Patterning: Strategies for Patterning Biomolecules with Dip‐Pen Nanolithography (Small 8/2011)

Dip‐pen nanolithography (DPN) is an atomic force microscopy (AFM)‐based lithography technique, which has the ability to fabricate patterns with a feature size down to approximately 15 nm using both top‐down and bottom‐up approaches. DPN utilizes the water meniscus formed between an AFM tip and a substrate to transfer ink molecules onto surfaces. A major application of this technique is the fabrication of micro‐ and nano‐arrays of patterned biomolecules. To achieve this goal, a variety of chemical approaches has been used. This review concisely describes the development of DPN in the past decade and presents the related chemical strategies that have been reported to fabricate biomolecular paterns with DPN at micrometer and nanometer scale, classified into direct‐ and indirect DPN methodologies, discussing tip‐functionalization strategies as well.     

Dip‐Pen‐Nanolithographic Patterning of Metallic,Semiconductor, and Metal Oxide Nanostructures on Surfaces

Dip‐pen nanolithography (DPN) is a powerful method to pattern nanostructures on surfaces by the controlled delivery of an “ink” coating the tip of an atomic force microscope upon scanning and contacting with surfaces. The growing interest in the use of nanoparticles as structural and functional elements for the fabrication of nanodevices suggests that the DPN‐stimulated patterning of nanoparticles on surfaces might be a useful technique to assemble hierarchical architectures of nanoparticles that could pave methodologies for functional nanocircuits or nanodevices. This Review presents different methodologies for the nanolithographic patterning of metallic, semiconductor, and metal oxide nanostructures on surfaces. The mechanisms involved in the formation of the nanostructures are discussed and the effects that control the dimensions of the resulting patterns are reviewed. The possible applications of the nanostructures are also addressed.

     

Room‐Temperature Triple‐Ligand Surface Engineering Synergistically Boosts Ink Stability,Recombination Dynamics,and Charge Injection toward EQE‐11.6% Perovskite QLEDs

Developing low‐cost and high‐quality quantum dots (QDs) or nanocrystals (NCs) and their corresponding efficient light‐emitting diodes (LEDs) is crucial for the next‐generation ultra‐high‐definition flexible displays. Here, there is a report on a room‐temperature triple‐ligand surface engineering strategy to play the synergistic role of short ligands of tetraoctylammonium bromide (TOAB), didodecyldimethylammonium bromide (DDAB), and octanoic acid (OTAc) toward “ideal” perovskite QDs with a high photoluminescence quantum yield (PLQY) of >90%, unity radiative decay in its intrinsic channel, stable ink characteristics, and effective charge injection and transportation in QD films, resulting in the highly efficient QD‐based LEDs (QLEDs). Furthermore, the QD films with less nonradiative recombination centers exhibit improved PL properties with a PLQY of 61% through dopant engineering in A‐site. The robustness of such properties is demonstrated by the fabrication of green electroluminescent LEDs based on CsPbBr₃ QDs with the peak external quantum efficiency (EQE) of 11.6%, and the corresponding peak internal quantum efficiency (IQE) and power efficiency are 52.2% and 44.65 lm W^?1, respectively, which are the most‐efficient perovskite QLEDs with colloidal CsPbBr₃ QDs as emitters up to now. These results demonstrate that the as‐obtained QD inks have a wide range application in future high‐definition QD displays and high‐quality lightings.     

EGaIn‐Assisted Room‐Temperature Sintering of Silver Nanoparticles for Stretchable,Inkjet‐Printed,Thin‐Film Electronics

Coating inkjet‐printed traces of silver nanoparticle (AgNP) ink with a thin layer of eutectic gallium indium (EGaIn) increases the electrical conductivity by six‐orders of magnitude and significantly improves tolerance to tensile strain. This enhancement is achieved through a room‐temperature “sintering” process in which the liquid‐phase EGaIn alloy binds the AgNP particles (≈100 nm diameter) to form a continuous conductive trace. Ultrathin and hydrographically transferrable electronics are produced by printing traces with a composition of AgNP‐Ga‐In on a 5 µm‐thick temporary tattoo paper. The printed circuit is flexible enough to remain functional when deformed and can support strains above 80% with modest electromechanical coupling (gauge factor ≈1). These mechanically robust thin‐film circuits are well suited for transfer to highly curved and nondevelopable 3D surfaces as well as skin and other soft deformable substrates. In contrast to other stretchable tattoo‐like electronics, the low‐cost processing steps introduced here eliminate the need for cleanroom fabrication and instead requires only a commercial desktop printer. Most significantly, it enables functionalities like “electronic tattoos” and 3D hydrographic transfer that have not been previously reported with EGaIn or EGaIn‐based biphasic electronics.     

