scholarly journals Light-sheet Microscopy for Three-dimensional Visualization of Human Immune Cells

10.3791/57651 ◽  
2018 ◽  
Author(s):  
Rouven Schoppmeyer ◽  
Renping Zhao ◽  
Markus Hoth ◽  
Bin Qu
Keyword(s):  
Immune Cells ◽  
Three Dimensional ◽  
Light Sheet ◽  
Light Sheet Microscopy ◽  
Human Immune

scholarly journals Three-dimensional quantification of twisting in the Arabidopsis petiole

2021 ◽  
Author(s):  
Yuta Otsuka ◽  
Hirokazu Tsukaya
Keyword(s):  
Cross Sections ◽  
Three Dimensional ◽  
Light Sheet ◽  
Underlying Mechanism ◽  
Two Dimensions ◽  
Observation Angle ◽  
Differential Growth ◽  
Light Sheet Microscopy ◽  
Definition Of ◽  
Twisting Angle

AbstractOrganisms have a variety of three-dimensional (3D) structures that change over time. These changes include twisting, which is 3D deformation that cannot happen in two dimensions. Twisting is linked to important adaptive functions of organs, such as adjusting the orientation of leaves and flowers in plants to align with environmental stimuli (e.g. light, gravity). Despite its importance, the underlying mechanism for twisting remains to be determined, partly because there is no rigorous method for quantifying the twisting of plant organs. Conventional studies have relied on approximate measurements of the twisting angle in 2D, with arbitrary choices of observation angle. Here, we present the first rigorous quantification of the 3D twisting angles of Arabidopsis petioles based on light sheet microscopy. Mathematical separation of bending and twisting with strict definition of petiole cross-sections were implemented; differences in the spatial distribution of bending and twisting were detected via the quantification of angles along the petiole. Based on the measured values, we discuss that minute degrees of differential growth can result in pronounced twisting in petioles.

scholarly journals Three-Dimensional Cross-Sectional Light-Sheet Microscopy Imaging of the Inflamed Mouse Gut

2017 ◽  
Vol 153 (4) ◽  
pp. 898-900 ◽  
Author(s):  
Sebastian Zundler ◽  
Anika Klingberg ◽  
Daniela Schillinger ◽  
Sarah Fischer ◽  
Clemens Neufert ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Light Sheet ◽  
Cross Sectional ◽  
Microscopy Imaging ◽  
Light Sheet Microscopy

scholarly journals Three Dimensional Fluorescence Imaging Using Multiple Light-Sheet Microscopy

PLoS ONE ◽  
2014 ◽  
Vol 9 (6) ◽  
pp. e96551 ◽  
Author(s):  
Kavya Mohan ◽  
Subhajit B. Purnapatra ◽  
Partha Pratim Mondal
Keyword(s):  
Fluorescence Imaging ◽  
Three Dimensional ◽  
Light Sheet ◽  
Light Sheet Microscopy

scholarly journals Actin-based protrusions of migrating neutrophils are intrinsically lamellar and facilitate direction changes

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Lillian K Fritz-Laylin ◽  
Megan Riel-Mehan ◽  
Bi-Chang Chen ◽  
Samuel J Lord ◽  
Thomas D Goddard ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Light Sheet ◽  
Time Lapse ◽  
Three Dimensions ◽  
Cortical Actin ◽  
Linear Arrangement ◽  
Collagen Matrices ◽  
Flat Surfaces ◽  
Actin Networks ◽  
Light Sheet Microscopy

Leukocytes and other amoeboid cells change shape as they move, forming highly dynamic, actin-filled pseudopods. Although we understand much about the architecture and dynamics of thin lamellipodia made by slow-moving cells on flat surfaces, conventional light microscopy lacks the spatial and temporal resolution required to track complex pseudopods of cells moving in three dimensions. We therefore employed lattice light sheet microscopy to perform three-dimensional, time-lapse imaging of neutrophil-like HL-60 cells crawling through collagen matrices. To analyze three-dimensional pseudopods we: (i) developed fluorescent probe combinations that distinguish cortical actin from dynamic, pseudopod-forming actin networks, and (ii) adapted molecular visualization tools from structural biology to render and analyze complex cell surfaces. Surprisingly, three-dimensional pseudopods turn out to be composed of thin (<0.75 µm), flat sheets that sometimes interleave to form rosettes. Their laminar nature is not templated by an external surface, but likely reflects a linear arrangement of regulatory molecules. Although we find that Arp2/3-dependent pseudopods are dispensable for three-dimensional locomotion, their elimination dramatically decreases the frequency of cell turning, and pseudopod dynamics increase when cells change direction, highlighting the important role pseudopods play in pathfinding.

scholarly journals Digital Spindle: A New Way to Explore Mitotic Functions by Whole Cell Data Collection and a Computational Approach

Cells ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 1255
Author(s):  
Norio Yamashita ◽  
Masahiko Morita ◽  
Hideo Yokota ◽  
Yuko Mimori-Kiyosue
Keyword(s):  
Cell Biology ◽  
Information Science ◽  
Three Dimensional ◽  
Light Sheet ◽  
Live Imaging ◽  
Three Dimensions ◽  
Complex Data ◽  
Spatiotemporal Resolution ◽  
Whole Cell ◽  
Light Sheet Microscopy

