scholarly journals Volumetric Lissajous Confocal Microscopy

2019 ◽  
Author(s):  
Takahiro Deguchi ◽  
Paolo Bianchini ◽  
Gemma Palazzolo ◽  
Michele Oneto ◽  
Alberto Diaspro ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Sampling Rate ◽  
Three Dimensional ◽  
Scanning Speed ◽  
Spatiotemporal Resolution ◽  
Volumetric Imaging ◽  
Excitation Beam ◽  
Spatial Changes ◽  
Processing Step ◽  
3D Information

AbstractDynamic biological systems present challenges to existing three-dimensional (3D) optical microscopes because of their continuous temporal and spatial changes. Most techniques are based on rigid architectures, as in confocal microscopy, where a laser beam is sequentially scanned at a predefined spatial sampling rate and pixel dwell time. Here, we developed volumetric Lissajous confocal microscopy to achieve unsurpassed 3D scanning speed with a tunable sampling rate. The system combines an acoustic liquid lens for continuous axial focus translation with a resonant scanning mirror. Accordingly, the excitation beam follows a dynamic Lissajous trajectory enabling sub-millisecond acquisitions of image series containing 3D information at a sub-Nyquist sampling rate. By temporal accumulation and/or advanced interpolation algorithms, volumetric imaging rate is selectable using a post-processing step at the desired spatiotemporal resolution for events of interest. We demonstrate multicolor and calcium imaging over volumes of tens of cubic microns with acquisition speeds up to 5 kHz.

scholarly journals A miniaturized optical tomography platform for volumetric imaging of engineered living systems

Lab on a Chip ◽  
2019 ◽  
Vol 19 (4) ◽  
pp. 550-561 ◽  
Author(s):  
Adem Polat ◽  
Shabir Hassan ◽  
Isa Yildirim ◽  
Luis Eduardo Oliver ◽  
Maryam Mostafaei ◽  
...  
Keyword(s):  
Optical Microscopy ◽  
Biological Sample ◽  
Optical Tomography ◽  
Three Dimensional ◽  
Living Systems ◽  
Volumetric Imaging ◽  
Non Invasive ◽  
3D Information

Volumetric optical microscopy approaches that enable acquisition of three-dimensional (3D) information from a biological sample are attractive for numerous non-invasive imaging applications.

scholarly journals Multi-color 4D superresolution light-sheet microscopy reveals organelle interactions at isotropic 100-nm resolution and sub-second timescales

2021 ◽  
Author(s):  
Peng Fei
Keyword(s):  
Dimensional Space ◽  
Three Dimensional ◽  
Light Sheet ◽  
Three Dimensions ◽  
Live Cells ◽  
Spatiotemporal Resolution ◽  
Volumetric Imaging ◽  
Light Sheet Microscopy ◽  
Intracellular Organelles ◽  
Microscopy Techniques

Long-term visualization of the dynamic organelle-organelle or protein-organelle interactions throughout the three-dimensional space of whole live cells is essential to better understand their functions, but this task remains challenging due to the limitations of existing three-dimensional fluorescence microscopy techniques, such as an insufficient axial resolution, low volumetric imaging rate, and photobleaching. Here, we present the combination of a progressive deep-learning superresolution strategy with a dual-ring-modulated SPIM design capable of visualizing the dynamics of intracellular organelles in live cells for hours at an isotropic spatial resolution of ~100 nm in three dimensions and a temporal resolution up to ~17 Hz. With a compelling spatiotemporal resolution, we substantially reveal the complex spatial relationships and interactions between the endoplasmic reticulum (ER) and mitochondria throughout live cells, providing new insights into ER-mediated mitochondrial division. We also localized the motion of Drp1 oligomers in three dimensions and observed Drp1-mediated mitochondrial branching for the first time.

