PHOEBE,a prototype scanning laser-feedback microscope for imaging biological cells in aqueous media

Based on the principle of laser-feedback interferometry (LFI), a laser-feedback microscope (LFM) has been constructed capable of providing an axial (z) resolution of a target surface topography of ～ 1 nm and a lateral (x, y) resolution of ～ 200 nm when used with a high-numerical-aperture oil-immersion microscope objective. LFI is a form of interferometry in which a laser's intensity is modulated by light re-entering the illuminating laser. Interfering with the light circulating in the laser resonant cavity, this back-reflected light gives information about an object's position and reflectivity. Using a 1-mW He–Ne (λ = 632·8 nm) laser, this microscope (PHOEBE) is capable of obtaining 256 × 256-pixel images over fields from (10 μm × 10 μm) to (120 μm × 120 μm) in ～ 30 s. An electromechanical feedback circuit holds the optical pathlength between the laser output mirror and a point on the scanned object constant; this allows two types of images (surface topography and surface reflectivity) to be obtained simultaneously. For biological cells, imaging can be accomplished using back-reflected light originating from small refractive-index changes (> 0·02) at cell membrane/water interfaces; alternatively, the optical pathlength through the cell interior can be measured point-by-point by growing or placing a cell suspension on a higher-reflecting substrate (glass or a silicon wafer). Advantages of the laser-feedback microscope in comparison to other confocal optical microscopes include: the simplicity of the single-axis interferometric design; the confocal property of the laser-feedback microscope (a virtual pinhole), which is achieved by the requirement that only light that re-enters the laser meeting the stringent frequency, spatial (TEM₀₀), and coherence requirements of the laser cavity resonator mode modulate the laser intensity; and the improved axial resolution, which is based on interferometric measurement of optical amplitude and phase rather than by use of a pinhole as in other types of confocal microscopes.     

Evaluation of nano-optical probe from scanning near-field optical microscope images

We studied a nanometre-sized optical probe in a scanning near-field optical microscope. The probe profile is determined by using a knife-edge method and a modulated transfer function evaluation method which uses nanometre-sized line-and-space tungsten patterns (with spaces 1 μm to 50 nm apart) on SiO₂ substrates. The aluminium-covered, pipette-pulled fibre probe used here has two optical probes: one with a large diameter (350 nm) and the other with a small diameter (10 nm). The small-diameter probe has an optical intensity ≈63 times larger than that of the large-diameter probe, but the power is about 1/25 of that of the large probe.     

Apparatus for vectorial Kerr confocal microscopy

We present a confocal microscopy setup that is able to record magneto-optical hysteresis cycles separating the in-plane and out-of-plane magnetization components. This apparatus is based on a modified commercial microscope, where the light beam has been deviated from the cylindrical symmetry axis of the objective lenses by inserting a translating plate in the optical path. The instrument allows for the magneto-optical imaging with a lateral resolution of 600 nm at λ = 635 nm light wavelength.     

Effects of defocus and algorithm on optical step height calibration

Defocus effects on step height measurements by interferometric microscopy are estimated using different algorithms to calculate the step height. The interferometric microscope here is a Mirau-type with a 20x objective and a numerical aperture (NA) of 0.4. Although the focus is adjusted within the range of the depth of focus, a defocus corresponding to 4 fringes (1.3 μm) distorts the measured profile into a curve with a radius of curvature of about 24 m. The effect of this distortion on step height determination by a basic ISO step height algorithm is estimated to be 0.11 nm/fringe. An algorithm developed by NIST and a second-order fitted ISO step-height algorithm show good robustness against defocus errors. Because of sample imperfection, a discrepancy of about 1 % exists among step-height values determined by the different algorithms at the optimum fringe contrast position. It has been pointed out theoretically that the NA correction factor varies with the difference of optical path length between the sample and the reference surface. However, in our case, the changes of optical path length do not change the NA correction factor by more than 0.3 %.     

Are electron tweezers possible?

