Ultrafast sensitivity-controlled and specific detection of extracellular vesicles using optical force with antibody-modified microparticles in a microflow system.

Extracellular vesicles (EVs), including nanoscale exosomes and ectosomes, hold promise as biomarkers that provide information about the cell of origin through their cargo of nucleic acids and proteins, both on their surface and within. Here, we develop a detection method of EVs based on light-induced acceleration of specific binding between their surface and antibody-modified microparticles, using a controlled microflow with three-dimensional analysis by confocal microscopy. Our method successfully detected 103-104 nanoscale EVs in liquid samples as small as a 500 nanoliters within 5 minutes, with the ability to distinguish multiple membrane proteins. Remarkably, we achieved the specific detection of EVs secreted from living cancer cell lines with high linearity, without the need for a time-consuming ultracentrifugation process that can take several hours. Furthermore, the detection range can be controlled by adjusting the action range of optical force using a defocused laser, consistent with the theoretical calculations. These findings demonstrate an ultrafast, sensitive, and quantitative approach for measuring biological nanoparticles, enabling innovative analyses of cell-to-cell communication and early diagnosis of various diseases, including cancer.

Light-Induced Condensation of Biofunctional Molecules around Targeted Living Cells to Accelerate Cytosolic Delivery.

The light-induced force and convection can be enhanced by the collective effect of electrons (superradiance and red shift) in high-density metallic nanoparticles, leading to macroscopic assembly of target molecules. We here demonstrate application of the light-induced assembly for drug delivery system with enhancement of cell membrane accumulation and penetration of biofunctional molecules including cell-penetrating peptides (CPPs) with superradiance-mediated photothermal convection. For induction of photothermal assembly around targeted living cells in cell culture medium, infrared continuous-wave laser light was focused onto high-density gold-particle-bound glass bottom dishes exhibiting plasmonic superradiance or thin gold-film-coated glass bottom dishes. In this system, the biofunctional molecules can be concentrated around the targeted living cells and internalized into them only by 100 s laser irradiation. Using this simple approach, we successfully achieved enhanced cytosolic release of the CPPs and apoptosis induction using a pro-apoptotic domain with a very low peptide concentration (nM level) by light-induced condensation.

Attogram-level light-induced antigen-antibody binding confined in microflow.

The analysis of trace amounts of proteins based on immunoassays and other methods is essential for the early diagnosis of various diseases such as cancer, dementia, and microbial infections. Here, we propose a light-induced acceleration of antigen-antibody reaction of attogram-level proteins at the solid-liquid interface by tuning the laser irradiation area comparable to the microscale confinement geometry for enhancing the collisional probability of target molecules and probe particles with optical force and fluidic pressure. This principle was applied to achieve a 102-fold higher sensitivity and ultrafast specific detection in comparison with conventional protein detection methods (a few hours) by omitting any pretreatment procedures; 47-750 ag of target proteins were detected in 300 nL of sample after 3 minutes of laser irradiation. Our findings can promote the development of proteomics and innovative platforms for high-throughput bio-analyses under the control of a variety of biochemical reactions.

Damage-free light-induced assembly of intestinal bacteria with a bubble-mimetic substrate.

Rapid evaluation of functions in densely assembled bacteria is a crucial issue in the efficient study of symbiotic mechanisms. If the interaction between many living microbes can be controlled and accelerated via remote assembly, a cultivation process requiring a few days can be ommitted, thus leading to a reduction in the time needed to analyze the bacterial functions. Here, we show the rapid, damage-free, and extremely dense light-induced assembly of microbes over a submillimeter area with the "bubble-mimetic substrate (BMS)". In particular, we successfully assembled 104-105 cells of lactic acid bacteria (Lactobacillus casei), achieving a survival rate higher than 95% within a few minutes without cultivation process. This type of light-induced assembly on substrates like BMS, with the maintenance of the inherent functions of various biological samples, can pave the way for the development of innovative methods for rapid and highly efficient analysis of functions in a variety of microbes.

Light-induced assembly of living bacteria with honeycomb substrate.