Nanobodies and Nanocrystals: Highly Sensitive Quantum Dot‐Based Homogeneous FRET Immunoassay for Serum‐Based EGFR Detection

Semiconductor quantum dot nanocrystals (QDs) for optical biosensing applications often contain thick polyethylene glycol (PEG)‐based coatings in order to retain the advantageous QD properties in biological media such as blood, serum or plasma. On the other hand, the application of QDs in Förster resonance energy transfer (FRET) immunoassays, one of the most sensitive and most common fluorescence‐based techniques for non‐competitive homogeneous biomarker diagnostics, is limited by such thick coatings due to the increased donor‐acceptor distance. In particular, the combination with large IgG antibodies usually leads to distances well beyond the common FRET range of approximately 1 to 10 nm. Herein, time‐gated detection of Tb‐to‐QD FRET for background suppression and an increased FRET range is combined with single domain antibodies (or nanobodies) for a reduced distance in order to realize highly sensitive QD‐based FRET immunoassays. The “(nano)²” immunoassay (combination of nanocrystals and nanobodies) is performed on a commercial clinical fluorescence plate reader and provides sub‐nanomolar (few ng/mL) detection limits of soluble epidermal growth factor receptor (EGFR) in 50 μL buffer or serum samples. Apart from the first demonstration of using nanobodies for FRET‐based immunoassays, the extremely low and clinically relevant detection limits of EGFR demonstrate the direct applicability of the (nano)^2‐ assay to fast and sensitive biomarker detection in clinical diagnostics.     

From Bioconjugation to Self‐Assembly in Nanobiotechnology: Quantum Dots Trapped and Stabilized by Toroid Protein Yoctowells

Conjugation of quantum dots (QDs) with proteins is applicable for biosensing and imaging in biomedical applications but also for the confined assembly of various nanostructures. Here, the conjugation of CdSe QDs ranging from 2.5 up to 9.5 nm with Hcp1, a yoctowell forming hexameric core protein of Pseudomonas aeruginosa, is described. Defined structures ranging from single QDs trapped within the donut‐shaped hexameric protein cavity over “globular” multi‐toroid architectures up to photonic wires of 850 nm in length and 9 nm in width are self‐assembled while preserving the native hexameric structure of the protein conserving the photoluminescence intensity of the QDs in aqueous solution for more than 3 month.     

Single‐Mode Lasing from Colloidal Water‐Soluble CdSe/CdS Quantum Dot‐in‐Rods

Core–shell CdSe/CdS nanocrystals are a very promising material for light emitting applications. Their solution‐phase synthesis is based on surface‐stabilizing ligands that make them soluble in organic solvents, like toluene or chloroform. However, solubility of these materials in water provides many advantages, such as additional process routes and easier handling. So far, solubilization of CdSe/CdS nanocrystals in water that avoids detrimental effects on the luminescent properties poses a major challenge. This work demonstrates how core–shell CdSe/CdS quantum dot‐in‐rods can be transferred into water using a ligand exchange method employing mercaptopropionic acid (MPA). Key to maintaining the light‐emitting properties is an enlarged CdS rod diameter, which prevents potential surface defects formed during the ligand exchange from affecting the photophysics of the dot‐in‐rods. Films made from water‐soluble dot‐in‐rods show amplified spontaneous emission (ASE) with a similar threshold (130 μJ/cm²) as the pristine material (115 μJ/cm²). To demonstrate feasibility for lasing applications, self‐assembled microlasers are fabricated via the “coffee‐ring effect” that display single‐mode operation and a very low threshold of ～10 μJ/cm². The performance of these microlasers is enhanced by the small size of MPA ligands, enabling a high packing density of the dot‐in‐rods.     

Dip-pen lithography using pens of different thicknesses

A laboratory method to produce AFM tips of different sizes has been developed based on laser irradiation of the commercial silicon nitride tips. A few shots of 60 mJ at 355 nm were found adequate to induce the desired bluntness from 40 nm to 500 nm in a controlled way. Dip-pen nanolithography (DPN) has been performed with the blunt tips using a colloidal ink consisting of Pd nanocrystals coated with polyvinyl pyrrolidone. The line patterns drawn bear a direct relation with the tip morphology, wider the tip, broader are the patterns, in general. The rate of deposition also increases with the tip dimension, but is not as much proportional for larger tips. The study highlights the potential ability of DPN in integrating nano and microelectronics.     

