Nano-optics with single quantum systems

This paper reviews the recent progress in using single quantum systems, here mainly single fluorescent molecules, as local probes for nano-optical field distributions. We start by discussing the role of the absorption cross-section for the spatial resolution attainable in such experiments and its behaviour for different environmental conditions. It is shown that the spatial distribution of field components in a high-numerical aperture laser focus can be mapped with high precision using single fluorescent molecules embedded in a thin polymer film on glass. With this proof-of-principle experiment as a starting point, the possibility of mapping strongly confined and enhanced nano-optical fields close to material structures, e.g. sharp metal tips, is discussed. The mapping of the spatial distribution of the enhanced field at an etched gold tip using a single molecule is presented as an example. Energy transfer effects and quenching are identified as possible artefacts in this context. Finally, it is demonstrated that the local quenching at a sharp metal structure nevertheless can be exploited as a novel contrast mechanism for ultrahigh-resolution optical microscopy with single-molecule sensitivity.     

In Vivo Volumetric Photoacoustic Molecular Angiography and Therapeutic Monitoring with Targeted Plasmonic Nanostars

Photoacoustic (PA) imaging promises deeper tissue penetration while maintaining rich optical contrast as compared to other high resolution optical imaging techniques. In this report, a near‐infrared pulse laser serves as the excitation source, and 128 ultrasonic transducers are spirally distributed on a hemispherical surface to receive PA signals for three‐dimensional (3D) image reconstruction. With these attributes, the unique modality produces an isotropic and homogeneous spatial resolution (～200 μm) with penetration depth of centimeters. Cyclic Arg‐Gly‐Asp (RGD) peptides conjugated plasmonic gold nanostars (RGD‐GNS) are designed to specifically target over‐expressed integrin α_vβ₃ on tumor neovasculature, enabling highly sensitive angiography and photothermal therapy (PTT). After the administration of RGD‐GNS, tumor angiogenesis is clearly imaged with enhanced contrast, and the growth of tumor is effectively inhibited by PTT after laser irradiation. This study suggest that the PA angiography with plasmonic RGD‐GNS can be applied as a triple functional platform for tumor diagnosis, PTT, and treatment monitoring. This PA technique offers deeper imaging depth with homogeneous resolution over existing optical imaging techniques for early diagnosis of tumor angiogenesis as well as on‐the‐spot nanotherapeutic evaluation.     

Automated, portable, low-cost bright-field and fluorescence microscope with autofocus and autoscanning capabilities

Optical microscopy is a simple, yet essential, imaging technology. Conventional laboratory-grade optical microscopes are bulky and costly, confining their use to within laboratory settings and restricting their accessibility in regions of limited resources. With the aim of overcoming these limitations, we have realized a portable, low-cost, and highly automated optical microscope that integrates mass-manufactured components, including light-emitting diodes, a web camera, optical disk drives, and a microcontroller. Our implementation is capable of bright-field and fluorescence imaging with micrometer-scale resolution and controlled mechanical actuation of both the lens and sample. We interface the lighting, image capture, and mechanical actuators of the microscope into a single software environment, enabling automation of common microscope operations, such as image focusing and large-area sample visualization. Combination of mechanical actuation and software automation into a compact, low-cost microscope system is an important initial step toward the goal of making optical microscopy universally accessible, portable, and easy to use.     

Wide-field fluorescence lifetime imaging with optical sectioning and spectral resolution applied to biological samples

Abstract. Wide-field fluorescence lifetime imaging with spectral resolution and optical sectioning has been performed to achieve five-dimensional fluorescence microscopy. Spectral filtering has been shown to have the potential to provide functional information about biological tissue by simultaneously measuring the spectral/lifetime signature of the sample. The potential to use multispectral imaging to separate cellular components spatially by their different emission wavelengths has also been demonstrated thus reducing artefacts in the calculated lifetime maps. The instrument is based on diode-pumped solid-state laser technology and an ultrafast gated optical image intensifier. Also reported is the use of a picosecond blue laser diode as the excitation source to produce a fluorescence lifetime microscope with a footprint of less than 0.25m².     

