S-polarized light-sheets improve resolution and light-efficiency in oblique plane microscopy (2303.14018v2)
Abstract: Oblique plane microscopy (OPM) offers 3D optically sectioned imaging with high spatial- and temporal-resolution while enabling conventional sample mounting. The technique uses a concatenation of three microscopes, two for remote focusing and a tilted tertiary microscope, often including an immersion objective, to image an oblique sample plane. This design induces Fresnel reflections and a reduced effective aperture, thus impacting the resolution and light efficiency of the system. Using vectorial diffraction simulations, the system performance was characterized based on illumination angle and polarization, signal to noise ratio, and refractive index of the tertiary objective immersion. We show that for samples with high fluorescent anisotropy, s-polarized light-sheets yield higher average resolution for all system configurations, as well as higher light-efficiency. We also provide a tool for performance characterization of arbitrary light-sheet imaging systems.
- Microscopy in 3d: a biologist’s toolbox. \JournalTitleTrends in cell biology 21, 682–691 (2011).
- Mappes, T. et al. The invention of immersion ultramicroscopy in 1912—the birth of nanotechnology? \JournalTitleAngewandte Chemie International Edition 51, 11208–11212 (2012).
- Orthogonal-plane fluorescence optical sectioning: Three-dimensional imaging of macroscopic biological specimens. \JournalTitleJournal of microscopy 170, 229–236 (1993).
- Fundamental reduction of the observation volume in far-field light microscopy by detection orthogonal to the illumination axis: confocal theta microscopy. \JournalTitleOptics Communications 111, 536–547 (1994).
- Stelzer, E. et al. A new tool for the observation of embryos and other large specimens: confocal theta fluorescence microscopy. \JournalTitleJournal of microscopy 179, 1–10 (1995).
- Chen, B.-C. et al. Lattice light-sheet microscopy: imaging molecules to embryos at high spatiotemporal resolution. \JournalTitleScience 346, 1257998 (2014).
- Selective plane illumination microscopy techniques in developmental biology. \JournalTitleDevelopment 136, 1963–1975, DOI: 10.1242/dev.022426 (2009).
- Santi, P. A. Light sheet fluorescence microscopy: a review. \JournalTitleJournal of Histochemistry & Cytochemistry 59, 129–138 (2011).
- Dunsby, C. Optically sectioned imaging by oblique plane microscopy. \JournalTitleOptics express 16, 20306–20316 (2008).
- Bouchard, M. B. et al. Swept confocally-aligned planar excitation (scape) microscopy for high-speed volumetric imaging of behaving organisms. \JournalTitleNature photonics 9, 113–119 (2015).
- Integrated one-and two-photon scanned oblique plane illumination (sopi) microscopy for rapid volumetric imaging. \JournalTitleOptics express 26, 13027–13041 (2018).
- Sparks, H. et al. Dual-view oblique plane microscopy (dopm). \JournalTitleBiomedical optics express 11, 7204–7220 (2020).
- Glaser, A. K. et al. A hybrid open-top light-sheet microscope for versatile multi-scale imaging of cleared tissues. \JournalTitleNature Methods 19, 613–619 (2022).
- Yang, B. et al. Daxi—high-resolution, large imaging volume and multi-view single-objective light-sheet microscopy. \JournalTitleNature methods 19, 461–469 (2022).
- Sapoznik, E. et al. A versatile oblique plane microscope for large-scale and high-resolution imaging of subcellular dynamics. \JournalTitleElife 9, e57681 (2020).
- An optical technique for remote focusing in microscopy. \JournalTitleOptics Communications 281, 880–887 (2008).
- Sensitivity of remote focusing microscopes to magnification mismatch. \JournalTitleJournal of Microscopy 288, 95–105 (2022).
- Andrewgyork/high_na_single_objective_ lightsheet:work-in-progress, DOI: 10.5281/zenodo.3376243 (2019).
- Yang, B. et al. Epi-illumination spim for volumetric imaging with high spatial-temporal resolution. \JournalTitleNature methods 16, 501–504 (2019).
- Lakowicz, J. R. Principles of fluorescence spectroscopy (Springer, 2006).
- Calculation of vectorial diffraction in optical systems. \JournalTitleJOSA A 35, 526–535 (2018).
- Gu, M. Advanced optical imaging theory, vol. 75 (Springer Science & Business Media, 2000).
- Debye, P. Das verhalten von lichtwellen in der n¨ahe eines brennpunktes oder einer brennlinie. \JournalTitleAnnals of Physics 335, 755–776 (1909).
- Fast focus field calculations. \JournalTitleOptics express 14, 11277–11291 (2006).
- Manton, J. Debye diffraction code. https://github.com/jdmanton/debye_diffraction_code (2022).
Sponsored by Paperpile, the PDF & BibTeX manager trusted by top AI labs.
Get 30 days freePaper Prompts
Sign up for free to create and run prompts on this paper using GPT-5.
Top Community Prompts
Collections
Sign up for free to add this paper to one or more collections.