From cells to organisms, every living system is three-dimensional (3D), but the performance of fluorescence microscopy has been largely limited when attempting to obtain an overview of systems’ dynamic processes in three dimensions. Recently, advanced light-sheet illumination technologies, allowing drastic improvement in spatial discrimination, volumetric imaging times, and phototoxicity/photobleaching, have been making live imaging to collect precise and reliable 3D information increasingly feasible. In particular, lattice light-sheet microscopy (LLSM), using an ultrathin light-sheet, enables whole-cell 3D live imaging of cellular processes, including mitosis, at unprecedented spatiotemporal resolution for extended periods of time. This technology produces immense and complex data, including a significant amount of information, raising new challenges for big image data analysis and new possibilities for data utilization. Once the data are digitally archived in a computer, the data can be reused for various purposes by anyone at any time. Such an information science approach has the potential to revolutionize the use of bioimage data, and provides an alternative method for cell biology research in a data-driven manner. In this article, we introduce examples of analyzing digital mitotic spindles and discuss future perspectives in cell biology.

scholarly journals Label-free and Multimodal Second Harmonic Generation Light Sheet Microscopy

2020 ◽  
Author(s):  
Niall Hanrahan ◽  
Simon I. R. Lane ◽  
Peter Johnson ◽  
Konstantinos Bourdakos ◽  
Christopher Brereton ◽  
...  
Keyword(s):  
Second Harmonic Generation ◽  
Harmonic Generation ◽  
Biological Samples ◽  
3D Imaging ◽  
Second Harmonic ◽  
Three Dimensional ◽  
Light Sheet ◽  
Optical Methods ◽  
Label Free ◽  
Light Sheet Microscopy

AbstractLight sheet microscopy (LSM) has emerged as one of most profound three dimensional (3D) imaging tools in the life sciences over the last decade. However, LSM is currently performed with fluorescence detection on one- or multi-photon excitation. Label-free LSM imaging approaches have been rather limited. Second Harmonic Generation (SHG) imaging is a label-free technique that has enabled detailed investigation of collagenous structures, including its distribution and remodelling in cancers and respiratory tissue, and how these link to disease. SHG is generally regarded as having only forward- and back-scattering components, apparently precluding the orthogonal detection geometry used in Light Sheet Microscopy. In this work we demonstrate SHG imaging on a light sheet microscope (SHG-LSM) using a rotated Airy beam configuration that demonstrates a powerful new approach to direct, without any further processing or deconvolution, 3D imaging of harmonophores such as collagen in biological samples. We provide unambiguous identification of SHG signals on the LSM through its wavelength and polarisation sensitivity. In a multimodal LSM setup we demonstrate that SHG and two-photon signals can be acquired on multiple types of different biological samples. We further show that SHG-LSM is sensitive to changes in collagen synthesis within lung fibroblast 3D cell cultures. This work expands on the existing optical methods available for use with light sheet microscopy, adding a further label-free imaging technique which can be combined with other detection modalities to realise a powerful multi-modal microscope for 3D bioimaging.

scholarly journals 3D Cell Neighbour Dynamics in Growing Pseudostratified Epithelia

2021 ◽  
Author(s):  
Harold F. Gómez ◽  
Mathilde S. Dumont ◽  
Leonie Hodel ◽  
Roman Vetter ◽  
Dagmar Iber
Keyword(s):  
Three Dimensional ◽  
Light Sheet ◽  
Cell Segmentation ◽  
Cross Sectional ◽  
Light Sheet Microscopy ◽  
Cell Organization ◽  
Epithelial Sheets ◽  
Epithelial Layers ◽  
3D Shapes ◽  
Lateral Cell

ABSTRACTDuring morphogenesis, epithelial sheets remodel into complex geometries. How cells dynamically organize their contact with neighbouring cells in these tightly packed tissues is poorly understood. We have used light-sheet microscopy of growing mouse embryonic lung explants, three-dimensional cell segmentation, and physical theory to unravel the principles behind 3D cell organization in growing pseudostratified epithelia. We find that cells have highly irregular 3D shapes and exhibit numerous neighbour intercalations along the apical-basal axis as well as over time. Despite the fluidic nature, the cell packing configurations follow fundamental relationships previously described for apical epithelial layers, i.e., Euler’s formula, Lewis’ law, and Aboav-Weaire’s law, at all times and across the entire tissue thickness. This arrangement minimizes the lateral cell-cell surface energy for a given cross-sectional area variability, generated primarily by the distribution and movement of nuclei. We conclude that the complex 3D cell organization in growing epithelia emerges from simple physical principles.

scholarly journals Three-dimensional tracking of plus-tips by lattice light-sheet microscopy permits the quantification of microtubule growth trajectories within the mitotic apparatus

2015 ◽  
Vol 20 (10) ◽  
pp. 101206 ◽  
Author(s):  
Norio Yamashita ◽  
Masahiko Morita ◽  
Wesley R. Legant ◽  
Bi-Chang Chen ◽  
Eric Betzig ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Light Sheet ◽  
Growth Trajectories ◽  
Mitotic Apparatus ◽  
Light Sheet Microscopy ◽  
Microtubule Growth
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