LivE imaging for in vivo cellular profiling

2016 ◽  
Author(s):  
Ralph Muller
Keyword(s):  
Live Imaging

scholarly journals Microfluidic Live-Imaging Technology to Perform Research Activities in 3D Models

2021 ◽  
Author(s):  
Arnaud Martino Capuzzo ◽  
Daniele Vigo
Keyword(s):  
Fluorescence Intensity ◽  
Regulatory Networks ◽  
3D Structure ◽  
Live Imaging ◽  
Two Dimensions ◽  
Live Cells ◽  
Straight Line Passing ◽  
3D Cultures ◽  
Research Activities

Morphological dissimilarity and its evolution over time are one of the most unexpected variations found when comparing cell cultures in 2D and 3D. Monolayer cells appear to flatten in the lower part of the plate, adhering to and spreading in the horizontal plane while not extending vertically. Consequently, cells developed in two dimensions have a forced apex-basal polarity. Co-cultivation and crosstalking between multiple cell types, which control development and formation in the in vivo counterpart, are possible in 3D cultures. With or without a scaffold matrix, 3D model culture may exhibit more in vivo-like morphology and physiology. 3D cultures mimic relevant physiological cellular processes, transforming them into one-of-a-kind drug screening platforms. The structures and dynamics of regulatory networks, which are increasingly studied with live-imaging microscopy, must be considered to help and guarantee the functional maintenance of a 3D structure. However, commercially available technologies that can be used for current laboratory needs are minimal, despite the need to make it easier to acquire cellular kinetics with high spatial and temporal resolution, in order to improve visual efficiency and, as a result, experimentation performance. The CELLviewer is a newly developed multi-technology instrument that integrates and synchronizes the work of various scientific disciplines. The aim of this study is to test the device using two different models: a single Jurkat cell and an MCF-7 spheroid. The two models are loaded into the microfluidic cartridge for each experiment after they have been grown and captured in time-lapse for a total of 4 hours. The samples used are tracked under the operation of the optics after adaptive autofocus, while slipping inside the cartridge chamber, and the 3D rotation was successfully obtained experimentally. The MitoGreen dye, a fluorescence marker selectively permeable to live cells, was then used to determine cell viability. To measure the model diameter, construct fluorescence intensity graphs along a straight line passing through the cell, and visualize the spatial fluorescence intensity distribution in 3D, ImageJ software was used.

scholarly journals In vivo live imaging of RNA polymerase II transcription factories in primary cells

2013 ◽  
Vol 27 (7) ◽  
pp. 767-777 ◽  
Author(s):  
A. Ghamari ◽  
M. P. C. van de Corput ◽  
S. Thongjuea ◽  
W. A. van Cappellen ◽  
W. van IJcken ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Live Imaging ◽  
Primary Cells ◽  
Transcription Factories ◽  
Rna Polymerase Ii Transcription

scholarly journals A coral-on-a-chip microfluidic platform enabling live-imaging microscopy of reef-building corals

2016 ◽  
Vol 7 (1) ◽  
Author(s):  
Orr H. Shapiro ◽  
Esti Kramarsky-Winter ◽  
Assaf R. Gavish ◽  
Roman Stocker ◽  
Assaf Vardi
Keyword(s):  
Live Imaging ◽  
Live Coral ◽  
Powerful Method ◽  
Microfluidic Platform ◽  
Experimental Platform ◽  
Micro Scale ◽  
Algal Symbionts ◽  
Spatio Temporal

Abstract Coral reefs, and the unique ecosystems they support, are facing severe threats by human activities and climate change. Our understanding of these threats is hampered by the lack of robust approaches for studying the micro-scale interactions between corals and their environment. Here we present an experimental platform, coral-on-a-chip, combining micropropagation and microfluidics to allow direct microscopic study of live coral polyps. The small and transparent coral micropropagates are ideally suited for live-imaging microscopy, while the microfluidic platform facilitates long-term visualization under controlled environmental conditions. We demonstrate the usefulness of this approach by imaging coral micropropagates at previously unattainable spatio-temporal resolutions, providing new insights into several micro-scale processes including coral calcification, coral–pathogen interaction and the loss of algal symbionts (coral bleaching). Coral-on-a-chip thus provides a powerful method for studying coral physiology in vivo at the micro-scale, opening new vistas in coral biology.

