Second harmonic generation microscopy for quantitative analysis of collagen fibrillar structure.

Second harmonic generation microscopy for quantitative analysis of collagen fibrillar structure.
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DOI:
10.1038/nprot.2012.009
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发表时间:
2012-03-08
期刊:
影响因子:
14.8
通讯作者:
--
中科院分区:
生物学1区
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二次谐波发生(SHG)显微镜已成为一个强大的模式,在各种组织的纤维胶原蛋白成像。由于其潜在的物理来源,它对胶原原纤维/纤维结构高度敏感,并且重要的是,对癌症,纤维化和结缔组织疾病等疾病中发生的变化高度敏感。我们讨论了如何SHG可以用来获得更多的组织中的胶原蛋白的组装结构信息比其他显微镜技术是可能的。我们首先提供了一个概述的最先进的SHG显微镜的物理背景,然后描述了需要进行激光扫描显微镜,使测量的光学修改。生物医学应用的关键方面是不同实验配置的能力和局限性。根据用户的经验水平,我们估计SHG仪器从其组成部分的设置和校准将需要2-4周。
Second-harmonic generation (SHG) microscopy has emerged as a powerful modality for imaging fibrillar collagen in a diverse range of tissues. Because of its underlying physical origin, it is highly sensitive to the collagen fibril/fiber structure, and, importantly, to changes that occur in diseases such as cancer, fibrosis and connective tissue disorders. We discuss how SHG can be used to obtain more structural information on the assembly of collagen in tissues than is possible by other microscopy techniques. We first provide an overview of the state of the art and the physical background of SHG microscopy, and then describe the optical modifications that need to be made to a laser-scanning microscope to enable the measurements. Crucial aspects for biomedical applications are the capabilities and limitations of the different experimental configurations. We estimate that the setup and calibration of the SHG instrument from its component parts will require 2–4 weeks, depending on the level of the user’s experience.
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