced Raman scattering (SERS).38
It is evident that the hard-template method is useful for the synthesis ofmonodisperse nanoparticles. In fact, it is currently the preferred method of synthesizing very high aspect ratio nano- objects.12 A wide range of chemistries can be applied, and it is straightforward to fabricate particles with multiple chemistries spatially resolved within the particles. One concern with this method is that as the particles are released upon dissolution of the template andmetal substrate (in the case of electrodeposition) the particle chemical composition must be compatible with the solvents used, usually acids and bases. Another drawback of this process is the limitations in nanoparticle shape;most nanopar- ticles have been synthesized with cylindrical geometries. Further- more, the issue of scalability is somewhat questionable because virtually all research has been conducted on relatively small scales (approximately 1 in.2), so whereas theoretically this approach can be high-throughput, this has yet to be truly developed. Among other factors, scale up will be dependent on the availability of large-area, high-density templates.
Microfluidic Particle Fabrication
One emerging strategy for the fabrication of particles with control over particle size and shape is fabrication with micro- fluidic devices. In amicrofluidic reactor,minute amounts of liquid
flow through micrometer-sized channels that can be designed to produce monodisperse emulsion droplets. These emulsion dro- plets are then solidified to form particles. Most methods for droplet formation in microfluidic devices are passive ones that rely on the liquid flow field deformations for droplet formation rather than mechanical actuation or moving parts.41 Passive droplet formation stems from several different device architec- tures such as multiphase flow at T junctions,42 flow-focusing channels,43 or coflow-based systems44 that exploit the laminar nature of fluid flow at the low Reynolds numbers associated with the small size of the channels. Because of the nature of these fluid dynamics, liquid-liquid interfaces for parallel flowing liquids experience only diffusion-based mixing and sharp boundaries are maintained, allowing for the formation of droplets with small size distributions.
The material of the droplets determines the method by which they can be solidified to form particles. Droplets can simply be cooled from a liquid to a room-temperature solid phase, or solvent can be evaporated from a dissolved polymer. Photo or thermal polymerization of droplets with reactive groups is an- other method of solidifying droplets. The solidification of dro- plets by means of a chemically driven polymerization reaction, such as the condensation of amines and carbonyl chlorides, has also been reported.45 Essentially, the droplets must be composed of a material that is capable of photochemical, thermal, or
Figure 1. (A)General scheme for nanorod synthesis via the deposition ofmaterials into a nanoporous template. Copyright 2006Wiley-VCH Verlag GmbH & Co. KGaA. Reproduced with permission from ref 12. (B) Bright-field optical micrographs of Au/Pt/Au rods. Copyright 1999 Wiley-VCH Verlag GmbH & Co. KGaA. Reproduced with permission from ref 39. (C) SEM images of Au-Ni multisegmented nanorods. Reprinted by permission fromMacmillan Publishers Ltd., ref 40, copyright 2009. (D) SEM image of Au-polypyrrole rods. From ref 35; reprinted with permission from AAAS.
(39) Martin, B. R.; Dermody, D. J.; et al. Adv. Mater. 1999, 11, 1021. (40) Banholzer, M. J.; Qin, L.; et al. Nat. Protoc. 2009, 4, 838.
(41) Christopher, G. F.; Anna, S. L. J. Phys. D: Appl. Phys. 2007, 40, R319. (42) Nisisako, T.; Torii, T.; et al. Lab Chip 2002, 2, 24. (43) Anna, S. L.; Bontoux, N.; Stone, H. A. Appl. Phys. Lett. 2003, 82, 364. (44) Kenis, P. J.; Ismagilov, R. F.; et al. Science 1999, 285, 83. (45) Cohen, I.; Li, H.; et al. Science 2001, 292(), 265.
DOI: 10.1021/la903890h ELangmuir XXXX, XXX(XX), XXX–XXX
Merkel et al. Invited Feature Article
chemical curing, or they must be readily melt- or solvent-proces- sable materials.
