Multifunctional carbon-nanotube cellular endoscopes

Glass micropipettes, atomic force microscope tips and nanoneedles can be used to interrogate cells, but these devices either have conical geometries that can damage cells during penetration or are incapable of continuous fluid handling. Here, we report a carbon-nanotube-based endoscope for interrogating cells, transporting fluids and performing optical and electrochemical diagnostics at the single organelle level. The endoscope, which is made by placing a multiwalled carbon nanotube (length, 50-60?μm) at the tip of a glass pipette, can probe the intracellular environment with a spatial resolution of ～100?nm and can also access organelles without disrupting the cell. When the nanotube is filled with magnetic nanoparticles, the endoscope can be remotely manoeuvered to transport nanoparticles and attolitre volumes of fluids to and from precise locations. Because they are mounted on conventional glass micropipettes, the endoscopes readily fit standard instruments, creating a broad range of opportunities for minimally invasive intracellular probing, drug delivery and single-cell surgery.     

Optically Controlled Pore Formation in Self‐Sealing Giant Porphyrin Vesicles

Efforts to develop self‐contained microreactors and artificial cells have been limited by difficulty in generating membranes that can be robustly and repeatedly manipulated to load and release cargo from phospholipid compartments. Here we describe a purely optical method to form pores in a membrane generated from porphyrin‐phospholipid conjugates electro‐assembled into microscale giant porphyrin vesicles and manipulated using confocal microscopy. The pores in the membrane resealed within a minute allowing for repeated pore formation with precise spatial and temporal control and optical gating to allow selective diffusion of biomolecules across the membrane. Temporal control of pore formation was illustrated by performing sequential DNA hybridization reactions. A biotin‐avidin based strategy was developed to selectively attach enzymes to the interior of the vesicle, demonstrating spatial control and the potential of giant porphyrin vesicles as versatile microreactors.     

Live-cell imaging of single receptor composition using zero-mode waveguide nanostructures

We exploit the optical and spatial features of subwavelength nanostructures to examine individual receptors on the plasma membrane of living cells. Receptors were sequestered in portions of the membrane projected into zero-mode waveguides. Using single-step photobleaching of green fluorescent protein incorporated into individual subunits, the resulting spatial isolation was used to measure subunit stoichiometry in α4β4 and α4β2 nicotinic acetylcholine and P2X2 ATP receptors. We also show that nicotine and cytisine have differential effects on α4β2 stoichiometry.     

Optical trapping and integration of semiconductor nanowire assemblies in water

Semiconductor nanowires have received much attention owing to their potential use as building blocks of miniaturized electrical, nanofluidic and optical devices. Although chemical nanowire synthesis procedures have matured and now yield nanowires with specific compositions and growth directions, the use of these materials in scientific, biomedical and microelectronic applications is greatly restricted owing to a lack of methods to assemble nanowires into complex heterostructures with high spatial and angular precision. Here we show that an infrared single-beam optical trap can be used to individually trap, transfer and assemble high-aspect-ratio semiconductor nanowires into arbitrary structures in a fluid environment. Nanowires with diameters as small as 20 nm and aspect ratios of more than 100 can be trapped and transported in three dimensions, enabling the construction of nanowire architectures that may function as active photonic devices. Moreover, nanowire structures can now be assembled in physiological environments, offering new forms of chemical, mechanical and optical stimulation of living cells.     

Plasmonic nearfield scanning probe with high transmission

Nearfield scanning optical microscopy (NSOM) offers a practical means of optical imaging, optical sensing, and nanolithography at a resolution below the diffraction limit of the light. However, its applications are limited due to the strong attenuation of the light transmitted through the subwavelength aperture. To solve this problem, we report the development of plasmonic nearfield scanning optical microscope with an efficient nearfield focusing. By exciting surface plasmons, plasmonic NSOM probes are capable of confining light into a 100 nm spot. We show by nearfield lithography experiments that the intensity at the near field is at least one order stronger than the intensity obtained from the conventional NSOM probes under the same illumination condition. Such a high efficiency can enable plasmonic NSOM as a practical tool for nearfield lithography, data storage, cellular visualization, and many other applications requiring efficient transmission with high resolution.     

Absorption coefficient imaging by near-field scanning optical microscopy in bacteria

We present a method for obtaining a position-dependent absorption coefficient from near-field scanning optical transmission microscopy. We show that the optical transmission intensity can be combined with the topography, resulting into an absorption coefficient that simplifies the analysis of different materials within a sample. The method is tested with the dye rhodamine 6G, and we show some analysis in biological samples such as bacteria KIebsiella pneumoniae and Pseudomonas aeruginosa. The calculated absorption coefficient images show important details of the bacteria, in particular for P. aeruginosa, in which membrane vesicles are clearly seen.     

