Fountain Pen-Inspired 3D Colloidal Assembly, Consisting of Metallic Nanoparticles on a Femtoliter Scale.

The 3D colloidal assemblies composed of nanoparticles (NPs) are closely associated with optical properties such as photonic crystals, localized surface plasmon resonance, and surface-enhanced Raman scattering. However, research on their fabrication remains insufficient. Here, the femtoliter volume of a 3D colloidal assembly is shown, using the evaporation of a fine fountain pen. A nano-fountain pen (NPF) with a micrometer-level tip inner diameter was adopted for the fine evaporation control of the ink solvent. The picoliters of the evaporation occurring at the NFP tip and femtoliter volume of the 3D colloidal assembly were analyzed using a diffusion equation. The shape of the 3D colloidal assembly was dependent on the evaporation regarding the accumulation time and tip size, and they exhibited random close packing. Using gold-, silver-, and platinum-NPs and mixing ratios of them, diverse 3D colloidal assemblies were formed. The spectra regarding a localized surface plasmon resonance of them were changed according to composition and mixing ratio. We expect that this could be widely applied as a simple fabrication tool in order to explore complex metamaterials constructed of nanoparticles, as this method is highly flexible in varying the shape as well as composition ratio of self-assembled structures.

Freestanding, Freeform Metamolecule Fibers Tailoring Artificial Optical Magnetism.

Metamolecule clusters support various unique types of artificial electromagnetism at optical frequencies. However, the technological challenges regarding the freeform fabrication of freestanding metamolecule clusters with programmed geometries and multiple compositions remain unresolved. Here, the freeform, freestanding raspberry-like metamolecule (RMM) fibers based on the directional guidance of a femtoliter meniscus are presented, resulting in the evaporative co-assembly of silica nanoparticles and gold nanoparticles with the aid of 3D nanoprinting. This method offers a facile and universal pathway to shape RMM fibers in 3D, enabling versatile manipulation of near- and far-field characteristics. In particular, the authors demonstrate the ability to decrease the scattering of the millimeter-scale RMM fiber in visible spectrum. In addition, the influence of electric and magnetic dipole modes on the directional scattering of RMM fibers is investigated. These experiments show that the magnetic response of an individual RMM can be controlled by adjusting the filling factor of gold nanoparticles. The authors anticipate that this method will allow for unrestricted design and realization of nanophotonic structures, surpassing the limitations of conventional fabrication processes.

Three-dimensional plasmonic nanoclusters driven by co-assembly of thermo-plasmonic nanoparticles and colloidal quantum dots.

Metallic nanoparticles that support localized surface plasmons have emerged as fundamental iconic building blocks for nanoscale photonics. Self-assembled clustering of plasmonic nanoparticles with controlled near-field interactions offers an interesting novel route to manipulate the electromagnetic fields at a subwavelength scale. Various bottom-up, self-assembly manners have been successfully devised to build plasmonic nanoparticle clusters displaying attractive optical properties. However, the incapability to configure on-demand architectures limits its practical reliability uses for scalable nanophotonic devices. Furthermore, a critical challenge has been addressing the accurate positioning of functional nanoparticles, including catalytic nanoparticles, dielectric nanoparticles, and quantum dots (QDs) in the clustered plasmonic hotspots. This work proposes a micropipette-based self-assembly method to fabricate three-dimensional architectures composed of colloidal clusters. The heterogeneous colloidal clusters comprising metallic nanoparticles and QDs are fabricated in one step by the micropipette-based self-assembly method. A plasmonic clustered pillar embedding QDs exhibited excellent photoluminescence characteristics compared to a collapsed pillar. The experimental and theoretical demonstration of the localized surface plasmon resonance and thermo-plasmonic properties of the colloidal clusters was performed.

Three-Dimensional Plasmonic Nanocluster-Driven Light-Matter Interaction for Photoluminescence Enhancement and Picomolar-Level Biosensing.

