Near-field optical microscope with a multiheight scanning imaging mode

A near-field scanning optical microscope has been developed to yield optical images with various gap distances between the probe and the sample surface. The microscope uses an apertureless metallic probe, the position of which is controlled by regulation of the tunneling-electron current from the probe to the sample and by computer-generated bias voltage. Experimental results of near-field optical imaging with the developed microscope at different gap distances are shown. Thirteen images at gap distances of 0 to 500nm demonstrate that the near-field image depends strongly on the gap distance. The imaging characteristics of a near-field imaging system are shown with the spatial-frequency spectra of images. Future investigation of the developed microscope is also discussed.     

多功能扫描质子核探针信号探测和数据获取系统

为了在一次扫描质子核探针实验中能获取多种物理信息,研制了多功能信号探测与数据获取系统。系统由组合探测器、多站多参量数据采集与束流扫描、样品台控制与显微观测三个子系统组成。组合探测器包括了Si(Li)X射线探测器,高纯Geγ射线探测器,以及Au(Si)面垒带电粒子探测器,可完成多种物理参数的同时获取。系统的核心数据采集与束流扫描系统基于NI公司的PXI-7852R数据采集卡和LabVIEW软件平台,具备了多个物理信号采集、多能谱显示以及束流扫描和二维成像等功能。六轴高精度真空样品台由计算机控制,可实现显微图像对样品的定位及对扫描区域的可视化选择。初步实验验证了该系统的可靠性与稳定性。     

Spectrally encoded confocal microscopy

An endoscope-compatible, submicrometer-resolution scanning confocal microscopy imaging system is presented. This approach, spectrally encoded confocal microscopy (SECM), uses a quasi-monochromatic light source and a transmission diffraction grating to detect the reflectivity simultaneously at multiple points along a transverse line within the sample. Since this method does not require fast spatial scanning within the probe, the equipment can be miniaturized and incorporated into a catheter or endoscope. Confocal images of an electron microscope grid were acquired with SECM to demonstrate the feasibility of this technique.     

Subdiffraction resolution in total internal reflection fluorescence microscopy with a grating substrate

We propose a fluorescence surface imaging system that presents a power of resolution beyond that of the diffraction limit without resorting to saturation effects or probe scanning. This is achieved by depositing the sample on an optimized periodically nanostructured substrate in a standard total internal reflection fluorescence microscope. The grating generates a high-spatial-frequency light grid that can be moved throughout the sample by changing the incident angle. An appropriate reconstruction procedure permits one to recover the fluorescence amplitude from the images obtained for various incidences. Simulations of this imaging system show that the resolution is not limited by diffraction but by the period of the grating.     

Microscopy refocusing and dark-field imaging by using a simple LED array

The condenser is one of the main components in most transmitted light compound microscopes. In this Letter, we show that such a condenser can be replaced by a programmable LED array to achieve greater imaging flexibility and functionality. Without mechanically scanning the sample or changing the microscope setup, the proposed approach can be used for dark-field imaging, bright-field imaging, microscopy sectioning, and digital refocusing. Images of a starfish embryo were acquired by using such an approach for demonstration.     

激光分光瞳共焦显微镜系统研制

为了满足生物类等样品对大工作距和高分辨率共焦显微镜的需求,将分光瞳技术与激光共焦显微技术结合应用到成像系统上。阐述了激光分光瞳共焦显微成像原理,首次成功搭建了相应的显微镜成像测量系统。理论分析和实验表明:分光瞳共焦显微技术独特的非共轴结构使系统的轴向分辨力是相同数值孔径物镜单轴系统的3倍以上,对理论高度为100nm的台阶样品进行成像测试,得到的样品三维形貌,成像质量良好。     

Multi-color miniature dual-axis confocal microscope for point-of-care pathology

We present a miniature microelectromechanical systems-based dual-axis confocal microscope capable of spatially coregistered fluorescence and reflectance imaging at multiple wavelengths. This device has a 10 mm diameter scan head with a 2 mm diameter tip for convenient use during surgery to guide tumor resection. The microscope has an adjustable focal depth of 20-200 micrometers and is capable of imaging with an axial resolution of 9 micrometers and in-plane resolution of 4 micrometers over a field of view of 450×450 micrometers. Simultaneous two-color imaging of individual optical sections is achieved by using a pair of grating-prism assemblies to compensate for chromatic dispersion in the 2 mm diameter gradient index relay lens at the distal tip of the device. Experimental measurements of the axial response of the microscope, as well as two-color images of a reflective bar target and fresh mouse brain tissues, demonstrate the performance of our device and its potential for multicolor in vivo optical sectioning microscopy.     