Some bacteria are recognized to produce useful substances and electric currents, offering a promising solution to environmental and energy problems. However, applications of high-performance microbial devices require a method to accumulate living bacteria into a higher-density condition in larger substrates. Here, we propose a method for the high-density assembly of bacteria (106 to 107 cells/cm2) with a high survival rate of 80 to 90% using laser-induced convection onto a self-organized honeycomb-like photothermal film. Furthermore, the electricity-producing bacteria can be optically assembled, and the electrical current can be increased by one to two orders of magnitude simply by increasing the number of laser irradiations. This concept can facilitate the development of high-density microbial energy conversion devices and provide new platforms for unconventional environmental technology.

Interparticle-Interaction-Mediated Anomalous Acceleration of Nanoparticles under Light-Field with Coupled Orbital and Spin Angular Momentum.

Spin-orbit interaction is a crucial issue in the field of nanoscale physics and chemistry. Here, we theoretically demonstrate that the spin angular momentum (SAM) can accelerate and decelerate the orbital motion of nanoparticles (NPs) via light-induced interparticle interactions by a circularly polarized optical vortex. The Laguerre-Gaussian beam as a conventional optical vortex with orbital angular momentum (OAM) induces the orbital and spinning motion of a trapped object depending on the spatial configuration. On the contrary, it is not clear whether circularly polarized light induces the orbital motion for the particles trapped off-axis. The present study reveals that the interparticle light-induced force due to the SAM enhances or weakens the orbital torque and modulates rotational dynamics depending on the number of NPs, where the rotation speed of NPs in the optical field with both positive SAM and OAM can be 4 times faster than that in the optical field with negative SAM and positive OAM. The obtained results will not only clarify the principle for the control of NPs based on OAM-SAM coupling via light-matter interaction but also contribute to the unconventional laser processing technique for nanostructures with various chiral symmetries.

Surfactant-Controlled Photothermal Assembly of Nanoparticles and Microparticles for Rapid Concentration Measurement of Microbes.

Light-induced heating on a solid-liquid interface can generate a vapor submillimeter bubble and fluid flow, which enables us to densely and rapidly assemble dispersoids into a desired position (photothermal assembly). Here, we revealed that the surface modulation of the light-induced bubble by a surfactant dominates the assembly dynamics of nanoparticles and microparticles as dispersoids, which results in highly efficient photothermal assembly under the surfactant-controlled fluid flow. This mechanism can facilitate the concentration measurement of small objects (microparticles, bacteria, viruses, etc.). Particularly, we found that the surfactant-controlled fluid flow and bubble enable high-density assembly of dispersoids and remarkable enhancement of assembly efficiency, achieving 10-20 times in comparison with the case of no surfactant. This result can extend the limit of measurable concentration by one order. Furthermore, this study revealed the influence of concentration, size, and constituent material of the dispersoids on the assembly efficiency for the improvement of measurement precision. These findings are crucial for laser-induced assembly for the rapid concentration measurement of various microbes without a cultivation process as bioanalysis, for the high-sensitivity detection of harmful particles, and for the colloidal lithography.

Macroscopically Anisotropic Structures Produced by Light-induced Solvothermal Assembly of Porphyrin Dimers.

Porphyrin-based molecules play an important role in natural biological systems such as photosynthetic antennae and haemoglobin. Recent organic chemistry provides artificial porphyrin-based molecules having unique electronic and optical properties, which leads to wide applications in material science. Here, we successfully produced many macroscopically anisotropic structures consisting of porphyrin dimers by light-induced solvothermal assembly with smooth evaporation in a confined volatile organic solvent. Light-induced fluid flow around a bubble on a gold nanofilm generated a sub-millimetre radial assembly of the tens-micrometre-sized petal-like structures. The optical properties of the petal-like structures depend on the relative angle between their growth direction and light polarisation, as confirmed by UV-visible extinction and the Raman scattering spectroscopy analyses, being dramatically different from those of structures obtained by natural drying. Thus, our findings pave the way to the production of structures and polycrystals with unique characteristics from various organic molecules.

Mechanism in External Field-mediated Trapping of Bacteria Sensitive to Nanoscale Surface Chemical Structure.