Silver‐Quantum‐Dot‐Modified MoO3 and MnO2 Paper‐Like Freestanding Films for Flexible Solid‐State Asymmetric Supercapacitors

Free‐standing paper‐like thin‐film electrodes have great potential to boost next‐generation power sources with highly flexible, ultrathin, and lightweight requirements. In this work, silver‐quantum‐dot‐ (2–5 nm) modified transition metal oxide (including MoO₃ and MnO₂) paper‐like electrodes are developed for energy storage applications. Benefitting from the ohmic contact at the interfaces between silver quantum dots and MoO₃ nanobelts (or MnO₂ nanowires) and the binder‐free nature and 0D/1D/2D nanostructured 3D network of the fabricated electrodes, substantial improvements on the electrical conductivity, efficient ionic diffusion, and areal capacitances of the hybrid nanostructure electrodes are observed. With this proposed strategy, the constructed asymmetric supercapacitors, with Ag quantum dots/MoO₃ “paper” as anode, Ag quantum dots/MnO₂ “paper” as cathode, and neutral Na₂SO₄/polyvinyl alcohol hydrogel as electrolyte, exhibit significantly enhanced energy and power densities in comparison with those of the supercapacitors without modification of Ag quantum dots on electrodes; present excellent cycling stability at different current densities and good flexibility under various bending states; offer possibilities as high‐performance power sources with low cost, high safety, and environmental friendly properties.     

Cover Picture: Surfactant‐Mediated Generation of Iso‐Oriented Dense and Mesoporous Crystalline Metal‐Oxide Layers (Adv. Mater. 14/2006)

For certain metal oxides (e.g., MoO₃) ordered mesoporous films can be obtained with the nanocrystals in the pore walls being uniformly oriented with respect to the substrate by applying evaporation‐induced self‐assembly followed by heating. This surprisingly facile process, described by Smarsly and co‐workers on p. 1827, works on different polar substrates (glass, metals, etc.) for oxides with anisotropic unit cells, based on the interaction with surfactants during nucleation (“soft‐epitaxy”).     

Low‐Density Lipoprotein‐Mimicking Nanoparticles for Tumor‐Targeted Theranostic Applications

This study introduces multifunctional lipid nanoparticles (LNPs), mimicking the structure and compositions of low‐density lipoproteins, for the tumor‐targeted co‐delivery of anti‐cancer drugs and superparamagnetic nanocrystals. Paclitaxel (4.7 wt%) and iron oxide nanocrystals (6.8 wt%, 11 nm in diameter) are co‐encapsulated within folate‐functionalized LNPs, which contain a cluster of nanocrystals with an overall diameter of about 170 nm and a zeta potential of about ‐40 mV. The folate‐functionalized LNPs enable the targeted detection of MCF‐7, human breast adenocarcinoma expressing folate receptors, in T₂‐weighted magnetic resonance images as well as the efficient intracellular delivery of paclitaxel. Paclitaxel‐free LNPs show no significant cytotoxicity up to 0.2 mg mL^?1, indicating the excellent biocompatibility of the LNPs for intracellular drug delivery applications. The targeted anti‐tumor activities of the LNPs in a mouse tumor model suggest that the low‐density lipoprotein‐mimetic LNPs can be an effective theranostic platform with excellent biocompatibility for the tumor‐targeted co‐delivery of various anti‐cancer agents.     

UV Raman spectroscopy of group IV nanocrystals embedded in a SiO<Subscript>2</Subscript> matrix

Nanostructures of both Ge nanocrystals formed by thermal oxidation of SiGe layers, and SiGe nanocrystals formed by crystallization of amorphous SiGe nanoparticles deposited by LPCVD have been analyzed by Raman spectroscopy. The nanostructures are formed on a silicon substrate. Raman spectra have been acquired with visible (514.5 nm) and UV (325 nm) excitation lines. When the amount of material is very small, as it has happens in these nanostructures, the visible line is not able to excite the characteristic peaks of the Ge or SiGe in the Raman spectrum; instead the Si second order spectrum of the substrate appears and it can be misinterpreted by attributing it to the Ge–Ge band associated with the nanocrystals. In this work, the use of UV excitation has been demonstrated to enhance the sensitivity respect to the conventional visible excitation, allowing the characteristic peaks of the Ge or SiGe nanocrystals to appear in the spectrum. We attributed this effect to the resonance effects.     