双成象单元扫描隧道显微镜与原子尺纳米计量技术

研制了双成象单元扫描隧道显微镜(STM),可同时对参考样品的原子晶格和被测样品扫描成象.计数原子晶格的数目,即可精确测定被测样品图象的尺度,以原子尺方式实现严格的纳米计量.本文介绍双成象单元的STM的原理和仪器系统,讨论原子尺纳米计量的可行性,给出被测样品图象的纳米计量结果.     

Wide-field coherent anti-Stokes Raman scattering microscopy with non-phase-matching illumination

We have developed and tested a wide-field coherent anti-Stokes Raman scattering (CARS) microscopy technique, which provides the simultaneous imaging of an extended illuminated area without scanning. This method is based on the non-phase-matching illumination of a sample and imaging of a CARS signal with a CCD camera using conventional microscope optics. We have identified a set of conditions on the illumination and imaging optics, as well as on sample preparation. Imaging of test objects proved high spatial resolution and chemical selectivity of this technique.     

Pixel super-resolution lensless in-line holographic microscope with hologram segmentation

We propose a resolution enhancement method for a lensless in-line holographic microscope(LIHM) by combining the hologram segmentation and pixel super-resolution(PSR) techniques. Our method is suitable for imaging specific target objects in samples, where the in-line hologram is disturbed by other objects in the samples. The resolution-enhancement capability of our method was proved by numerical simulations and imaging experiments while using a standard resolution target in a two-layer setup. We also applied our LIHM system to image the sample of living algae Euglena gracilis in water solution for further demonstration.     

利用扫描近场红外显微镜对化学样品组分进行成像研究

介绍了用北京自由电子激光为光源的扫描近场红外显微镜对化学样品组分进行的成像研究，通过所得到的近场微区图像，可以对样品在微区范围内的成分组成，混合的均匀程度等作出比较清晰的判断. 关键词： 自由电子激光 近场光学 扫描近场红外显微镜     

Tomographic imaging of a suspending single live cell using optical tweezer-combined full-field optical coherence tomography

We propose a label-free depth-resolved tomographic scheme for imaging a single live cell in fluid. This approach utilizes a modified time-domain full-field optical coherence tomography (FF-OCT) system combined with an optical tweezer technique. The optical trap for holding a moving specimen is made by tightly focusing a 1064 nm Q-switching pulsed laser beam with a 1.0 NA microscope objective in the sample arm of the FF-OCT part. By cosharing the probe for both systems, the optical actions of trapping and cellular resolution tomographic imaging could be achieved simultaneously. Feasibility of the combined system is demonstrated by imaging micron-sized polystyrene beads and a living suspension cell in medium.     

Time-resolved multispectral X-ray imaging with multi-channel Kirkpatrick-Baez microscope for plasma diagnostics at Shenguang-II laser facility

A time-resolved multispectral X-ray imaging approach with new version of multi-channel Kirkpatrick- Baez （KB） microscope is developed for laser plasma diagnostics at the kilo joule-class Shenguang-II laser facility （SG-II）. The microscope uses a total external reflection mirror in the sagittal direction and an array of multilayer mirrors in the tangential direction to obtain multiple individual high-resolution, high- throughput, and quasi-monochromatic X-ray images. The time evolution of the imploded target in multiple X-ray energy bands can be acquired when coupled with an X-ray streak camera. The experimental result of the time-resolved 2.5 and 3.0 keV dual-spectral self-emission imaging of the undoped CH shell target on SG-II is given.     

Pattern projection for subpixel resolved imaging in microscopy

In this paper, we present a new approach providing super resolved images exceeding the geometrical limitation given by the detector pixel size of the imaging camera. The concept involves the projection of periodic patterns on top of the sample, which are then investigated under a microscope. Combining spatial scanning together with proper digital post-processing algorithm yields the improved geometrical resolution enhancement. This new method is especially interesting for microscopic imaging when the resolution of the detector is lower than the resolution due to diffraction.     