scholarly journals 3D Local in vivo Environment (LivE) imaging for single cell protein analysis of bone tissue

2016 ◽  
Vol 2 (1) ◽  
pp. 449-453 ◽  
Author(s):  
Carly Taylor ◽  
Ariane Scheuren ◽  
Andreas Trüssel ◽  
Ralph Müller
Keyword(s):  
Bone Tissue ◽  
Bone Remodelling ◽  
Three Dimensional ◽  
Relevant Information ◽  
Live Imaging ◽  
Bone Diseases ◽  
Cell Protein ◽  
Cellular Expression ◽  
3D Network

AbstractThe molecular processes behind pathological bone remodelling seen in diseases such as osteoporosis are unclear. However, a recently developed methodological platform known as Local in vivo Environment (LivE) imaging has been used to link cellular expression data to the local remodelling and mechanical environment in 2D sections of bone tissue. The method therefore can be used to give insight into which proteins are important for pathological bone remodelling. However, the cells within bone tissue exist as a 3D network. Therefore extension of LivE to accommodate 3D data may provide additional physiologically relevant information that is not possible to determine using 2D analysis alone. This will have implications for the further understanding of the cellular basis that underlies bone diseases such as osteoporosis. Here the LivE imaging technique is expanded to incorporate data from cells in a three dimensional manner via a serial sectioning technique. The methodological steps involved in the LivE imaging approach are defined and the optimisation steps performed are explained in detail.

scholarly journals Tissue fluidity mediated by adherens junction dynamics promotes planar cell polarity-driven ommatidial rotation

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Nabila Founounou ◽  
Reza Farhadifar ◽  
Giovanna M. Collu ◽  
Ursula Weber ◽  
Michael J. Shelley ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Cell Polarity ◽  
Planar Cell Polarity ◽  
Adherens Junction ◽  
Live Imaging ◽  
Cellular Migration ◽  
Imaging Analysis ◽  
Ommatidial Rotation ◽  
Eye Morphogenesis

AbstractThe phenomenon of tissue fluidity—cells’ ability to rearrange relative to each other in confluent tissues—has been linked to several morphogenetic processes and diseases, yet few molecular regulators of tissue fluidity are known. Ommatidial rotation (OR), directed by planar cell polarity signaling, occurs during Drosophila eye morphogenesis and shares many features with polarized cellular migration in vertebrates. We utilize in vivo live imaging analysis tools to quantify dynamic cellular morphologies during OR, revealing that OR is driven autonomously by ommatidial cell clusters rotating in successive pulses within a permissive substrate. Through analysis of a rotation-specific nemo mutant, we demonstrate that precise regulation of junctional E-cadherin levels is critical for modulating the mechanical properties of the tissue to allow rotation to progress. Our study defines Nemo as a molecular tool to induce a transition from solid-like tissues to more viscoelastic tissues broadening our molecular understanding of tissue fluidity.

scholarly journals In vivo live imaging of postnatal neural stem cells

Development ◽  
2021 ◽  
Vol 148 (18) ◽  
Author(s):  
Alina Marymonchyk ◽  
Sarah Malvaut ◽  
Armen Saghatelyan
Keyword(s):  
Stem Cells ◽  
Neural Stem Cells ◽  
In Vivo Imaging ◽  
Live Imaging ◽  
Future Directions ◽  
Environmental Signals ◽  
New Information ◽  
Imaging Results

ABSTRACT Neural stem cells (NSCs) are maintained in specific regions of the postnatal brain and contribute to its structural and functional plasticity. However, the long-term renewal potential of NSCs and their mode of division remain elusive. The use of advanced in vivo live imaging approaches may expand our knowledge of NSC physiology and provide new information for cell replacement therapies. In this Review, we discuss the in vivo imaging methods used to study NSC dynamics and recent live-imaging results with respect to specific intracellular pathways that allow NSCs to integrate and decode different micro-environmental signals. Lastly, we discuss future directions that may provide answers to unresolved questions regarding NSC physiology.

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