A super-oscillatory lens optical microscope for subwavelength imaging

The past decade has seen an intensive effort to achieve optical imaging resolution beyond the diffraction limit. Apart from the Pendry-Veselago negative index superlens, implementation of which in optics faces challenges of losses and as yet unattainable fabrication finesse, other super-resolution approaches necessitate the lens either to be in the near proximity of the object or manufactured on it, or work only for a narrow class of samples, such as intensely luminescent or sparse objects. Here we report a new super-resolution microscope for optical imaging that beats the diffraction limit of conventional instruments and the recently demonstrated near-field optical superlens and hyperlens. This non-invasive subwavelength imaging paradigm uses a binary amplitude mask for direct focusing of laser light into a subwavelength spot in the post-evanescent field by precisely tailoring the interference of a large number of beams diffracted from a nanostructured mask. The new technology, which--in principle--has no physical limits on resolution, could be universally used for imaging at any wavelength and does not depend on the luminescence of the object, which can be tens of micrometres away from the mask. It has been implemented as a straightforward modification of a conventional microscope showing resolution better than λ/6.     

Single cell matrix-assisted laser desorption/ionization mass spectrometry imaging

Application of mass spectrometry imaging (MS imaging) analysis to single cells was so far restricted either by spatial resolution in the case of matrix-assisted laser desorption/ionization (MALDI) or by mass resolution/mass range in the case of secondary ion mass spectrometry (SIMS). In this study we demonstrate for the first time the combination of high spatial resolution (7 μm pixel), high mass accuracy (<3 ppm rms), and high mass resolution (R = 100?000 at m/z = 200) in the same MS imaging measurement of single cells. HeLa cells were grown directly on indium tin oxide (ITO) coated glass slides. A dedicated sample preparation protocol was developed including fixation with glutaraldehyde and matrix coating with a pneumatic spraying device. Mass spectrometry imaging measurements with 7 μm pixel size were performed with a high resolution atmospheric-pressure matrix-assisted laser desorption/ionization (AP-MALDI) imaging source attached to an Exactive Orbitrap mass spectrometer. Selected ion images were generated with a bin width of Δm/z = ±0.005. Selected ion images and optical fluorescence images of HeLa cells showed excellent correlation. Examples demonstrate that a lower mass resolution and a lower spatial resolution would result in a significant loss of information. High mass accuracy measurements of better than 3 ppm (root-mean-square) under imaging conditions provide confident identification of imaged compounds. Numerous compounds including small metabolites such as adenine, guanine, and cholesterol as well as different lipid classes such as phosphatidylcholine, sphingomyelin, diglycerides, and triglycerides were detected and identified based on a mass spectrum acquired from an individual spot of 7 μm in diameter. These measurements provide molecularly specific images of larger metabolites (phospholipids) in native single cells. The developed method can be used for a wide range of detailed investigations of metabolic changes in single cells.     

High frequency ultrasound imaging with optical arrays

Dynamically focused and steered high frequency ultrasound imaging systems require arrays with fine element spacing, wide bandwidths, and large apertures. However, these characteristics are difficult to achieve at frequencies greater than 30 MHz using conventional array construction methods. Optical schemes offer a solution. Focused laser beams incident on a suitable surface can generate and detect acoustic radiation. Precisely controlling the position and size of the beams defines points of transmission and detection, making it possible for pulse-echo image formation by synthetic aperture methods. An optical detection array was built, relying on a conventional piezoelectric transducer as an ultrasound source. The detection system, with near optimal resolution over a wide depth of field, demonstrates the potential for high frequency array implementation using optical techniques. A possible application is in pathology, where 2-D or 3-D fine resolution pulse-echo imaging can be performed in situ without the need for biopsies.     