Intracellular recordings of action potentials by an extracellular nanoscale field-effect transistor

The ability to make electrical measurements inside cells has led to many important advances in electrophysiology. The patch clamp technique, in which a glass micropipette filled with electrolyte is inserted into a cell, offers both high signal-to-noise ratio and temporal resolution. Ideally, the micropipette should be as small as possible to increase the spatial resolution and reduce the invasiveness of the measurement, but the overall performance of the technique depends on the impedance of the interface between the micropipette and the cell interior, which limits how small the micropipette can be. Techniques that involve inserting metal or carbon microelectrodes into cells are subject to similar constraints. Field-effect transistors (FETs) can also record electric potentials inside cells, and because their performance does not depend on impedance, they can be made much smaller than micropipettes and microelectrodes. Moreover, FET arrays are better suited for multiplexed measurements. Previously, we have demonstrated FET-based intracellular recording with kinked nanowire structures, but the kink configuration and device design places limits on the probe size and the potential for multiplexing. Here, we report a new approach in which a SiO2 nanotube is synthetically integrated on top of a nanoscale FET. This nanotube penetrates the cell membrane, bringing the cell cytosol into contact with the FET, which is then able to record the intracellular transmembrane potential. Simulations show that the bandwidth of this branched intracellular nanotube FET (BIT-FET) is high enough for it to record fast action potentials even when the nanotube diameter is decreased to 3 nm, a length scale well below that accessible with other methods. Studies of cardiomyocyte cells demonstrate that when phospholipid-modified BIT-FETs are brought close to cells, the nanotubes can spontaneously penetrate the cell membrane to allow the full-amplitude intracellular action potential to be recorded, thus showing that a stable and tight seal forms between the nanotube and cell membrane. We also show that multiple BIT-FETs can record multiplexed intracellular signals from both single cells and networks of cells.     

Optical detection of evanescent ultrasound waves in water

Acoustical perturbation by targets smaller than the wavelength can generate evanescent waves, which decay with the distance of propagation. By putting such targets immediately under the free surface of water, the diffracted acoustical field can excite the surface membrane before evanescence, and it produces a static ripple because of the radiation pressure. A collimated beam of light reflects at the perturbed surface, and it becomes modulated in phase. In this work we show experimental results where we achieve an optical image of the acoustical evanescent waves produced by an array-like target. Arising from the theory, we present a numerical calculus of the optical image produced by the ultrasonic field diffracted from the target in order to verify the experimental results. With this method, we look for a spatial resolution smaller than acoustical wavelength, for normal incidence and plane waves. In our experimental device, we use a sound wavelength in water greater than 1.5 mm, generated by a PZT transducer. We can resolve an array of 1.0 mm of periodicity.     

汽车制动主缸补偿孔形位尺寸检测双远心光学系统研究

为了解决制动主缸补偿孔的成像问题,设计了一种远心内窥成像光学系统。在设计中采用工业内窥镜来延长成像系统的视距,它的后端与CCD摄像机相连接,并通过内窥镜的前后移动来捕捉补偿孔的图像信息。该系统由于将内窥镜的成像光路设计成一个像方远心光路与物方远心光路合一的光路,较好地解决制动主缸补偿孔图像因为虚焦而造成的检测误差问题,从而提高了系统的检测精度。实验结果表明,系统的测量精度达到0.01 mm。     

被动式胶囊内窥镜在小肠内运动的力学分析

将蠕动过程中的小肠简化为不可压缩弹性薄膜,结合旋转壳体的薄膜应力平衡方程,研究小肠蠕动推进被动式胶囊内窥镜的力学行为。对推导公式进行了数值分析,分析表明：在小肠蠕动过程中,小肠对于胶囊内窥镜的推力与其直径的平方成正比;综合考虑患者感受、检查成功性,胶囊内窥镜的直径应该设计为7～15 mm;在假设胶囊行进速度为常数时,粘滞阻力和其直径与长度乘积呈线性关系,但数值较小。     

Extraordinary optical transmission brightens near-field fiber probe

Near-field scanning optical microscopy (NSOM) offers high optical resolution beyond the diffraction limit for various applications in imaging, sensing, and lithography; however, for many applications the very low brightness of NSOM aperture probes is a major constraint. Here, we report a novel NSOM aperture probe that gives a 100× higher throughput and 40× increased damage threshold than conventional near-field aperture probes. These brighter probes facilitate near-field imaging of single molecules with apertures as small as 45 nm in diameter. We achieve this improvement by nanostructuring the probe and by employing a novel variant of extraordinary optical transmission, relying solely on a single aperture and a coupled waveguide. Comprehensive electromagnetic simulations show good agreement with the measured transmission spectra. Due to their significantly increased throughput and damage threshold, these resonant configuration probes provide an important step forward for near-field applications.     