Laser Scanning Confocal Microscopy and Three-Dimesnsional Reconstruction of the Mitotic Spindles of Unequally Dividing Embryonic Cells

1990 ◽  
Vol 48 (3) ◽  
pp. 166-167
Author(s):  
J. Holy ◽  
G. Schatten
Keyword(s):  
Confocal Microscopy ◽  
Mitotic Spindle ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Laser Scanning Confocal Microscopy ◽  
Two Dimensions ◽  
Embryonic Cells ◽  
Mitotic Spindles ◽  
Cleavage Division ◽  
Scanning Confocal Microscopy

One of the classic limitations of light microscopy has been the fact that three dimensional biological events could only be visualized in two dimensions. Recently, this shortcoming has been overcome by combining the technologies of laser scanning confocal microscopy (LSCM) and computer processing of microscopical data by volume rendering methods. We have employed these techniques to examine morphogenetic events characterizing early development of sea urchin embryos. Specifically, the fourth cleavage division was examined because it is at this point that the first morphological signs of cell differentiation appear, manifested in the production of macromeres and micromeres by unequally dividing vegetal blastomeres.The mitotic spindle within vegetal blastomeres undergoing unequal cleavage are highly polarized and develop specialized, flattened asters toward the micromere pole. In order to reconstruct the three-dimensional features of these spindles, both isolated spindles and intact, extracted embryos were fluorescently labeled with antibodies directed against either centrosomes or tubulin.

Analyzing the Impact of X-Ray Tomography on the Reliability of Integrated Circuits

2015 ◽  
Author(s):  
Halit Dogan ◽  
Md Mahbub Alam ◽  
Navid Asadizanjani ◽  
Sina Shahbazmohamadi ◽  
Domenic Forte ◽  
...  
Keyword(s):  
Integrated Circuits ◽  
Integrated Circuit ◽  
3D Imaging ◽  
Three Dimensional ◽  
Serial Sectioning ◽  
Promising Technique ◽  
Flash Memories ◽  
X Ray ◽  
3D Information ◽  
The Impact

Abstract X-ray tomography is a promising technique that can provide micron level, internal structure, and three dimensional (3D) information of an integrated circuit (IC) component without the need for serial sectioning or decapsulation. This is especially useful for counterfeit IC detection as demonstrated by recent work. Although the components remain physically intact during tomography, the effect of radiation on the electrical functionality is not yet fully investigated. In this paper we analyze the impact of X-ray tomography on the reliability of ICs with different fabrication technologies. We perform a 3D imaging using an advanced X-ray machine on Intel flash memories, Macronix flash memories, Xilinx Spartan 3 and Spartan 6 FPGAs. Electrical functionalities are then tested in a systematic procedure after each round of tomography to estimate the impact of X-ray on Flash erase time, read margin, and program operation, and the frequencies of ring oscillators in the FPGAs. A major finding is that erase times for flash memories of older technology are significantly degraded when exposed to tomography, eventually resulting in failure. However, the flash and Xilinx FPGAs of newer technologies seem less sensitive to tomography, as only minor degradations are observed. Further, we did not identify permanent failures for any chips in the time needed to perform tomography for counterfeit detection (approximately 2 hours).

scholarly journals High-speed volumetric two-photon fluorescence imaging of neurovascular dynamics

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Jiang Lan Fan ◽  
Jose A. Rivera ◽  
Wei Sun ◽  
John Peterson ◽  
Henry Haeberle ◽  
...  
Keyword(s):  
Blood Flow ◽  
High Speed ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Fluorescence Signal ◽  
Imaging Method ◽  
Spatiotemporal Resolution ◽  
Two Photon

AbstractUnderstanding the structure and function of vasculature in the brain requires us to monitor distributed hemodynamics at high spatial and temporal resolution in three-dimensional (3D) volumes in vivo. Currently, a volumetric vasculature imaging method with sub-capillary spatial resolution and blood flow-resolving speed is lacking. Here, using two-photon laser scanning microscopy (TPLSM) with an axially extended Bessel focus, we capture volumetric hemodynamics in the awake mouse brain at a spatiotemporal resolution sufficient for measuring capillary size and blood flow. With Bessel TPLSM, the fluorescence signal of a vessel becomes proportional to its size, which enables convenient intensity-based analysis of vessel dilation and constriction dynamics in large volumes. We observe entrainment of vasodilation and vasoconstriction with pupil diameter and measure 3D blood flow at 99 volumes/second. Demonstrating high-throughput monitoring of hemodynamics in the awake brain, we expect Bessel TPLSM to make broad impacts on neurovasculature research.

