scholarly journals Live cell tracking of symmetry break in actin cytoskeleton triggered by abrupt changes in micromechanical environments

2015 ◽  
Vol 3 (12) ◽  
pp. 1539-1544 ◽  
Author(s):  
S. Inoue ◽  
V. Frank ◽  
M. Hörning ◽  
S. Kaufmann ◽  
H. Y. Yoshikawa ◽  
...  
Keyword(s):  
Symmetry Breaking ◽  
Actin Cytoskeleton ◽  
Live Cell Imaging ◽  
Cell Tracking ◽  
Cell Imaging ◽  
Live Cell ◽  
Abrupt Changes ◽  
Stimulus Responsive ◽  
Responsive Hydrogels ◽  
Symmetry Break

Stimulus responsive hydrogels and live cell imaging allow for the quantitative parameterization of symmetry breaking in remodelling actin cytoskeleton.

scholarly journals Rapid Morphological and Cytoskeletal Response to Microgravity in Human Primary Macrophages

2019 ◽  
Vol 20 (10) ◽  
pp. 2402 ◽  
Author(s):  
Cora Sandra Thiel ◽  
Svantje Tauber ◽  
Beatrice Lauber ◽  
Jennifer Polzer ◽  
Christian Seebacher ◽  
...  
Keyword(s):  
Actin Cytoskeleton ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Imaging ◽  
Live Cell ◽  
Cellular Structures ◽  
Microgravity Environment ◽  
Confocal Laser ◽  
Human Macrophages ◽  
Imaging Experiment

The FLUMIAS (Fluorescence-Microscopic Analyses System for Life-Cell-Imaging in Space) confocal laser spinning disk fluorescence microscope represents a new imaging capability for live cell imaging experiments on suborbital ballistic rocket missions. During the second pioneer mission of this microscope system on the TEXUS-54 suborbital rocket flight, we developed and performed a live imaging experiment with primary human macrophages. We simultaneously imaged four different cellular structures (nucleus, cytoplasm, lysosomes, actin cytoskeleton) by using four different live cell dyes (Nuclear Violet, Calcein, LysoBrite, SiR-actin) and laser wavelengths (405, 488, 561, and 642 nm), and investigated the cellular morphology in microgravity (10−4 to 10−5 g) over a period of about six minutes compared to 1 g controls. For live imaging of the cytoskeleton during spaceflight, we combined confocal laser microscopy with the SiR-actin probe, a fluorogenic silicon-rhodamine (SiR) conjugated jasplakinolide probe that binds to F-actin and displays minimal toxicity. We determined changes in 3D cell volume and surface, nuclear volume and in the actin cytoskeleton, which responded rapidly to the microgravity environment with a significant reduction of SiR-actin fluorescence after 4–19 s microgravity, and adapted subsequently until 126–151 s microgravity. We conclude that microgravity induces geometric cellular changes and rapid response and adaptation of the potential gravity-transducing cytoskeleton in primary human macrophages.

scholarly journals Live cell imaging reveals actin-cytoskeleton-induced self-association of the actin-bundling protein WLIM1

2014 ◽  
Vol 127 (6) ◽  
pp. 1357-1357
Author(s):  
C. Hoffmann ◽  
D. Moes ◽  
M. Dieterle ◽  
K. Neumann ◽  
F. Moreau ◽  
...  
Keyword(s):  
Actin Cytoskeleton ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Self Association

scholarly journals Live cell imaging reveals actin-cytoskeleton-induced self-association of the actin-bundling protein WLIM1

2013 ◽  
Vol 127 (3) ◽  
pp. 583-598 ◽  
Author(s):  
C. Hoffmann ◽  
D. Moes ◽  
M. Dieterle ◽  
K. Neumann ◽  
F. Moreau ◽  
...  
Keyword(s):  
Actin Cytoskeleton ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Self Association

scholarly journals Dendritic spines on GABAergic neurons respond to cholinergic signaling in theCaenorhabditis elegansmotor circuit

2019 ◽  
Author(s):  
Andrea Cuentas-Condori ◽  
Ben Mulcahy ◽  
Siwei He ◽  
Sierra Palumbos ◽  
Mei Zhen ◽  
...  
Keyword(s):  
Actin Cytoskeleton ◽  
Motor Neurons ◽  
Dendritic Spines ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Spine Density ◽  
C Elegans ◽  
Cholinergic Signaling ◽  
Spine Morphogenesis

SUMMARYDendritic spines are specialized postsynaptic structures that detect and integrate presynaptic signals. The shape and number of dendritic spines are regulated by neural activity and correlated with learning and memory. Most studies of spine function have focused on the mammalian nervous system. However, spine-like protrusions have been previously reported in invertebrates, suggesting that the experimental advantages of smaller model organisms could be exploited to study the biology of dendritic spines. Here, we document the presence of dendritic spines inCaenorhabditis elegansmotor neurons. We used super-resolution microscopy, electron microscopy, live-cell imaging and genetic manipulation to show that GABAergic motor neurons display functional dendritic spines. Our analysis revealed salient features of dendritic spines: (1) A key role for the actin cytoskeleton in spine morphogenesis; (2) Postsynaptic receptor complexes at the tips of spines in close proximity to presynaptic active zones; (3) Localized postsynaptic calcium transients evoked by presynaptic activity; (4) The presence of endoplasmic reticulum and ribosomes; (5) The regulation of spine density by presynaptic activity. These studies provide a solid foundation for a new experimental paradigm that exploits the power ofC. elegansgenetics and live-cell imaging for fundamental studies of dendritic spine morphogenesis and function.HIGHLIGHTS-Spines inC. elegansGABAergic motor neurons are enriched in actin cytoskeleton.-Spines are dynamic structures.-Spines display Ca++transients coupled with presynaptic activation.-Spine density is regulated during development and is modulated by actin dynamics and cholinergic signaling.

