scholarly journals Light Sheet Microscopy Using Single Objective Lens

2017 ◽  
Vol 57 (4) ◽  
pp. 200-201
Author(s):  
Sadao OTA
Keyword(s):  
Light Sheet ◽  
Objective Lens ◽  
Light Sheet Microscopy ◽  
Single Objective

scholarly journals High-Resolution, Large Field-of-View, and Multi-View Single Objective Light-Sheet Microscopy

2020 ◽  
Author(s):  
Bin Yang ◽  
Alfred Millett-Sikking ◽  
Merlin Lange ◽  
Ahmet Can Solak ◽  
Hirofumi Kobayashi ◽  
...  
Keyword(s):  
High Resolution ◽  
Light Sheet ◽  
Field Of View ◽  
Large Field ◽  
Sample Rotation ◽  
Light Sheet Microscopy ◽  
Spatio Temporal ◽  
Large Living ◽  
Single Objective ◽  
Imaging Speed

Light-sheet microscopy has become the preferred method for long-term imaging of large living samples because of its low photo-invasiveness and good optical sectioning capabilities. Unfortunately, refraction and scattering often pose obstacles to light-sheet propagation and limit imaging depth. This is typically addressed by imaging multiple complementary views to obtain high and uniform image quality throughout the sample. However, multi-view imaging often requires complex multi-objective configurations that complicate sample mounting, or sample rotation that decreases imaging speed. Recent developments in single-objective light-sheet microscopy have shown that it is possible to achieve high spatio-temporal resolution with a single objective for both illumination and detection. Here we describe a single-objective light-sheet microscope that achieves: (i) high-resolution and large field-of-view imaging via a custom remote focusing objective; (ii) simpler design and ergonomics by remote placement of coverslips; (iii) fast volumetric imaging by means of light-sheet stabilised stage scanning – a novel scanning modality that extends the imaging volume without compromising imaging speed nor quality; (iv) multi-view imaging by means of dual orthogonal light-sheet illumination. Finally, we demonstrate the speed, field of view and resolution of our novel instrument by imaging zebrafish tail development.

Single-objective multiphoton light-sheet microscopy for tumor organoid screening

2019 ◽  
Author(s):  
Eric D. Borrego ◽  
Trung D. Nguyen ◽  
Tim Yeh ◽  
Yen-Liang Liu ◽  
Sarah Bi ◽  
...  
Keyword(s):  
Light Sheet ◽  
Light Sheet Microscopy ◽  
Single Objective

scholarly journals Single Objective Light-Sheet Microscopy for High-Speed Whole-Cell 3D Super-Resolution

2017 ◽  
Vol 112 (3) ◽  
pp. 187a ◽  
Author(s):  
Marjolein B.M. Meddens ◽  
Sheng Liu ◽  
Patrick S. Finnegan ◽  
Thayne L. Edwards ◽  
Conrad D. James ◽  
...  
Keyword(s):  
High Speed ◽  
Super Resolution ◽  
Light Sheet ◽  
Whole Cell ◽  
Light Sheet Microscopy ◽  
Single Objective

scholarly journals MEMS enabled miniaturized light-sheet microscopy with all optical control

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Spyridon Bakas ◽  
Deepak Uttamchandani ◽  
Hiroshi Toshiyoshi ◽  
Ralf Bauer
Keyword(s):  
Microelectromechanical Systems ◽  
Light Sheet ◽  
Objective Lens ◽  
Clinical Settings ◽  
Optical Elements ◽  
Light Sheet Microscopy ◽  
All Optical ◽  
Excitation Laser ◽  
Cost Efficient ◽  
Tunable Lenses

AbstractWe have designed and implemented a compact, cost-efficient miniaturised light-sheet microscopy system based on optical microelectromechanical systems scanners and tunable lenses. The system occupies a footprint of 20 × 28 × 13 cm3 and combines off-the-shelf optics and optomechanics with 3D-printed structural and optical elements, and an economically costed objective lens, excitation laser and camera. All-optical volume scanning enables imaging of 435 × 232 × 60 µm3 volumes with 0.25 vps (volumes per second) and minimum lateral and axial resolution of 1.0 µm and 3.8 µm respectively. An open-top geometry allows imaging of samples on flat bottomed holders, allowing integration with microfluidic devices, multi-well plates and slide mounted samples, with applications envisaged in biomedical research and pre-clinical settings.

scholarly journals High-efficiency, large-area lattice light-sheet generation by dielectric metasurfaces

Nanophotonics ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 4043-4051
Author(s):  
Fenghua Shi ◽  
Jing Wen ◽  
Dangyuan Lei
Keyword(s):  
Spatial Resolution ◽  
Temporal Resolution ◽  
High Efficiency ◽  
Numerical Aperture ◽  
Light Sheet ◽  
Field Of View ◽  
Objective Lens ◽  
High Temporal Resolution ◽  
Light Sheet Microscopy ◽  
Illumination Efficiency

AbstractLattice light-sheet microscopy (LLSM) was developed for long-term live-cell imaging with ultra-fine three-dimensional (3D) spatial resolution, high temporal resolution, and low photo-toxicity by illuminating the sample with a thin lattice-like light-sheet. Currently available schemes for generating thin lattice light-sheets often require complex optical designs. Meanwhile, limited by the bulky objective lens and optical components, the light throughput of existing LLSM systems is rather low. To circumvent the above problems, we utilize a dielectric metasurface of a single footprint to replace the conventional illumination modules used in the conventional LLSM and generate a lattice light-sheet with a ~3-fold broader illumination area and a significantly leveraged illumination efficiency, which consequently leads to a larger field of view with a higher temporal resolution at no extra cost of the spatial resolution. We demonstrate that the metasurface can manipulate spatial frequencies of an input laser beam in orthogonal directions independently to break the trade-off between the field of view and illumination efficiency of the lattice light-sheet. Compared to the conventional LLSM, our metasurface module serving as an ultra-compact illumination component for LLSM at an ease will potentially enable a finer spatial resolution with a larger numerical-aperture detection objective lens.