Plasmonic nanoparticle clusters promise to support unique engineered electromagnetic responses at optical frequencies, realizing a new concept of devices for nanophotonic applications. However, the technological challenges associated with the fabrication of three-dimensional nanoparticle clusters with programmed compositions remain unresolved. Here, we present a novel strategy for realizing heterogeneous structures that enable efficient near-field coupling between the plasmonic modes of gold nanoparticles and various other nanomaterials via a simple three-dimensional coassembly process. Quantum dots embedded in the plasmonic structures display ∼56 meV of a blue shift in the emission spectrum. The decay enhancement factor increases as the total contribution of radiative and nonradiative plasmonic modes increases. Furthermore, we demonstrate an ultracompact diagnostic platform to detect M13 viruses and their mutations from femtoliter volume, sub-100 pM analytes. This platform could pave the way toward an effective diagnosis of diverse pathogens, which is in high demand for handling pandemic situations.

Neural mechanism mimetic selective electronic nose based on programmed M13 bacteriophage.

The electronic nose is a reliable practical sensor device that mimics olfactory organs. Although numerous studies have demonstrated excellence in detecting various target substances with the help of ideal models, biomimetic approaches still suffer in practical realization because of the inability to mimic the signal processing performed by olfactory neural systems. Herein, we propose an electronic nose based on the programable surface chemistry of M13 bacteriophage, inspired by the neural mechanism of the mammalian olfactory system. The neural pattern separation (NPS) was devised to apply the pattern separation that operates in the memory and learning process of the brain to the electronic nose. We demonstrate an electronic nose in a portable device form, distinguishing polycyclic aromatic compounds (harmful in living environment) in an atomic-level resolution (97.5% selectivity rate) for the first time. Our results provide practical methodology and inspiration for the second-generation electronic nose development toward the performance of detection dogs (K9).

Polarization Angle Dependence of Optical Gain in a Hybrid Structure of Alexa-Flour 488/M13 Bacteriophage.

We measured optical modal gain of a dye-virus hybrid structure using a variable stripe length method, where Alexa-fluor-488 dye was coated on a virus assembly of M13 bacteriophage. Inspired by the structural periodicity of the wrinkle-like virus assembly, the edge emission of amplified spontaneous emission was measured for increasing excited optical stripe length, which was aligned to be either parallel or perpendicular to the wrinkle alignment. We found that the edge emission showed a strong optical anisotropy, and a spectral etalon also appeared in the gain spectrum. These results can be attributed to the corrugated structure, which causes a similar effect to a DFB laser, and we also estimated effective cavity lengths.

A DNA-derived phage nose using machine learning and artificial neural processing for diagnosing lung cancer.

There is a growing interest in electronic nose-based diagnostic systems that are fast and portable. However, existing technologies are suitable only for operation in the laboratory, making them difficult to apply in a rapid, non-face-to-face, and field-suitable manner. Here, we demonstrate a DNA-derived phage nose (D2pNose) as a portable respiratory disease diagnosis system requiring no pretreatment. D2pNose was produced based on phage colour films implanted with DNA sequences from mammalian olfactory receptor cells, and as a result, it possesses the comprehensive reactivity of these cells. The manipulated surface chemistry of the genetically engineered phages was verified through a correlation analysis between the calculated and the experimentally measured reactivity. Breaths from 31 healthy subjects and 31 lung cancer patients were collected and exposed to D2pNose without pretreatment. With the help of deep learning and neural pattern separation, D2pNose has achieved a diagnostic success rate of over 75% and a classification success rate of over 86% for lung cancer based on raw human breath. Based on these results, D2pNose can be expected to be directly applicable to other respiratory diseases.

Fabrication of Chiral M13 Bacteriophage Film by Evaporation-Induced Self-Assembly.

Biomacromolecules are likely to undergo self-assembly and show specific collective behavior concentrated in the medium. Although the assembly procedures have been studied for unraveling their mysteries, there are few cases to directly demonstrate the collective behavior and phase transition process in dynamic systems. In the contribution, the drying process of M13 droplet is investigated, and can be successfully simulated by a doctor blade coating method. The morphologies in the deposited film are measured by atomic force microscopy and the liquid crystal phase development is captured in real time using polarized optical microscope. Collective behaviors near the contact line are characterized by the shape of meniscus curve and particle movement velocity. With considering rheological properties and flow, the resultant chiral film is used to align gold nanorods, and this approach can suggest a way to use M13 bacteriophage as a scaffold for the multi-functional chiral structures.

