scholarly journals Selective volume illumination microscopy offers synchronous volumetric imaging with high contrast

2018 ◽  
Author(s):  
Thai V. Truong ◽  
Daniel B. Holland ◽  
Sara Madaan ◽  
Andrey Andreev ◽  
Josh V. Troll ◽  
...  
Keyword(s):  
Light Field ◽  
Image Data ◽  
Light Sheet ◽  
Beating Heart ◽  
Wide Field ◽  
Axial Resolution ◽  
3D Images ◽  
Volumetric Imaging ◽  
Volume Of Interest ◽  
Image Volume

AbstractLight field microscopy provides an efficient means to collect 3D images in a single acquisition, as its plenoptic detection captures an extended image volume in one snapshot. The ability of light field microscopy to simultaneously capture image data from a volume of interest, such as a functioning brain or a beating heart, is compromised by inadequate contrast and effective resolution, due, in large part, to light scattering by the tissue. Surprisingly, a major contribution to the image degradation is the signal scattered into the volume of interest by the typical wide-field illumination that excites the sample region outside the volume of interest. Here, we minimize this degradation by employing selective volume illumination, using a modified light sheet approach to illuminate preferentially the volume of interest. This minimizes the unavoidable background generated when extraneous regions of the sample are illuminated, dramatically enhancing the contrast and effective resolution of the captured and reconstructed images. Light Field Selective Volume Illumination Microscopy (LF-SVIM, SVIM for short) dramatically improves the performance of light field microscopy, and offers an unprecedented combination of synchronous z-depth coverage, lateral and axial resolution, and imaging speed.

scholarly journals High-contrast, synchronous volumetric imaging with selective volume illumination microscopy

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Thai V. Truong ◽  
Daniel B. Holland ◽  
Sara Madaan ◽  
Andrey Andreev ◽  
Kevin Keomanee-Dizon ◽  
...  
Keyword(s):  
Light Field ◽  
Sample Volume ◽  
Wide Field ◽  
High Contrast ◽  
Background Signal ◽  
Low Contrast ◽  
Volumetric Imaging ◽  
Larval Zebrafish ◽  
Volume Of Interest ◽  
Symbiotic Partner

AbstractLight-field fluorescence microscopy uniquely provides fast, synchronous volumetric imaging by capturing an extended volume in one snapshot, but often suffers from low contrast due to the background signal generated by its wide-field illumination strategy. We implemented light-field-based selective volume illumination microscopy (SVIM), where illumination is confined to only the volume of interest, removing the background generated from the extraneous sample volume, and dramatically enhancing the image contrast. We demonstrate the capabilities of SVIM by capturing cellular-resolution 3D movies of flowing bacteria in seawater as they colonize their squid symbiotic partner, as well as of the beating heart and brain-wide neural activity in larval zebrafish. These applications demonstrate the breadth of imaging applications that we envision SVIM will enable, in capturing tissue-scale 3D dynamic biological systems at single-cell resolution, fast volumetric rates, and high contrast to reveal the underlying biology.

scholarly journals 3D Interferometric Lattice Light-Sheet Imaging

2020 ◽  
Author(s):  
Bin Cao ◽  
Guanshi Wang ◽  
Jieru Li ◽  
Alexandros Pertsinidis
Keyword(s):  
Single Molecule ◽  
Super Resolution ◽  
Light Sheet ◽  
Detection Sensitivity ◽  
Live Cells ◽  
Background Suppression ◽  
Axial Resolution ◽  
Volumetric Imaging ◽  
Spatio Temporal ◽  
And Function

Understanding cellular structure and function requires live-cell imaging with high spatio-temporal resolution and high detection sensitivity. Direct visualization of molecular processes using single-molecule/super-resolution techniques has thus been transformative. However, extracting the highest-resolution 4D information possible from weak and dynamic fluorescence signals in live cells remains challenging. For example, some of the highest spatial resolution methods, e.g. interferometric (4Pi) approaches1-6 can be slow, require high peak excitation intensities that accelerate photobleaching or suffer from increased out-of-focus background. Selective-plane illumination (SPIM)7-12 reduces background, but most implementations typically feature modest spatial, especially axial, resolution. Here we develop 3D interferometric lattice light-sheet (3D-iLLS) imaging, a technique that overcomes many of these limitations. 3D-iLLS provides, by virtue of SPIM, low light levels and photobleaching, while providing increased background suppression and significantly improved volumetric imaging/sectioning capabilities through 4Pi interferometry. We demonstrate 3D-iLLS with axial resolution and single-particle localization precision down to <100nm (FWHM) and <10nm (1σ) respectively. 3D-iLLS paves the way for a fuller elucidation of sub-cellular phenomena by enhanced 4D resolution and SNR live imaging.