scholarly journals Single-Objective Multiphoton Light-sheet Microscopy for Lung Cancer Organoid Screening

2020 ◽  
Vol 118 (3) ◽  
pp. 308a
Author(s):  
Trung D. Nguyen ◽  
Yen-Liang Liu ◽  
Dat Nguyen ◽  
Yuan-I Chen ◽  
Yu-An Kuo ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Light Sheet ◽  
Light Sheet Microscopy ◽  
Single Objective

scholarly journals Crossbill: an open access single objective light-sheet microscopy platform

2021 ◽  
Author(s):  
Manish Kumar ◽  
Sandeep Kishore ◽  
David McLean ◽  
Yevgenia Kozorovitskiy
Keyword(s):  
Open Access ◽  
Light Sheet ◽  
Time Lapse ◽  
Volumetric Imaging ◽  
Open Hardware ◽  
Light Sheet Microscopy ◽  
Multiple Alternative ◽  
Optical Configuration ◽  
Time Lapse Imaging ◽  
Single Objective

We present an open access scanned oblique plane microscopy platform Crossbill. It combines a new optical configuration, open hardware assembly, a systematic alignment protocol, and dedicated control software to provide a compact, versatile, high resolution single objective light-sheet microscopy platform. The demonstrated configuration yields the most affordable sub-micron resolution oblique plane microscopy system to date. We add galvanometer enabled tilt-invariant lateral scan for multi-plane, multi-Hz volumetric imaging capability. A precision translation stage extends stitched field of view to centimeter scale. The accompanying open software is optimized for Crossbill and can be easily extended to include alternative configurations. Using Crossbill, we demonstrate large volume structural fluorescence imaging with sub-micron lateral resolution in zebrafish and mouse brain sections. Crossbill is also capable of multiplane functional imaging, and time-lapse imaging. We suggest multiple alternative configurations to extend Crossbill to diverse microscopy applications.

scholarly journals Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution

2016 ◽  
Vol 7 (6) ◽  
pp. 2219 ◽  
Author(s):  
Marjolein B. M. Meddens ◽  
Sheng Liu ◽  
Patrick S. Finnegan ◽  
Thayne L. Edwards ◽  
Conrad D. James ◽  
...  
Keyword(s):  
High Speed ◽  
Super Resolution ◽  
Light Sheet ◽  
Whole Cell ◽  
Light Sheet Microscopy ◽  
Single Objective

scholarly journals Light sheet microscopy for everyone? Experience of building an OpenSPIM to study flatworm development

2016 ◽  
Author(s):  
Johannes Girstmair ◽  
Anne Zakrzewski ◽  
François Lapraz ◽  
Mette Handberg-Thorsager ◽  
Pavel Tomancak ◽  
...  
Keyword(s):  
Open Access ◽  
Open Source ◽  
Early Development ◽  
Thin Sheet ◽  
Light Sheet ◽  
Time Lapse ◽  
Objective Lens ◽  
Light Sheet Microscopy ◽  
Model Animal ◽  
Standard Design

BACKGROUND: Selective plane illumination microscopy (SPIM a type of light sheet microscopy) involves focusing a thin sheet of laser light through a specimen at right angles to the objective lens. As only the thin section of the specimen at the focal plane of the lens is illuminated, out of focus light is naturally absent and toxicity due to light (phototoxicity) is greatly reduced enabling longer term live imaging. OpenSPIM is an open access platform (Pitrone et al. 2013 and OpenSPIM.org) created to give new users step by step instructions on building a basic configuration of a SPIM microscope, which can in principle be adapted and upgraded to each laboratorys own requirements and budget. Here we describe our own experience with the process of designing, building, configuring and using an OpenSPIM for our research into the early development of the polyclad flatworm Maritigrella crozieri, a non model animal. RESULTS: Our OpenSPIM builds on the standard design with the addition of two colour laser illumination for simultaneous detection of two probes/molecules and dual sided illumination, which provides more even signal intensity across a specimen. Our OpenSPIM provides high resolution 3d images and time lapse recordings, and we demonstrate the use of two colour lasers and the benefits of two color dual-sided imaging. We used our microscope to study the development of the embryo of the polyclad flatworm Maritigrella crozieri. The capabilities of our microscope are demonstrated by our ability to record the stereotypical spiral cleavage pattern of Maritigrella with high speed multi view time lapse imaging. 3D and 4D (3D and time) reconstruction of early development from these data is possible using image registration and deconvolution tools provided as part of the open source Fiji platform. We discuss our findings on the pros and cons of a self built microscope. CONCLUSIONS: We conclude that home built microscopes, such as an OpenSPIM, together with the available open source software, such as MicroManager and Fiji, make SPIM accessible to anyone interested in having continuous access to their own light-sheet microscope. However, building an OpenSPIM is not without challenges and an open access microscope is a worthwhile, if significant, investment of time and money. Multi view 4D microscopy is more challenging than we had expected. We hope that our gained experience during this project will help future OpenSPIM users with similar ambitions.

Sign in / Sign up

Export Citation Format

Share Document