Active Self‐Assembly of Train‐Shaped DNA Nanostructures via Catalytic Hairpin Assembly Reactions

Biomolecular self‐assembly is a powerful approach for fabricating supramolecular architectures. Over the past decade, a myriad of biomolecular assemblies, such as self‐assembly proteins, lipids, and DNA nanostructures, have been used in a wide range of applications, from nano‐optics to nanoelectronics and drug delivery. The method of controlling when and where the self‐assembly starts is essential for assembly dynamics and functionalization. Here, train‐shaped DNA nanostructures are actively self‐assembled using DNA tiles as artificial “carriages,” hairpin structures as “couplers,” and initiators of catalytic hairpin assembly (CHA) reactions as “wrenches.” The initiator wrench can selectively open the hairpin couplers to couple the DNA tile carriages with high product yield. As such, DNA nanotrains are actively prepared with two, three, four, or more carriages. Furthermore, by flexibly modifying the carriages with “biotin seats” (biotin‐modified DNA tiles), streptavidin “passengers” are precisely arranged in corresponding seats. The applications of the CHA‐triggered self‐assembly mechanism are also extended for assembling the large DNA origami dimer. With the creation of 1D architectures established, it is thought that this CHA‐triggered self‐assembly mechanism may provide a new element of control for complex autonomous assemblies from a variety of starting materials with specific sites and times.     

Strategies for patterning biomolecules with dip-pen nanolithography

Dip-pen nanolithography (DPN) is an atomic force microscopy (AFM)-based lithography technique, which has the ability to fabricate patterns with a feature size down to approximately 15 nm using both top-down and bottom-up approaches. DPN utilizes the water meniscus formed between an AFM tip and a substrate to transfer ink molecules onto surfaces. A major application of this technique is the fabrication of micro- and nano-arrays of patterned biomolecules. To achieve this goal, a variety of chemical approaches has been used. This review concisely describes the development of DPN in the past decade and presents the related chemical strategies that have been reported to fabricate biomolecular patterns with DPN at micrometer and nanometer scale, classified into direct- and indirect DPN methodologies, discussing tip-functionalization strategies as well.     

Light‐Induced Buckles Localized by Polymeric Inks Printed on Bilayer Films

Buckling instabilities generate microscale features in thin films in a facile manner. Buckles can form, for example, by heating a metal/polymer film stack on a rigid substrate. Thermal expansion differences of the individual layers generate compressive stress that causes the metal to buckle over the entire surface. The ability to dictate and confine the location of buckle formation can enable patterns with more than one length scale, including hierarchical patterns. Here, sacrificial “ink” patterned on top of the film stack localizes the buckles via two mechanisms. First, stiff inks suppress buckles such that only the non‐inked regions buckle in response to infrared light. The metal in the non‐inked regions absorbs the infrared light and thus gets sufficiently hot to induce buckles. Second, soft inks that absorb light get hot faster than the non‐inked regions and promote buckling when exposed to visible light. The exposed metal in the non‐inked regions reflects the light and thus never get sufficiently hot to induce buckles. This second method works on glass substrates, but not silicon substrates, due to the superior thermal insulation of glass. The patterned ink can be removed, leaving behind hierarchical patterns consisting of regions of buckles among non‐buckled regions.     

Progression in the Fountain Pen Approach: From 2D Writing to 3D Free‐Form Micro/Nanofabrication

The fountain pen approach, as a means for transferring materials to substrates, has shown numerous incarnations in recent years for creating 2D micro/nanopatterns and even generating 3D free‐form nanostructures using a variety of material “inks”. While the idea of filled reservoirs used to deliver material to a substrate via a capillary remains unchanged since antiquity, the advent of precise micromanipulation systems and functional material “inks” allows the extension of this mechanism to more high‐tech applications. Herein, the recent growth in meniscus guided fountain pen approaches for benchtop micro/nanofabrication, which has occurred in the last decade, is discussed. Particular attention is given to the theory, equipment, and experimentation encompassing this unique direct writing approach. A detailed exploration of the diverse ink systems and functional device applications borne from this strategy is put forth to reveal its rapid expansion to a broad range of scientific and engineering disciplines. As such, this informative review is provided for researchers considering adoption of this recent advancement of a familiar technology.     