Molecular imprinting technique enables the selective binding of nanoscale target molecules to a polymer film, within which their chemical structure is transcribed. Here, we report the successful production of mixed bacterial imprinted film (BIF) from several food poisoning bacteria by the simultaneous imprinting of their nanoscale surface chemical structures (SCS), and provide highly selective trapping of original micron-scale bacteria used in the production process of mixed BIF even for multiple kinds of bacteria in real samples. Particularly, we reveal the rapid specific identification of E. coli group serotypes (O157:H7 and O26:H11) using an alternating electric field and a quartz crystal microbalance. Furthermore, we have performed the detailed physicochemical analysis of the specific binding of SCS and molecular recognition sites (MRS) based on the dynamic Monte Carlo method under taking into account the electromagnetic interaction. The dielectrophoretic selective trapping greatly depends on change in SCS of bacteria damaged by thermal treatment, ultraviolet irradiation, or antibiotic drugs, which can be well explained by the simulation results. Our results open the avenue for an innovative means of specific and rapid detection of unknown bacteria for food safety and medicine from a nanoscale viewpoint.

Review: Novel sensing strategies for bacterial detection based on active and passive methods driven by external field.

For sustainable human life, biosensing systems for contaminants or disease-causing bacteria are crucial for food security, environmental improvement, and disease prevention. With an aim of enhancing the sensitivity and detection speed, many researchers have developed efficient detection methods for target bacteria. In this review, we discuss recent topics related to active and passive bacterial detection methods, including (1) optical approaches with unique functional nano- and micro-structures, and (2) electrical approaches involving mechanical modulation and electrochemical reactions. Particularly, we discuss the prospects in the development of label-free, rapid, and highly sensitive biosensors based on active detection principles with light-induced dynamics, in conjunction with dielectrophoresis-induced selective trapping.

Optical Trap-Mediated High-Sensitivity Nanohole Array Biosensors with Random Nanospikes.

We clarify an unconventional principle of the light-driven operation of a biosensor for enhanced sensitivity with the help of random nanospikes added to the surface of a nanohole array. Such a system is capable of optically guiding viruses and trapping them in the vicinity of a highly sensitive site by an anomalous light-induced force arising from random-nanospike-modulated extraordinary optical transmission and the plasmonic mirror image in a virus as a dielectric submicron object. In particular, after guiding the viruses near the apex of nanospikes, there are conditions where the spectral peak shift of extraordinary optical transmission can be greatly increased and reach several hundred nanometers in comparison with that of a conventional nanohole array without random nanospikes. These results will allow for the development of a simple, rapid, and highly sensitive virus detection method based on optical trapping with the help of random-nanospike-modulated extraordinary optical transmission, facilitating convenient medical diagnosis and food inspection.

Submillimetre Network Formation by Light-induced Hybridization of Zeptomole-level DNA.

Macroscopic unique self-assembled structures are produced via double-stranded DNA formation (hybridization) as a specific binding essential in biological systems. However, a large amount of complementary DNA molecules are usually required to form an optically observable structure via natural hybridization, and the detection of small amounts of DNA less than femtomole requires complex and time-consuming procedures. Here, we demonstrate the laser-induced acceleration of hybridization between zeptomole-level DNA and DNA-modified nanoparticles (NPs), resulting in the assembly of a submillimetre network-like structure at the desired position with a dramatic spectral modulation within several minutes. The gradual enhancement of light-induced force and convection facilitated the two-dimensional network growth near the air-liquid interface with optical and fluidic symmetry breakdown. The simultaneous microscope observation and local spectroscopy revealed that the assembling process and spectral change are sensitive to the DNA sequence. Our findings establish innovative guiding principles for facile bottom-up production via various biomolecular recognition events.

Catalytic Activities for Glucose Oxidation of Au/Pd Bimetallic Nanoparticles Prepared via Simultaneous NaBH4 Reduction.

Au/Pd bimetallic nanoparticles (BNPs) were prepared by simultaneous reduction method using NaBH4 as a reducing reagent. The effects of particle size, electronic structure and composition upon the catalytic activities of the BNPs for aerobic glucose oxidation were investigated. The PVP-protected Au/Pd BNPs of about 2.0 nm in diameter synthesized via rapid injection of NaBH4 possessed a high catalytic activity for aerobic glucose oxidation. The catalytic activity of BNPs with the Au/Pd atomic ratio of 60/40 was more than two times higher than that of Au nanoparticles (NPs) though the latter were smaller. This can be ascribed to the presence of negatively charged Au atoms arisen from electron donation from neighboring Pd atoms via electronic charge transfer. In contrast, Au/Pd BNPs synthesized via dropwise addition of NaBH4 into the starting solution and having the large mean particle sizes, showed a low catalytic activity.

