Clearer view for TIRF and oblique illumination microscopy

Reto Fiolka

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

In Total Internal Reflection Fluorescence (TIRF) microscopy, the sample is illuminated with an evanescent field that yields a thin optical section. However, its widefield detection has no rejection mechanism against out-of-focus blur from scattered light that can compromise TIRF images. Here I demonstrate that via structured illumination, out-of-focus blur can be effectively suppressed in TIRF microscopy, yielding strikingly clearer images. The same mechanism can also be applied to oblique illumination schemes that extend the reach of TIRF microscopy beyond the basal surface of the cell. The two imaging modes are used to image a biosensor, clathrin coated vesicles and the actin cytoskeleton in different cell types with improved contrast.

Original languageEnglish (US)
Pages (from-to)29556-29567
Number of pages12
JournalOptics Express
Volume24
Issue number26
DOIs
StatePublished - Dec 26 2016

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illumination
microscopy
fluorescence
cells
bioinstrumentation
rejection

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics

Cite this

Clearer view for TIRF and oblique illumination microscopy. / Fiolka, Reto.

In: Optics Express, Vol. 24, No. 26, 26.12.2016, p. 29556-29567.

Research output: Contribution to journalArticle

Fiolka, Reto. / Clearer view for TIRF and oblique illumination microscopy. In: Optics Express. 2016 ; Vol. 24, No. 26. pp. 29556-29567.
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