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Divergent excitation two photon microscopy for 3D random access mesoscale imaging at single cell resolution

View ORCID ProfileFK Janiak, View ORCID ProfileP Bartel, View ORCID ProfileMR Bale, View ORCID ProfileT Yoshimatsu, E Komulainen, View ORCID ProfileM Zhou, View ORCID ProfileK Staras, View ORCID ProfileLL Prieto-Godino, View ORCID ProfileT Euler, View ORCID ProfileM Maravall, View ORCID ProfileT Baden
doi: https://doi.org/10.1101/821405
FK Janiak
1Sussex Neuroscience, School of Life Sciences, University of Sussex, UK
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  • For correspondence: f.k.janiak@sussex.ac.uk t.baden@sussex.ac.uk
P Bartel
1Sussex Neuroscience, School of Life Sciences, University of Sussex, UK
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MR Bale
1Sussex Neuroscience, School of Life Sciences, University of Sussex, UK
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T Yoshimatsu
1Sussex Neuroscience, School of Life Sciences, University of Sussex, UK
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E Komulainen
1Sussex Neuroscience, School of Life Sciences, University of Sussex, UK
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M Zhou
1Sussex Neuroscience, School of Life Sciences, University of Sussex, UK
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K Staras
1Sussex Neuroscience, School of Life Sciences, University of Sussex, UK
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LL Prieto-Godino
2The Francis Crick Institute, London, UK
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T Euler
3Institute of Ophthalmic Research, University of Tuebingen, Germany
4Centre for Integrative Neuroscience, University of Tuebingen, Germany
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M Maravall
1Sussex Neuroscience, School of Life Sciences, University of Sussex, UK
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T Baden
1Sussex Neuroscience, School of Life Sciences, University of Sussex, UK
3Institute of Ophthalmic Research, University of Tuebingen, Germany
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  • For correspondence: f.k.janiak@sussex.ac.uk t.baden@sussex.ac.uk
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ABSTACT

In neuroscience, diffraction limited two-photon (2P) microscopy is a cornerstone technique that permits minimally invasive optical monitoring of neuronal activity. However, most conventional 2P microscopes impose significant constraints on the size of the imaging field-of-view and the specific shape of the effective excitation volume, thus limiting the scope of biological questions that can be addressed and the information obtainable. Here, employing ‘divergent beam optics’ (DBO), we present an ultra-low-cost, easily implemented and flexible solution to address these limitations, offering a several-fold expanded three-dimensional field of view that also maintains single-cell resolution. We show that this implementation increases both the space-bandwidth product and effective excitation power, and allows for straight-forward tailoring of the point-spread-function. Moreover, rapid laser-focus control via an electrically tunable lens now allows near-simultaneous imaging of remote regions separated in three dimensions and permits the bending of imaging planes to follow natural curvatures in biological structures. Crucially, our core design is readily implemented (and reversed) within a matter of hours, and fully compatible with a wide range of existing 2P customizations, making it highly suitable as a base platform for further development. We demonstrate the application of our system for imaging neuronal activity in a variety of examples in mice, zebrafish and fruit flies.

Footnotes

  • https://github.com/BadenLab/DBOscope

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted October 29, 2019.
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Divergent excitation two photon microscopy for 3D random access mesoscale imaging at single cell resolution
FK Janiak, P Bartel, MR Bale, T Yoshimatsu, E Komulainen, M Zhou, K Staras, LL Prieto-Godino, T Euler, M Maravall, T Baden
bioRxiv 821405; doi: https://doi.org/10.1101/821405
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Divergent excitation two photon microscopy for 3D random access mesoscale imaging at single cell resolution
FK Janiak, P Bartel, MR Bale, T Yoshimatsu, E Komulainen, M Zhou, K Staras, LL Prieto-Godino, T Euler, M Maravall, T Baden
bioRxiv 821405; doi: https://doi.org/10.1101/821405

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