A confocal beam scanning white-light microscope

We report on a confocal beam scanning microscope utilizing a continuous Xe short-arc lamp operating in the visible spectrum with unprecedented radiance. Measurements of lateral and vertical resolution will be presented and compared with those of an equivalent scanning laser microscope. Resolution of the white-light microscope is equivalent to that of the scanning laser microscope. White-light microscope images positively stand out from those of the scanning laser microscope by their lack of artefacts caused by interference.     

A hybrid scanning force and light microscope for surface imaging and three-dimensional optical sectioning in differential interference contrast

The design of a scanned-cantilever-type force microscope is presented which is fully integrated into an inverted high-resolution video-enhanced light microscope. This set-up allows us to acquire thin optical sections in differential interference contrast (DIC) or polarization while the force microscope is in place. Such a hybrid microscope provides a unique platform to study how cell surface properties determine, or are affected by, the three-dimensional dynamic organization inside the living cell. The hybrid microscope presented in this paper has proven reliable and versatile for biological applications. It is the only instrument that can image a specimen by force microscopy and high-power DIC without having either to translate the specimen or to remove the force microscope. Adaptation of the design features could greatly enhance the suitability of other force microscopes for biological work.     

Real-time image quality assessment with mixed Lagrange time delay estimation autoregressive (MLTDEAR) model

A proposal to assess the quality of scanning electron microscope images using mixed Lagrange time delay estimation technique is presented. With optimal scanning electron microscope scan rate information, online images can be quantified and improved. The online quality assessment technique is embedded onto a scanning electron microscope frame grabber card for real‐time image processing. Different images are captured using scanning electron microscope and a database is built to optimally choose filter parameters. An optimum choice of filter parameters is obtained. With the optimum choice of scan rate, noise can be removed from real‐time scanning electron microscope images without causing any sample contamination or increasing scanning time.     

LED光源和CCD在荧光显微镜上的应用研究

荧光显微镜是常用的医用光学仪器之一,分布在各大医院科研、病理、检验等部门。光源是荧光显微镜的主要组成部分,使用新型的LED光源替代传统的高压汞灯,以及把数码相机改为CCD摄像装置,使现有的荧光显微镜上了一个等级,达到现代荧光显微镜的水平。     

Measurement of cellular elastic properties by acoustic microscopy

The acoustic microscope is used to investigate the elastic properties of living biological cells. A quantitative model is developed relating acoustic microscope image contrast to cellular elastic properties. Cytoplasmic acoustic attentuation is measured by focusing the acoustic microscope on the surface of the underlying substrate. Cytoplasmic acoustic impedance is measured by focusing the acoustic microscope on the top surface of the cell. The model allows the acoustic microscope to give quantitative information about cellular elasticity on a subcellular scale.     

Quantitative DNA measurements in an instrument combining scanning electron microscopy and light microscopy

An instrument for combined scanning electron microscopy (SEM) and light microscopy (LM) to which a photometer unit is attached is described. A special stage in the vacuum chamber of a scanning electron microscope incorporates light microscope optics (objective and condenser) designed for transmission and epi-illumination fluorescence LM. An optical bridge connects these optics to a light microscope, without objective and condenser. The possibility of performing quantitative DNA measurements in this combined microscope (the LM/SEM) was tested using preparations of either chicken erythrocytes, human lymphocytes, or mouse liver cells. The cells were fixed, brought on a cover-glass, quantitatively stained for DNA, dehydrated, and critical point dried (CPD). After mounting the cells were coated with gold. The specimens were brought into the vacuum chamber of the combined microscope and individual cells were studied with SEM and LM. Simultaneously DNA measurements were performed by means of the photometer unit attached to the microscope. It is shown in this study that DNA measurements of cells in the combined microscope give similar results when compared to DNA measurements of embedded cells performed with a conventional fluorescence microscope. Furthermore, it is shown that although the gold layer covering the LM/SEM specimens weakens the fluorescence signal, it does not interfere with the DNA measurements.     

