Tracking Gene Expression via Light Sheet Microscopy and Computer Vision in Living Organisms

2018 ◽  
Author(s):  
Eli Buckner ◽  
Chanae Ottley ◽  
Cranos Williams ◽  
Angels de Luis Balaguer ◽  
Charles E. Melvin ◽  
...  
Keyword(s):  
Gene Expression ◽  
Computer Vision ◽  
Light Sheet ◽  
Light Sheet Microscopy ◽  
Living Organisms

scholarly journals Hydrogel encapsulation of living organisms for long-term microscopy

2017 ◽  
Author(s):  
Kyra Burnett ◽  
Eric Edsinger ◽  
Dirk R. Albrecht
Keyword(s):  
Light Sheet ◽  
C Elegans ◽  
Light Sheet Microscopy ◽  
Confined Spaces ◽  
Single Cell Imaging ◽  
Volumetric Images ◽  
Living Organisms ◽  
Resolution Single ◽  
Hydrogel Encapsulation

AbstractImaging living organisms at high spatial resolution requires effective and innocuous immobilization. Long-term imaging, across development or behavioral states, places further demands on sample mounting with minimal perturbation of the organism. Here we present a simple and inexpensive method for rapid encapsulation of small animals of any developmental stage within a photocrosslinked polyethylene glycol (PEG) hydrogel, gently restricting movement within their confined spaces. Immobilized animals maintained a normal, uncompressed morphology in a hydrated environment and could be exposed to different aqueous chemicals. We focus in particular on the nematode C. elegans, an organism that is typically viewed with paralyzing reagents, nanobeads, adhesives, or microfluidic traps. The hydrogel is optically clear, non-autofluorescent, and nearly index-matched with water for use with light-sheet microscopy. We captured volumetric images of optogenetically-stimulated responses in multiple sensory neurons over 14 hours using a diSPIM light-sheet microscope, and immobilized worms were recoverable and viable after 24 hours encapsulation. We further imaged living pygmy squid hatchlings to demonstrate size scalability, characterized immobilization quality for various crosslinking parameters and identified paralytic-free conditions suitable for high-resolution single cell imaging. PEG hydrogel encapsulation enables continuous observation for hours of small living organisms, from yeast to zebrafish, and is compatible with multiple microscope mounting geometries.

Adaptive light-sheet microscopy for long-term, high-resolution imaging in living organisms

2016 ◽  
Vol 34 (12) ◽  
pp. 1267-1278 ◽  
Author(s):  
Loïc A Royer ◽  
William C Lemon ◽  
Raghav K Chhetri ◽  
Yinan Wan ◽  
Michael Coleman ◽  
...  
Keyword(s):  
High Resolution ◽  
Light Sheet ◽  
High Resolution Imaging ◽  
Light Sheet Microscopy ◽  
Resolution Imaging ◽  
Living Organisms

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

scholarly journals Effect of captopril on post-infarction remodelling visualized by light sheet microscopy and echocardiography

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Urmas Roostalu ◽  
Louise Thisted ◽  
Jacob Lercke Skytte ◽  
Casper Gravesen Salinas ◽  
Philip Juhl Pedersen ◽  
...  
Keyword(s):  
Morphological Changes ◽  
Light Sheet ◽  
Automated Image Analysis ◽  
Border Zone ◽  
Vascular Density ◽  
Mouse Heart ◽  
Imaging Method ◽  
Light Sheet Microscopy ◽  
Post Surgery ◽  
Captopril Treatment

AbstractAngiotensin converting enzyme inhibitors, among them captopril, improve survival following myocardial infarction (MI). The mechanisms of captopril action remain inadequately understood due to its diverse effects on multiple signalling pathways at different time periods following MI. Here we aimed to establish the role of captopril in late-stage post-MI remodelling. Left anterior descending artery (LAD) ligation or sham surgery was carried out in male C57BL/6J mice. Seven days post-surgery LAD ligated mice were allocated to daily vehicle or captopril treatment continued over four weeks. To provide comprehensive characterization of the changes in mouse heart following MI a 3D light sheet imaging method was established together with automated image analysis workflow. The combination of echocardiography and light sheet imaging enabled to assess cardiac function and the underlying morphological changes. We show that delayed captopril treatment does not affect infarct size but prevents left ventricle dilation and hypertrophy, resulting in improved ejection fraction. Quantification of lectin perfused blood vessels showed improved vascular density in the infarct border zone in captopril treated mice in comparison to vehicle dosed control mice. These results validate the applicability of combined echocardiographic and light sheet assessment of drug mode of action in preclinical cardiovascular research.

The Light-Sheet Microscopy

2021 ◽  
Author(s):  
Rolf Theodor Borlinghaus
Keyword(s):  
Light Sheet ◽  
Light Sheet Microscopy

Scattering‐Based Light‐Sheet Microscopy for Rapid Cellular Imaging of Fresh Tissue

2020 ◽  
Author(s):  
Christopher D. Nguyen ◽  
Patrick K. O'Neal ◽  
Nachiket Kulkarni ◽  
Eric Yang ◽  
Dongkyun Kang
Keyword(s):  
Light Sheet ◽  
Cellular Imaging ◽  
Fresh Tissue ◽  
Light Sheet Microscopy

scholarly journals Light sheet fluorescence microscopy of optically cleared brains for studying the glymphatic system

2020 ◽  
Vol 40 (10) ◽  
pp. 1975-1986
Author(s):  
Nicholas B Bèchet ◽  
Tekla M Kylkilahti ◽  
Bengt Mattsson ◽  
Martina Petrasova ◽  
Nagesh C Shanbhag ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Fluid Transport ◽  
Light Sheet ◽  
Brain Parenchyma ◽  
Sleep State ◽  
Glymphatic System ◽  
Light Sheet Microscopy ◽  
Highly Active ◽  
Quantitative Analyses ◽  
Light Sheet Fluorescence Microscopy

Fluid transport in the perivascular space by the glia-lymphatic (glymphatic) system is important for the removal of solutes from the brain parenchyma, including peptides such as amyloid-beta which are implicated in the pathogenesis of Alzheimer’s disease. The glymphatic system is highly active in the sleep state and under the influence of certain of anaesthetics, while it is suppressed in the awake state and by other anaesthetics. Here we investigated whether light sheet fluorescence microscopy of whole optically cleared murine brains was capable of detecting glymphatic differences in sleep- and awake-mimicking anaesthesia, respectively. Using light-sheet imaging of whole brains, we found anaesthetic-dependent cerebrospinal fluid (CSF) influx differences, including reduced tracer influx along tertiary branches of the middle cerebral artery and reduced influx along dorsal and anterior penetrating arterioles, in the awake-mimicking anaesthesia. This study establishes that light sheet microscopy of optically cleared brains is feasible for quantitative analyses and can provide images of the entire glymphatic system in whole brains.

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