Shapes produced in these channels have been limited to spheres, cylinders, and spherically derived shapes such as discs and ellipsoids. More complex and chemically anisotropic shapes such as hemispheres46 and core-shell46 and janus particles18 were produced by confining the droplets with the geometry of the device and with coflowing laminar streams, though these remain derivatives of the original spherical shape. Microfluidic techni- ques have readily produced particles in the size range of 10- 100μm, though reductionmuchbelow this size scale has remained a limitation of this technology.15
Recently, theDoyle grouphas developed photolithographically coupled approaches that have produced more complex shapes by the use of a photolithographic mask.20,47 In continuous- flow lithography (CFL), a flowing stream of monomer in a polydimethylsiloxane (PDMS) channel was exposed in mask- defined shapes with short bursts of UV light. Particles were cross linked by UV light exposure almost instantaneously and did not plug up the channel as a result of the oxygen-mediated inhibition of polymerization at PDMS surfaces, which left a thin layer of unpolymerized monomer at the channel walls.48 The output of particles was continuous, and the shape was defined by the limits of the lithographic mask (Figure 2). Exposure across several fluid streams resulted in the formation of Janus-type particles with two or more chemically distinct regions on each particle.20 CFL systems have some limits to resolution and speed, however, because higher flow speeds and smaller particle sizes caused blurring or smearing of the particles formed in the moving stream of polymerizable fluid. Higher resolution has been achieved with
the use of stop-flow lithography (SFL), a process during which the flow of monomer was stopped briefly during each exposure. This process enhanced the resolution of the lithographic techni- que compared to CFL but was limited somewhat by the time taken by the flowing liquid to stop and restart. Complex shapes have been formed at high resolution on the micrometer scale, including a range of concave- and convex-sided pyramids, rec- tangles, and spheres as well as rectangular toroids, all with multiple, chemically distinct phases possible.47
Microfluidic methods have demonstrated their versatility in terms of particle shape and chemical anisotropy of the particles produced. The emergence of CFL and SFL techniques have increased the resolution of this technique, though there are material limitations; particles made in this way have been limited to polymerizable fluids, and solvent choice has been limited by the swelling of the devicematerial, PDMS, inmany solvents. Another limitation has been the size of particles that can be produced with these methods. Particles on the order of 3-100 μm were readily fabricated, but submicrometer particle sizes remain a challenge for microfluidic technology. The scalability of this technique is somewhat in question, though the autonomous nature of the technology makes it a tempting target. In a simple CFL device, a throughput of near 100 particles/s was reported.20 Simply stack- ing many devices together has been used to scale up the produc- tion of particles, whereas the simplest strategy has been seen as impractical because of the complications involved in linking all of the required tubes and pumps required for multiple devices.49
With typical flow rates in the range of 1000 μL/h for droplet- based reactors, an industrially useful output of 1 L of material/h would require 1000 reactors chained together, which seems feasible. More complicated devices, which have as many as 128 droplet-producing cross junctions, have been fabricated to
Figure 2. (A) Schematic used for continuous-flow lithography and (B)microparticles of varying shapes fabricatedwith thismethod, adapted by permission fromMacmillan Publishers Ltd., ref 20, copyright 2006. (C) (Right) Schematic of the channel and flow configuration used for the formation of the bicolored Janus particles shown on the left, reproduced with permission from ref 50, copyright 2006Wiley-VCHVerlag GmbH & Co. KGaA.
(46) Nie, Z.; Xu, S.; Seo, M; Lewis, P. C.; Kumacheva, E. J. Am. Chem. Soc. 2005, 127, 8058. (47) Dendukuri, D.; Gu, S. S.; et al. Lab Chip 2007, 7, 818. (48) Decker, C.; Jenkins, A. D. Macromolecules 1985, 18, 1241. (49) Joanicot, M.; Ajdari, A. Science 2005, 309, 887.
F DOI: 10.1021/la903890h Langmuir XXXX, XXX(XX), XXX–XXX
Invited Feature Article Merkel et al.
facilitate scale up and reduce the number of devices needed for the mass production of particles.16,50
Spherical particles prepared by bottom-up methods such as microemulsion can be manipulated to produce anisotropic shapes. This particle-stretching method initially was used to deform polystyrene (PS) particles that were embedded in a poly- vinyl alcohol (PVA) film.Uponheating to a temperature atwhich the particles could deform, the filmwas stretched and then rapidly quenched, yielding ellipsoidal particles upon dissolution of the PVA fim.51
Recently, Champion et al. reported the formation of complex shapes via a similar method (Figure 3). In this method, as before, (PS) particleswere embedded in aPVA film. ThePS particles were liquefied by the addition of heat or solvent, allowing the PS to flow, filling void spaces in the surrounding film, or to stretch with the film as a result of hydrogen-bonding-based attraction to the surrounding film. The manipulation of the PS particles required exposure to temperatures on the order of 140 C or organic solvents (toluene), which places some limits on the encapsulation of cargo by these particles. Carefulmodifications to the stretching protocol allowed for the selective formation of over 20 distinct particle shapes upon solidification of the particles by cooling or the removal of solvent and subsequent release from the PVA film.52
This particle-stretchingmethod has also been demonstrated on particles composed of PLGA, a hydrophobic polymer that has utility in drug-delivery applications. PLGA particles were de- formed from their initial spherical shape at much lower tempera-
tures than possible with PS particles because of the lower glass- transition temperature of this material (40-60 C for PLGA vs 90 C for PS). Particles processed in this way were able to encapsulate a fluorescently labeled protein (66 kDaFITC-labeled bovine serum albumin) with no leaching of the protein during stretching of the particles, suggesting that the processingmethods weremild enough to accommodate many such types of biological cargo.5
One distinct advantage of the particle-stretching technique has been the ability to compare differently shaped particles with identical volumes by preparing different particles shaped from the same stock of spherical particles. Material compatibility is a concern for this method, and the bulk of the literature deals with PS- and PLGA-based particles, though one can imagine that manymaterials could bemanipulated in this way by adjusting the conditions used to fluidize the particles and the material in which the particles are embedded. This method is versatile with respect to particle size, and spherical PS particles with diameters from 60 nm to 100 μm have been subjected to these stretching proto- cols. The scalability of the method remains to be seen because particles have been produced only on a laboratory scale. Particle yields of 108-1010 particles per stretching apparatus have been reported,5 though thicker or stacked films and larger stretching mechanisms could increase throughput. The polydispersity of the particles prepared by stretching methods has reflected the poly- dispersity in the spherical particles before stretching. Careful selection of a spherical particle source could lead to the formation of more monodisperse populations of stretched particles.