scholarly journals Elastic transformation of histological slices allows precise co-registration with microCT data sets for a refined virtual histology approach

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jonas Albers ◽  
Angelika Svetlove ◽  
Justus Alves ◽  
Alexander Kraupner ◽  
Francesca di Lillo ◽  
...  
Keyword(s):  
Three Dimensional ◽  
High Specificity ◽  
Histopathological Analysis ◽  
Specific Information ◽  
Data Sets ◽  
Virtual Histology ◽  
Novel Approach ◽  
Cell Tissue ◽  
Immuno Histochemistry ◽  
3D Information

AbstractAlthough X-ray based 3D virtual histology is an emerging tool for the analysis of biological tissue, it falls short in terms of specificity when compared to conventional histology. Thus, the aim was to establish a novel approach that combines 3D information provided by microCT with high specificity that only (immuno-)histochemistry can offer. For this purpose, we developed a software frontend, which utilises an elastic transformation technique to accurately co-register various histological and immunohistochemical stainings with free propagation phase contrast synchrotron radiation microCT. We demonstrate that the precision of the overlay of both imaging modalities is significantly improved by performing our elastic registration workflow, as evidenced by calculation of the displacement index. To illustrate the need for an elastic co-registration approach we examined specimens from a mouse model of breast cancer with injected metal-based nanoparticles. Using the elastic transformation pipeline, we were able to co-localise the nanoparticles to specifically stained cells or tissue structures into their three-dimensional anatomical context. Additionally, we performed a semi-automated tissue structure and cell classification. This workflow provides new insights on histopathological analysis by combining CT specific three-dimensional information with cell/tissue specific information provided by classical histology.

Two-Photon Excitation Imaging of Glucose Metabolism in Living Tissue

1997 ◽  
Vol 3 (S2) ◽  
pp. 305-306
Author(s):  
David W. Piston
Keyword(s):  
Confocal Microscopy ◽  
Collection Efficiency ◽  
Three Dimensional ◽  
Spatial Filter ◽  
Input Power ◽  
Photon Absorption ◽  
Focal Volume ◽  
Two Photon ◽  
Photon Excitation ◽  
Two Photon Excitation

Two-photon excitation microscopy (TPEM) provides attractive advantages over confocal microscopy for three-dimensionally resolved fluorescence imaging and photochemistry. It provides three-dimensional resolution and eliminates background equivalent to an ideal confocal microscope without requiring a confocal spatial filter, whose absence enhances fluorescence collection efficiency. This results in inherent submicron optical sectioning by excitation alone. In practice, TPEM is made possible by the very high local instantaneous intensity provided by a combination of diffraction-limited focusing of a single laser beam in the microscope and the temporal concentration of 100 femtosecond pulses generated by a mode-locked laser. Resultant peak excitation intensities are 106 times greater than the CW intensities used in confocal microscopy, but the pulse duty cycle of 10−5 limits the average input power to less than 10 mW, only slightly greater than the power normally used in confocal microscopy. Because of the intensity-squared dependence of the two-photon absorption, the excitation is limited to the focal volume.

scholarly journals 3D monitoring of the surface slippage effect on micro-particle sedimentation by digital holographic microscopy

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Majid Panahi ◽  
Ramin Jamali ◽  
Vahideh Farzam Rad ◽  
Mojtaba Khorasani ◽  
Ahamd Darudi ◽  
...  
Keyword(s):  
Slip Length ◽  
Particle Interaction ◽  
Three Dimensional ◽  
Digital Holographic Microscopy ◽  
Particle Sedimentation ◽  
Micro Particles ◽  
Slippage Effect ◽  
Holographic Microscopy ◽  
3D Information ◽  
Experimental Findings

