Gaussian vs. Bessel light-sheets: performance analysis in live large sample imaging

Sascha L. Reidt; Ricardo B. C. Correia; Mark Donnachie; Cornelis J. Weijer; Michael P. MacDonald

doi:10.1117/12.2277324

25 August 2017 Gaussian vs. Bessel light-sheets: performance analysis in live large sample imaging

Sascha L. Reidt, Ricardo B. C. Correia, Mark Donnachie, Cornelis J. Weijer, Michael P. MacDonald

Proceedings Volume 10347, Optical Trapping and Optical Micromanipulation XIV; 1034709 (2017) https://doi.org/10.1117/12.2277324
Event: SPIE Nanoscience + Engineering, 2017, San Diego, California, United States

Abstract

Lightsheet fluorescence microscopy (LSFM) has rapidly progressed in the past decade from an emerging technology into an established methodology. This progress has largely been driven by its suitability to developmental biology, where it is able to give excellent spatial-temporal resolution over relatively large fields of view with good contrast and low phototoxicity. In many respects it is superseding confocal microscopy. However, it is no magic bullet and still struggles to image deeply in more highly scattering samples. Many solutions to this challenge have been presented, including, Airy and Bessel illumination, 2-photon operation and deconvolution techniques. In this work, we show a comparison between a simple but effective Gaussian beam illumination and Bessel illumination for imaging in chicken embryos. Whilst Bessel illumination is shown to be of benefit when a greater depth of field is required, it is not possible to see any benefits for imaging into the highly scattering tissue of the chick embryo.

Citation Download Citation

Sascha L. Reidt, Ricardo B. C. Correia, Mark Donnachie, Cornelis J. Weijer, and Michael P. MacDonald "Gaussian vs. Bessel light-sheets: performance analysis in live large sample imaging", Proc. SPIE 10347, Optical Trapping and Optical Micromanipulation XIV, 1034709 (25 August 2017); https://doi.org/10.1117/12.2277324

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