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

Sascha Reidt, Ricardo Bango Da Cunha Correia, Mark Donnachie, Cornelis Weijer, Michael MacDonald

Research output: Chapter in Book/Report/Conference proceedingConference contribution

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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.
Original languageEnglish
Title of host publicationProceedings of SPIE
Subtitle of host publicationOptical Trapping and Optical Micromanipulation XIV
EditorsKishan Dholakia, Gabriel C. Spalding
Place of PublicationUnited States of America
PublisherSPIE-International Society for Optical Engineering
Pages1-10
Number of pages10
Volume10347
ISBN (Electronic)9781510611528
ISBN (Print)9781510611511
DOIs
Publication statusPublished - 25 Aug 2017
EventSPIE NanoScience + Engineering: Optical Trapping and Optical Micromanipulation XIV - San Diego, United States
Duration: 6 Aug 201710 Aug 2017

Publication series

NameProceedings of SPIE
PublisherSPIE - International Society for Optical Engineering
Volume10347
ISSN (Print)0277-786X
ISSN (Electronic)1995-756X

Conference

ConferenceSPIE NanoScience + Engineering
CountryUnited States
CitySan Diego
Period6/08/1710/08/17

Keywords

  • Lightsheet fluorescence microscopy
  • Bessel beams
  • DSLM
  • SPIM
  • chick embryos

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    Reidt, S., Bango Da Cunha Correia, R., Donnachie, M., Weijer, C., & MacDonald, M. (2017). Gaussian vs. Bessel light-sheets: performance analysis in live large sample imaging. In K. Dholakia, & G. C. Spalding (Eds.), Proceedings of SPIE: Optical Trapping and Optical Micromanipulation XIV (Vol. 10347, pp. 1-10). [1034709] (Proceedings of SPIE; Vol. 10347). SPIE-International Society for Optical Engineering. https://doi.org/10.1117/12.2277324