Photolithography, the workhorse of the semiconductor indus- try for high-volume manufacturing,53 has also been directly applied to the fabrication of discrete monodisperse colloidal particles.23,54,55 There are four major steps in the process. The first is the spin coating of a sacrificial layer, followed by a photoresist layer on a silicon wafer. The particle will be composed of the photoresist material, so the thickness of that layer will determine the particle height. Second, the photoresist is exposed to UV light through a mask with the specific pattern of interest. This exposure step can be carried out in an automated UV lithographic projection exposure system, or stepper.23 In the case of a negative photoresist, the exposed areas of the photoresist are rendered insoluble after postexposure baking steps because of cross linking. The third step is the development of the photoresist, where an organic solvent dissolves the unexposed regions leaving discrete photoresist particles on the sacrificial layer. The final step is the dissolution of the sacrificial layer to release the particles. A simple illustration of this process is shown in Figure 4A.
Both Mason et al.23 and Stroock et al.24 have reported using this type of approach to generate highly monodisperse, shape- specific colloidal particles. Stroock et al.56 demonstrated the ability to tailor the surface roughness of the particles bymodifying the development conditions (Figure 4B).Mason et al.23 illustrated the shape versatility of this process by generating particles with a wide range of shapes, including all 26 letters of the English alphabet. The fabricationof the lithographically derived particles, or LithoParticles as coined by Mason, was also found to be amenable to the incorporation of both organic dyes and nano- particles such as iron oxide. It was even feasible to fabricate
Figure 3. (A) Two general schemes used in stretching spherical particles to different shapes. In scheme A, particles are liquefied prior to stretching the film, and in scheme B, the film has stretched prior to particle liquidification. (B) Polystyrene particles prepared via the method shown in scheme A. (C) Polystyrene particles prepared via scheme B. Figure adapted from ref 52, copyright 2007 National Academy of Sciences, U.S.A.
(50) Nisisako, T.; Torii, T.; et al. Adv. Mater. 2006, 18, 1152. (51) Ho, C. C.; Keller, A.; Odell, J. A.; Ottewill, R. H. Colloid Polym. Sci. 1993,
271, 469. (52) Champion, J. A.; Katare, Y. K.; et al. Proc. Natl. Acad. Sci. U.S.A. 2007,
(53) Geissler, M.; Xia, Y. Adv. Mater. 2004, 16, 1249. (54) Jang, J.-H.; Ullal, C. K.; et al. Nano Lett. 2007, 7, 647. (55) Moon, J. H.; Kim, A. J.; et al. Adv. Mater. 2007, 19, 2508. (56) Badaire, S.; Cottin-Bizonne, C.; et al. Langmuir 2008, 24, 11451.
DOI: 10.1021/la903890h GLangmuir XXXX, XXX(XX), XXX–XXX
Merkel et al. Invited Feature Article
hybrid bilayer Janus particles through the successive exposure of two different masks. Even though this photolithographic process has been optimized for photoresists, it is possible to use other UV-reactive systems. For example, microparticles composed of methacrylated hyperbranched polyglycerols were fabricated di- rectly using photolithography.57
There are many clear advantages to this type of processing: there is excellent shape fidelity down to the submicrometer level, and the process can be high-throughput. For example, if a stepper with an automated wafer-handling system is used, it is possible to expose one 5 in. wafer every minute to yield about 108 particles/ min.23 (This estimate does not, however, consider the postexpo- sure bake, development and release steps.) It was envisioned that by incorporating all of the equipment necessary for making the particles into a robotically automated track system a continuous rate of 108 particles/min is achievable.23 However, this creates a major stumbling block to using this process as a large-scale particle-fabrication technology because the enormous capital expenditure required to run such an expensive lithographic exposure system continuously is extremely daunting. Another major issue with this type of processing is the limitations in the chemistries that can be used in particle fabrication. These issues have led to the development of alternative routes to LithoParticle production that take advantage of templating, where only the initial reusable template requires photolithography.58
Other direct photolithographic approaches to particle fabrica- tion include the use of interference lithography, where the con- structive and destructive interference of multiple laser beams at the surface of a photoresist generates patternswithout the use of a mask.59 This is a benefit because high-resolution masks can be fairly expensive to fabricate. Usually, the projected patterns are limited to regularly spaced lines or dots;59 this strategy has been used to fabricate cylindrical particles with a varying aspect ratio from0.4 to 10.55However, Thomas and co-workers54 have shown that this process can be used to fabricate multivalent 3D colloidal particles (Figure 4C). These complex shapes are extremely diffi- cult to fabricate by other top-down fabrication processes.