Binary Superlattices from {Mo132} Polyoxometalates and Maghemite Nanocrystals: Long‐Range Ordering and Fine‐Tuning of Dipole Interactions

In the present article, the successful coassembly of spherical 6.2 nm maghemite (γ‐Fe₂O₃) nanocrystals and giant polyoxometalates (POMs) such as 2.9 nm {Mo₁₃₂} is demonstrated. To do so, colloidal solutions of oleic acid‐capped γ‐Fe₂O₃ and long‐chain alkylammonium‐encapsulated {Mo₁₃₂} dispersed in chloroform are mixed together and supported self‐organized binary superlattices are obtained upon the solvent evaporation on immersed substrates. Both electronic microscopy and small angles X‐ray scattering data reveal an AB‐type structure and an enhanced structuration of the magnetic nanocrystals (MNCs) assembly with POMs in octahedral interstices. Therefore, {Mo₁₃₂} acts as an efficient binder constituent for improving the nanocrystals ordering in 3D films. Interestingly, in the case of didodecyldimethylammonium (C₁₂)‐encapsulated POMs, the long‐range ordered binary assemblies are obtained while preserving the nanocrystals magnetic properties due to weak POMs–MNCs interactions. On the other hand, POMs of larger effective diameter can be employed as spacer blocks for MNCs as shown by using {Mo₁₃₂} capped with dioctadecyldimethylammonium (C₁₈) displaying longer chains. In that case, it is shown that POMs can also be used for fine‐tuning the dipolar interactions in γ‐Fe₂O₃ nanocrystal assemblies.     

Hierarchical nanostructures of PbTiO3 through mesocrystal formation

Novel hierarchical self-assembled structures; bur-like PbTiO3 nanostructures were made by self-assembly of PbTiO3 nanocrystals under hydrothermal conditions using sodium dodecylbenzene sulfonate surfactant. The bur-like nanostructures exhibit a unique geometrical shape with cores of agglomerated nanocrystals and outershells of nanorods. The nanorods were between 30 nm and 100 nm in diameter and from several hundred nm up to 2 microm in length. We demonstrate that these nanostructures are formed in a two step process where agglomeration of PbTiO3 nanoparticles into microspheres occurs in a first step, followed by assembly of cube-shaped nanoparticle building blocks into PbTiO3 mesocrystals in a second step. The mesocrystals continuously grow into nanorods from the surface of the microspheres acting as a substrate.     

Programmed Nanoparticle‐Loaded Nanoparticles for Deep‐Penetrating 3D Cancer Therapy

Tumors are 3D, composed of cellular agglomerations and blood vessels. Therapies involving nanoparticles utilize specific accumulations due to the leaky vascular structures. However, systemically injected nanoparticles are mostly uptaken by cells located on the surfaces of cancer tissues, lacking deep penetration into the core cancer regions. Herein, an unprecedented strategy, described as injecting “nanoparticle‐loaded nanoparticles” to address the long‐lasting problem is reported for effective surface‐to‐core drug delivery in entire 3D tumors. The “nanoparticle‐loaded nanoparticle” is a silica nanoparticle (≈150 nm) with well‐developed, interconnected channels (diameter of ≈30 nm), in which small gold nanoparticles (AuNPs) (≈15 nm) with programmable DNA are located. The nanoparticle (AuNPs)‐loaded nanoparticles (silica): (1) can accumulate in tumors through leaky vascular structures by protecting the inner therapeutic AuNPs during blood circulation, and then (2) allow diffusion of the AuNPs for penetration into the entire surface‐to‐core tumor tissues, and finally (3) release a drug triggered by cancer‐characteristic pH gradients. The hierarchical “nanoparticle‐loaded nanoparticle” can be a rational design for cancer therapies because the outer large nanoparticles are effective in blood circulation and in protection of the therapeutic nanoparticles inside, allowing the loaded small nanoparticles to penetrate deeply into 3D tumors with anticancer drugs.     

Ink‐on‐Probe Hydrodynamics in Atomic Force Microscope Deposition of Liquid Inks

The controlled deposition of attolitre volumes of liquids may engender novel applications such as soft, nano‐tailored cell‐material interfaces, multi‐plexed nano‐arrays for high throughput screening of biomolecular interactions, and localized delivery of reagents to reactions confined at the nano‐scale. Although the deposition of small organic molecules from an AFM tip, known as dip‐pen nanolithography (DPN), is being continually refined, AFM deposition of liquid inks is not well understood, and is often fraught with inconsistent deposition rates. In this work, the variation in feature‐size over long term printing experiments for four model inks of varying viscosity is examined. A hierarchy of recurring phenomena is uncovered and there are attributed to ink movement and reorganisation along the cantilever itself. Simple analytical approaches to model these effects, as well as a method to gauge the degree of ink loading using the cantilever resonance frequency, are described. In light of the conclusions, the various parameters which need to be controlled in order to achieve uniform printing are dicussed. This work has implications for the nanopatterning of viscous liquids and hydrogels, encompassing ink development, the design of probes and printing protocols.