用家用数码照相机直接经显微镜目镜摄取高质量的数码图像

在生物学和医学的科研、教学中，往往需要使用生物显微镜和解剖镜。而与之相配套的照相装置，是保存、发表和教学展示光学显微图像的必备设备。一般的显微照相装置需要配有专用的光学适配装置，用于特定型号的显微镜，拥有成套设备的成本较高。本文探讨了使用普通家用数码照相机直接经生物显微镜目镜，摄取高质量、高分辨率数码光学显微图像的可行性和影响因素。     

自制"显微镜照相云台"

<生物学通报>(2007-12.P49)登载曹西南等的"用家用数码照相机直接经显微镜目镜摄取高质量的数码图像".该文阐述了可用家用数码照相机从显微镜目镜摄取高质量的数码图像.     

数码摄像机在显微镜教学中的应用

利用数码摄像机将显微镜内的显微图像转变成视频信号,再传输到电视机或电脑中,通过电视机或投影机银幕展示给学生观察,实现非数码显微镜关于显微图像的演示和贮存功能。     

用于动物分类学研究领域的显微成像新技术

UV-C光学共聚焦显微图像系统集成了光学和数字图像分析技术，特别适用于采集非平面样品（高度变化范围超过了显微镜影深范围）的图像，它彻底解决了传统光学显微镜成像时尚倍数与大景深不能共存的问题。     

用于动物分类学研究领域的显微成像新技术

UV-C光学共聚焦显微图像系统集成了光学和数字图像分析技术，特别适用于采集非平面样品（高度变化范围超过了显微镜景深范围）的图像，它彻底解决了传统光学显微镜成像时高倍数与大景深不能共存的问题，使样品的不同高低部位都能清晰成像，可以获得像电镜图像一样的巨大景深和精致的细节，     

用于动物分类学研究领域的显微成像新技术

UV-C光学共聚焦昱微图像系统集成了光学和数字图象分析技术,特别适用于采集非平面样品（高度变化范围超过了显微镜景深范围）的图象,它彻底解决了传统光学显微镜成像时高倍数与大景深不能共存的问题,使样品的不同高低部位都能清晰成像,可以获得像电镜图像一样的巨大景深和精致的细节,     

用激光扫描共聚焦显微镜观察雪松花粉和花粉管

为更直观地观察和显示花粉和花粉管中细胞结构及其细胞核的状态与行为。雪松花粉和花粉管经卡诺液固定,分别以埃氏苏木精、曙红、Hoechst 33243单染和曙红-Hoechst 33342双染后,用冬青油整体透明,在激光扫描共聚焦显微镜下观察。4种染色法观察效果不同;以曙红-Hoechst 33342双染的样品观察效果最佳,在紫外光激发下清晰地显示出细胞核,在488 nm激光激发下不仅能清晰看到花粉和花粉管壁结构,且能分辨管细胞、柄细胞及体细胞的结构特点和空间位置关系。建立了一种快速简便的适于在激光扫描共聚焦显微镜下观察花粉和花粉管中成员细胞结构及其细胞核的状态、行为的制片技术;激光扫描共聚焦显微镜具有独特的共轭成像装置、连续光学扫描、图像三维重组和多通道检测等功能,极好地展示了雪松花粉和花粉管的结构特点,相比于传统的光学显微镜和荧光显微镜,其观察到的图像更清晰、更直观、更具立体感。     

全新的教学方式——Motic数码互动实验室

数码互动实验室系统是一种全新的教学方式,通过先进的数码、网络技术．提供清晰的多画面实时显示和丰富的交互手段。通过此系统教师只需一台电脑就可以同时控制学生端多台数码显微镜,并可对每一台数码显微镜的实时图像进行单独调整,及时掌握学生实验的最新动态,并给予指导。     

全新的教学方式——Motic数码互动实验室

<正>数码互动实验室系统是一种全新的教学方式,通过先进的数码、网络技术.提供清晰的多画面实时显示和丰富的交互手段。通过此系统教师只需一台电脑就可以同时控制学生端多台数码显微镜.并可对每一台数码显微镜的实时图像进行单独调整,及时掌握学生实验的最新动态.并给予指导。数码互动实验室以其声像并茂.师生互动的教学特点,改变了以往传统教学方     

全新的教学方式——Motic数码互动实验室

<正>数码互动实验室系统是一种全新的教学方式,通过先进的数码、网络技术,提供清晰的多画面实时显示和丰富的交互手段。通过此系统教师只需一台电脑就可以同时控制学生端多台数码显微镜,并可对每一台数码显微镜的实时图像进行单独调整,及时掌握学生实验的最新动态,并给予指导。     

Real time imaging of single fluorophores on moving actin with an epifluorescence microscope.