A versatile multipurpose scanning probe microscope

A combined scanning probe microscope has been developed that allows simultaneous operation as a non‐contact/tapping mode atomic force microscope, a scattering near‐field optical microscope, and a scanning tunnelling microscope on conductive samples. The instrument is based on a commercial optical microscope. It operates with etched tungsten tips and exploits a tuning fork detection system for tip/sample distance control. The system has been tested on a p‐doped silicon substrate with aluminium depositions, being able to discriminate the two materials by the electrical and optical images with a lateral resolution of 130 nm.     

傅立叶变换红外显微镜新技术及应用

FT-IR显微镜技术是采用FT-IR光谱仪与光学显微镜联用的新型系统。红外显微镜技术是一种非破坏性技术,仅需要少量的样品,这些优势使得红外显微镜被理想地用于数量有限的样品。这里将论述红外显微镜对一种特殊法庭样品一头发纤维的分析。红外显微镜对于法庭科学是一个很重要的工具。     

Unique laser-scanning optical microscope for low-temperature imaging and spectroscopy

Low-temperature optical characterization of single quantum nanostructures can reveal detailed information on structure-dependent properties of these materials. We describe the development of a unique laser-scanning optical microscope capable of low-temperature single molecule/particle imaging and spectroscopy. Making use of the magnification of a microscope objective, the laser- scanning scheme of the present microscope allows for high-repeatability imaging over large sample areas. The microscope is utilized to measure the low-temperature Raman scattering spectra of individual single-walled carbon nanotubes and single molecule fluorescence spectra of conjugated polymers. The developed low-temperature microscope can be applied to study a wide array of nanomaterials at a single particle level.     

Design and performance of a high-resolution photoelectron microscope

The design of a high-resolution photoelectron microscope (photoelectron emission microscope) is described. It is an oil-free ultrahigh-vacuum instrument utilizing electrostatic electron optics. New designs are presented for a specimen translator, cathode stage, aperture stop control, electrostatic hexapole stigmator, beam shutter, and camera system. These components could also be used in a low-energy electron microscope (LEEM). The theoretical resolution of this instrument is 5 nm for UV illumination near the photoemission threshold. The photoelectron microscope is now in operation at the University of Oregon, and it is achieving results within a factor of two of this design limit.     

A novel high-throughput scanning microscope for label-free detection of protein and small-molecule chemical microarrays

We describe a novel scanning optical microscope based on a polarization-modulated nulling ellipsometry. The new microscope employs a combination of scanning mirror and sample translation and thus enables high-throughput label-free detection of biomolecular microarrays with more than 10 000 protein or small-molecule targets. For illustration, we show the image of a 2760-spot protein microarray on a functionalized glass slide obtained with such a microscope. The new scanning microscope is also capable of determining, in parallel, the real-time binding kinetics of multiple molecular species under aqueous conditions.     

Reflected-light differential-interference microscopy: principles,use and image interpretation

The reflected-light differential-interference microscope is a simple optical instrument easy to use and adjust, which reveals surface contours in variations of light intensity or in colour with a clarity and sensitivity not previously achieved. It can reveal new structures or display familiar tissue and cellular architecture in new and refreshing aspects. This type of microscope can provide insights into surface characteristics in metallurgy, microelectronics, biology, etc. Understanding how the differential-interference microscope works, how to control it and how to interpret the images produced, is essential to using the microscope fruitfully.     

Novel instrument for surface plasmon polariton tracking in space and time

We describe the realization of a phase-sensitive and ultrafast near-field microscope, optimized for investigation of surface plasmon polariton propagation. The apparatus consists of a homebuilt near-field microscope that is incorporated in Mach-Zehnder-type interferometer which enables heterodyne detection. We show that this microscope is able to measure dynamical properties of both photonic and plasmonic systems with phase sensitivity.     

A compact sensor head for simultaneous scanning force and near-field optical microscopy

A compact sensor head based on scanning force microscopy (SFM) using cantilever probes has been developed. The idea is to replace the microscope objective of a conventional optical microscope by this compact module and turn the optical microscope into a scanning force and near-field optical microscope with subwavelength resolution. We describe our concept and present initial results showing images of the object’s optical properties and surface topography recorded simultaneously.     