scholarly journals Acoustic light-sheet microscopy

2021 ◽  
Author(s):  
Stefan Wunderl ◽  
Ayumu Ishijima ◽  
Etsuo Susaki ◽  
Zihui Xu ◽  
Hong Song ◽  
...  
Keyword(s):  
Light Propagation ◽  
High Spatial Resolution ◽  
Single Cells ◽  
Acoustic Pulse ◽  
Light Sheet ◽  
Wide Field ◽  
Axial Resolution ◽  
3D Objects ◽  
Light Sheet Microscopy ◽  
The Brain

Light-sheet imaging of 3D objects with high spatial resolution remains an open challenge because of the trade-off between field-of-view (FOV) and axial resolution originating from the diffraction of light. We developed acoustic light-sheet microscopy (acoustic LSM), which actively manipulates the light propagation inside a large sample to obtain wide-field microscopic images deep inside a target. By accurately coupling a light-sheet illumination pulse into a planar acoustic pulse, the light-sheet can be continuously guided over large distances. We imaged a fluorescence-labeled transparent mouse brain for the FOVs of 19.3 x 12.4 mm2 and 9.7 x 5.9 mm2 with resolved microstructures and single cells deep inside the brain. Acoustic LSM creates new opportunities for the application of light-sheet in the field of industry to basic science.

scholarly journals Three-photon light-sheet fluorescence microscopy

2018 ◽  
Author(s):  
Adriá Escobet-Montalbán ◽  
Federico M. Gasparoli ◽  
Jonathan Nylk ◽  
Pengfei Liu ◽  
Zhengyi Yang ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Imaging Modality ◽  
Light Sheet ◽  
Wide Field ◽  
Excitation Light ◽  
Volumetric Imaging ◽  
Cellular Spheroids ◽  
Light Sheet Microscopy ◽  
Photon Excitation ◽  
Light Sheet Fluorescence Microscopy

We present the first demonstration of three-photon excitation light-sheet fluorescence microscopy. Light-sheet fluorescence microscopy in single- and two-photon modes has emerged as a powerful wide-field, low photo-damage technique for fast volumetric imaging of biological samples. We extend this imaging modality to the three-photon regime enhancing its penetration depth. Our present study uses a standard conventional femtosecond pulsed laser at 1000 nm wavelength for the imaging of 450 µm diameter cellular spheroids. In addition, we show, experimentally and through numerical simulations, the potential advantages in three-photon light-sheet microscopy of using propagation-invariant Bessel beams in preference to Gaussian beams.

scholarly journals Instantaneous isotropic volumetric imaging of fast biological processes

2018 ◽  
Author(s):  
Nils Wagner ◽  
Nils Norlin ◽  
Jakob Gierten ◽  
Gustavo de Medeiros ◽  
Bálint Balázs ◽  
...  
Keyword(s):  
Blood Flow ◽  
Light Field ◽  
Flow Imaging ◽  
Biological Processes ◽  
Motion Artefacts ◽  
3D Images ◽  
Volumetric Imaging ◽  
Blood Flow Imaging ◽  
Cellular Resolution ◽  
Simultaneous Acquisition

AbstractCapturing highly dynamic biological processes at sub-cellular resolution is a recurring challenge in biology. Here we show that combining selective volume illumination with simultaneous acquisition of orthogonal light-fields yields 3D images with high, isotropic spatial resolution and free of reconstruction artefacts, thereby overcoming current limitations of light-field microscopy implementations. We demonstrate Medaka heart and blood flow imaging with single-cell resolution and free of motion artefacts at volume rates up to 200Hz.

scholarly journals Recurrent neural network-based volumetric fluorescence microscopy

2021 ◽  
Vol 10 (1) ◽  
Author(s):  
Luzhe Huang ◽  
Hanlong Chen ◽  
Yilin Luo ◽  
Yair Rivenson ◽  
Aydogan Ozcan
Keyword(s):  
Neural Network ◽  
Image Reconstruction ◽  
Fluorescence Microscopy ◽  
Recurrent Network ◽  
Sample Volume ◽  
Depth Of Field ◽  
Objective Lens ◽  
Wide Field ◽  
Volumetric Imaging ◽  
3D Image Reconstruction

