Widefield light sheet microscopy using an Airy beam combined with deep-learning super-resolution

AbstractImaging across length scales and in depth has been an important pursuit of widefield optical imaging. This promises to reveal fine cellular detail within a widefield snapshot of a tissue sample. Current advances often sacrifice resolution through selective sub-sampling to provide a wide field of view in a reasonable time scale. We demonstrate a new avenue for recovering high-resolution images from sub-sampled data in light-sheet microscopy using deep-learning super-resolution. We combine this with the use of a widefield Airy beam to achieve high-resolution imaging over extended fields of view and depths. We characterise our method on fluorescent beads as test targets. We then demonstrate improvements in imaging amyloid plaques in a cleared brain from a mouse model of Alzheimer’s disease, and in excised healthy and cancerous colon and breast tissues. This development can be widely applied in all forms of light sheet microscopy to provide a two-fold increase in the dynamic range of the imaged length scale. It has the potential to provide further insight into neuroscience, developmental biology and histopathology.

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Achromatic terahertz Airy beam generation with dielectric metasurfaces

Nanophotonics ◽

10.1515/nanoph-2020-0536 ◽

2020 ◽

Vol 0 (0) ◽

Author(s):

Qingqing Cheng ◽

Juncheng Wang ◽

Ling Ma ◽

Zhixiong Shen ◽

Jing Zhang ◽

...

Keyword(s):

Biomedical Imaging ◽

Frequency Dispersion ◽

Light Sheet ◽

Photonic Devices ◽

Self Healing ◽

Beam Focusing ◽

Light Sheet Microscopy ◽

Airy Beam ◽

Potential Applications ◽

Airy Beams

AbstractAiry beams exhibit intriguing properties such as nonspreading, self-bending, and self-healing and have attracted considerable recent interest because of their many potential applications in photonics, such as to beam focusing, light-sheet microscopy, and biomedical imaging. However, previous approaches to generate Airy beams using photonic structures have suffered from severe chromatic problems arising from strong frequency dispersion of the scatterers. Here, we design and fabricate a metasurface composed of silicon posts for the frequency range 0.4–0.8 THz in transmission mode, and we experimentally demonstrate achromatic Airy beams exhibiting autofocusing properties. We further show numerically that a generated achromatic Airy-beam-based metalens exhibits self-healing properties that are immune to scattering by particles and that it also possesses a larger depth of focus than a traditional metalens. Our results pave the way to the realization of flat photonic devices for applications to noninvasive biomedical imaging and light-sheet microscopy, and we provide a numerical demonstration of a device protocol.

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Circumvention of common labeling artifacts using secondary nanobodies

10.1101/818351 ◽

2019 ◽

Author(s):

Shama Sograte-Idrissi ◽

Thomas Schlichthaerle ◽

Carlos J. Duque-Afonso ◽

Mihai Alevra ◽

Sebastian Strauss ◽

...

Keyword(s):

Super Resolution ◽

Light Sheet ◽

Common Procedure ◽

Tissue Samples ◽

Localization Accuracy ◽

Light Sheet Microscopy ◽

Large Size ◽

Resolution Imaging ◽

Single Domain Antibodies ◽

Secondary Antibodies

AbstractThe most common procedure to reveal the location of specific (sub)cellular elements in biological samples is via immunostaining followed by optical imaging. This is typically performed with target-specific primary antibodies (1.Abs), which are revealed by fluorophore-conjugated secondary antibodies (2.Abs). However, at high resolution this methodology can induce a series of artifacts due to the large size of antibodies, their bivalency, and their polyclonality. Here we use STED and DNA-PAINT super-resolution microscopy or light sheet microscopy on cleared tissue to show how monovalent secondary reagents based on camelid single-domain antibodies (nanobodies; 2.Nbs) attenuate these artifacts. We demonstrate that monovalent 2.Nbs have four additional advantages: 1) they increase localization accuracy with respect to 2.Abs; 2) they allow direct pre-mixing with 1.Abs before staining, reducing experimental time, and enabling the use of multiple 1.Abs from the same species; 3) they penetrate thick tissues efficiently; and 4) they avoid the artificial clustering seen with 2.Abs both in live and in poorly fixed samples. Altogether, this suggests that 2.Nbs are a valuable alternative to 2.Abs, especially when super-resolution imaging or staining of thick tissue samples are involved.

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Deep-learning on-chip light-sheet microscopy enabling video-rate volumetric imaging of dynamic biological specimens

Lab on a Chip ◽

10.1039/d1lc00475a ◽

2021 ◽

Author(s):

Xiaopeng Chen ◽

Junyu Ping ◽

Yixuan Sun ◽

Chengqiang Yi ◽

Sijian Liu ◽

...

Keyword(s):

Deep Learning ◽

Light Scattering ◽

Light Sheet ◽

High Contrast ◽

Spatiotemporal Resolution ◽

Volumetric Imaging ◽

Light Sheet Microscopy ◽

High Spatiotemporal Resolution ◽

On Chip

Volumetric imaging of dynamic signals in a large, moving, and light-scattering specimen is extremely challenging, owing to the requirement on high spatiotemporal resolution and difficulty in obtaining high-contrast signals. Here...

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A polymer gel index-matched to water enables diverse applications in fluorescence microscopy

10.1101/2020.10.04.324996 ◽

2020 ◽

Author(s):

Xiaofei Han ◽

Yijun Su ◽

Hamilton White ◽

Kate M. O’Neill ◽

Nicole Y. Morgan ◽

...

Keyword(s):

Surface Passivation ◽

Super Resolution ◽

Light Sheet ◽

Polymer Gel ◽

Structured Illumination ◽

Structured Illumination Microscopy ◽

Neural Structure ◽

Uv Curable ◽

Isotropic Resolution ◽

Light Sheet Microscopy

AbstractWe demonstrate diffraction-limited and super-resolution imaging through thick layers (tens-hundreds of microns) of BIO-133, a biocompatible, UV-curable, commercially available polymer with a refractive index (RI) matched to water. We show that cells can be directly grown on BIO-133 substrates without the need for surface passivation and use this capability to perform extended time-lapse volumetric imaging of cellular dynamics 1) at isotropic resolution using dual-view light-sheet microscopy, and 2) at super-resolution using instant structured illumination microscopy. BIO-133 also enables immobilization of 1) Drosophila tissue, allowing us to track membrane puncta in pioneer neurons, and 2) Caenorhabditis elegans, which allows us to image and inspect fine neural structure and to track pan-neuronal calcium activity over hundreds of volumes. Finally, BIO-133 is compatible with other microfluidic materials, enabling optical and chemical perturbation of immobilized samples, as we demonstrate by performing drug and optogenetic stimulation on cells and C. elegans.

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