scholarly journals Capturing volumetric dynamics at high speed in the brain by confocal light field microscopy

2020 ◽  
Author(s):  
Zhenkun Zhang ◽  
Lu Bai ◽  
Lin Cong ◽  
Peng Yu ◽  
Tianlei Zhang ◽  
...  
Keyword(s):  
High Speed ◽  
Light Field ◽  
Prey Capture ◽  
Vascular System ◽  
Imaging Techniques ◽  
Three Dimensions ◽  
Proper Function ◽  
Imaging Method ◽  
Volumetric Imaging ◽  
Neural Activities

AbstractNeural network performs complex computations through coordinating collective neural dynamics that are fast and in three-dimensions. Meanwhile, its proper function relies on its 3D supporting environment, including the highly dynamic vascular system that drives energy and material flow. Better understanding of these processes requires methods to capture fast volumetric dynamics in thick tissue. This becomes challenging due to the trade-off between speed and optical sectioning capability in conventional imaging techniques. Here we present a new imaging method, confocal light field microscopy, to enable fast volumetric imaging deep into brain. We demonstrated the power of this method by recording whole brain calcium transients in freely swimming larval zebrafish and observed behaviorally correlated activities on single neurons during its prey capture. Furthermore, we captured neural activities and circulating blood cells over a volume ⌀ 800 μm × 150 μm at 70 Hz and up to 600 μm deep in the mice brain.

scholarly journals Fast Objective Coupled Planar Illumination Microscopy

2018 ◽  
Author(s):  
Cody Greer ◽  
Timothy E. Holy
Keyword(s):  
Fluorescence Microscopy ◽  
High Speed ◽  
Imaging Techniques ◽  
Light Sheet ◽  
Frame Rate ◽  
Three Dimensions ◽  
Two Photon Microscopy ◽  
Image Sharing ◽  
Scanning Rate ◽  
Fast Light

Among optical imaging techniques light sheet fluorescence microscopy stands out as one of the most attractive for capturing high-speed biological dynamics unfolding in three dimensions. The technique is potentially millions of times faster than point-scanning techniques such as two-photon microscopy. However current-generation light sheet microscopes are limited by volume scanning rate and/or camera frame rate. We present speed-optimized Objective Coupled Planar Illumination (OCPI) microscopy, a fast light sheet technique that avoids compromising image quality or photon efficiency. We increase volume scanning rate to 40 Hz for volumes up to 700 µm thick and introduce Multi-Camera Image Sharing (MCIS), a technique to scale imaging rate by parallelizing acquisition across cameras. Finally, we demonstrate fast calcium imaging of the larval zebrafish brain and find a heartbeat-induced artifact that can be removed by filtering when the imaging rate exceeds 15 Hz. These advances extend the reach of fluorescence microscopy for monitoring fast processes in large volumes.

The kinematics and mechanism of prey capture in the African pig-nosed frog (Hemisus marmoratum): description of a radically divergent anuran tongue.

1995 ◽  
Vol 198 (9) ◽  
pp. 2025-2040 ◽  
Author(s):  
D Ritter ◽  
K Nishikawa
Keyword(s):  
Feeding Behavior ◽  
Strong Evidence ◽  
Muscle Function ◽  
High Speed ◽  
Prey Capture ◽  
Three Dimensions ◽  
Muscle Denervation ◽  
Genioglossus Muscle ◽  
Pulling Forces ◽  
Hydraulic Mechanism

High-speed videography and muscle denervation experiments were used to quantify the feeding kinematics of Hemisus marmoratum and to test hypotheses of muscle function. The feeding behavior of H. marmoratum, which feeds on ants and termites, differs radically from that of other frogs that have been studied. During feeding in H. marmoratum, the tongue 'telescopes' straight out of the mouth, as opposed to the 'flipping' tongue trajectory observed in most other frogs. At the time of prey contact, two lateral lobes of tissue at the tongue tip envelop the prey. These lateral lobes are capable of applying significant pulling forces to the prey and the tongue is, therefore, described as prehensile. The trajectory of the tongue can be adjusted throughout protraction so that the frog can 'aim' its tongue in all three dimensions; distance, azimuth and elevation. Bilateral denervation of the genioglossus muscles results in a complete lack of tongue protraction, indicating that the genioglossus muscle is the main tongue protractor in H. marmoratum, as in other frogs. Thus, H. marmoratum provides strong evidence of functional conservatism of the genioglossus muscle within anurans. Bilateral denervation of the hyoglossus muscle indicates that although the hyoglossus is involved in several aspects of normal tongue retraction, including the prehensile capability of the tongue tip, it is not necessary for tongue retraction. Unilateral denervation of the genioglossus muscle causes significant deviation of the tongue towards the denervated side, providing evidence for a mechanism of lateral tongue aiming. On the basis of the kinematics of prey capture, the anatomy of the tongue and the results of the denervation experiments, we propose that H. marmoratum uses a hydraulic mechanism to protract its tongue.

