在厚的不均匀样品中成像深而清晰

J. Andilla, Omar E. Olarte, R. Aviles-Espinosa, P. Loza-Álvarez
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引用次数: 0

摘要

获取大样本内部深处的图像是当前生物学应用中最需要的改进之一。吸收、散射和光学像差是这类样品中遇到的主要困难。自适应光学已从天文学中引入,用于处理样品引起的光学像差。非线性显微镜和SPIM被认为是深入样品成像的有趣选择。特别是,由于其低光漂白特性,光片显微镜为获取信息开辟了新的机会,例如在长时间内进行大型3D成像。在这项工作中,我们对自适应光学在线性和非线性模式下的荧光显微镜应用进行了概述。然后,我们将重点讨论两个正交光路的光片显微镜结构,这意味着光学校正方面的新要求。我们将看到在薄片显微镜中出现的不同问题,特别是在成像大型和非平面样品时。最后,我们将研究等地斑块问题。
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Imaging deep and clear in thick inhomogeneous samples
Acquisition of images deep inside large samples is one of the most demanded improvements that current biology applications ask for. Absorption, scattering and optical aberrations are the main difficulties encountered in these types of samples. Adaptive optics has been imported form astronomy to deal with the optical aberrations induced by the sample. Nonlinear microscopy and SPIM have been proposed as interesting options to image deep into a sample. Particularly, light-sheet microscopy, due to its low photo bleaching properties, opens new opportunities to obtain information for example in long time lapses for large 3D imaging. In this work, we perform an overview of the application of adaptive optics to the fluorescence microscopy in linear and non-linear modalities. Then we will focus in the light-sheet microscopy architecture of two orthogonal optical paths which implies new requirements in terms of optical correction. We will see the different issues that appear in light-sheet microscopy particularly when imaging large and non-flat samples. Finally, we will study the problem of the isoplanetic patches.
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