scholarly journals Image quality guided smart rotation improves coverage in microscopy

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Jiaye He ◽  
Jan Huisken
Keyword(s):  
Image Quality ◽  
Light Sheet ◽  
Field Of View ◽  
Optical Performance ◽  
Sample Rotation ◽  
Optical Instruments ◽  
Selective Plane Illumination Microscopy ◽  
Data Volume ◽  
Optical Paths ◽  
Additional Hardware

AbstractFluorescence microscopy is an essential tool for biological discoveries. There is a constant demand for better spatial resolution across a larger field of view. Although strides have been made to improve the theoretical resolution and speed of the optical instruments, in mesoscopic samples, image quality is still largely limited by the optical properties of the sample. In Selective Plane Illumination Microscopy (SPIM), the achievable optical performance is hampered by optical degradations encountered in both the illumination and detection. Multi-view imaging, either through sample rotation or additional optical paths, is a popular strategy to improve sample coverage. In this work, we introduce a smart rotation workflow that utilizes on-the-fly image analysis to identify the optimal light sheet imaging orientations. The smart rotation workflow outperforms the conventional approach without additional hardware and achieves a better sample coverage using the same number of angles or less and thereby reduces data volume and phototoxicity.

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.

scholarly journals Transverse Chromatic Aberrations in Virtual Reality Devices

Frameless ◽  
2019 ◽  
Vol 1 (1) ◽  
pp. 1-4
Author(s):  
Ryan Beams ◽  
◽  
Wei-Chung Cheng ◽  
Andrea S. Kim ◽  
Aldo Badano ◽  
...  
Keyword(s):  
Virtual Reality ◽  
Image Quality ◽  
Relative Size ◽  
Chromatic Aberration ◽  
Field Of View ◽  
Test Results ◽  
Optical Performance ◽  
Color Changes ◽  
Head Mounted Display ◽  
Chromatic Aberrations

We demonstrate a method for measuring the transverse chromatic aberration (TCA) in a virtual reality head-mounted display (VR HMD). This procedure was used to characterize the optical performance of the Oculus Go VR HMD. Results show a measurable TCA for angles larger than approximately 6◦ from the center of the field of view. TCA can be thought of as a wavelength dependent magnification, and as a result, the relative size of objects vary based on the rendering color. In addition, this leads to color changes in the image due to mixing with neighboring pixels, which impacts image quality. The test results for the Oculus Go show promise for characterizing TCA across different HMDs.

Polar coordinate coherent passive acoustic beamforming for the modifiable improvement of image quality and field of view

2021 ◽  
Author(s):  
Chunqi Li ◽  
Harry R. Clegg ◽  
Thomas M. Carpenter ◽  
Luzhen Nie ◽  
Steven Freear ◽  
...  
Keyword(s):  
Image Quality ◽  
Field Of View ◽  
Polar Coordinate ◽  
Passive Acoustic

scholarly journals Long-term in toto cell tracking using lightsheet microscopy of the zebrafish tailbud

2019 ◽  
Vol 3 ◽  
pp. 163
Author(s):  
Timothy Fulton ◽  
Martin O. Lenz ◽  
Leila Muresan ◽  
Toby Andrews ◽  
Courtney Lancaster ◽  
...  
Keyword(s):  
Temporal Resolution ◽  
Cell Tracking ◽  
Image Acquisition ◽  
Light Sheet ◽  
Field Of View ◽  
Image Capture ◽  
Acquisition Period ◽  
Global Movement ◽  
Microscope Stage

In toto light-sheet imaging allows the tracking of entire growing tissues with high spatial and temporal resolution for many hours. However, this technology requires a sample to be immobilised to ensure that the tissue of interest remains within the field of view throughout the image acquisition period. We have developed a method of mounting and image capture for long-term light-sheet imaging of a growing zebrafish tailbud from the 18 somite stage through to the end of somitogenesis. By tracking the global movement of the tailbud during image acquisition and feeding this back to the microscope stage, we are able to ensure that the growing tissue remains within the field of view throughout image acquisition. Here, we present three representative datasets of embryos in which all nuclei are labelled and tracked until the completion of somitogenesis.

On the use of image quality measures of multiviews in light sheet fluorescence 3D microscopy

2018 ◽  
Author(s):  
Carole Frindel ◽  
Charlotte Riviere ◽  
Rosa Huaman ◽  
Andrea BASSI ◽  
David Rousseau
Keyword(s):  
Image Quality ◽  
Quality Measures ◽  
Light Sheet ◽  
3D Microscopy ◽  
Image Quality Measures

scholarly journals In Vivo Cell Tracking and Cleared Tissue Imaging with Extended Field of View Selective Plane Illumination Microscopy

2019 ◽  
Vol 116 (3) ◽  
pp. 566a
Author(s):  
Leonardo A. Saunders ◽  
Devin Pace ◽  
Arianna Gentile ◽  
Dominik Stich ◽  
Angeles B. Ribera ◽  
...  
Keyword(s):  
Cell Tracking ◽  
Field Of View ◽  
Tissue Imaging ◽  
Selective Plane Illumination Microscopy ◽  
Extended Field ◽  
Extended Field Of View

Enlarged field of view based on Schwartz modulation for light sheet fluorescence microscopy in deep tissue

2020 ◽  
Vol 45 (17) ◽  
pp. 4851
Author(s):  
Xiaobin Xu ◽  
Jiajia Chen ◽  
Biwei Zhang ◽  
Limeng Huang ◽  
Yao Zheng ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Light Sheet ◽  
Field Of View ◽  
Deep Tissue ◽  
Light Sheet Fluorescence Microscopy

The Field Emission Gun Scanning Electron Microscopehigh Resolution at Low Beam Energies

1994 ◽  
Vol 332 ◽  
Author(s):  
David C Joy
Keyword(s):  
Electron Microscope ◽  
High Resolution ◽  
Field Emission ◽  
State Of The Art ◽  
Design Criteria ◽  
Nanometer Scale ◽  
Optical Performance ◽  
Bulk Specimen ◽  
Optical Instruments ◽  
Scanning Electron

ABSTRACTThe scanning electron microscope (SEM) is the most widely used, as well as the most versatile, of electron-optical instruments. In all of the imaging modes of the instrument it is now possible to achieve a spatial resolution on the nanometer scale from a bulk specimen, provided that the electron-optical performance of the instrument is of the necessary quality. In practice this means that a field emission gun (FEG) as well as a highly excited probe forming lens must be used. With state-of-the-art instrumentation, image resolutions as good as 4 - 5 nm at lkeV, and lnm at 20keV, are now achievable. This paper outlines the design criteria for a high resolution FEG SEM and discusses the type of image information available. The performance achieved is compared to that of other types of microscope that offer similar capabilities.

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