Fresnel diffraction model for the point spread of a laser light profile microscope (LPM)

Light profile microscopy is a new method of thin film imaging in which a microscope forms an image of a laser beam intersecting a thin layer along its depth axis. Emission (luminescence or scatter) from the beam volume is transmitted through a cross sectional view surface and used by a microscope to form an image of the beam propagating through the sample. Fresnel diffraction theory is used in this work to derive a point spread model for a microscope operating in this configuration, assuming that the emission from the source beam is completely incoherent. This theory illustrates the dependence of the image resolution on the laser beam radius, which determines the effective thickness of the system object seen by the microscope. The effect of relative displacement of the source beam center from the object plane of the microscope is evaluated. The present theory shows that radial resolution may be evaluated by means of an object focus envelope and a focal energy distribution computed for the objective lens. PACS 42.25Fx; 07.60Pb     

Numerical correction of coherence gate in full-field swept-source interference microscopy

A big problem in low-coherence interference microscopy is the degradation of the coherence signal caused by shift of the angular and temporal spectrum gates. It limits the depth of field in confocal optical coherence microscopy and degrades images of sample inner structure in most interference microscopy techniques. To overcome this problem we propose numerical correction of the coherence gate in application to full-field swept-source interference microscopy. The proposed technique allows three-dimensional sample imaging without mechanical movement of the microscope components and is also capable of determining separately the geometrical thickness and the refractive index of the sample layers, when the sample contains a transversal pattern. The applicability of the proposed technique is verified with numerical simulation.     

HeLa细胞突起中微丝束的纳米分辨荧光成像

在Matlab编程环境下模拟了单分子定位显微的纳米分辨成像,在传统实验参数条件下,对不同间隔分子带模型进行了模拟成像.模拟结果表明,单分子定位显微方法可以区分中心相隔20 nm的两个分子带.同时,也分析了不同像元大小对单分子定位精度的影响.此外,通过单分子定位显微方法在IX71倒置荧光显微镜上实现了纳米分辨,系统极限分辨率,即半高全宽为48 nm.在该系统上,获得了HeLa细胞突起中微丝束结构的纳米分辨图像.从重构获得的图像中可以看到微丝束的直径为75—200 nm.     

KBA显微镜成像系统的成像模拟及X光成像实验的研究

KBA显微镜是一种非轴对称、非共轴的掠入射成像系统。其结构复杂,调节精度要求很高,在实际成像实验操作中难以掌握其成像特性。利用光学设计软件模拟其成像,对系统的调节和成像分析提供有益的参考。利用光学设计软件ZEMAX模拟了KBA显微镜对点源的成像过程,给出了KBA显微镜成像系统的焦深约为1 mm,景深为50 mm左右。并且由模拟可知,掠入射角对成像的影响很大。对像素尺寸约10μm的探测设备,模拟得出KBA成像系统的空间分辨力上限为3μm左右。基于星光Ⅱ装置对周期为20μm的网格靶成像,获得了KBA显微镜较为清晰的X光图像。该项工作为进一步开展掠入射成像系统的研究奠定了基础。     

Programmable Colored Illumination Microscopy (PCIM): A practical and flexible optical staining approach for microscopic contrast enhancement

Programmable colored illumination microscopy (PCIM) has been proposed as a flexible optical staining technique for microscopic contrast enhancement. In this method, we replace the condenser diaphragm of a conventional microscope with a programmable thin film transistor-liquid crystal display (TFT-LCD). By displaying different patterns on the LCD, numerous established imaging modalities can be realized, such as bright field, dark field, phase contrast, oblique illumination, and Rheinberg illuminations, which conventionally rely on intricate alterations in the respective microscope setups. Furthermore, the ease of modulating both the color and the intensity distribution at the aperture of the condenser opens the possibility to combine multiple microscopic techniques, or even realize completely new methods for optical color contrast staining, such as iridescent dark-field and iridescent phase-contrast imaging. The versatility and effectiveness of PCIM is demonstrated by imaging of several transparent colorless specimens, such as unstained lung cancer cells, diatom, textile fibers, and a cryosection of mouse kidney. Finally, the potentialities of PCIM for RGB-splitting imaging with stained samples are also explored by imaging stained red blood cells and a histological section.     

深紫外激光光发射与热发射电子显微镜在热扩散阴极研究中的应用

对热扩散阴极表面微区发射状态进行原位观察和分析一直是热阴极研究的重要课题.本文着重介绍深紫外激光光发射电子/热发射电子显微镜的基本原理及其在热扩散阴极研究中的典型实例.系统配备了高温激活所用的加热装置,样品可被加热至1400℃.系统具有光发射电子、阴极热发射电子、光发射电子和阴极热发射电子联合三种电子成像模式.应用表明,对于热扩散阴极而言,深紫外激光光发射电子像适于呈现阴极表面的微观结构形貌;热发射电子像适于反映阴极表面的本征热电子发射及均匀性;光电子和热电子联合成像适于对阴极表面的有效发射点做出精确定位.