Voltammetric detection and profiling of isoprenoid quinones hydrophobically transferred from bacterial cells.

We have developed a novel bacterial detection technique by desiccating a bacterial suspension deposited on an electrode. It was also found that the use of an indium-tin-oxide (ITO) electrode dramatically improved the resolution of the voltammogram, allowing us to observe two pairs of redox peaks, each assigned to the adsorption of isoprenoid ubiquinone (UQn) and menaquinone (MKn), which were present in the bacterial cell envelopes, giving midpeak potentials of -0.015 and -0.25 V versus Ag|AgCl|saturated KCl| at pH 7.0, respectively. Most of the microorganisms classified in both the Gram-negative and -positive bacteria gave well-defined redox peaks, demonstrating that this procedure made the detection of the quinones possible without solvent extraction. It has been demonstrated that the present technique can be used not only for the detection of bacteria, but also for profiling of the isoprenoid quinones, which play important roles in electron and proton transfer in microorganisms. In this respect, the present technique provides a much more straightforward way than the solvent extraction in that one sample can be prepared in 1 min by heat evaporation of a suspension containing the targeted bacteria, which has been applied on the ITO electrode.

Electrochemical evaluation of poly(3,4-ethylenedioxythiophene) films doped with bacteria based on viability analysis.

To immobilize viable bacteria on an electrode, we present a novel and straightforward technique that relies on the negative ζ-potentials of bacteria for insertion into conducting polymers as dopants. In the present study, we conducted an electrochemical polymerization of poly(3,4-ethylenedioxythiophene) (PEDOT) doped with various gram-negative bacteria, including Pseudomonas aeruginosa, Escherichia coli, and Shewanella oneidensis. The PEDOT film doped with bacteria indicated a typical redox response, high conductivity, and electrochemical stability. Fluorescence microscopy confirmed that approximately 90% of the bacteria incorporated into the PEDOT film at >0.5 µm in thickness were viable.

A high performance photothermal film with spherical shell-type metallic nanocomposites for solar thermoelectric conversion.

A photothermal film (PTF) with densely assembled gold nanoparticle-fixed beads on a polymer substrate is fabricated. Remarkably, a temperature rise higher than 40 °C is achieved in the PTF with only 100 seconds of artificial solar irradiation, and the output power of the thermoelectric device was enhanced to be one order higher than that without PTF. These results will pioneer a rapid solar thermoelectric device.

Development of an observation platform for bacterial activity using polypyrrole films doped with bacteria.

In our study, various bacteria, including Gram-negative (Pseudomonas aeruginosa, Escherichia coli, Acinetobacter calcoaceticus, Serratia marcescens, Shewanella oneidensis) and Gram-positive (Bacillus subtilis) bacteria, were straightforwardly immobilized into the conducting polymers (CPs) by electrochemical deposition. The doping state of bacteria in the polymer films (polypyrrole and poly(3,4-ethylenedioxythiophene)) varied according to the polymerization conditions. The viability of bacteria in the polymers and of those adsorbed on various substrates was evaluated. The activity of bacteria doped on the polymer film was evaluated by cyclic voltammetry in a thin-layer cell.

Fluorescent carbon nanowires made by pyrolysis of DNA nanofibers and plasmon-assisted emission enhancement of their fluorescence.

We report on a facile method for preparing fluorescent carbon nanowires (CNWs) with pyrolysis of highly aligned DNA nanofibers as carbon sources. Silver nanoparticle (AgNP)-doped CNWs were also produced using pyrolysis of DNA nanofibers with well-attached AgNPs, indicating emission enhancement assisted by localized plasmon resonances.

Construction of nanoantennas on the bacterial outer membrane.

We demonstrate a simple manipulation of gold nanoparticles that creates a structure-dependent nanometer-scale antenna on the surface of bacteria. Our studies illuminate the concept of the "effective use of light" based on the absorption and emission of light by antennas formed on bacteria.