Optical bioelectronic nose of outstanding sensitivity and selectivity toward volatile organic compounds implemented with genetically engineered bacteriophage: Integrated study of multi-scale computational prediction and experimental validation.

Genetic engineering of a bacteriophage is a promising way to develop a highly functional biosensor. Almost countless configurational degree of freedom in the manipulation, considerable uncertainty and cost involved with the approach, however, have been huddles for the objective. In this paper, we demonstrate rapidly responding optical biosensor with high selectivity toward gaseous explosives with genetically engineered phages. The sensors are equipped with peptide sequences in phages optimally interacting with the volatile organic compounds (VOCs) in visible light regime. To overcome the conventional issues, we use extensive utilization of empirical calculations to construct a large database of 8000 tripeptides and screen the best for electronic nose sensing performance toward nine VOCs belonging to three chemical classes. First-principles density functional theory (DFT) calculations unveil underlying correlations between the chemical affinity and optical property change on an electronic band structure level. The computational outcomes are validated by in vitro experimental design and testing of multiarray sensors using genetically modified phage implemented with five selected tripeptide sequences and wild-type phages. The classification success rates estimated from hierarchical cluster analysis are shown to be very consistent with the calculations. Our optical biosensor demonstrates a 1 ppb level of sensing resolution for explosive VOCs, which is a substantial improvement over conventional biosensor. The systematic interplay of big data-based computational prediction and in situ experimental validation can provide smart design principles for unconventionally outstanding biosensors.

Programmable self-assembly of M13 bacteriophage for micro-color pattern with a tunable colorization.

Over the last decade, the M13 bacteriophage has been used widely in various applications, such as sensors, bio-templating, and solar cells. The M13 colorimetric sensor was developed to detect toxic gases to protect the environment, human health, and national security. Recent developments in phage-based colorimetric sensor technologies have focused on improving the sensing characteristics, such as the sensitivity and selectivity on a large scale. On the other hand, few studies have examined precisely controllable micro-patterning techniques in phage-based self-assembly. This paper developed a color patterning technique through self-assembly of the M13 bacteriophages. The phage was self-assembled into a nanostructure through precise temperature control at the meniscus interface. Furthermore, barcode color patterns could be fabricated using self-assembled M13 bacteriophage on micrometer scale areas by manipulating the grooves on the SiO2 surface. The color patterns exhibited color tunability based on the phage nano-bundles reactivity. Overall, the proposed color patterning technique is expected to be useful for preparing new color sensors and security patterns.

Large-Area Virus Coated Ultrathin Colorimetric Sensors with a Highly Lossy Resonant Promoter for Enhanced Chromaticity.

Acclimatable colors in response to environmental stimuli, which are naturally endowed with some living things, can provide an opportunity for humans to recognize hazardous substances without taking empirical risks. Despite efforts to create artificial responsive colors, realistic applications in everyday life require an immediate/distinct colorimetric realization with wide chromatic selectivity. A dynamically responsive virus (M-13 phage)-based changeable coloring strategy is presented with a highly lossy resonant promoter (HLRP). An ultrathin M-13 phage layer for rapid response to external stimuli displays colorimetric behavior, even in its subtle swelling with strong resonances on HLRP, which is modeled using the complex effective refractive index. Optimal designs of HLRP for several material combinations allow selective chromatic responsivity from the corresponding wide color palette without modification of the dynamic responsive layer. As a practical demonstration, the spatially designed colorimetric indicator, which is insensitive/sensitive to external stimuli, provides an intuitive perception of environmental changes with hidden/revealed patterns. Furthermore, the proposed colorimetric sensor is tested by exposure to various volatile organic chemicals and endocrine disrupting chemicals for versatile detectability, and is fabricated in a wafer-scale sample for large-area scalability.

M13 Bacteriophage-Assisted Morphological Engineering of Crack-Based Sensors for Highly Sensitive and Wide Linear Range Strain Sensing.

