scholarly journals Isotropic Light-Sheet Microscopy and Automated Cell Lineage Analyses to Catalogue Caenorhabditis elegans Embryogenesis with Subcellular Resolution

10.3791/59533 ◽  
2019 ◽  
Author(s):  
Leighton H. Duncan ◽  
Mark W. Moyle ◽  
Lin Shao ◽  
Titas Sengupta ◽  
Richard Ikegami ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Cell Lineage ◽  
Light Sheet ◽  
Light Sheet Microscopy ◽  
Subcellular Resolution

scholarly journals Imaging plant germline differentiation within Arabidopsis flowers by light sheet microscopy

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Sona Valuchova ◽  
Pavlina Mikulkova ◽  
Jana Pecinkova ◽  
Jana Klimova ◽  
Michal Krumnikl ◽  
...  
Keyword(s):  
Higher Plants ◽  
Three Dimensional ◽  
Cell Lineage ◽  
Light Sheet ◽  
Three Dimensional Model ◽  
Light Sheet Microscopy ◽  
Meiotic Chromosomes ◽  
Short Period ◽  
Germline Differentiation ◽  
Subcellular Resolution

In higher plants, germline differentiation occurs during a relatively short period within developing flowers. Understanding of the mechanisms that govern germline differentiation lags behind other plant developmental processes. This is largely because the germline is restricted to relatively few cells buried deep within floral tissues, which makes them difficult to study. To overcome this limitation, we have developed a methodology for live imaging of the germ cell lineage within floral organs of Arabidopsis using light sheet fluorescence microscopy. We have established reporter lines, cultivation conditions, and imaging protocols for high-resolution microscopy of developing flowers continuously for up to several days. We used multiview imagining to reconstruct a three-dimensional model of a flower at subcellular resolution. We demonstrate the power of this approach by capturing male and female meiosis, asymmetric pollen division, movement of meiotic chromosomes, and unusual restitution mitosis in tapetum cells. This method will enable new avenues of research into plant sexual reproduction.

scholarly journals Imaging plant germline differentiation within Arabidopsis flower by light sheet microscopy

2019 ◽  
Author(s):  
Sona Valuchova ◽  
Pavlina Mikulkova ◽  
Jana Pecinkova ◽  
Jana Klimova ◽  
Michal Krumnikl ◽  
...  
Keyword(s):  
Higher Plants ◽  
Three Dimensional ◽  
Cell Lineage ◽  
Light Sheet ◽  
Three Dimensional Model ◽  
Light Sheet Microscopy ◽  
Meiotic Chromosomes ◽  
Short Period ◽  
Germline Differentiation ◽  
Subcellular Resolution

AbstractIn higher plants, germline differentiation occurs during a relatively short period within developing flowers. Understanding of the mechanisms that govern germline differentiation lags behind other plant developmental processes. This is largely because the germline is restricted to relatively few cells buried deep within floral tissues, which makes them difficult to study. To overcome this limitation, we have developed a methodology for live imaging of the germ cell lineage within floral organs of Arabidopsis using light sheet fluorescence microscopy. We have established reporter lines, cultivation conditions, and imaging protocols for high-resolution microscopy of developing flowers continuously for up to several days. We used multiview imagining to reconstruct a three-dimensional model of a flower at subcellular resolution. We demonstrate the power of this approach by capturing male and female meiosis, asymmetric pollen division, movement of meiotic chromosomes, and unusual restitution mitosis in tapetum cells. This method will enable new avenues of research into plant sexual reproduction.

scholarly journals Minutes-timescale 3D isotropic imaging of entire organs at subcellular resolution by content-aware compressed-sensing light-sheet microscopy

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Chunyu Fang ◽  
Tingting Yu ◽  
Tingting Chu ◽  
Wenyang Feng ◽  
Fang Zhao ◽  
...  
Keyword(s):  
Compressed Sensing ◽  
Light Sheet ◽  
System Level ◽  
Projection Neurons ◽  
Long Distance ◽  
Rapid Acquisition ◽  
Light Sheet Microscopy ◽  
Macro Scale ◽  
Subcellular Resolution ◽  
Content Aware

