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 MEMS enabled miniaturized light-sheet microscopy with all optical control

2021 ◽  
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 (MEMS) 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 340 × 190 × 60 µm3 volumes with 0.25 vps and minimum lateral and axial resolution of 0.9 µm and 2.95 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.

Mechanical Design of the One Million Volt U.S. Steel Microscope Column

1967 ◽  
Vol 25 ◽  
pp. 268-269
Author(s):  
J. R. Fairbanks
Keyword(s):  
Mechanical Design ◽  
Concrete Slab ◽  
Objective Lens ◽  
Optical Elements ◽  
Structural Support ◽  
All Optical ◽  
The One ◽  
Frame Construction ◽  
Type 304 ◽  
Reference Plate

A unitized frame construction is employed as structural support for the one million volt column components. In this way alignment between the high voltage accelerator and the objective lens is maintained, and parallelism between all optical elements is held while permitting manual traverse of the other lenses. The frame, a 700 lb., type 304 stainless casting is suspended from a reference plate, forty inches in diameter by two inches thick. The reference plate is supported by an arched tripod with legs set in a concrete slab isolated from the building proper. (Figure 1) An expandable vacuum seal is located in each of the five shelves of the frame, providing a vacuum seal when expanded and clearance for lens removal when retracted.

scholarly journals Three-Dimensional Microscopy by Milling with Ultraviolet Excitation

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Jiaming Guo ◽  
Camille Artur ◽  
Jason L. Eriksen ◽  
David Mayerich
Keyword(s):  
Three Dimensional ◽  
Light Sheet ◽  
Clinical Settings ◽  
Three Dimensional Imaging ◽  
Image Stack ◽  
Light Sheet Microscopy ◽  
Ultraviolet Excitation ◽  
Tissue Changes ◽  
Block Face ◽  
Core Facilities

Abstract Analysis of three-dimensional biological samples is critical to understanding tissue function and the mechanisms of disease. Many chronic conditions, like neurodegenerative diseases and cancers, correlate with complex tissue changes that are difficult to explore using two-dimensional histology. While three-dimensional techniques such as confocal and light-sheet microscopy are well-established, they are time consuming, require expensive instrumentation, and are limited to small tissue volumes. Three-dimensional microscopy is therefore impractical in clinical settings and often limited to core facilities at major research institutions. There would be a tremendous benefit to providing clinicians and researchers with the ability to routinely image large three-dimensional tissue volumes at cellular resolution. In this paper, we propose an imaging methodology that enables fast and inexpensive three-dimensional imaging that can be readily integrated into current histology pipelines. This method relies on block-face imaging of paraffin-embedded samples using deep-ultraviolet excitation. The imaged surface is then ablated to reveal the next tissue section for imaging. The final image stack is then aligned and reconstructed to provide tissue models that exceed the depth and resolution achievable with modern three-dimensional imaging systems.

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 Enhanced Light Sheet Elastic Scattering Microscopy by Using a Supercontinuum Laser

2019 ◽  
Vol 2 (3) ◽  
pp. 57 ◽  
Author(s):  
Diego Di Battista ◽  
David Merino ◽  
Giannis Zacharakis ◽  
Pablo Loza-Alvarez ◽  
Omar E. Olarte
Keyword(s):  
Elastic Scattering ◽  
Spatial Scales ◽  
Scattered Light ◽  
Spatial Coherence ◽  
Light Sheet ◽  
Structural Features ◽  
Light Sheet Microscopy ◽  
Excitation Laser ◽  
Polarization Filtering ◽  
Microscopy Techniques

Light sheet fluorescence microscopy techniques have revolutionized biological microscopy enabling low-phototoxic long-term 3D imaging of living samples. Although there exist many light sheet microscopy (LSM) implementations relying on fluorescence, just a few works have paid attention to the laser elastic scattering source of contrast available in every light sheet microscope. Interestingly, elastic scattering can potentially disclose valuable information from the structure and composition of the sample at different spatial scales. However, when coherent scattered light is detected with a camera sensor, a speckled intensity is generated on top of the native imaged features, compromising their visibility. In this work, we propose a novel light sheet based optical setup which implements three strategies for dealing with speckles of elastic scattering images: (i) polarization filtering; (ii) reducing the temporal coherence of the excitation laser light; and, (iii) reducing the spatial coherence of the light sheet. Finally, we show how these strategies enable pristine light-sheet elastic-scattering imaging of structural features in challenging biological samples avoiding the deleterious effects of speckle, and without relying on, but complementing, fluorescent labelling.

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.

scholarly journals Achromatic terahertz Airy beam generation with dielectric metasurfaces

Nanophotonics ◽  
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.

scholarly journals Single-Molecule Light-Sheet Microscopy with Local Nanopipette Delivery

2021 ◽  
Vol 93 (8) ◽  
pp. 4092-4099
Author(s):  
Bing Li ◽  
Aleks Ponjavic ◽  
Wei-Hsin Chen ◽  
Lee Hopkins ◽  
Craig Hughes ◽  
...  
Keyword(s):  
Single Molecule ◽  
Light Sheet ◽  
Light Sheet Microscopy
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