Flat-field illumination for quantitative fluorescence imaging | Zendy

Ian Khaw | Zendy; Benjamin Croop | Zendy; Jialei Tang | Zendy; Anna Möhl | Zendy; Ulrike Fuchs | Zendy; Kyu Young Han | Zendy

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Flat-field illumination for quantitative fluorescence imaging

Author(s) -

Ian Khaw,

Benjamin Croop,

Jialei Tang,

Anna Möhl,

Ulrike Fuchs,

Kyu Young Han

Publication year - 2018

Publication title -

optics express

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.394

H-Index - 271

ISSN - 1094-4087

DOI - 10.1364/oe.26.015276

Subject(s) - optics , microscopy , materials science , coherence (philosophical gambling strategy) , total internal reflection fluorescence microscope , fluorescence lifetime imaging microscopy , gaussian beam , optical coherence tomography , gaussian , fluorescence , physics , beam (structure) , quantum mechanics

The uneven illumination of a Gaussian profile makes quantitative analysis highly challenging in laser-based wide-field fluorescence microscopy. Here we present flat-field illumination (FFI) where the Gaussian beam is reshaped into a uniform flat-top profile using a high-precision refractive optical component. The long working distance and high spatial coherence of FFI allows us to accomplish uniform epi and TIRF illumination for multi-color single-molecule imaging. In addition, high-throughput borderless imaging is demonstrated with minimal image overlap.

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