AbstractRapid 3D imaging of entire organs and organisms at cellular resolution is a recurring challenge in life science. Here we report on a computational light-sheet microscopy able to achieve minute-timescale high-resolution mapping of entire macro-scale organs. Through combining a dual-side confocally-scanned Bessel light-sheet illumination which provides thinner-and-wider optical sectioning of deep tissues, with a content-aware compressed sensing (CACS) computation pipeline which further improves the contrast and resolution based on a single acquisition, our approach yields 3D images with high, isotropic spatial resolution and rapid acquisition over two-order-of-magnitude faster than conventional 3D microscopy implementations. We demonstrate the imaging of whole brain (~400 mm3), entire gastrocnemius and tibialis muscles (~200 mm3) of mouse at ultra-high throughput of 5~10 min per sample and post-improved subcellular resolution of ~ 1.5 μm (0.5-μm iso-voxel size). Various system-level cellular analyses, such as mapping cell populations at different brain sub-regions, tracing long-distance projection neurons over the entire brain, and calculating neuromuscular junction occupancy across whole muscle, are also readily accomplished by our method.

scholarly journals Minutes-timescale 3D isotropic imaging of entire organs at subcellular resolution by content-aware compressed-sensing light-sheet microscopy

2019 ◽  
Author(s):  
Chunyu Fang ◽  
Tingting Chu ◽  
Tingting Yu ◽  
Yujie Huang ◽  
Yusha Li ◽  
...  
Keyword(s):  
Compressed Sensing ◽  
Spatial Resolution ◽  
Light Sheet ◽  
System Level ◽  
Projection Neurons ◽  
Long Distance ◽  
Light Sheet Microscopy ◽  
Macro Scale ◽  
Subcellular Resolution ◽  
Content Aware

AbstractInstant 3D imaging of entire organs and organisms at cellular resolution is a recurring challenge in life science. Here we report on a computational light-sheet microscopy able to achieve minute-timescale mapping of entire macro-scale organs at high spatial resolution, thereby overcoming the throughput limit of current 3D microscopy implementations. Through combining a dual-side confocally-scanned Bessel light-sheet illumination which provides thinner-and-wider optical sectioning of deep tissues, with a content-aware compressed sensing (CACS) computation pipeline which further improves the contrast and resolution based on a single acquisition, our method yields 3D images with high, isotropic spatial resolution and rapid acquisition improved by two-orders of magnitude. We demonstrate the imaging of whole brain (∼400 mm3), entire gastrocnemius and tibialis muscles (∼200 mm3) of mouse at subcellular resolution (0.5-μm isovoxel) and ultra-high throughput of 5∼10 minutes per sample. Various system-level cellular analyses, such as mapping cell populations at different brain sub-regions, tracing long-distance projection neurons over the entire brain, and calculating neuromuscular junction occupancy across whole muscle, were also readily enabled by our method.

scholarly journals Isoflurane Exposure in Juvenile Caenorhabditis elegans Causes Persistent Changes in Neuron Dynamics

2020 ◽  
Vol 133 (3) ◽  
pp. 569-582 ◽  
Author(s):  
Gregory S. Wirak ◽  
Christopher V. Gabel ◽  
Christopher W. Connor
Keyword(s):  
Caenorhabditis Elegans ◽  
Early Adulthood ◽  
Light Sheet ◽  
Loss Of Function ◽  
Behavioral Impairment ◽  
Mtor Inhibition ◽  
Late Adulthood ◽  
C Elegans ◽  
Light Sheet Microscopy ◽  
Activity Dynamics