Imprint Lithography Methods
Imprint-lithography-based methods have emerged in recent years as promising techniques for the scalable fabrication of
shape-specific colloidal particles. Soft-lithography-based techni- ques, which were pioneered by Whitesides et al. in the late 1990s, begin with the use of a rigid template that is used to fabricate an elastomericmold, which can then be used to replicate the shape of the original template in a molding or stamping process.61 Imprint methods were first envisioned as a high-resolution, low-cost alternative to traditional photolithograpic techniques for micro- and nanoscale pattern replication rather than the fabrication of distinct particles. Particle fabrication has been a challenge due in large part to the persistent formation of a residual layer ofmolded material, called a flash layer, that connects the potential parti- cles.62 Much attention has been directed toward the removal or avoidance of this flash layer with the goal of particle fabrication. Step and flash imprint lithography (S-FIL) uses a rigid, UV- transparent template as a mold material, creating an embossed film of particles from aUV-curablematerial. The interconnecting flash layer is removed by an etching step, with the dissolution of a sacrificial layer yielding distinct particles.63 Removal of the flash layer via an oxygen plasma etch may be undesirable for some particle compositions or cargos, though etch times from 20 to 90 s have been reported for effective particle release, and the biological activity of an encapsulated protein was maintained after 20 s of etching.63
Utilization of the traditionalmoldmaterial for soft lithography applications, a cross-linked poly(dimethyl siloxane) (PDMS) elastomer, typically results in particles connected by such a flash layer. Recently, microparticles have been fabricated with PDMS molds without the formation of a flash layer by the careful application of photocurable acrylate resins as well as aqueous and organic solutions of polymers to the molds.64 Surface treat- ment can increase or decrease the hydrophilicity of the mold, facilitating mold filling, mold dewetting, and particle release.65
PRINT technology (Figure 5), which utilizes a fluorinated elastomeric mold in an imprint lithographic technique, allows for the facile fabrication of distinct particles without the forma- tion of a flash layer while maintaining many of the advantages of the soft lithography technique while avoiding some of the difficulties associated with PDMS.66
Figure 4. (A) Simple scheme showing thephotolithographic fabricationof colloidal particles, adaptedwithpermission fromref 24, copyright 2007 American Chemical Society. (B) Cylindrical photoresist particles engineered with rough side walls (particle radius= 520 nm, height= 875nm), adaptedwithpermission from ref 56, copyright 2008AmericanChemical Society. (C) 4-Valent and6-valent nanoparticles fabricated using interference lithography (scale bars represent 300 nm), adapted with permission from ref 60, copyright 2007 American Chemical Society.
(57) Oudshoorn, M. H. M.; Penterman, R.; et al. Langmuir 2007, 23, 11819. (58) Hernandez, C. J.; Kun, Z.; et al. Soft Mater 2007, 5, 13. (59) Gates, B. D.; Xu, Q.; et al. Chem. Rev. 2005, 105, 1171. (60) Jang, J.-H.; Ullal, C. K.; et al. Nano Lett. 2007, 7, 647.
(61) Xia, Y.; Whitesides, G. M. Angew. Chem., Int. Ed. 1998, 34, 550. (62) Dumond, J. L. H. Y. Adv. Mater. 2008, 20, 1291. (63) Glangchai, L. C.; Caldorera-Moore, M.; et al. J. Controlled Release 2008,
125, 263. (64) Guan, J.; Ferrell, N.; et al. Biomaterials 2006, 27, 4034. (65) Moran, I. W. C.; Dalton, F.; Jhaveri, S. B.; Carter, K. R. Soft Matter 2008,
4, 168. (66) Rolland, J. P.; Maynor, B. W.; et al. J. Am. Chem. Soc. 2005, 127, 10096.
H DOI: 10.1021/la903890h Langmuir XXXX, XXX(XX), XXX–XXX
Invited Feature Article Merkel et al.
Particle Replication in Nonwetting Templates (PRINT)
Although a number of published manuscripts are available describing a wide variety of particles fabricated using the PRINT technique (Figure 6),27,66,68-72 the details and rationale behind particle fabrication can be confusing. With this in mind, the goal of this section is to revisit past particle-fabrication methods to examine the current state of the art of the PRINT technique, to provide a clear picture of how the technique works, and to look at why the PRINT technique can be readily scaled up.