Three Dimensional Imaging of LIGA-Made Microcomponents

2004 ◽  
Vol 126 (4) ◽  
pp. 813-821 ◽  
Author(s):  
Douglas Chinn ◽  
Peter Ostendorp ◽  
Mike Haugh ◽  
Russell Kershmann ◽  
Thomas Kurfess ◽  
...  
Keyword(s):  
Computer Aided Design ◽  
High Speed ◽  
Three Dimensional ◽  
Point Clouds ◽  
Three Dimensions ◽  
Cad Model ◽  
Volumetric Imaging ◽  
Data Registration ◽  
Liga Technology ◽  
Coordinate Metrology

Nickel and nickel-alloy microparts sized on the order of 5–1000 microns have been imaged in three dimensions using a new microscopic technique, Digital Volumetric Imaging (DVI). The gears were fabricated using Sandia National Laboratories’ LIGA technology (lithography, molding, and electroplating). The images were taken on a microscope built by Resolution Sciences Corporation by slicing the gear into one-micron thin slices, photographing each slice, and then reconstructing the image with software. The images were matched to the original CAD (computer aided design) model, allowing LIGA designers, for the first time, to see visually how much deviation from the design is induced by the manufacturing process. Calibration was done by imaging brass ball bearings and matching them to the CAD model of a sphere. A major advantage of DVI over scanning techniques is that internal defects can be imaged to very high resolution. In order to perform the metrology operations on the microcomponents, high-speed and high-precision algorithms are developed for coordinate metrology. The algorithms are based on a least-squares approach to data registration the {X,Y,Z} point clouds generated from the component surface onto a target geometry defined in a CAD model. Both primitive geometric element analyses as well as an overall comparison of the part geometry are discussed. Initial results of the micromeasurements are presented in the paper.

scholarly journals Fast objective coupled planar illumination microscopy

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Cody J. Greer ◽  
Timothy E. Holy
Keyword(s):  
Fluorescence Microscopy ◽  
High Speed ◽  
Imaging Techniques ◽  
Light Sheet ◽  
Three Dimensions ◽  
Planar Imaging ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Larval Zebrafish ◽  
Fast Light

Abstract Among optical imaging techniques light sheet fluorescence microscopy is one of the most attractive for capturing high-speed biological dynamics unfolding in three dimensions. The technique is potentially millions of times faster than point-scanning techniques such as two-photon microscopy. However light sheet microscopes are limited by volume scanning rate and/or camera speed. We present speed-optimized Objective Coupled Planar Illumination (OCPI) microscopy, a fast light sheet technique that avoids compromising image quality or photon efficiency. Our fast scan system supports 40 Hz imaging of 700 μm-thick volumes if camera speed is sufficient. We also address the camera speed limitation by introducing Distributed Planar Imaging (DPI), a scaleable technique that parallelizes image acquisition across cameras. Finally, we demonstrate fast calcium imaging of the larval zebrafish brain and find a heartbeat-induced artifact, removable when the imaging rate exceeds 15 Hz. These advances extend the reach of fluorescence microscopy for monitoring fast processes in large volumes.

scholarly journals HiLo Based Line Scanning Temporal Focusing Microscopy for High-Speed, Deep Tissue Imaging

Membranes ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 634
Author(s):  
Ruheng Shi ◽  
Yuanlong Zhang ◽  
Tiankuang Zhou ◽  
Lingjie Kong
Keyword(s):  
High Speed ◽  
Three Dimensions ◽  
Tissue Imaging ◽  
Deep Penetration ◽  
High Temporal Resolution ◽  
Structured Illumination ◽  
Volumetric Imaging ◽  
Temporal Focusing ◽  
Line Scanning

