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1. 发明申请

WO2006123967A2 FLUORESCENT NANOSCOPY METHOD 审中-公开
标题翻译：荧光纳米科技
公开(公告)号：WO2006123967A2
公开(公告)日：2006-11-23
申请号：PCT/RU2006000231
申请日：2006-05-05
申请人： KLIMOV ANDREY ALEXEEVICH , KLIMOV DMITRY ANDREEVICH , KLIMOV EVGENIY ANDREEVICH , KLIMOVA TATIANA VITALYEVNA
发明人： KLIMOV ANDREY ALEXEEVICH , KLIMOV DMITRY ANDREEVICH , KLIMOV EVGENIY ANDREEVICH , KLIMOVA TATIANA VITALYEVNA
CPC分类号： G01N21/6428 , G01N21/6458 , G01N21/6486 , H04N7/18 , H04N13/0275 , Y10S977/84 , Y10S977/868 , Y10S977/869 , Y10S977/881 , Y10T436/143333 , Y10T436/25
摘要： The invention relates to analysis of object dyed with fluorescent colouring agents carried out with the aid of a fluorescent microscope which is modified in terms of improved resolving power and called a nanoscope thereafter. The inventive method is carried out with the aid of a microscope comprising an optical system for visualising and projecting a sample image to a video camera which records and digitises images of individual fluorescence molecules and nanoparticles at a low noise, a computer for recording and processing images, a sample holder arranged in front of an object lens, a fluorescent radiation exciting source and a set of replaceable suppression filter for separating the sample fluorescent light. The invention is characterised in that the separately fluorescing visible molecules and nanoparticles are periodically formed in different object parts, the laser produces the oscillation thereof which is sufficient for recording the non-overlapping images of said molecules and nanoparticles and for decolouring already recorded fluorescent molecules, wherein tens of thousands of pictures of recorded individual molecule and nanoparticle images (in the form of stains whose diameter is of the order of a fluorescent light wavelength multiplied by a microscope amplification) are processed by a computer for searching the co-ordinates of the stain centres and building the object image according to millions of calculated stain center co-ordinates corresponding to the co-ordinates of the individual fluorescent molecules and nanoparticles. Said invention makes it possible to obtain a two- and three-dimensional image with a resoling power better than 20 nm and to record a colour image by dyeing proteins, nucleic acids and lipids with different colouring agents.
摘要翻译：本发明涉及使用荧光显色剂进行荧光着色剂染色的物体的分析，所述荧光显微镜在分辨能力提高方面被修改，此后称为纳米镜。本发明的方法借助于包括用于将样本图像可视化并投影到以低噪声记录和数字化各个荧光分子和纳米颗粒的图像的摄像机的光学系统的显微镜进行，用于记录和处理图像的计算机，配置在物镜前面的样品架，荧光放射线激励源和用于分离样品荧光的一组可更换抑制滤光片。本发明的特征在于，在不同的对象部分中周期性地形成分开的荧光可见分子和纳米颗粒，激光产生其足以记录所述分子和纳米颗粒的非重叠图像并且用于脱色已经记录的荧光分子的振荡，其中记录的单个分子和纳米颗粒图像（其直径为荧光波长乘以显微镜放大的污渍的形式）的数万张照片由计算机处理，用于搜索染色体的坐标根据与各个荧光分子和纳米颗粒的坐标相对应的数百万计算的染色中心坐标来建立对象图像。本发明使得可以获得具有优于20nm的分辨率的二维和三维图像，并通过用不同着色剂对蛋白质，核酸和脂质进行染色来记录彩色图像。

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