AbstractVolumetric imaging of samples using fluorescence microscopy plays an important role in various fields including physical, medical and life sciences. Here we report a deep learning-based volumetric image inference framework that uses 2D images that are sparsely captured by a standard wide-field fluorescence microscope at arbitrary axial positions within the sample volume. Through a recurrent convolutional neural network, which we term as Recurrent-MZ, 2D fluorescence information from a few axial planes within the sample is explicitly incorporated to digitally reconstruct the sample volume over an extended depth-of-field. Using experiments on C. elegans and nanobead samples, Recurrent-MZ is demonstrated to significantly increase the depth-of-field of a 63×/1.4NA objective lens, also providing a 30-fold reduction in the number of axial scans required to image the same sample volume. We further illustrated the generalization of this recurrent network for 3D imaging by showing its resilience to varying imaging conditions, including e.g., different sequences of input images, covering various axial permutations and unknown axial positioning errors. We also demonstrated wide-field to confocal cross-modality image transformations using Recurrent-MZ framework and performed 3D image reconstruction of a sample using a few wide-field 2D fluorescence images as input, matching confocal microscopy images of the same sample volume. Recurrent-MZ demonstrates the first application of recurrent neural networks in microscopic image reconstruction and provides a flexible and rapid volumetric imaging framework, overcoming the limitations of current 3D scanning microscopy tools.

scholarly journals Optical Navigation Sensor for Runway Relative Positioning of Aircraft during Final Approach

Sensors ◽  
2021 ◽  
Vol 21 (6) ◽  
pp. 2203
Author(s):  
Antal Hiba ◽  
Attila Gáti ◽  
Augustin Manecy
Keyword(s):  
Optical Sensors ◽  
Inertial Sensor ◽  
Image Data ◽  
Flight Test ◽  
Image Features ◽  
Wide Field ◽  
Test Results ◽  
Wide Field Of View ◽  
Final Approach ◽  
Marker Detection

Precise navigation is often performed by sensor fusion of different sensors. Among these sensors, optical sensors use image features to obtain the position and attitude of the camera. Runway relative navigation during final approach is a special case where robust and continuous detection of the runway is required. This paper presents a robust threshold marker detection method for monocular cameras and introduces an on-board real-time implementation with flight test results. Results with narrow and wide field-of-view optics are compared. The image processing approach is also evaluated on image data captured by a different on-board system. The pure optical approach of this paper increases sensor redundancy because it does not require input from an inertial sensor as most of the robust runway detectors.

scholarly journals Light sheet-excited spontaneous Raman imaging of a living fish by optical sectioning in a wide field Raman microscope

2012 ◽  
Vol 20 (15) ◽  
pp. 16195 ◽  
Author(s):  
Yusuke Oshima ◽  
Hidetoshi Sato ◽  
Hiroko Kajiura-Kobayashi ◽  
Tetsuaki Kimura ◽  
Kiyoshi Naruse ◽  
...  
Keyword(s):  
Light Sheet ◽  
Raman Imaging ◽  
Wide Field ◽  
Optical Sectioning

scholarly journals Tracking cells in epithelial acini by light sheet microscopy reveals proximity effects in breast cancer initiation

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Ashna Alladin ◽  
Lucas Chaible ◽  
Lucia Garcia del Valle ◽  
Reither Sabine ◽  
Monika Loeschinger ◽  
...  
Keyword(s):  
Single Cells ◽  
Image Data ◽  
Integrated Approach ◽  
Light Sheet ◽  
Mammary Epithelial ◽  
Proximity Effects ◽  
Transformed Cells ◽  
Light Sheet Microscopy ◽  
Primary Mouse ◽  
Longitudinal Imaging

Cancer clone evolution takes place within tissue ecosystem habitats. But, how exactly tumors arise from a few malignant cells within an intact epithelium is a central, yet unanswered question. This is mainly due to the inaccessibility of this process to longitudinal imaging together with a lack of systems that model the progression of a fraction of transformed cells within a tissue. Here, we developed a new methodology based on primary mouse mammary epithelial acini, where oncogenes can be switched on in single cells within an otherwise normal epithelial cell layer. We combine this stochastic breast tumor induction model with inverted light-sheet imaging to study single-cell behavior for up to four days and analyze cell fates utilizing a newly developed image-data analysis workflow. The power of this integrated approach is illustrated by us finding that small local clusters of transformed cells form tumors while isolated transformed cells do not.

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