Positively answering the question in the title, we demonstrate in this work single electron beam trapping and steering of 20–300 nm solid Al nanoparticles generated inside opaque submicron-sized molten Al–Si eutectic alloy spheres. Imaging of solid nanoparticles and liquid alloy in real time was performed using energy filtering in an analytical transmission electron microscope (TEM). Energy-filtering TEM combined with valence electron energy-loss spectroscopy enabled us to investigate in situ nanoscale transformations of the internal structure, temperature dependence of plasmon losses, and local electronic and optical properties under melting and crystallization of individual binary alloy particles. For particles below 20 nm in size, enhanced vibrations of the dynamic solid–liquid interface due to instabilities near the critical threshold were observed just before melting. The obtained results indicate that focused electron beams can act as a tool for manipulation of metal nanoparticles by transferring linear and angular mechanical momenta. Such thermally assisted electron tweezers can be utilized for touchless manipulation and processing of individual nano-objects and potentially for fabrication of assembled nanodevices with atomic level sensitivity and lateral resolution provided by modern electron optical systems. This is by three orders of magnitude better than for light microscopy utilized in conventional optical tweezers. New research directions and potential applications of trapping and tracking of nano-objects by focused electron beams are outlined.     

光学隐蔽深度测量系统的小型化设计

为了保障航行器水下航行时对可见光波段光学隐蔽的需求,以及实时测量其所在位置的光学隐蔽深度,设计了小型化可见光波段的光学隐蔽深度测量系统。根据光学隐蔽深度模型,优化设计了海水上行辐照度、海水下行辐照度、海水漫衰减系数、海水体衰减系数的测量方法。优化设计后的系统有21个测量通道,测量光谱为390~667nm,工作深度可达50m。进行了海上试验,试验结果显示:在天气良好的条件下,水下航行器在520~560nm可见光波段内存在暴露窗口,同时测得特征长度为0.75m的模型的光学隐蔽深度为3.5m。得到的结果表明,设计的小型化可见光波段光学隐蔽深度测量系统可以实现光学隐蔽深度的测量,测量装置具有体积小、重量轻、系统性能稳定等优点,适用于水下航行器搭载,可为下一步实现可自主升降的光学隐蔽深度测量系统设计提供理论技术支撑。     

采用多层膜偏光器的EUV实验室Farady旋光测量装置

开发了用于实验室的ＥＵＶ（５０～７０ｅＶ）波段的Ｆａｒａｄａｙ 旋光测量装置。其中光源采用了激光等离子体光源。起偏器和检偏器均采用了入射角为准Ｂｒｅｗ ｓｔｅｒ角的反射式Ａｌ／ＹＢ６多层膜。利用４ 块Ｓｍ －Ｃｏ 永久磁铁在被测样品处生成了０．８２Ｔ的磁场。对强磁材料Ｎｉ的Ｍ２,３吸收端进行了Ｆａｒａｄａｙ 旋光测量,测量结果在６５．３ｅＶ处为－ ２．７°／２１ｎｍ 。该结果与我们在同步辐射光源上测量的结果相符合。这是ＥＵＶ 波段首次利用激光等离子体光源成功进行的磁光效应测量实验。     

激光共振电离质谱计用光入射系统的研制

针对NINT3000质谱计的结构特点,利用高斯光学理论,推导出双透镜系统光学参数传递方程,并以此为依据设计出一套光学系统和光路调整装置,实现了激光束在离子源腔内的精确定位和聚集,焦斑半径为160μm,调节精度为10μm。目前,该系统已经安装在激光共振电离质谱计上,并实现了镥的单色双光子共振电离。     