In 2004, Rolland et al. reported the synthesis of a new perfluoropolyether (PFPE)-based elastomer,73 a material that proved to be more effective than PDMS as a template for soft- lithography-based particle fabrication. Photocurable PFPE (Fluorocur, Liquidia Technologies Inc., Research Triangle Park, NC) was synthesized by end capping a fluorinated PFPE diol (Solvay) with isocyanatoethyl methacrylate to form PFPE di- methacrylate. The end-functionalized fluorinated oil had a posi- tive spreading coefficient, allowing it to completely wet and envelop the delicate details of a master template. Curing of the oil was accomplishedunderUV light (365 nm,∼10000-15000mJ/ cm2) after the addition of a photoinitiator. The resulting PFPE elastomers have three distinct advantages over silicone templates for use in soft lithography. First, the low surface energy of the
elastomer (8-10 dyn/cm)74 facilitates the removal of an excess flash layer. Other lithographic methods typically require an etching step to remove this layer, complicating the scale up of these methods.75 Second, PFPE was observed to be compatible with a number of organic solvents that were incompatible with silicone molds.73 It was found that whereas both Sylgard 184 (a siliconematerial) andPFPEmolds exhibited negligible swelling in the presence of water and methanol, Sylgard 184 molds were permeable to toluene and dichloromethane. An examination of several other common solvents and polymerizable organics showed a similar trend, with PDMS coupons generally showing a higher retention of these organic fluids (Table 2). With the elimination of swelling, master-to-replica fidelity was improved and the reproduction of much smaller feature sizes was possible. In 2006,Maynor et al. showed that a variety of nanoscale artifacts could be reproduced using the PRINT technique, including 60 nm adenoviruses, 45 nm polymeric toroidal micelles, and single- walled carbon nanotubes with heights as low as 2 nm.74 Third, the highly fluorinated nature of the mold (similar to that of Teflon) facilitated the removal of particles from the mold. In contrast, other soft lithography methods required surface mod- ification of the molds to facilitate particle removal.65
Initially, mold fabrication was performed by drop casting 20-30 mL of Fluorocur resin onto a 6-8-in.-diameter patterned silicon master template that was then cured photochemically in a UV oven.66 Molds were released from the template by slowly peeling them back from the silicon wafer using PTFE tweezers so as not to scratch the fragile surface features. Because of the delicate nature and high cost associated with using large quan- tities of Fluorocur resin for the thick molds, the scale of particle production was limited. With this in mind, a proprietary roll-to- roll system capable of producing up to 1000 ft.2/day of PFPE molds on a flexible poly(ethylene terephthalate) (PET) backing was recently engineered by Liquidia Technologies.25 The new process yields mechanically robust molds and uses a reduced amount of Fluorocur resin, making the process less costly and highly scalable.
The filling of any patternedmold is a process that relies heavily on the surface energies of the materials in play. To fill the cavities effectively, a preparticle solution should have the ability to partially wet the surface of the mold. Solutions capable of wetting the surface of a mold are drawn into the cavities of the mold by capillary forces. During the fabricating of embossed films, the removal of excess solution is not a concern, but in particle fabrication, in which individual structures rather than one mono- lith are desired, the management of excess preparticle solution is crucial. After a solid embossed film is created, the step and flash imprint lithography (S-FIL) method, recently commercialized by Molecular Imprints, Inc., removes the flash layer using a “break- through” etch to produce individual surface features. For high- throughput particle fabrication, however, it is desirable to eliminate any etching steps because they are time-intensive and not amenable to a continuous process. Using the PRINT method, the fabrication of discrete particles was made possible by the removal of excess solution prior to the solidification of particles in the mold. Excess preparticle solution was removed by first laminating the filled mold with ahigh-surface-energymaterial (i.e., PET, surface energy=42.1 mN/m)77 and then by peeling back the laminate material. Capillary
Figure 5. General schematic representation of the PRINT pro- cess. (A) A preparticle solution (red) is distributed evenly in the elastomeric mold (green) by a roller (black) with a high-surface- energy polymer sheet, which wicks away excess solution. (B) The particle solution is solidified in themold. (C) Particles are removed from the mold with a harvesting film (yellow). (D) Dissolution of the film yields free-flowing particles in solution.
