Flat-Field Illumination For Quantitative Fluorescence Imaging

Abstract

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.

Publication Date

6-11-2018

Publication Title

Optics Express

Volume

26

Issue

12

Number of Pages

15276-15288

Document Type

Article

Personal Identifier

scopus

DOI Link

https://doi.org/10.1364/OE.26.015276

Socpus ID

85048362122 (Scopus)

Source API URL

https://api.elsevier.com/content/abstract/scopus_id/85048362122

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