Optical fringe effects at prism borders in human tooth enamel sections

When ground sections of mammalian teeth are viewed under the light microscope each enamel prism appears to be surrounded by a prism sheath about 0.5 μm] wide. Seen by the electron microscope these sheaths are at most 0.1 μm wide. Because this dimension is beyond the resolving power of the light microscope it is difficult to explain the origin of the optical image. Optical fringes are frequently seen at the borders of transversely cut enamel prisms. The widths of these fringes have been measured under different optical conditions. It is concluded that the fringes are produced due to interference between direct light rays and those reflected from prism borders. A requisite for clear fringe production is that the reflectance of the prism border should be high. This high reflectance could only be achieved if prisms are separated from each other by material having a refractive index which differs significantly from that of the prisms themselves. It is therefore suggested that prisms are separated by a protein layer. It is then possible to explain why prism sheaths appear 0.5 μm wide under the light microscope. Study of mammalian tooth enamel by light microscopy suggests to the observer that the 5 μm wide rods (generally referred to as prisms) which constitute the enamel are each partially surrounded by a ‘sheath’ about 0.5 μm wide. Micro-radiographs indicate that the sheath is considerably less mineralized than the body of the prism (Glas, 1965). In contrast, from direct observation of enamel sections by electron microscopy, it has been concluded that no prism sheath exists and that prisms are separated by an interface which is bordered on its two sides by differently orientated crystals (Helmcke 1960, 1967; Ronnholm, 1962; Meckel, Griebstein & Neal, 1965). Between these two extremes are some electron microscope observations appearing to demonstrate the presence of a 0.1 μm wide membrane-like prism sheath devoid of inorganic crystals (Frank & Nalbandian, 1967) or an irregular region of microporosity where the different orientation of adjacent crystals results in a packing defect (Johnson, 1967). Any enamel section observed with the electron microscope may well have been distorted during its preparation for examination. Shrinkage due to dehydration is one of the most likely artifacts. Because the amount of the distortion cannot be known with accuracy it is difficult to decide which of the above electron-microscope appearances is closest to the actual structure of the prism boundary during life. It has recently been observed that when viewed end-on, the borders of prisms frequently appear as light striations (Osborn, 1968 and in press). In the present investigation the widths of these light striations have been measured under different optical conditions on an enamel section which was maintained in a hydrated condition. It was thought that this data on the optical properties of the hydrated prism boundary could be used to predict the structure of the boundary in vivo. A similar shrinkage is to be expected in sections examined with the light microscope because these have usually been dehydrated prior to mounting. It is therefore reasonable to try to relate the 0.5 μm wide optical image seen with the light microscope to the two different structures observed with the electron microscope. If no prism sheath exists it might be argued that the refractive index variation between the borders of adjacent prisms could in some way account for the production of the optical image. However, because this variation is probably less than 0.02 (Fremlin & Mathieson 1964) it is not clear how it could be used to explain the high contrast of the optical image. Furthermore, even if a 0.1 μm wide prism sheath exists, it is significantly thinner than the wavelength of visible light; again it is not clear how its presence results in the production of an image in the light microscope.     

基于LabVIEW的光纤傅里叶变换光谱仪数据处理技术

为了提高编程效率与增强光谱仪数据分析处理能力,结合LabVIEW软件在信号处理方面的优势,利用虚拟仪器技术实现了光纤傅里叶变换光谱仪干涉图处理与光谱复原。采用均匀抽样算法,利用由光纤Mach-Zehnder干涉仪输出的参考光干涉图过零点取样,实现对测试光干涉图的等光程间隔采样,消除了由压电陶瓷非线性光程调制引起的误差。采用求摸法复原出光谱图,自动消除了相位误差。利用LabVIEW软件完成了对宽带ASE光源的干涉图数据采集、显示与处理并最终得到光谱图。结果表明,与光栅光谱仪测得的光谱相比,光谱线型趋于一致。     

Evaluation of nano-optical probe from scanning near-field optical microscope images

We studied a nanometre-sized optical probe in a scanning near-field optical microscope. The probe profile is determined by using a knife-edge method and a modulated transfer function evaluation method which uses nanometre-sized line-and-space tungsten patterns (with spaces 1 microm to 50 nm apart) on SiO2 substrates. The aluminium-covered, pipette-pulled fibre probe used here has two optical probes: one with a large diameter (350 nm) and the other with a small diameter (10 nm). The small-diameter probe has an optical intensity approximately 63 times larger than that of the large-diameter probe, but the power is about 1/25 of that of the large probe.     