Dielectrophoretic assembly of insulinoma cells and fluorescent nanosensors into three-dimensional pseudo-islet constructs

Dielectrophoretic forces, generated by radio-frequency voltages applied to micromachined, transparent, indium tin oxide electrodes, have been used to condense suspensions of insulinoma cells (BETA-TC-6 and INS-1) into a 10times10 array of three-dimensional cell constructs. Some of these constructs, measuring ~150 mum in diameter, 120 mum in height and containing around 1000 cells, were of the same size and cell density as a typical islet of Langerhans. With the dielectrophoretic force maintained, these engineered cell constructs were able to withstand mechanical shock and fluid flow forces. Reproducibility of the process required knowledge of cellular dielectric properties, in terms of membrane capacitance and membrane conductance, which were obtained by electrorotation measurements. The ability to incorporate fluorescent nanosensors, as probes of cellular oxygen and pH levels, into these 'pseudo-islets' was also demonstrated. The footprint of the 10times10 array of cell constructs was compatible with that of a 1536 microtitre plate, and thus amenable to optical interrogation using automated plate reading equipment.     

Visualization of individual single-walled carbon nanotubes by fluorescent polymer wrapping

Manipulating optical properties of single-walled nanotubes (SWNTs) is necessary for the development of nanoscale optical devices and probes for biomedical research. In life sciences it will make possible the direct observation of SWNTs inside living cells using optical microscopes. In the nanotechnology field it will enable the development of nanosensors with fluorescent reporting. However, the direct fluorescent labeling of SWNTs is obstructed by their strong light quenching qualities. Besides, chemical functionalization of SWNTs needed for the covalent attachment of fluorescent dyes could change favorable properties of nanotubes. Here we report that optical properties of SWNTs can be manipulated without their covalent modification by wrapping them with fluorescently labeled polymer poly(vinylpyrrolidone) (PVP-1300). Fluorescent PVP-1300 forms a monomolecular approximately 2.5 nm thick layer coiling around individual SWNTs and nanotube bundles. PVP casing is fluorescent although it is only several nanometers thick. This makes individual SWNTs observable by a fluorescent microscope. The spare polymer strands left over after wrapping around the relatively shorter nanotubes form junctions between SWNTs tying them together into new configurations, primarily Y- and psi-type junctions. The ability to use a single fluorescent polymer strand to fasten nanotubes together can be useful in assembly of nanotube-made devices. In PVP-covered SWNTs multiple fluorophores are attached to each single nanotube making them unique composite fluorophores attractive as parts of biological fluorescent probes and in the development of the new materials in photonics and nanotechnology.     

Design and characterization of coherent integrated fiber-optic imaging probes

An integrated fiber-optic probe comprising a short length of multimode fiber that is fusion spliced to a monomode optical fiber has been fabricated for imaging and nonimaging applications. The fiber probe, typically 250 μm in diameter, can deliver a focused Gaussian spot approximately 25 μm in diameter at a distance of approximately 500 μm from the tip. Two off-the-shelf graded-index multimode fibers have been used in the fabrication of imaging and nonimaging probes. These integrated probes have considerably improved the spatial resolution of backscatter lensless fiber probes being utilized in the dynamic light-scattering characterization of colloidal suspension.     

Design and characterization of a liquid-crystal spatial light modulator for a polarization-insensitive optical space switch

We report preliminary results concerning a free-space optical switch between single-mode fibers with a ferroelectric liquid-crystal (FLC) spatial light modulator (SLM). In particular, we show experimentally that such a device can operate in a polarization-insensitive manner. The influence of the geometrical and physical features of the FLC SLM on the overall performance of the optical fiber switch are also discussed.     