Relatively simple modifications of an ordinary epifluorescence microscope have greatly reduced its background luminescence, allowing continuous and real time imaging of single fluorophores in an aqueous medium. Main modifications were changing the excitation light path and setting an aperture stop so that stray light does not scatter inside the microscope. A simple and accurate method using actin filaments is presented to establish the singularity of the observed fluorophores. It was possible, at the video rate of 30 frames/s, to image individual tetramethylrhodamine fluorophores bound to actin filaments sliding over heavy meromyosin. The successful imaging of moving fluorophores demonstrates that conventional microscopes may become a routine tool for studying dynamic interactions among individual biomolecules in physiological environments.     

High-performance confocal system for microscopic or endoscopic applications

We designed a high-performance confocal system that can be easily adapted to an existing light microscope or coupled with an endoscope for remote imaging. The system employs spatially and temporally patterned illumination produced by one of several mechanisms, including a micromirror array video projection device driven by a computer video source or a microlens array scanned by a piezo actuator in the microscope illumination path. A series of subsampled "component" video images are acquired from a solid-state video camera. Confocal images are digitally reconstructed using "virtual pinhole" synthetic aperture techniques applied to the collection of component images. Unlike conventional confocal techniques that raster scan a single detector and illumination point, our system samples multiple locations in parallel, with particular advantages for monitoring fast dynamic processes. We compared methods of patterned illumination and confocal image reconstruction by characterizing the point spread function, contrast, and intensity of imaged objects. Sample 3D reconstructions include a diatom and a Golgi-stained nerve cell collected in transmission.     

Dual-view microscopy with a single camera: real-time imaging of molecular orientations and calcium

A new microscope technique, termed "W" (double view video) microscopy, enables simultaneous observation of two different images of an object through a single video camera or by eye. The image pair may, for example, be transmission and fluorescence, fluorescence at different wavelengths, or mutually perpendicular components of polarized fluorescence. Any video microscope can be converted into a dual imager by simple insertion of a small optical device. The continuous appearance of the dual image assures the best time resolution in existing and future video microscopes. As an application, orientations of actin protomers in individual, moving actin filaments have been imaged at the video rate. Asymmetric calcium influxes into a cell exposed to an intense electric pulse have also been visualized.     

一种目视激光显微镜

本文介绍一种目视激光显微镜。该装置采用白炽灯和激光做光源。通过调压器衬底亮度可以调整到零。由于激光的高亮度和强相干性,与普通显微镜相比,该显微镜具有景深长,分辨率高,层次丰富的特点。使用该显微镜时能实现镜象的假色彩编码,且镜象具有立体感。文中报导了该显微镜的原理和使用效果。     

Video image processing greatly enhances contrast, quality, and speed in polarization-based microscopy

Video cameras with contrast and black level controls can yield polarized light and differential interference contrast microscope images with unprecedented image quality, resolution, and recording speed. The theoretical basis and practical aspects of video polarization and differential interference contrast microscopy are discussed and several applications in cell biology are illustrated. These include: birefringence of cortical structures and beating cilia in Stentor, birefringence of rotating flagella on a single bacterium, growth and morphogenesis of echinoderm skeletal spicules in culture, ciliary and electrical activity in a balancing organ of a nudibranch snail, and acrosomal reaction in activated sperm.     

UV microbeam irradiations of the mitotic spindle I. The UV-microbeam apparatus

Summary We describe the assembly of a UV microbeam microscope based on a Zeiss IM35 inverted microscope. The important UV transmitting elements are standard UV epifluorescence attachments available from Zeiss; the main modification involves fitting an adjustable slit in place of the field diaphragm. We describe how to align and focus the UV source for optimal irradiations. Our current version of this machine is also fitted with a monochromator and using monochromatic UV light, we can reproduceably create Areas of Reduced Birefringence in spindle fibres with ca. 2–3 s irradiations, while continually observing the fibres. The microscope is stable and easy to set up, allowing many consecutive experiments to be done, including multiple irradiations on the one cell. In conjunction with video image processing techniques, the cells can be observed continuously using polarising, Nomarski or other optical systems. Some preliminary observations demonstrating the versatility of the machine are described.Abbrevations ARB areas of reduced birefringence - MT microtubules - UV ultraviolet     