High-speed, optical-sectioning imaging is highly desired in biomedical studies, as most bio-structures and bio-dynamics are in three-dimensions. Compared to point-scanning techniques, line scanning temporal focusing microscopy (LSTFM) is a promising method that can achieve high temporal resolution while maintaining a deep penetration depth. However, the contrast and axial confinement would still be deteriorated in scattering tissue imaging. Here, we propose a HiLo-based LSTFM, utilizing structured illumination to inhibit the fluorescence background and, thus, enhance the image contrast and axial confinement in deep imaging. We demonstrate the superiority of our method by performing volumetric imaging of neurons and dynamical imaging of microglia in mouse brains in vivo.

scholarly journals View-channel-depth light-field microscopy: real-time volumetric reconstruction of biological dynamics by deep learning

2018 ◽  
Author(s):  
Zhaoqiang Wang ◽  
Lanxin Zhu ◽  
Hao Zhang ◽  
Guo Li ◽  
Chengqiang Yi ◽  
...  
Keyword(s):  
High Speed ◽  
Light Field ◽  
Three Dimensional ◽  
Image Sequences ◽  
Biological Processes ◽  
Channel Depth ◽  
Volumetric Imaging ◽  
C Elegans ◽  
Volumetric Reconstruction ◽  
Zebrafish Heart

AbstractLight-field microscopy has emerged as a technique of choice for high-speed volumetric imaging of fast biological processes. However, artefacts, non-uniform resolution, and a slow reconstruction speed have limited its full capabilities for in toto extraction of the dynamic spatiotemporal patterns in samples. Here, we combined a view-channel-depth (VCD) neural network with light-field microscopy to mitigate these limitations, yielding artefact-free three-dimensional image sequences with uniform spatial resolution and three-order-higher video-rate reconstruction throughput. We imaged neuronal activities across moving C. elegans and blood flow in a beating zebrafish heart at single-cell resolution with volume rates up to 200 Hz.

High Speed Micro Holographic PIV Measurements of Microorganisms

2008 ◽  
Author(s):  
M. Zarzecki ◽  
F. J. Diez
Keyword(s):  
Velocity Field ◽  
High Speed ◽  
Fourier Transforms ◽  
Three Dimensional ◽  
Three Dimensions ◽  
Imaging Method ◽  
Time Resolved ◽  
Feeding Methods ◽  
Holographic Particle Image Velocimetry ◽  
Three Components

Holographic particle image velocimetry (PIV) is a novel application of holography that allows for tracking of small particle sized objects in a small volume. Whereas regular PIV allows for the two in-plane components of the velocity field to be measured, and stereoscopic PIV allows for the three-components of the velocity field to be measured in a thin plane, holographic PIV allows for the three-components of the velocity to be measured for each individual particle present in the measuring volume, thus allowing to fully resolve fluid flows that are inherently 3D in nature. There are many examples of three dimensional flows in nature including turbulence flows, but another very interesting application very well suited for this technique involves tracking living microorganisms in order to study their motion and their means of propulsion. As part of this research a micro organism was tracked in three dimensions using a high speed microscopic holographic imaging method. The ability to track organisms in 3D allows better understanding and characterizing of their behavior including their propulsion methods, their feeding methods and their interaction with each other. The time resolved holograms were reconstructed in Matlab using Fast Fourier Transforms. A laser pointer was used as a source of coherent light, and a high speed PIV camera (Photron APX Ultima) was used to capture the images. A beam expander was used to increase the diameter of the laser beam allowing for a larger tracking area. Results with this system will show the trajectories in 3D of microorganisms as well as the three components of the velocity field showing the interaction of the organisms with their environment.

scholarly journals High-speed large-scale 4D activities mapping of moving C. elegans by deep-learning-enabled light-field microscopy on a chip

2021 ◽  
Author(s):  
Tingting Zhu ◽  
Lanxin Zhu ◽  
Yi Li ◽  
Xiaopeng Chen ◽  
Mingyang He ◽  
...  
Keyword(s):  
Deep Learning ◽  
High Speed ◽  
Large Scale ◽  
Light Field ◽  
Wild Type ◽  
C Elegans ◽  
Neural Activities ◽  
Biological Studies ◽  
Different Types ◽  
Field Imaging

