Integrated one- and two-photon scanned oblique plane illumination (SOPi) microscopy for rapid volumetric imaging

Manish Kumar, Sandeep Kishore, Jordan Nasenbeny, David L. McLean, Yevgenia Kozorovitskiy*

*Corresponding author for this work

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

Versatile, sterically accessible imaging systems capable of in vivo rapid volumetric functional and structural imaging deep in the brain continue to be a limiting factor in neuroscience research. Towards overcoming this obstacle, we present integrated one- and two-photon scanned oblique plane illumination (SOPi, /sōpī/) microscopy which uses a single front-facing microscope objective to provide light-sheet scanning based rapid volumetric imaging capability at subcellular resolution. Our planar scan-mirror based optimized light-sheet architecture allows for non-distorted scanning of volume samples, simplifying accurate reconstruction of the imaged volume. Integration of both one-photon (1P) and two-photon (2P) light-sheet microscopy in the same system allows for easy selection between rapid volumetric imaging and higher resolution imaging in scattering media. Using SOPi, we demonstrate deep, large volume imaging capability inside scattering mouse brain sections and rapid imaging speeds up to 10 volumes per second in zebrafish larvae expressing genetically encoded fluorescent proteins GFP or GCaMP6s. SOPi’s flexibility and steric access makes it adaptable for numerous imaging applications and broadly compatible with orthogonal techniques for actuating or interrogating neuronal structure and activity.

Original languageEnglish (US)
Pages (from-to)13027-13041
Number of pages15
JournalOptics Express
Volume26
Issue number10
DOIs
StatePublished - May 14 2018

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics

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