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首页> 外文期刊>Journal of Microscopy >Axial phase-darkfield-contrast (apdc), a new technique for variable optical contrasting in light microscopy
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Axial phase-darkfield-contrast (apdc), a new technique for variable optical contrasting in light microscopy

机译：轴向相位-暗场对比度（apdc），一种用于光学显微镜中可变光学对比的新技术

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摘要

Axial phase-darkfield-contrast (APDC) has been developed as an illumination technique in light microscopy which promises significant improvements and a higher variability in imaging of several transparent 'problem specimens'. With this method, a phase contrast image is optically superimposed on an axial darkfield image so that a partial image based on the principal zeroth order maximum (phase contrast) interferes with an image, which is based on the secondary maxima (axial darkfield). The background brightness and character of the resulting image can be continuously modulated from a phase contrast-dominated to a darkfield-dominated character. In order to achieve this illumination mode, normal objectives for phase contrast have to be fitted with an additional central light stopper needed for axial (central) darkfield illumination. In corresponding condenser light masks, a small perforation has to be added in the centre of the phase contrast providing light annulus. These light modulating elements are properly aligned when the central perforation is congruent with the objective's light stop and the light annulus is conjugate with the phase ring. The breadth of the condenser light annulus and thus the intensity of the phase contrast partial image can be regulated with the aperture diaphragm. Additional contrast effects can be achieved when both illuminating light components are filtered at different colours. In this technique, the axial resolution (depth of field) is significantly enhanced and the specimen's three-dimensional appearance is accentuated with improved clarity as well as fine details at the given resolution limit. Typical artefacts associated with phase contrast and darkfield illumination are reduced in our methods.
机译：轴向相位-暗场对比度（APDC）已被开发为光学显微镜中的照明技术，有望显着改善并提高多个透明“问题标本”成像的可变性。利用这种方法，将相位对比图像光学地叠加在轴向暗场图像上，使得基于主零阶最大值（相位对比）的部分图像与基于次要最大值（轴向暗场）的图像发生干涉。最终图像的背景亮度和字符可以从相位对比为主的字符转换为暗场为主的字符。为了实现这种照明模式，必须为相位对比的正常目标安装轴向（中央）暗场照明所需的附加中央挡光板。在相应的聚光器光罩中，必须在相衬的中心添加一个小孔，以提供光环空。当中心孔眼与物镜的挡光片相吻合并且光环与相位环共轭时，这些光调制元件将正确对齐。聚光光环的宽度以及由此相衬部分图像的强度可以通过孔径光阑来调节。当两个照明光分量以不同的颜色过滤时，可以实现额外的对比度效果。在此技术中，轴向分辨率（景深）得到了显着提高，并且在给定分辨率极限下，标本的三维外观更加清晰，细节也更加清晰。在我们的方法中，减少了与相位对比和暗场照明相关的典型伪像。

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