Scanning near-field cathodoluminescence investigations

By implementing a scanning near-field optical microscope into the analysis chamber of a scanning electron microscope, the light emitted due to cathodoluminescence can be locally detected in the near-field using tapered, coated optical fibers. In addition to the ability to perform contactless measurements of local diffusion lengths, the achievable spatial resolution can be increased to about 50 nanometers.     

Towards phonon photonics: scattering-type near-field optical microscopy reveals phonon-enhanced near-field interaction

Diffraction limits the spatial resolution in classical microscopy or the dimensions of optical circuits to about half the illumination wavelength. Scanning near-field microscopy can overcome this limitation by exploiting the evanescent near fields existing close to any illuminated object. We use a scattering-type near-field optical microscope (s-SNOM) that uses the illuminated metal tip of an atomic force microscope (AFM) to act as scattering near-field probe. The presented images are direct evidence that the s-SNOM enables optical imaging at a spatial resolution on a 10 nm scale, independent of the wavelength used (λ=633 nm and 10 μm). Operating the microscope at specific mid-infrared frequencies we found a tip-induced phonon-polariton resonance on flat polar crystals such as SiC and Si₃N₄. Being a spectral fingerprint of any polar material such phonon-enhanced near-field interaction has enormous applicability in nondestructive, material-specific infrared microscopy at nanoscale resolution. The potential of s-SNOM to study eigenfields of surface polaritons in nanostructures opens the door to the development of phonon photonics—a proposed infrared nanotechnology that uses localized or propagating surface phonon polaritons for probing, manipulating and guiding infrared light in nanoscale devices, analogous to plasmon photonics.     

The effect of swelling on the interference microscopy of fixed biological material

Measurements with the interference microscope show the optical-path difference relative to the medium of many biological objects after histological processing to be less in water than would be expected on the basis of measurements made in other media. This anomalous optical-path difference is found also with sections of gelatin (autoradiographic stripping film), but not if the gelatin is mounted on the slide in such a way that swelling is constrained to a direction parallel to the optical axis of the microscope. Swelling in water has been demonstrated in a variety of formalin-fixed and de-fatted tissues. It is suggested that the anomalous optical-path differences in water are due to swelling causing a change in the lateral dimensions of the object, and hence loss of material from the optical path. The fact that the anomaly has also been found with scanning and integrating interference microscopes is discussed, and it is suggested that the validity of such methods needs re-examination.     

光纤熔接机高清显微物镜光学系统设计

在光纤熔接过程中,为了实现光纤高质量熔接,需要一个高清显微物镜来确保纤芯的准确对准。运用Zemax软件设计一款用于光纤纤芯对准的显微物镜,该物镜由6片透镜组成,放大率为8倍,数值孔径为0.25,工作距为13.4mm,共轭距为85mm,以CCD作为图像接收器件。显微物镜采用正向光路进行优化设计,正向光路设计的显微物镜更能贴近实际使用状态,能够更加清晰准确地检测到纤芯位置。该物镜工作波长为486~656nm,具有工作距离长、共轭距短、精度高等特点。     

Integration of a high‐NA light microscope in a scanning electron microscope

We present an integrated light‐electron microscope in which an inverted high‐NA objective lens is positioned inside a scanning electron microscope (SEM). The SEM objective lens and the light objective lens have a common axis and focal plane, allowing high‐resolution optical microscopy and scanning electron microscopy on the same area of a sample simultaneously. Components for light illumination and detection can be mounted outside the vacuum, enabling flexibility in the construction of the light microscope. The light objective lens can be positioned underneath the SEM objective lens during operation for sub‐10 μm alignment of the fields of view of the light and electron microscopes. We demonstrate in situ epifluorescence microscopy in the SEM with a numerical aperture of 1.4 using vacuum‐compatible immersion oil. For a 40‐nm‐diameter fluorescent polymer nanoparticle, an intensity profile with a FWHM of 380 nm is measured whereas the SEM performance is uncompromised. The integrated instrument may offer new possibilities for correlative light and electron microscopy in the life sciences as well as in physics and chemistry.     

