Even illumination in total internal reflection fluorescence microscopy using laser light

被引：46
|
作者：
Fiolka, R. [1 ]
Belyaev, Y. [1 ]
Ewers, H. [2 ]
Stemmer, A. [1 ]
机构：
[1] ETH, Nanotechnol Grp, CH-8092 Zurich, Switzerland
[2] ETH, Inst Biochem, CH-8093 Zurich, Switzerland
来源：
MICROSCOPY RESEARCH AND TECHNIQUE | 2008年 / 71卷 / 01期
关键词：
total internal reflection fluorescence (TIRF) microscopy; evanescent fields; illumination; intereference fringes; coherence;
D O I：
10.1002/jemt.20527
中图分类号：
R602 [外科病理学、解剖学]; R32 [人体形态学];
学科分类号：
100101 ;
摘要：
In modern fluorescence microscopy, lasers are a widely used source of light, both for imaging in total internal reflection and epi-illumination modes. In wide-field imaging, scattering of highly coherent laser light due to imperfections in the light path typically leads to nonuniform illumination of the specimen, compromising image analysis. We report the design and construction of an objective-launch total internal reflection fluorescence microscopy system with excellent evenness of specimen illumination achieved by azimuthal rotation of the incoming illuminating laser beam. The system allows quick and precise changes of the incidence angle of the laser beam and thus can also be used in an epifluorescence mode.
引用
收藏
页码：45 / 50
页数：6
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