We report a novel fusion of microfluidics and light-field microscopy, to achieve high-speed 4D (space + time) imaging of moving C. elegans on a chip. Our approach combines automatic chip-based worm loading / compartmentalization / flushing / reloading with instantaneous deep-learning light-field imaging of moving worm. Taken together, we realized intoto image-based screening of wild-type and uncoordinated-type worms at a volume rate of 33 Hz, with sustained observation of 1 minute per worm, and overall throughput of 42 worms per hour. With quickly yielding over 80000 image volumes that four-dimensionally visualize the dynamics of all the worms, we can quantitatively analyse their behaviours as well as the neural activities, and correlate the phenotypes with the neuron functions. The different types of worms can be readily identified as a result of the high-throughput activity mapping. Our approach shows great potential for various lab-on-a-chip biological studies, such as embryo sorting and cell growth assays.

scholarly journals High-speed multifocus phase imaging in thick tissue

2021 ◽  
Author(s):  
Sheng Xiao ◽  
Shuqi Zheng ◽  
Jerome Mertz
Keyword(s):  
High Speed ◽  
Imaging Techniques ◽  
Index Of Refraction ◽  
Phase Imaging ◽  
Biological Processes ◽  
Thick Sample ◽  
Volumetric Imaging ◽  
Phase Microscopy ◽  
Large Numbers ◽  
Qualitative Phase

Phase microscopy is widely used to image unstained biological samples. However, most phase imaging techniques require transmission geometries, making them unsuited for thick sample applications. Moreover, when applied to volumetric imaging, phase imaging generally requires large numbers of measurements, often making it too slow to capture live biological processes with fast 3D index-of-refraction variations. By combining oblique back-illumination microscopy and a z-splitter prism, we perform phase imaging that is both epi-mode and multifocus, enabling high-speed 3D phase imaging in thick, scattering tissues with a single camera. We demonstrate here 3D qualitative phase imaging of blood flow in chick embryos over a field of view of 546 × 546 × 137 μm3 at speeds up to 47 Hz.

Five-Dimensional Microscopy using Widefield-Deconvolution: Practical Considerations and Biological Applications

1996 ◽  
Vol 54 ◽  
pp. 268-269
Author(s):  
W.F. Marshall ◽  
K. Oegema ◽  
J. Nunnari ◽  
A.F. Straight ◽  
D.A. Agard ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
High Speed ◽  
Laser Scanning ◽  
Ccd Camera ◽  
Image Plane ◽  
Time Lapse ◽  
Laser Scanning Confocal Microscopy ◽  
Three Dimensions ◽  
Excitation Light ◽  
Cooled Ccd

The ability to image cells in three dimensions has brought about a revolution in biological microscopy, enabling many questions to be asked which would be inaccessible without this capability. There are currently two major methods of three dimensional microscopy: laser-scanning confocal microscopy and widefield-deconvolution microscopy. The method of widefield-deconvolution uses a cooled CCD to acquire images from a standard widefield microscope, and then computationally removes out of focus blur. Using such a scheme, it is easy to acquire time-lapse 3D images of living cells without killing them, and to do so for multiple wavelengths (using computer-controlled filter wheels). Thus, it is now not only feasible, but routine, to perform five dimensional microscopy (three spatial dimensions, plus time, plus wavelength).Widefield-deconvolution has several advantages over confocal microscopy. The two main advantages are high speed of acquisition (because there is no scanning, a single optical section is acquired at a time by using a cooled CCD camera) and the use of low excitation light levels Excitation intensity can be much lower than in a confocal microscope for three reasons: 1) longer exposures can be taken since the entire 512x512 image plane is acquired in parallel, so that dwell time is not an issue, 2) the higher quantum efficiently of a CCD detect over those typically used in confocal microscopy (although this is expected to change due to advances in confocal detector technology), and 3) because no pinhole is used to reject light, a much larger fraction of the emitted light is collected. Thus we can typically acquire images with thousands of photons per pixel using a mercury lamp, instead of a laser, for illumination. The use of low excitation light is critical for living samples, and also reduces bleaching. The high speed of widefield microscopy is also essential for time-lapse 3D microscopy, since one must acquire images quickly enough to resolve interesting events.

Sign in / Sign up

Export Citation Format

Share Document