Computer-controlled optical scanning tile microscope

A new type of computer-controlled optical scanning, high-magnification imaging system with a large field of view is described that overcomes the commonly believed incompatibility of achieving both high magnification and a large field of view. The new system incorporates galvanometer scanners, a CCD camera, and a high-brightness LED source for the fast acquisition of a large number of a high-resolution segmented tile images with a magnification of 800x for each tile. The captured segmented tile images are combined to create an effective enlarged view of a target totaling 1.6 mm x 1.2 mm in area. The speed and sensitivity of the system make it suitable for high-resolution imaging and monitoring of a small segmented area of 320 microm x 240 microm with 4 microm resolution. Each tile segment of the target can be zoomed up without loss of the high resolution. This new microscope imaging system gives both high magnification and a large field of view. This microscope can be utilized in medicine, biology, semiconductor inspection, device analysis, and quality control.     

用于纳米级表面形貌测量的光学显微测头

为了满足纳米级表面形貌样板的高精度非接触测量需求,研制了一种高分辨力光学显微测头。以激光全息单元为光源和信号拾取器件,利用差动光斑尺寸变化探测原理,建立了微位移测量系统,结合光学显微成像系统,形成了高分辨力光学显微测头。将该测头应用于纳米三维测量机,对台阶高度样板和一维线间隔样板进行了测量实验。结果表明：该光学显微测头结合纳米三维测量机可实现纳米级表面形貌样板的可溯源测量,具有扫描速度快、测量分辨力高、结构紧凑和非接触测量等优点,对解决纳米级表面形貌测量难题具有重要实用价值。     

Wide-Field and Real-Time Super-Resolution Optical Imaging By Titanium Dioxide Nanoparticle-Assembled Solid Immersion Lens

Super-resolution optical imaging techniques can break the optical diffraction limit, thus providing unique opportunities to visualize the microscopic world at the nanoscale. Although near-field optical microscopy techniques have been proven to achieve significantly improved imaging resolution, most near-field approaches still suffer from a narrow field of view (FOV) or difficulty in obtaining wide-field images in real time, which may limit their widespread and diverse applications. Here, the authors experimentally demonstrate an optical microscope magnification and image enhancement approach by using a submillimeter-sized solid immersion lens (SIL) assembled by densely-packed 15 nm TiO₂ nanoparticles through a silicone oil two-step dehydration method. This TiO₂ nanoparticle-assembled SIL can achieve both high transparency and high refractive index, as well as sufficient mechanical strength and easy-to-handle size, thus providing a fast, wide-field, real-time, non-destructive, and low-cost solution for improving the quality of optical microscopic observation of a variety of samples, including nanomaterials, cancer cells, and living cells or bacteria under conventional optical microscopes. This study provides an attractive alternative to simplify the fabrication and applications of high-performance SILs.     

FPGA Implementation of a Digital Sequential Phase-Shift Stroboscope for In-Plane Vibration Measurements With Subpixel Accuracy

This paper describes a sequential phase-shift stroboscope and a subpixel imaging system that is able to measure with high resolution weak in-plane harmonic motions, such as those of microelectromechanical systems (MEMS). The synchronization of both MEMS stimulus and light strobe can be obtained by either using the camera output frame signal or driving the external trigger of the camera, depending on the operation principle of the camera. Experimental results concern a small silicon cantilever resonator excited in its first and second vibration modes and a quartz tuning fork. Two optical configurations have been developed. The first one is based on the use of a macroobjective for large fields of view with micrometer resolution displacement measurement, and the second is based on an optical microscope for weak amplitude vibration measurements with a resolution down to a few nanometers. The whole logic unit of the stroboscopic signal generator is implemented into a low-cost field-programmable gate array, thus offering high flexibility.      