AbstractIn several phenomena in biology and industry, it is required to understand the comprehensive behavior of sedimenting micro-particles in fluids. Here, we use the numerical refocusing feature of digital holographic microscopy (DHM) to investigate the slippage effect on micro-particle sedimentation near a flat wall. DHM provides quantitative phase contrast and three-dimensional (3D) imaging in arbitrary time scales, which suggests it as an elegant approach to investigate various phenomena, including dynamic behavior of colloids. 3D information is obtained by post-processing of the recorded digital holograms. Through analysis of 3D trajectories and velocities of multiple sedimenting micro-particles, we show that proximity to flat walls of higher slip lengths causes faster sedimentation. The effect depends on the ratio of the particle size to (1) the slip length and (2) its distance to the wall. We corroborate our experimental findings by a theoretical model which considers both the proximity and the particle interaction to a wall of different hydrophobicity in the hydrodynamic forces.

scholarly journals Virtual Archaeology of Death and Burial: A Procedure for Integrating 3D Visualization and Analysis in Archaeothanatology

2021 ◽  
Vol 7 (1) ◽  
pp. 540-555
Author(s):  
Hayley L. Mickleburgh ◽  
Liv Nilsson Stutz ◽  
Harry Fokkens
Keyword(s):  
Spatial Information ◽  
3D Visualization ◽  
Rapid Development ◽  
Three Dimensional ◽  
3D Models ◽  
The Body ◽  
Test Case ◽  
Archaeology Of Death ◽  
Human Burials ◽  
3D Information

Abstract The reconstruction of past mortuary rituals and practices increasingly incorporates analysis of the taphonomic history of the grave and buried body, using the framework provided by archaeothanatology. Archaeothanatological analysis relies on interpretation of the three-dimensional (3D) relationship of bones within the grave and traditionally depends on elaborate written descriptions and two-dimensional (2D) images of the remains during excavation to capture this spatial information. With the rapid development of inexpensive 3D tools, digital replicas (3D models) are now commonly available to preserve 3D information on human burials during excavation. A procedure developed using a test case to enhance archaeothanatological analysis and improve post-excavation analysis of human burials is described. Beyond preservation of static spatial information, 3D visualization techniques can be used in archaeothanatology to reconstruct the spatial displacement of bones over time, from deposition of the body to excavation of the skeletonized remains. The purpose of the procedure is to produce 3D simulations to visualize and test archaeothanatological hypotheses, thereby augmenting traditional archaeothanatological analysis. We illustrate our approach with the reconstruction of mortuary practices and burial taphonomy of a Bell Beaker burial from the site of Oostwoud-Tuithoorn, West-Frisia, the Netherlands. This case study was selected as the test case because of its relatively complete context information. The test case shows the potential for application of the procedure to older 2D field documentation, even when the amount and detail of documentation is less than ideal.

scholarly journals Small footprint optoelectrodes using ring resonators for passive light localization

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Vittorino Lanzio ◽  
Gregory Telian ◽  
Alexander Koshelev ◽  
Paolo Micheletti ◽  
Gianni Presti ◽  
...  
Keyword(s):  
State Of The Art ◽  
Three Dimensional ◽  
Network Activity ◽  
Ring Resonators ◽  
Spatiotemporal Resolution ◽  
Optical Ring Resonators ◽  
Light Localization ◽  
Order Of Magnitude ◽  
Small Footprint

AbstractThe combination of electrophysiology and optogenetics enables the exploration of how the brain operates down to a single neuron and its network activity. Neural probes are in vivo invasive devices that integrate sensors and stimulation sites to record and manipulate neuronal activity with high spatiotemporal resolution. State-of-the-art probes are limited by tradeoffs involving their lateral dimension, number of sensors, and ability to access independent stimulation sites. Here, we realize a highly scalable probe that features three-dimensional integration of small-footprint arrays of sensors and nanophotonic circuits to scale the density of sensors per cross-section by one order of magnitude with respect to state-of-the-art devices. For the first time, we overcome the spatial limit of the nanophotonic circuit by coupling only one waveguide to numerous optical ring resonators as passive nanophotonic switches. With this strategy, we achieve accurate on-demand light localization while avoiding spatially demanding bundles of waveguides and demonstrate the feasibility with a proof-of-concept device and its scalability towards high-resolution and low-damage neural optoelectrodes.

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