Fabrication of an atomic resolution low cost STM-SNOM hybrid probe

We derived a simple method to fabricate STM-SNOM hybrid probes obtained from commercial cheap communication optical fibers. The tips are fabricated by a methodology that combines two well-known techniques: the selective attack by a buffered solution and the protected layer chemical etching, in a single new one-step technique. The tailored probes are then sputtered by metal and mounted on a STM setup. The usual difficulties of integrating the optical fiber in the STM head are solved originally with a particular home made mount described in details. We will show that the resulting probes reach atomic resolution on both vertical and horizontal scale, and that the optical imaging is free of artifacts and satisfactory with a lateral resolution in the order of lamda/20, as far as we know the finest resolution obtained with a system based on a hybrid fiber probe. We believe that our methodology is very interesting for its simplicity of realization and for the good resolving power in both SNOM and STM modes.     

Tumor detection strategy using ZnO light-emitting nanoprobes

Traditional methods of detecting cancer cells, such as fluorescence, have their limits and can hardly be used for identification during tumor resection. Here we report an alternative tumor detection technology using ZnO nanorods bonded to antibodies as cancer cell probes. Our experiment shows that antibodies toward epidermal growth factor receptor (EGFR) can be connected to ZnO nanorods and to EGFR receptors of SCC (squamous cell carcinoma). The cancer cell can be recognized by the naked eye or an optical microscope with the help of purple light emission from ZnO/EGFR antibody probes. On the other hand, for cells with less EGFR expression, in our case Hs68, no purple light was observed as the probes were washed off. From the photoluminescent spectra, the peak intensity ratio between the purple light (from ZnO at the wavelength 377 nm) and the green band (from the autofluorescence of cells) is much higher with the presence in SCC, as compared with Hs68. The ZnO/EGFR antibody probes have the potential to be applied to surgery for real-time tumor cell identification. The cancer cells will be excised with the help of purple light emission.     

A design method of spatiotemporal optical pulse using level-set based time domain topology optimization

Recently, two emerging areas of photonics research, ultrafast photonics, and nanophotonics have started to come together. One of the main problems in this field is the precise control of spatial and temporal profiles of the optical pulses. In this paper, we propose a design method for user-specified spatiotemporal optical pulses using a level set-based time-domain topology optimization method. In the proposed method, the optimization problem is formulated based on time domain Maxwell equations so that the spatiotemporal optical pulses can be treated directly. The objective function is defined using the envelope information of the pulses, and an efficient way to calculate this information, based on calculations of the complex electromagnetic field, is introduced. A level set-based topology optimization method is applied to obtain optimized configurations. Using the proposed method, the spatiotemporal user-specified pulse profiles can be designed by modifying the structural details of the nanostructures through which the pulses propagate. As a simple example, we demonstrate that the optimized structures focus optical pulses into a single or multiple focal points with a user-specified pulse-width. The results show that the proposed method is able to design highly controlled spatiotemporal optical pulses by engineering the nanophotonic structure.     

Vectorial modeling of near-field imaging with uncoated fiber probes: transfer function and resolving power

Using exact 3D vectorial simulations of radiation coupling into uncoated dielectric fiber probes, we calculate amplitude transfer functions for conical single-mode fiber tips at the light wavelength of 633 nm. The coupling efficiency of glass fiber tips is determined in a wide range of spatial frequencies of the incident radiation for opening angles varying from 30 degrees to 120 degrees . The resolution in near-field imaging with these tips is considered for field distributions limited in both direct and spatial-frequency space. The characteristics of the transfer functions describing the relation between probed optical fields and near-field images are analyzed in detail. The importance of utilizing a perfectly sharp tip is also examined.     

Cell uptake and intracellular visualization using quantum dots or nuclear localization signal-modified quantum dots with gold nanoparticles as quenchers

Understanding and controlling the interactions between nanoscale objects and living cells is of great importance for diagnostic imaging and therapeutic applications. Quantum dots (QDs) have remarkable optical characteristics, such as uniquely feature bright, photostable, tunable and narrow fluorescence emissions, as well as broad absorption spectra. Here we report a platform of using quantum dots to investigate the cell uptake and the interactions between nanoscale objects and cells. QDs are uptaken by BHK cells easily through endocytosis. We could clearly differentiate the QDs outside the cell or inside the cell by quenching the QDs with similar sized gold nanoparticles and reduce the noise of fluorescent image. Microscopic images show that QDs are homogeneously distributed within the whole cell except the nucleus. However, unmodified QDs could not penetrate the nuclear membrane and move into the nucleus. Coupling QDs with Nuclear Localization Signal (NLS, CGGGPKKKRKVGG) can significantly enhance the translocation amount of QDs into the cell and cell nucleus. This method combined with microscopy imaging system can visualize the particle delivery routes and provide valuable information in the drug/gene delivery and tumor diagnosis.