Despite their extraordinary mechanosensitivities, most channel-like crack-based strain sensors are limited by their poor levels of stretchability and linearity. This work presents a simple yet efficient way of modulating the cracking structure of thin metal films on elastomers to facilitate the development of high-performance wearable strain sensors. A net-shaped crack structure based on a thin platinum (Pt) film can be produced by coating an elastomer surface with M13 bacteriophages (phages) and consequently engineering the surface strain upon stretching. This process produces a Pt-on-phage (PoP) strain sensor that simultaneously exhibits high levels of stretchability (24%), sensitivity (maximum gauge factor ≈ 845.6 for 20-24%), and linearity (R2 ≈ 0.988 up to 20%). In addition, the sensor performance can be further modulated by either changing the phage coating volume or adding a silver nanowire coating to the PoP sensor film. The balanced strain-sensing performance, combined with fast response times and high levels of mechanical flexibility and operational stability, enables the devices to detect a wide range of human motions in real time after being attached to various body parts. Furthermore, PoP-based strain sensors can be usefully extended to detect more complex multidimensional strains through further strain engineering on a cross-patterned PoP film.

Hierarchical Cluster Analysis of Medical Chemicals Detected by a Bacteriophage-Based Colorimetric Sensor Array.

M13 bacteriophage-based colorimetric sensors, especially multi-array sensors, have been successfully demonstrated to be a powerful platform for detecting extremely small amounts of target molecules. Colorimetric sensors can be fabricated easily using self-assembly of genetically engineered M13 bacteriophage which incorporates peptide libraries on its surface. However, the ability to discriminate many types of target molecules is still required. In this work, we introduce a statistical method to efficiently analyze a huge amount of numerical results in order to classify various types of target molecules. To enhance the selectivity of M13 bacteriophage-based colorimetric sensors, a multi-array sensor system can be an appropriate platform. On this basis, a pattern-recognizing multi-array biosensor platform was fabricated by integrating three types of sensors in which genetically engineered M13 bacteriophages (wild-, RGD-, and EEEE-type) were utilized as a primary building block. This sensor system was used to analyze a pattern of color change caused by a reaction between the sensor array and external substances, followed by separating the specific target substances by means of hierarchical cluster analysis. The biosensor platform could detect drug contaminants such as hormone drugs (estrogen) and antibiotics. We expect that the proposed biosensor system could be used for the development of a first-analysis kit, which would be inexpensive and easy to supply and could be applied in monitoring the environment and health care.

Experimental and numerical evaluation of a genetically engineered M13 bacteriophage with high sensitivity and selectivity for 2,4,6-trinitrotoluene.

Selective and sensitive detection of desired targets is very critical in sensor design. Here, we report a genetically engineered M13 bacteriophage-based sensor system evaluated by quantum mechanics (QM) calculations. Phage display is a facile way to develop the desired peptide sequences, but the resulting sequences can be imperfect peptides for binding of target molecules. A TNT binding peptide (WHW) carrying phage was self-assembled to fabricate thin films and tested for the sensitive and selective surface plasmon resonance-based detection of TNT molecules at the 500 femtomole level. SPR studies performed with the WHW peptide and control peptides (WAW, WHA, AHW) were well-matched with those of the QM calculations. Our combined method between phage engineering and QM calculation will significantly enhance our ability to design selective and sensitive sensors.

Improvement of High Affinity and Selectivity on Biosensors Using Genetically Engineered Phage by Binding Isotherm Screening.

The genetically engineered M13 bacteriophage (M13 phage), developed via directed evolutionary screening process, can improve the sensitivity of sensors because of its selective binding to a target material. Herein, we propose a screening method to develop a selective and sensitive bioreporter for toxic material based on genetically engineered M13 phage. The paraquat (PQ)-binding M13 phage, developed by directed evolution, was used. The binding affinities of the PQ-binding M13 phage to PQ and similar molecules were analyzed using isothermal titration calorimetry (ITC). Based on the isotherms measured by ITC, binding affinities were calculated using the one-site binding model. The binding affinity was 5.161 × 10-7 for PQ, and 3.043 × 10-7 for diquat (DQ). The isotherm and raw ITC data show that the PQ-binding M13 phage does not selectively bind to difenzoquat (DIF). The phage biofilter experiment confirmed the ability of PQ-binding M13 bacteriophage to bind PQ. The surface-enhanced Raman scattering (SERS) platform based on the bioreporter, PQ-binding M13 phage, exhibited 3.7 times the signal intensity as compared with the wild-type-M13-phage-coated platform.

