Journal
MICROSCOPY RESEARCH AND TECHNIQUE
Volume 71, Issue 1, Pages 45-50Publisher
WILEY
DOI: 10.1002/jemt.20527
Keywords
total internal reflection fluorescence (TIRF) microscopy; evanescent fields; illumination; intereference fringes; coherence
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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.
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