Tilt-invariant scanned oblique plane illumination microscopy for large-scale volumetric imaging

Kumar, Manish; Kozorovitskiy, Yevgenia

doi:10.1364/ol.44.001706

Cited by 33 publications

(32 citation statements)

References 20 publications

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“…As compared to the previously reported results (9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22), the achievable FOV (5.8 mm×4.9 mm×0.7 mm) is an order of magnitude higher, the largest FOV in OPM so far to the best of our knowledge.…”

Section: Discussionsupporting

confidence: 42%

“…As the issue of unparallel light-sheets lying in the scanning geometry of a typical OPM, the tiltinvariant scanning method (19) can be adopted in future improvements. Another major issue of OPM is the light loss in the remote imaging system.…”

Section: Discussionmentioning

confidence: 99%

“…To increase the light collecting efficiency and improve the resolution, water immersion technique can be introduced in the remote imaging system (18). A tilt-invariant scanned OPM (SoPi) was investigated (19); two-photon excitation technique was adopted (20,21); and simultaneous multimodal 3D imaging with optical coherence tomography (OCT) was demonstrated (22).…”

Section: Introductionmentioning

confidence: 99%

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Wide field-of-view volumetric imaging by a mesoscopic scanning oblique plane microscopy with switchable objective lens

Shao

Kılıç

Yin

et al. 2020

Preprint

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AbstractConventional light sheet fluorescence microscopy (LSFM), or selective plane illumination microscopy (SPIM), enables high resolution 3D imaging over a large volume by using two orthogonally aligned objective lenses to decouple excitation and emission. The recent development of oblique plane microscopy (OPM) simplifies LSFM design with only one single objective lens, by using off-axis excitation and remote focusing. However, most reports on OPM has a limited microscopic field of view (FOV), typically within 1×1 mm2. Our goal is to overcome the limitation with a new variant of OPM to achieve mesoscopic FOV. We implemented an optical design of mesoscopic scanning OPM to allow using low numerical aperture (NA) objective lens. The angle of the intermediate image before the remote focusing system was increased by a demagnification under Scheimpflug condition such that the light collecting efficiency in the remote focusing system was significantly improved. We characterized the 3D resolutions and FOV by imaging fluorescence microspheres, and demonstrated the volumetric imaging on intact whole zebrafish larvae, mouse cortex, and multiple Caenorhabditis elegans (C. elegans). We demonstrate a mesoscopic FOV up to ~6× 5×0.6 mm3 volumetric imaging, the largest reported FOV by OPM so far. The angle of the intermediate image plane is independent of the magnification. As a result, the system is highly versatile, allowing simple switching between different objective lenses with low (10x, NA 0.3) and median NA (20x, NA 0.5). Detailed microvasculature in zebrafish larvae, mouse cortex, and neurons in C. elegans are clearly visualized in 3D. The proposed mesoscopic scanning OPM allows using low NA objective such that centimeter-level FOV volumetric imaging can be achieved. With the extended FOV, simple sample mounting protocol, and the versatility of changeable FOVs/resolutions, our system will be ready for the varieties of applications requiring in vivo volumetric imaging over large length scales.

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Section: Discussionsupporting

confidence: 42%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Wide field-of-view volumetric imaging by a mesoscopic scanning oblique plane microscopy with switchable objective lens

Shao

Kılıç

Yin

et al. 2020

Preprint

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“…To combine the best features of the previous two arrangements, we introduced a plane mirror based scanned oblique plane illumination (SOPi) microscopy [17,18]. Our work focused on optical ray tracing simulations to obtain an optimized scan geometry to resolve the tilt variance Fig.…”

Section: Introductionmentioning

confidence: 99%

“…(b) Second arrangement uses a refractive transmission scanning window to perform constant tilt lateral scan [16]. (c) Third arrangement uses a plane mirror scanner to aim for constant tilt lateral scan [17][18][19][20][21]. MO: microscope objective.…”

Section: Introductionmentioning

confidence: 99%

Tilt (in)variant lateral scan in oblique plane microscopy: a geometrical optics approach

Kumar¹,

Kozorovitskiy²

2020

Biomed. Opt. Express

Self Cite

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Oblique plane microscopy (OPM) is a single objective light-sheet microscopy that performs three dimensional (3D) imaging by axial scan of the generated light-sheet. Recently, multiple techniques for lateral scanning of the generated light-sheet in OPM have emerged. However, their suitability for geometrically distortion free 3D imaging, which essentially requires a constant tilt light-sheet scan, has not been evaluated. In this work, we use a geometrical optics approach and derive analytical relationship for the amount of tilt variance in planar mirror based scanned oblique plane illumination (SOPi) arrangement. We experimentally validate the derived relationship and use it to arrive at an optimized scanner geometry and to understand its associated limitations. We discuss the effects of scanning on optical aberrations and 3D field of view in optimized, tilt invariant, lateral scanning OPM systems. We also provide experimental strategies enabling precise scanner alignment for tilt invariance, as well as an open source platform for rapid design of new oblique light-sheet microscopes.

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Cellular resolution imaging of neuronal activity across space and time in the mammalian brain

Clough

Chen

2019

Current Opinion in Biomedical Engineering

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Tilt-invariant scanned oblique plane illumination microscopy for large-scale volumetric imaging

Cited by 33 publications

References 20 publications

Wide field-of-view volumetric imaging by a mesoscopic scanning oblique plane microscopy with switchable objective lens

Wide field-of-view volumetric imaging by a mesoscopic scanning oblique plane microscopy with switchable objective lens

Tilt (in)variant lateral scan in oblique plane microscopy: a geometrical optics approach

Cellular resolution imaging of neuronal activity across space and time in the mammalian brain

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