生物显微镜变倍物镜的光学设计

生物显微镜也可以像体视显微镜一样进行连续变倍观察,提出了生物显微镜变倍物镜的光学设计方法,介绍了设计前要求考虑的各项技术指标,以解决物镜像质和结构紧凑之间的矛盾,最终给出了一个变倍比达到10:1的生物显微镜变倍物镜的设计实例.     

软X射线显微术和光谱显微术

X射线显微术可直接在水环境中对胶体颗粒尺寸范围内的颗粒进行高分辨率成像,将该项技术与高分辨率光谱相结合,还可用于光谱显微研究。其中,常用的两种X射线显微镜是透射显微镜和扫描透射显微镜,文中示出了它们的装置图。由于X射线显微镜能迅速拍下一物体的高分辨率图像,所以,作为一种分析仪器,扫描X射线显微镜更适合作光谱显微研究。作为形态学目视化的一个示例,本文用一台透射X射线显微镜拍摄了粘土和土壤样品的图像。根据X射线图像进行的低温层析实验所获得的图像得到了有关细菌构成的显微生存环境以及其它土壤胶状体的3D结构信息。对扫描透射X射线显微镜拍摄的一系列图像进行分析,得到了土壤样品的形貌特性和化学特性。     

现代显微成像技术综述

概述了光学宽视场显微镜、共聚焦显微镜、超分辨率显微镜中所应用的现代显微成像技术,对各种传统和先进的显微成像原理进行了总结。光学宽视场显微镜最常用的显微技术有明场成像、暗场成像、相衬成像、偏光成像、微分干涉(DIC)成像、调制对比成像和荧光成像。相衬成像中根据不同的成像结构还有切趾相衬成像。微分干涉除了传统的偏振光照明还有圆偏振光照明(C-DIC)和专用于塑料的微分干涉(PlasDIC)。共聚焦显微镜随着计算机技术和制造技术的发展而有了巨大的发展。除了传统的共聚焦荧光显微镜以外,还有连续反斯托克斯拉曼散射(CARS)共聚焦、多光子共聚焦和白光共聚焦。超分辨率显微镜中主要介绍了受激辐射淬灭(STED)技术和紧随基态淬灭显微技术的单分子返回(GSDIM)技术。     

Zero-loss energy filtering under low-dose conditions using a post-column energy filter

Electron cryomicroscopy combined with energy filtering can be performed under low-dose conditions using a post-column energy filter. The microscope combined with the filter is set up such that it can be used with similar ease as a conventional microscope, the main difference being that all filter and microscope control is performed through a central computer and images are recorded with a cooled slow-scan CCD camera. The microscope can also still be used for regular imaging on film as without the filter. Owing to the 18 times post-magnification of the filter, the microscope normally has to be operated at a small magnification, e.g. 3000×, and the beam has to be contracted to a small spot, e.g. 5 mm, in the plane of the microscope viewing screen. Computer control allows one to perform a variety of tasks automatically, such as autofocusing, thickness measurements, most-probable-loss imaging, CCD spot-scanning and tomography. The gain in contrast due to zero-loss energy filtering is analysed using visual inspection, power spectra and Fourier ring correlation. The thickness range for ice-embedded specimens in which a filter at 120 kV is most useful appears to be between 100 and 300 nm.     

Angular distribution of polarized photon-pairs in a scattering medium with a Zeeman laser scanning confocal microscope

A novel confocal microscope designed for use with turbid media is proposed. We use a Zeeman laser as the light source. Based on the properties of two‐frequency polarized photon‐pairs and the common‐path feature of polarized photon‐pairs with heterodyne detection employed in the proposed confocal microscope, three gatings (spatial filtering gating, polarization gating and spatial coherence gating) are thus simultaneously incorporated in the microscope. Experimental results for the angular distribution of polarized photon‐pairs in a scattering medium indicate that polarization gating and spatial coherence gating preclude the detection of multiply scattered photons, whereas the pinhole selects the least scattered photon‐pairs. Thus, better performance for axial resolution than can be obtained with a conventional confocal microscope is demonstrated experimentally. In addition, the proposed microscope is able to either look deeper into a turbid medium or work with a denser medium; furthermore, the axial resolution is improved.