Rappels technologiques,biais méthodologiques possibles et conditions de mesure

Computer-aided sperm analysis (CASA) was developed in the last decade to overcome the problems related to the inherent subjectivity of manual semen analysis. A typical CASA system includes a video camera, a microscope equiped with phase optics and a warming stage, a microcomputer with hardware and software dedicated to motion analysis, a monitor and a printer. The image of the sperm cells under the microscope is transformed into discrete pixels producing a voltage proportional to the intensity of the light as light strikes the camera’s CCD array. In standard video technology, the first field (set of rows; odd lines) is scanned in one sixtieth of a second and remains illuminated, then the alternate set of rows (even lines) is scanned in the next one sixtieth of a second to display the complete video frame which requires one thirthieth of a second. The first step called digitization is the encoding of video pixels as numbers following a gray level scale between white and black. From this process the detection of the sperm heads in the field is performed and repeated on a field by field basis. The centroïd coordinates of the sperm heads in the field are calculated and sequentially transmitted to the hard disk. A path finder algorithm connects the centroïds through time allowing the reconstruction of trajectories and the calculation of various parameters. Many factors modulating CASA measurements can invalidate the results of analysis. Consequently, a standardization of CASA is necessary. Various guidelines for accurate analysis are proposed. In summary, basic knowledge on sperm physiology, CASA technology and rigorous standardized conditions of analysis are required for providing reliable and accurate results because CASA systems are not ready-to-use “robots”. CASA is useful for routine semen analysis and essential for the measurement of sperm kinematics for clinical as well as for research purposes.     

Comparison of absorption measurements of DNA stain content by utilizing video and scanning image cytometers

After staining with the Feulgen reaction, the DNA stain contents of 155 mouse bone marrow cells and 22 adjacent chicken erythrocytes were measured by absorption image cytometry by utilizing two different systems--a scanning cytometer and a video cytometer. In the scanning cytometer (M85 microdensitometer, Vickers Instruments, Malden, MA), a spot of light was scanned across the cell. In the video cytometer (TAS Plus, E. Leitz, Rockleigh, NJ), the microscope field, which may contain several nuclei, was imaged onto a Plumbicon video camera. With each system, cells were scanned, digitized into their elementary pixels, and analyzed to determine their integrated absorbance. Comparison of the DNA stain contents of the same G0/G1 bone marrow cells and chicken erythrocytes, as measured by video and scanning cytometry, showed that both techniques gave comparable results; scanning cytometry is more precise. The coefficients of variation of the measurements for the G0/G1 bone marrow cells and for the chicken erythrocytes were 5.9% and 7.0%, respectively, when measured by video cytometry at the absorption peak (584 nm), compared to 4.1% and 3.5%, respectively, for the same cells when measured by scanning cytometry off the absorption peak (615 nm). The video-based measurements were relatively lower than the scanning measurements for darkly stained cells; this suggests that glare and other optical errors which increase with stain darkness caused greater systematic errors in the video cytometer than they did in the scanning cytometer.     

Image analysis method for the rapid counting of Saccharomyces cerevisiae cells

An image analysis system which incorporates a microscope, video camera, monitor, and Apple computer and which uses image area to count Saccharomyces cerevisiae cells is described and evaluated. Yeast cell suspensions of densities of up to 100 X 10(6) cells per ml can be counted when viewed in the counting chamber of a hemacytometer. The yeast image area measured depends upon the light intensity used to illuminate the yeast cells, the sharpness of the image focused on the monitor, and the grey level selected when scanning the digitized image on the monitor of the Apple computer, all of which can be controlled. The image area also depends upon the yeast strain and medium in which the culture is grown, but it is not affected by the concentration of sugar or ethanol in which the yeast cells are suspended. Yeast growth measured by image analysis can be calibrated to give results similar to those obtained with hemacytometer counting. Yeast cells can be counted in the presence of high cell densities of bacteria by adjusting the grey level at which the digitized image is scanned.     

Image analysis method for the rapid counting of Saccharomyces cerevisiae cells.

An image analysis system which incorporates a microscope, video camera, monitor, and Apple computer and which uses image area to count Saccharomyces cerevisiae cells is described and evaluated. Yeast cell suspensions of densities of up to 100 X 10(6) cells per ml can be counted when viewed in the counting chamber of a hemacytometer. The yeast image area measured depends upon the light intensity used to illuminate the yeast cells, the sharpness of the image focused on the monitor, and the grey level selected when scanning the digitized image on the monitor of the Apple computer, all of which can be controlled. The image area also depends upon the yeast strain and medium in which the culture is grown, but it is not affected by the concentration of sugar or ethanol in which the yeast cells are suspended. Yeast growth measured by image analysis can be calibrated to give results similar to those obtained with hemacytometer counting. Yeast cells can be counted in the presence of high cell densities of bacteria by adjusting the grey level at which the digitized image is scanned.