High-resolution monochromatic x-ray imaging system based on spherically bent crystals

We have developed an improved x-ray imaging system based on spherically curved crystals. It is designed and used for diagnostics of targets ablatively accelerated by the Nike KrF laser. A spherically curved quartz crystal (d = .?, R = mm) has been used to produce monochromatic backlit images with the He-like Si resonance line (1865 eV) as the source of radiation. The spatial resolution of the x-ray optical system is 1.7 mum in selected places and 2-3 mum over a larger area. Time-resolved backlit monochromatic images of polystyrene planar targets driven by the Nike facility have been obtained with a spatial resolution of 2.5 mum in selected places and 5 mum over the focal spot of the Nike laser.     

New opportunities in micro- and macro-attenuated total reflection infrared spectroscopic imaging: spatial resolution and sampling versatility

New opportunities exist to obtain chemical images using attenuated total reflection infrared (ATR-IR) spectroscopy. This paper shows the feasibility of obtaining FT-IR images with a spatial resolution of at least 3-4 microm using a Ge ATR objective coupled with an infrared microscope. The improved spatial resolution compared to FT-IR images obtained by the transmission method is due to the high refractive index of the ATR crystal, which gives a high numerical aperture and hence, a higher spatial resolution. FT-IR imaging with a conventional diamond ATR accessory has been investigated. This is the first time that FT-IR imaging is reported using such a versatile accessory based on a diamond ATR crystal. These results showed that a spatial resolution up to 13 microm can be achieved without the use of infrared microscope objectives. One advantage of the diamond element is that it allows pressure to be applied and hence, good contact to be obtained over the whole field of view.     

一种基于光拍频的连续激光调制光谱测量方法

针对F-P干涉仪分析连续激光调制光谱的局限性,本文提出了一种基于光拍频的连续激光调制光谱的测量方法,该方法可实现调制频率低达千赫兹的连续激光调制光谱的测量.本方法以光电转换理论为基础,利用频谱分析仪测得参考光与连续调制光谱的拍频信号,然后通过相应的数学计算得到连续调制光谱的各个光频的相对电场强度,从而实现连续调制光谱的分析.本文在理论建模与分析的基础上,利用该方法对半导体激光器出射激光经电光调制器调制产生的调制光谱进行了测量,测量结果与F-P干涉仪测量结果一致,验证了该方法的可行性.     

Carbon Nanotubes as an Ultrafast Emitter with a Narrow Energy Spread at Optical Frequency

Ultrafast electron pulses, combined with laser‐pump and electron‐probe technologies, allow ultrafast dynamics to be characterized in materials. However, the pursuit of simultaneous ultimate spatial and temporal resolution of microscopy and spectroscopy is largely subdued by the low monochromaticity of the electron pulses and their poor phase synchronization to the optical excitation pulses. Field‐driven photoemission from metal tips provides high light‐phase synchronization, but suffers large electron energy spreads (3–100 eV) as driven by a long wavelength laser (>800 nm). Here, ultrafast electron emission from carbon nanotubes (≈1 nm radius) excited by a 410 nm femtosecond laser is realized in the field‐driven regime. In addition, the emitted electrons have great monochromaticity with energy spread as low as 0.25 eV. This great performance benefits from the extraordinarily high field enhancement and great stability of carbon nanotubes, superior to metal tips. The new nanotube‐based ultrafast electron source opens exciting prospects for extending current characterization to sub‐femtosecond temporal resolution as well as sub‐nanometer spatial resolution.     

Suppression of nonlinear frequency sweep in an optical frequency-domain reflectometer by use of Hilbert transformation

A compensation technique for reducing the effect of nonlinear optical frequency swept in an optical frequency-domain reflectometer (OFDR) is proposed. The instantaneous sweep optical frequency of an OFDR laser source is directly obtained by analysis of the interference signal from an auxiliary interferometer with a Hilbert transformation. Beating OFDR data from a main interferometer are regenerated with respect to the measured instantaneous optical frequency. We show that this technique dramatically improves the spatial resolution of a conventional OFDR and can be applied to an optical frequency-domain medical imaging system to eliminate the problem of a nonlinear frequency sweep effect.     