(67) Zhang, H.; Nunes, J. K.; et al. New J. Phys. 2009, 11, 075018. (68) Gratton, S. E.; Napier, M. E.; et al. Pharm. Res. 2008, 25, 2845. (69) Gratton, S. E.; Pohlhaus, P. D.; et al. J. Controlled Release 2007, 121, 10. (70) Gratton, S. E.; Ropp, P. A.; et al. Proc. Natl. Acad. Sci. U.S.A. 2008, 105,
11613. (71) Herlihy, K. P.; Nunes, J.; et al. Langmuir 2008, 24, 8421. (72) Petros, R. A.; Ropp, P. A.; et al. J. Am. Chem. Soc. 2008, 130, 5008. (73) Rolland, J. P.; Van Dam, R. M.; et al. J. Am. Chem. Soc. 2004, 126, 2322. (74) Maynor, B. W.; Larue, I.; et al. Small 2007, 3, 845.
(75) Xia, Y.; Rogers, J. A.; et al. Chem. Rev. 1999, 99, 1823. (76) Orgel, R.; Merkel, T. J. Unpublished work, 2008. (77) Smithson, R.; McClure, D.; et al. Thin Solid Films 1997, 307, 110.
DOI: 10.1021/la903890h ILangmuir XXXX, XXX(XX), XXX–XXX
Merkel et al. Invited Feature Article
forces trap the liquid in the mold whereas excess solution adheres preferentially to the high-surface-energy material and is wicked away, leaving isolated reservoirs of preparticle solution, thus elim- inating the need for a scum-removal step.
Like surface energy, other factors such as film uniformity and film thickness of the preparticle solution play important roles in effective large-scale particle fabrication. Uniform films were drawn on a high-surface-energy sheet using a metal Mayer rod with grooved surface features and a mechanical film coating instrument (R.D. Specialties Inc.). Film deposition by a Mayer rod is very amenable to a continuous roll-to-roll process, making
it possible to easily lay thousands of feet of uniform film on a substrate. The film thickness was dictated by both the grooved feature size of the Mayer rod and the concentration of the preparticle solution.
Unlike patterning on a solid substrate or with a rigid pat- terned template, which can be time-consuming, patterning of the preparticle solution can be done in a continuous fashion when the template and preparticle solution are both on a flexible backing. Other particle fabrication methods, such as S-FIL, involve a step-by-step procedure in which a master template is brought into contact with a solid substrate in the presence of a preparticle solution, a step that can damage the master template if not performed with care. In the PRINT process, after a uni- form film was cast on PET as described above, the PET was laminated onto an empty mold using an in-house single-nip lamination system. Lamination conditions (pressure, speed, tem- perature, and humidity) were optimized for each new class of particles, theoretically allowing the continuous fabrication of hundreds of feet of laminated mold. Monomer-based prepar- ticle films that flowed freely into the cavities of the mold and had low vapor pressure were laminated at room temperature.71
Volatile monomers and other small-molecule systems required that the lamination temperature be reduced and that the lamina- tion take place in a glovebag or another controlled environment. Polymeric films such as poly(ethylene glycol) (PEG) or poly- (lactic-co-glycolic acid) (PLGA) sometimes required heat to raise the temperature of the film to above the glass-transition tem- perature of the polymer, allowing the polymer to flow into the mold.
Figure 6. Examples of size, shape, composition, cargo, and surface chemistry control with the PRINT process: (A) 80 nm 2000 nm wormlike cross-linked poly(ethylene glycol) (PEG) nanoparticles on a harvesting layer,with the inset showing anAFM image of a harvested layer; (B) 3 μm toroids with roughened top surfaces made from cross-linked PEG; (C) hexnut-shaped cross-linked PEGmicroparticles; (D) siRNA-loaded 200 nmalbuminnanoparticles harvested onto amedical adhesive; (E) boomerang-shaped cross-linkedPEGmicroparticles on a harvesting layer; (F) 80 nm 360 nmcross-linkedPEGparticles on a polymeric harvesting layer; (G-I) 2mm 2mm 6mmcross-linked PEG rectangular prisms showing increasing chemical anisotropy [(G)monophasic, (H) biphasic, and (I) triphasic]. ImageD is reprintedwith permission from ref 27, copyright 2008 American Chemical Society. Images H and I are reproduced with permission from ref 67, copyright 2009 IOP Publishing Ltd.
Table 2. Comparison of Swelling Behavior, Reported as the Percent Increase in the Weight of Coupons of Cross-Linked Elastomeric
Materials after Soaking Overnighta
PDMS (wt % uptake)
PFPE (wt % uptake)
water 0.37 ( 0.04 0.39 ( 0.16 hexane 109.82 ( 1.41 1.72 ( 0.25 THF 145.46 ( 1.68 6.95 ( 0.08 DMSO 2.36 ( 0.50 1.98 ( 0.17 isopropanol 17.65 ( 2.44 2.38 ( 0.06 acetone 20.15 ( 1.05 4.81 ( 0.06 poly(ethylene glycol)
methyl ether 1.04 ( 0.45 0.36 ( 0.07
2-hydroxyethyl acrylate 1.45 ( 0.23 1.03 ( 0.22 N-vinyl-2-pyrrolidone 5.55 ( 0.31 2.34 ( 0.27 PEGtriacrylate 1.40 ( 0.06 0.31 ( 0.05
aTraditional soft lithography mold material, poly(dimethylsiloxane) (PDMS), and PRINT mold material, perfluoropolyether (PFPE), were compared with regard to several solvents and polymerizable mono- mers.76
J DOI: 10.1021/la903890h Langmuir XXXX, XXX(XX), XXX–XXX
Invited Feature Article Merkel et al.
