Axicon-based Bessel beams for flat-field illumination in total internal reflection fluorescence microscopy

Schreiber B, Elsayad K, Heinze KG (2017)

Publication Type: Journal article

Publication year: 2017

Journal

Optics Letters Optical Society of America

Book Volume: 42

Pages Range: 3880-3883

Journal Issue: 19

DOI: 10.1364/OL.42.003880

Abstract

Total internal reflection fluorescence microscopy (TIRF-M) provides low-invasive high-contrast surface imaging with optical sectioning of typically 100–200 nm. Thus, TIRF-M has become an established tool for imaging surfaces, including cell membranes. For TIRF-M, a homogenous evanescent field of excitation over the whole field of view is generally desired for quantitative microscopy; however, this is not necessarily straightforward to generate with Gaussian beams. In recent years, several improvements on TIRF-M have been developed that have addressed non-uniform scattering fringes and other artifacts. Here, we introduce a cost-effective TIRF setup with a very low degree of complexity and no moving parts, which provides a flattop-like excitation profile. The setup uses a tunable laser ring zoom focus system to generate a full 360° TIRF illumination. Two axicon lenses and one focus lens allow for generation and control of the ring diameter to tune the TIRF excitation angle. We show that 360° laser illumination in combination with a radial polarizer will generate an evanescent Bessel-beam excitation field that exhibits a flattop intensity over an extended part of the field of view, and demonstrate the advantages of this axicon-based Bessel beam illumination for live-cell imaging.

Involved external institutions

Julius-Maximilians-Universität Würzburg DE Germany (DE) Vienna Biocenter VBC AT Austria (AT)

How to cite

APA:

Schreiber, B., Elsayad, K., & Heinze, K.G. (2017). Axicon-based Bessel beams for flat-field illumination in total internal reflection fluorescence microscopy. Optics Letters, 42(19), 3880-3883. https://doi.org/10.1364/OL.42.003880

MLA:

Schreiber, Benjamin, Kareem Elsayad, and Katrin G. Heinze. "Axicon-based Bessel beams for flat-field illumination in total internal reflection fluorescence microscopy." Optics Letters 42.19 (2017): 3880-3883.

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