Background Animal studies demonstrate that anesthetic exposure during neurodevelopment can lead to persistent behavioral impairment. The changes in neuronal function underlying these effects are incompletely understood. Caenorhabditis elegans is well suited for functional imaging of postanesthetic effects on neuronal activity. This study aimed to examine such effects within the neurocircuitry underlying C. elegans locomotion. Methods C. elegans were exposed to 8% isoflurane for 3 h during the neurodevelopmentally critical L1 larval stage. Locomotion was assessed during early and late adulthood. Spontaneous activity was measured within the locomotion command interneuron circuitry using confocal and light-sheet microscopy of the calcium-sensitive fluorophore GCaMP6s. Results C. elegans exposed to isoflurane demonstrated attenuation in spontaneous reversal behavior, persisting throughout the animal’s lifespan (reversals/min: untreated early adulthood, 1.14 ± 0.42, vs. isoflurane-exposed early adulthood, 0.83 ± 0.55; untreated late adulthood, 1.75 ± 0.64, vs. isoflurane-exposed late adulthood, 1.14 ± 0.68; P = 0.001 and 0.006, respectively; n > 50 animal tracks/condition). Likewise, isoflurane exposure altered activity dynamics in the command interneuron AVA, which mediates crawling reversals. The rate at which AVA transitions between activity states was found to be increased. These anesthetic-induced effects were more pronounced with age (off-to-on activity state transition time (s): untreated early adulthood, 2.5 ± 1.2, vs. isoflurane-exposed early adulthood, 1.9 ± 1.3; untreated late adulthood, 4.6 ± 3.0, vs. isoflurane-exposed late adulthood, 3.0 ± 2.4; P = 0.028 and 0.008, respectively; n > 35 traces acquired from more than 15 animals/condition). Comparable effects were observed throughout the command interneuron circuitry, indicating that isoflurane exposure alters transition rates between behavioral crawling states of the system overall. These effects were modulated by loss-of-function mutations within the FoxO transcription factor daf-16 and by rapamycin-mediated mechanistic Target of Rapamycin (mTOR) inhibition. Conclusions Altered locomotive behavior and activity dynamics indicate a persistent effect on interneuron dynamics and circuit function in C. elegansafter developmental exposure to isoflurane. These effects are modulated by a loss of daf-16 or mTOR activity, consistent with a pathologic activation of stress-response pathways. Editor’s Perspective What We Already Know about This Topic What This Article Tells Us That Is New

scholarly journals Multi-view light-sheet imaging and tracking with the MaMuT software reveals the cell lineage of a direct developing arthropod limb

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Carsten Wolff ◽  
Jean-Yves Tinevez ◽  
Tobias Pietzsch ◽  
Evangelia Stamataki ◽  
Benjamin Harich ◽  
...  
Keyword(s):  
Limb Development ◽  
Expression Patterns ◽  
Cell Lineage ◽  
Light Sheet ◽  
Limb Bud ◽  
Spatial Modulation ◽  
Spatiotemporal Resolution ◽  
Cell Behaviors ◽  
Light Sheet Microscopy ◽  
Anterior Posterior

During development, coordinated cell behaviors orchestrate tissue and organ morphogenesis. Detailed descriptions of cell lineages and behaviors provide a powerful framework to elucidate the mechanisms of morphogenesis. To study the cellular basis of limb development, we imaged transgenic fluorescently-labeled embryos from the crustacean Parhyale hawaiensis with multi-view light-sheet microscopy at high spatiotemporal resolution over several days of embryogenesis. The cell lineage of outgrowing thoracic limbs was reconstructed at single-cell resolution with new software called Massive Multi-view Tracker (MaMuT). In silico clonal analyses suggested that the early limb primordium becomes subdivided into anterior-posterior and dorsal-ventral compartments whose boundaries intersect at the distal tip of the growing limb. Limb-bud formation is associated with spatial modulation of cell proliferation, while limb elongation is also driven by preferential orientation of cell divisions along the proximal-distal growth axis. Cellular reconstructions were predictive of the expression patterns of limb development genes including the BMP morphogen Decapentaplegic.

scholarly journals Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution

2019 ◽  
Vol 16 (11) ◽  
pp. 1109-1113 ◽  
Author(s):  
Tonmoy Chakraborty ◽  
Meghan K. Driscoll ◽  
Elise Jeffery ◽  
Malea M. Murphy ◽  
Philippe Roudot ◽  
...  
Keyword(s):  
Light Sheet ◽  
Light Sheet Microscopy ◽  
Subcellular Resolution

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.

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