Broadband FUV imaging spectrometer: advanced design with a single toroidal uniform-line-space grating

Performances of a far-ultraviolet (FUV) imaging spectrometer in an advanced design are presented with a toroidal uniform-line-space (TULS) grating. It provides high spatial resolution and spectral resolution for a broadband and a wide field of view. A particular analysis for the grating aberrations, including all the high-order coefficients neglected by previous existing designs, was generated for indicating their significance. The analysis indicates that these high-order off-axis aberrations would have a remarkable influence on the design results. The transcendental equations composed of these aberration coefficients do not have analytic solutions in algebra. To solve the problem, the past designs always do some simplified calculation which only suits a narrow field of view and waveband. Thus, the optimization of the genetic algorithm is introduced to propose reasonable ranges of optical parameters. Then ZEMAX software is used to obtain the final optical system from these ranges. By comparing different design results of the same example, our advanced TULS design performs better than conventional TULS design and spherical varied-line-space grating design, and as well as the toroidal varied-line-space design. It is demonstrated that aberrations are minimized when the TULS design is operated by our method. The advanced design is low-cost, easy to fabricate, and more suitable for FUV observations.     

Imaging spectrograph for interstellar shocks: a narrowband imaging payload for the far ultraviolet

We present an imaging spectrometer developed for narrowband imaging at 1035 A with high (approximately 1-arc sec) spatial resolution over a modest field of view (approximately 5 arc min). The instrument is based on a conventional Gregorian telescope with aberration-corrected holographic rulings on the secondary optic. These aberration-correcting rulings enable stigmatic imaging in diffracted light with a minimum number of optical elements, thereby maintaining a high system efficiency. The capabilities of this instrument allow us to map the distribution of UV-emitting material in the hot (approximately 300,000 K) plasma from shocks in supernova remnants. Although this design is optimized for imaging near 1035 A, the basic concept can be applied to provide narrowband imaging or long-slit imaging spectroscopy at any wavelength. In addition, a larger field of view is possible with a corresponding loss in spatial resolution.     

High-spatial-resolution sub-surface imaging using a laser-based acoustic microscopy technique

Scanning acoustic microscopy techniques operating at frequencies in the gigahertz range are suitable for the elastic characterization and interior imaging of solid media with micrometer-scale spatial resolution. Acoustic wave propagation at these frequencies is strongly limited by energy losses, particularly from attenuation in the coupling media used to transmit ultrasound to a specimen, leading to a decrease in the depth in a specimen that can be interrogated. In this work, a laser-based acoustic microscopy technique is presented that uses a pulsed laser source for the generation of broadband acoustic waves and an optical interferometer for detection. The use of a 900-ps microchip pulsed laser facilitates the generation of acoustic waves with frequencies extending up to 1 GHz which allows for the resolution of micrometer-scale features in a specimen. Furthermore, the combination of optical generation and detection approaches eliminates the use of an ultrasonic coupling medium, and allows for elastic characterization and interior imaging at penetration depths on the order of several hundred micrometers. Experimental results illustrating the use of the laser-based acoustic microscopy technique for imaging micrometer-scale subsurface geometrical features in a 70-μm-thick single-crystal silicon wafer with a (100) orientation are presented.     

Short-coherence digital microscopy by use of a lensless holographic imaging system

An optical system based on short-coherence digital holography suitable for three-dimensional (3D) microscopic investigations is described. The light source is a short-coherence laser, and the holograms are recorded on a CCD sensor. The interference (hologram) occurs only when the path lengths of the reference and the object beam are matched within the coherence length of the laser. The image of the part of the sample that matches the reference beam is reconstructed by numerical evaluation of the hologram. The advantages of the method are high numerical aperture (this means high spatial resolution), detection of the 3D shape, and a lensless imaging system. Experimental results are presented.