Even illumination in total internal reflection fluorescence microscopy using laser light

R. Fiolka, Y. Belyaev, H. Ewers, A. Stemmer

Research output: Contribution to journalArticlepeer-review

39 Scopus citations

Abstract

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 non-uniform 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.

Original languageEnglish (US)
Pages (from-to)45-50
Number of pages6
JournalMicroscopy Research and Technique
Volume71
Issue number1
DOIs
StatePublished - Jan 1 2008

Keywords

  • Coherence
  • Evanescent fields
  • Illumination
  • Intereference fringes
  • Total internal reflection fluorescence (TIRF) microscopy

ASJC Scopus subject areas

  • Anatomy
  • Histology
  • Instrumentation
  • Medical Laboratory Technology

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