Bioinspired M-13 bacteriophage-based photonic nose for differential cell recognition.

A bioinspired M-13 bacteriophage-based photonic nose was developed for differential cell recognition. The M-13 bacteriophage-based photonic nose exhibits characteristic color patterns when phage bundle nanostructures, which were genetically modified to selectively capture vapor phase molecules, are structurally deformed. We characterized the color patterns of the phage bundle nanostructure in response to cell proliferation via several biomarkers differentially produced by cells, including hydrazine, o-xylene, ethylbenzene, ethanol and toluene. A specific color enables the successful identification of different types of molecular and cellular species. Our sensing technique utilized the versatile M-13 bacteriophage as a building block for fabricating bioinspired photonic crystals, which enables ease of fabrication and tunable selectivity through genetic engineering. Our simple and versatile bioinspired photonic nose could have possible applications in sensors for human health and national security, food discrimination, environmental monitoring, and portable and wearable sensors.

Correction: Bioinspired M-13 bacteriophage-based photonic nose for differential cell recognition.

[This corrects the article DOI: 10.1039/C6SC02021F.].

Identification of Endocrine Disrupting Chemicals using a Virus-Based Colorimetric Sensor.

A simple and portable colorimetric sensor based on M13 bacteriophage (phage) was devised to identify a class of endocrine disrupting chemicals, including benzene, phthalate, and chlorobenzene derivatives. Arrays of structurally and genetically modified M13 bacteriophage were fabricated so as to produce a biomimetic colorimetric sensor, and color changes in the phage arrays in response to several benzene derivatives were characterized. The sensor was also used to classify phthalate and chlorobenzene derivatives as representatives of endocrine disrupting chemicals. The characteristic color patterns obtained on exposure to various benzene derivatives enabled similar chemical structures in the vapor phase to be classified. Our sensing approach based on the use of a genetically surface modified M13 bacteriophage offers a promising platform for portable, simple environmental monitors that could be extended for use in numerous application areas, including food monitoring, security monitoring, explosive risk assessment, and point of care testing.

Biomimetic self-templating optical structures fabricated by genetically engineered M13 bacteriophage.

Here, we describe a highly sensitive and selective surface plasmon resonance sensor system by utilizing self-assembly of genetically engineered M13 bacteriophage. About 2700 copies of genetically expressed peptide copies give superior selectivity and sensitivity to M13 phage-based SPR sensor. Furthermore, the sensitivity of the M13 phage-based SPR sensor was enhanced due to the aligning of receptor matrix in specific direction. Incorporation of specific binding peptide (His Pro Gln: HPQ) gives M13 bacteriophage high selectivity for the streptavidin. Our M13 phage-based SPR sensor takes advantage of simplicity of self-assembly compared with relatively complex photolithography techniques or chemical conjugations. Additionally, designed structure which is composed of functionalized M13 bacteriophage can simultaneously improve the sensitivity and selectivity of SPR sensor evidently. By taking advantages of the genetic engineering and self-assembly, we propose the simple method for fabricating novel M13 phage-based SPR sensor system which has a high sensitivity and high selectivity.

Virus based Full Colour Pixels using a Microheater.

Mimicking natural structures has been received considerable attentions, and there have been a few practical advances. Tremendous efforts based on a self-assembly technique have been contributed to the development of the novel photonic structures which are mimicking nature's inventions. We emulate the photonic structures from an origin of colour generation of mammalian skins and avian skin/feathers using M13 phage. The structures can be generated a full range of RGB colours that can be sensitively switched by temperature and substrate materials. Consequently, we developed an M13 phage-based temperature-dependent actively controllable colour pixels platform on a microheater chip. Given the simplicity of the fabrication process, the low voltage requirements and cycling stability, the virus colour pixels enable us to substitute for conventional colour pixels for the development of various implantable, wearable and flexible devices in future.