Although the majority of top-down particle-fabrication pro- cesses rely on photochemical curing, particle solidification using PRINT can be quickly accomplished by a variety of different methods. Monomer systems incorporating a photochemical in- itiator such as 1-hydroxycylohexyl phenyl ketone or 2,2-diethoxy- acetophenone (HCPK and DEAP, Aldrich) were cured in a UV chamber.72 Some monomeric preparticle systems that were sensitive to light were instead cured thermally using a tempera- ture-sensitive initiator such as 2,2-azobisisobutyronitrile (AIBN, Aldrich). Polymeric systems cast from a solvent were dried under vacuum, and neat polymeric systems were dropped below their glass-transition temperatures. Small-molecule- and protein-based “biological” systems that were cast from a solution were typically lyophilized.27
With the PRINT process, particle solidification, regardless of the chemical process, was typically performed in one of two ways: either close-faced, with a PET sheet laminated to the surface of the filled mold, or open-faced,70 with the filled mold exposed to a N2 purge. Solidified particles trapped in the mold or bound to a harvesting layer of PET could be harvested immediately or were wound up and stored until harvesting at a later date.
With the PRINT method, removing particles from the mold was initially performed by physical agitation of the particles with the sharp end of a glass slide.69 A number of problems were associated with this harvesting technique. First, this method was found to damage the surface of themold, creating large chunks of PFPE mold that would have to be filtered out later. Second, scraping dry particles could cause them to aerosolize. Third, the harvesting process was not scalable. New harvesting techniques have been developed that are more gentle, safe, and scalable. If solidified with an open face as noted above, the particles in the mold could be laminated to an adhesive layer such as polyvinyl pyrrolidone (PVP) or cyanoacrylate on a flexible or rigid backing such as PET or glass slides. Once the adhesive has solidified, the mold is peeled away from the substrate, revealing an array of free- standing particles. The adhesive can then be dissolved and the particles collected in solution. If the particles are solidified with a closed face (laminated to PET), then they will adhere to PET once the mold is peeled back. The free-standing particles can then be harvested by lamination to another adhesive layer or by physical agitation with a soft rubber cell scrapper in the presence of a solvent. As with film fabrication, mold filling, and particle solidification, particle harvesting as a continuous process is possible using the PRINT method.
Regardless of the fabrication method, after being harvested, particles obtained from top-down techniques generally require some form of purification to remove debris or residual com- pounds from the fabrication process and to prepare particles for their intended use. As mentioned above, when using AAO as a template, particle fragmentation often occurred.14 Even particles that required only minimal manipulation by physical stretching on a sheet of PVArequiredmultiple rinsing steps to remove excess PVA from solution.52 Fortunately, a wide variety of purification methods are currently available, and it is usually possible to find at least one method that fits well with a particle-fabrication technique. Although all purification methods have their own distinct advantages and disadvantages, some are intrinsically slow
(on the basis of batch-to-batch processing) and others are much faster (on the basis of automated continuous processes).
Dialysis (>100 000MWCO, Float-a-Lyzer, SpectrumLabora- tories, Inc.) was originally used for PRINT particle fabrication to remove impurities such as the sol fraction and to exchange buffer solutions. This method was found to be a relatively slow process, and particles were often physisorbed to the membrane surface. In a typical purification, spent solvent had to be exchanged for clean solvent at least three times over the course of 1 to 2 days.
Centrifugation is perhaps the simplest method of particle purification, and it is widely used as a quick method to rinse particles and separate variously sized populations of particles gravimetrically.68 In centrifugation, particles are spun out of solution under high centrifugal force. As the particle size de- creases, the centrifugal speed and the time required to pellet out particles effectively increases. The particle yield can be reduced when centrifugation is halted prematurely. As such, the purifica- tion of small, stable particles of less than a few hundred nano- meters in diameter can be quite time-consuming, requiring hours or days to pellet a sample fully. Other problems including irreversible aggregation may be encountered when particles are centrifuged for long periods of time (>1 h) at high speed (14 000 rpm/20000 G). Additionally, resuspending pelleted particles by vigorous mixing and/or sonication can damage particles com- posed of less robust matrix materials or with high aspect ratios and fragile cargo, such as proteins and other biologically active cargo, and should be performed with care.
