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Sevenfold improvement of axial resolution in 3D wide-field microscopy using two objective-lenses

机译:三维宽场显微镜轴向分辨率的七倍性镜片用两个物镜镜头

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A weakness of standard 3D microscopies - both confocal and widefield$PLU@deconvolution - is that their resolution is substantially worse in the axial direction than in the lateral plane. We describe two new widefield techniques with substantially improved axial resolution that actually exceeds the lateral resolution. As is well known, the resolution is related to the angle over which the objective lens collects light. In our first technique, light is collected over an enlarged set of angles by using two objective lenses on opposite sides of the sample. The two image beams are combined coherently on the same CCD camera. Interference between the beams yields new, previously inaccessible sample information. The second technique applies a similar concept to the illumination light in fluorescence microscopy. Light from an extended, spatially incoherent light source - such as a standard arc lamp - is split and directed through the two opposing objective lenses so as to create a narrow interference fringe at the focal plane in the sample. This spatial structure in the excitation light yields access to new sample information. The two techniques can easily be used together; the combined technique promises an axial resolution improvement of a factor of seven over standard widefield microscopy.
机译:标准3D显微术的一个缺点 - 共焦和广角$ PLU @去卷积 - 是它们的分辨率是在轴向方向上比在横向平面内显着恶化。我们描述与实际超过横向分辨率显着改善的轴向分辨率两个新的宽视野的技术。如众所周知的,分辨率是与在其上的物镜收集光的角度。在我们的第一技术中,光被通过使用对样品的相对侧上的两个物镜收集的放大组角度。两个图像光束相同的CCD照相机上相干组合。之间干涉光产量新,以前无法进入的样本信息。第二种技术适用类似的概念,以在荧光显微镜的照明光。光从扩展,空间非相干的光源 - 比如标准弧光灯 - 被分割,并通过两个对置的物镜引导以便在样品中的焦平面创建一个窄干涉条纹。在激发光的产量这种空间结构获得新的样品的信息。两种技术可以很容易地被一起使用;将合并的技术有望七个比标准宽视场显微镜的因素的轴向分辨率的改善。

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