Comparing electron tomography and HRTEM slicing methods as tools to measure the thickness of nanoparticles

Nanoparticles’ morphology is a key parameter in the understanding of their thermodynamical, optical, magnetic and catalytic properties. In general, nanoparticles, observed in transmission electron microscopy (TEM), are viewed in projection so that the determination of their thickness (along the projection direction) with respect to their projected lateral size is highly questionable. To date, the widely used methods to measure nanoparticles thickness in a transmission electron microscope are to use cross-section images or focal series in high-resolution transmission electron microscopy imaging (HRTEM “slicing”). In this paper, we compare the focal series method with the electron tomography method to show that both techniques yield similar particle thickness in a range of size from 1 to 5 nm, but the electron tomography method provides better statistics since more particles can be analyzed at one time. For this purpose, we have compared, on the same samples, the nanoparticles thickness measurements obtained from focal series with the ones determined from cross-section profiles of tomograms (tomogram slicing) perpendicular to the plane of the substrate supporting the nanoparticles. The methodology is finally applied to the comparison of CoPt nanoparticles annealed ex situ at two different temperatures to illustrate the accuracy of the techniques in detecting small particle thickness changes.     

Analysis of fluorescence from algae fossils of the Neoproterozoic Doushantuo formation of China by confocal laser scanning microscope

Chinese algae fossils can provide unique information about the evolution of the early life. Thin sections of Neoproterozoic algae fossils, from Guizhou, China, were studied by confocal laser scanning microscopy, and algae fossils were fluorescenced at different wavelengths when excited by laser light of 488 nm, 476 nm, and 568 nm wavelength. When illuminated by 488 nm laser light, images of the algae fossils were sharper and better defined than when illuminated by 476 nm and 568 nm laser light. The algae fossils fluoresce at a wide range of emission wavelengths. The three-dimensional images of the fluorescent algae fossils were compared with the transmission images taken by light microscope. We found that the fluorescence image of the confocal laser scanning microscope in a single optical section could pass for the transmission image taken by a light microscope. We collected images at different sample depths and made a three-dimensional reconstruction of the algae fossils. And on the basis of the reconstruction of the three-dimensional fluorescent images, we conclude that the two algae fossils in our present study are red algae.     

In situ multipurpose time-resolved spectrometer for monitoring nanoparticle generation in a high-pressure fluid

We developed a multipurpose time-resolved spectrometer for studying the dynamics of nanoparticles generated by pulsed-laser ablation (PLA) in a high-pressure fluid. The apparatus consists of a high-pressure optical cell and three spectrometers for in situ measurements. The optical cell was designed for experiments at temperatures up to 400 K and pressures up to 30 MPa with fluctuations within ±0.1% h(-1). The three spectrometers were used for the following in situ measurements at high pressures: (i) transient absorption spectrum measurements from 350 to 850 nm to investigate the dynamics of nanoparticle generation from nanoseconds to milliseconds after laser irradiation, (ii) absorption spectrum measurements from 220 to 900 nm to observe the time evolution of nanoparticles from seconds to hours after laser ablation, and (iii) dynamic light scattering measurements to track nanoparticles with sizes from 10 nm to 10 μm in the time range from seconds to hours after laser ablation. By combining these three spectrometers, we demonstrate in situ measurements of gold nanoparticles generated by PLA in supercritical fluids. This is the first report of in situ time-resolved measurements of the dynamics of nanoparticles generated in a supercritical fluid.     

真空紫外辐照对Lumogen薄膜损伤及光学性能的影响

为研究Lumogen(C22H16N2O6)薄膜在真空紫外波段的光致发光特性及辐照损伤,采用热阻蒸发法,以氟化镁为基底制备Lumogen薄膜.使用真空紫外荧光光谱仪、原子力显微镜(AFM)、扫描电子显微镜(SEM)、紫外?可见分光光度计等仪器分别对薄膜的光致发光特性、荧光强度衰减变化、表面形貌、透过率等进行测试与表征....