Centrifugal filtration using 0.1-μm-cutoff poly(vinylidene fluoride) (PVDF) membranes (Microcon-MC,Millipore) proved to be an efficient alternative to standard centrifugation. Lower speeds are required for normal filtration; therefore, aggregation is typically reversible. Unfortunately, once spun down, particles are partially adhered to PVDF membrane. The strength of particle adhesion to the membrane depends strongly on both the chem- istry of the particles and that of the membrane. The removal of particles from the membrane was found to be optimal when the membrane was physically removed from its housing and soni- cated in solution. Although a variety of polymeric membranes and pore sizes are available, making this technique adaptable to a variety of particle-fabrication methods, particle loss is often dramatic (as much as a 50% loss of mass) and the batch process with repeated rinsing can be slow and therefore is not ideal for high-throughput particle fabrication.
Filtration using indirect or tangential flow filtration (TFF) systems appeared to be the most promising approach to par- ticle purification. In TFF filtration systems such as KrosFlo (SpectrumLaboratories, Inc.), particle solutions traveled tangen- tially to the membrane. Both diafiltration and concentration are possible without changing particles from one setup to another. Particle loss by adsorption to the membrane and to surfaces was reduced because particles were not forced to interact directly with the membrane or multiple vials and/or pipet tips. Stirred-cell filtration (Millipore), another indirect filtrationmethod, was also examined. Although this method of purification appears to be promising, inadequate stirring often leads to filters clogged with particles, creating prolonged filtration times. Additionally, if the cell is allowed to run dry, then a dramatic loss of particle yield is observed because of irreversible adhesion of the particles to the filter membrane.
Other methods of particle purification are also under develop- ment. Magnetic purification, for example, is currently being used for particles containing magnetic cargo. Although it was strictly limited to magnetic particles, this method, using magnetic col- umns (Minisep, Miltenyi Biotec), appeared to be promising for
DOI: 10.1021/la903890h KLangmuir XXXX, XXX(XX), XXX–XXX
Merkel et al. Invited Feature Article
purification and other functionalized particle applications. In addition to magnetic filtration, flow-field fractionation (FFF) is of interest for the future purification and separation of particles because particle aggregation is not as prevalent as in other centrifugal techniques.
One unique feature of template-based particle fabrication is that solidified particles, either in themold or on a harvesting layer of PET, are easily subjected to a number of regiospecific chemical modifications.67 The chemical modification of particles while in the mold made it possible to manipulate one face of the particle. Once the particles were removed from the mold, particle modi- fication on a harvesting layer allowed the manipulation of the remaining sides of each particle. This is an exciting development for a number of fields of research, including nano- and micro- robotics78,79 and targeted drug delivery.67 Ongoing research in our laboratory is focused on the directed cellular uptake of regiomodified particles as well as the beginnings of shape-specific building blocks for directed assembly in smart fluids and micro- robotics applications.
There is a growing and widening demand for monodisperse, shape-specific colloidal particles in both the academic and in- dustrial sectors. As yet, no standard particle-fabrication method has arisen to fulfill this unmet need. We have identified and examined several candidate top-down particle-fabrication meth- ods, each ofwhich is capable of producing shape-specific particles composed of a range of materials with the highest potential for
scalability. In particular, we have scrutinized some of the materi- als and methods used in PRINT particle fabrication, modifica- tion, and purification, many of which may be of general use for particles derived by other methods.
Each of the particle-fabrication methods discussed herein possesses inherent strengths and weaknesses in their versatility and capabilities. Hard-template methods seem to be ideally suited to the formation of high-aspect-ratio particles though other shapes have not been demonstrated. Particle-stretching methods have the advantage ofmodifyingprefabricated spherical particles, which can be more readily mass produced, to achieve nonsphe- rical shapes. Microfluidic methods have, perhaps, been the most broadly developed of those mentioned here and have been demonstrated to have excellent scalability and versatility. Photo- lithographicmethods offer excellent resolution of particle size and shape and have scalability comparable to that seen in the semiconductor industry, though the cost of equipment required for such facilities remains a hurdle to mass particle production. The PRINT method offers nanometer-scale resolution with control over particle size and shape and scalability in the form of a reel-to-reel production process. Of these methods, only the PRINT and microfluidic methods have demonstrated their scal- ability, though hard-template, photolithographic, and particle- stretching methods are all scalable in theory.
Acknowledgment. We acknowledge the STC program of the NSF (CHE-9876674), the Carolina Center for Cancer Nanotech- nology Excellence (NIH-U54-CA119343), ONR (N00014-07-1- 0260), DARPA (07-4627), and Liquidia Technologies, Inc. for funding. Liquidia Technologies was recently launched by J.M.D. and colleagues to commercialize breakthroughs from his labora- tory, including micro- and nanofluidics, soft lithography, and PRINT technologies.
(78) Glotzer, S. C.; Solomon, M. Nat. Mater. 2007, 6, 557. (79) Sundararajan, S.; Lammert, P. E.; et al. Nano Lett. 2008, 8, 1271.