scholarly journals Accurate cell tracking and lineage construction in live-cell imaging experiments with deep learning

2019 ◽  
Author(s):  
Erick Moen ◽  
Enrico Borba ◽  
Geneva Miller ◽  
Morgan Schwartz ◽  
Dylan Bannon ◽  
...  
Keyword(s):  
Deep Learning ◽  
Single Cell ◽  
Live Cell Imaging ◽  
Cell Tracking ◽  
Cell Imaging ◽  
Single Cells ◽  
Live Cell ◽  
Imaging Data ◽  
Cell Nuclei ◽  
Data Types

AbstractLive-cell imaging experiments have opened an exciting window into the behavior of living systems. While these experiments can produce rich data, the computational analysis of these datasets is challenging. Single-cell analysis requires that cells be accurately identified in each image and subsequently tracked over time. Increasingly, deep learning is being used to interpret microscopy image with single cell resolution. In this work, we apply deep learning to the problem of tracking single cells in live-cell imaging data. Using crowdsourcing and a human-in-the-loop approach to data annotation, we constructed a dataset of over 11,000 trajectories of cell nuclei that includes lineage information. Using this dataset, we successfully trained a deep learning model to perform cell tracking within a linear programming framework. Benchmarking tests demonstrate that our method achieves state-of-the-art performance on the task of cell tracking with respect to multiple accuracy metrics. Further, we show that our deep learning-based method generalizes to perform cell tracking for both fluorescent and brightfield images of the cell cytoplasm, despite having never been trained on those data types. This enables analysis of live-cell imaging data collected across imaging modalities. A persistent cloud deployment of our cell tracker is available at http://www.deepcell.org.

scholarly journals Single cell tracking based on Voronoi partition via stable matching

2020 ◽  
Author(s):  
Young Hwan Chang ◽  
Jeremy Linsley ◽  
Josh Lamstein ◽  
Jaslin Kalra ◽  
Irina Epstein ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Tracking ◽  
Cell Imaging ◽  
Stable Matching ◽  
Live Cell ◽  
High Temporal Resolution ◽  
Imaging Data ◽  
Matching Problem ◽  
Voronoi Partition ◽  
Over Time

AbstractLive-cell imaging is an important technique to study cell migration and proliferation as well as image-based profiling of drug perturbations over time. To gain biological insights from live-cell imaging data, it is necessary to identify individual cells, follow them over time and extract quantitative information. However, since often biological experiment does not allow the high temporal resolution to reduce excessive levels of illumination or minimize unnecessary oversampling to monitor long-term dynamics, it is still a challenging task to obtain good tracking results with coarsely sampled imaging data. To address this problem, we consider cell tracking problem as “stable matching problem” and propose a robust tracking method based on Voronoi partition which adapts parameters that need to be set according to the spatio-temporal characteristics of live cell imaging data such as cell population and migration. We demonstrate the performance improvement provided by the proposed method using numerical simulations and compare its performance with proximity-based tracking and nearest neighbor-based tracking.

scholarly journals Actin chromobody imaging reveals sub-organellar actin dynamics

2019 ◽  
Author(s):  
Cara R. Schiavon ◽  
Tong Zhang ◽  
Bing Zhao ◽  
Leonardo Andrade ◽  
Melissa Wu ◽  
...  
Keyword(s):  
Cell Migration ◽  
Fluorescence Microscopy ◽  
Actin Cytoskeleton ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Fluorescent Protein ◽  
Live Cell ◽  
Actin Dynamics ◽  
Spatiotemporal Resolution ◽  
In Cells

AbstractThe actin cytoskeleton plays multiple critical roles in cells, from cell migration to organelle dynamics. The small and transient actin structures regulating organelle dynamics are difficult to detect with fluorescence microscopy. We developed an approach using fluorescent protein-tagged actin nanobodies targeted to organelle membranes to enable live cell imaging of sub-organellar actin dynamics with unprecedented spatiotemporal resolution. These probes reveal that ER-associated actin drives fission of multiple organelles including mitochondria, endosomes, lysosomes, peroxisomes, and the Golgi.

Live cell imaging and single cell tracking of mesenchymal stromal cellsin vitro

2016 ◽  
pp. 323-346
Author(s):  
James A. Cornwell ◽  
Maria Z. Gutierrez ◽  
Richard P. Harvey ◽  
Robert E. Nordon
Keyword(s):  
Single Cell ◽  
Live Cell Imaging ◽  
Cell Tracking ◽  
Cell Imaging ◽  
Live Cell
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