Expansion Microscopy Of C Elegans
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Expansion Microscopy Of C Elegans
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Author : Chih-Chieh Jay Yu
language : en
Publisher:
Release Date : 2020
Expansion Microscopy Of C Elegans written by Chih-Chieh Jay Yu and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2020 with categories.
Expansion microscopy (ExM) enables 3-D, nanoscale-precise imaging of biological specimens by isotropic swelling of hydrogel-embedded, chemically processed tissue. Such capability raises the question of whether nanoscale mapping of biomolecules could be performed in an entire organism, which would allow super-resolution-mediated in situ analyses, such as digital quantification of biomolecules and mapping of synaptic contacts, to be performed within the context of an entire nervous system. The nematode Caenorhabditis elegans could be a suitable model for such organism-wide analyses, due to its tractable physical size, deterministic cell lineage, ease of genetic control, and well-established literature. However, C. elegans is enclosed in a chemically impermeable and mechanically tough cuticle, which could hinder the deployment of ExM. In this thesis, we present a strategy, expansion of C. elegans (ExCel), to expand fixed, cuticle-enclosed intact animals of C. elegans. ExCel enables simultaneous readout of fluorescent proteins, RNAs, DNA locations, and anatomical structures at resolutions of ~65-75 nm (3.3-3.8x linear expansion). We also developed epitope-preserving ExCel, which enables imaging of endogenous proteins stained by antibodies, and iterative ExCel, which enables imaging of fluorescent proteins at a ~25-nm resolution (20x linear expansion). We demonstrate the utility of the ExCel toolbox for multiplexed imaging of multiple molecular types, for mapping synaptic proteins, for identifying previously unreported proteins at cell junctions, and for gene expression analysis in multiple individual neurons of the same animal. In addition to ExCel, we discuss two other ExM-related technologies, including tetragel, which is a highly homogeneous hydrogel network that improves the nanoscale isotropy of biological ultrastructure expanded by ExM, and stochastic arrangement of reporters in clusters (STARC), which is a strategy for recording neuronal activity at a subneurite-level resolution, in densely labeled neuronal populations. Taken together, the work presented in this thesis extends the capabilities of ExM, and lays the foundation for a comprehensive, functionally and structurally informed analysis of an entire organism, which could reveal new insights in neuroscience, organismal development, and systems biology.
Expansion Microscopy To Visualize Cell Cycle Events In The Abbreviated Cell Cycle In Caenorhabditis Elegans Roundworm Germline Stem Cells
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Author : Taylor Campbell
language : en
Publisher:
Release Date : 2021
Expansion Microscopy To Visualize Cell Cycle Events In The Abbreviated Cell Cycle In Caenorhabditis Elegans Roundworm Germline Stem Cells written by Taylor Campbell and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2021 with Cell cycle categories.
The abbreviated cell cycle is a poorly understood form of the cell cycle essential for genome stability in stem cells. To visualize cell-cycle events in the abbreviated cell cycle, we optimized a protocol for expansion microscopy in stem cells of the reproductive system of the model organism C. elegans. Optimizations included development of new mounting techniques and modifications to DNA staining methods. These optimizations produced brighter samples that were easier to find and focus on at the microscope. These optimizations, combined with future labeling of DNA replication, will expand our understanding of how cells with an abbreviated cell cycle maintain genome stability.
Tools For Connectomics In C Elegans
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Author : Nicholas Craig Barry
language : en
Publisher:
Release Date : 2018
Tools For Connectomics In C Elegans written by Nicholas Craig Barry and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2018 with categories.
Efforts to model computation in biological neural networks require knowledge of the structure of the network, the dynamics that play across it, and a network simple enough to be tractable to our incipient analyses. The simplicity of the 302-node nervous system of the nematode C. elegans and its transparency have made it an attractive model organism in neuroscience for several decades. Indeed, Caenorhabditis elegans has long been touted as the only species for which the connectome is known, reconstructed by hand from electron micrographs. However, while the number and identity of neurons in C. elegans appears fixed across animals, the variability in the connections between them has not been sufficiently characterized by the above efforts, which examined only a handful of animals and required many years of human labor. Such a characterization, and, moreover, an ability to accurately assess shifts in these neural graphs on timescales compatible with the pace and statistical rigor of scientific research would significantly accelerate efforts to understand neural computation. This thesis lays the groundwork for the development of such a framework. The expansion microscopy tissue preparation platform provided the basis for the set of experiments described within, in which strategies for molecular annotation of C. elegans and the subsequent protocols for readout are examined.
Caenorhabditis Elegans
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Author : Henry F. Epstein
language : en
Publisher: Academic Press
Release Date : 1995
Caenorhabditis Elegans written by Henry F. Epstein and has been published by Academic Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 1995 with Caenorhabditis elegans categories.
The first of its kind, this laboratory handbook emphasizes diverse methods and technologies needed to investigate C. elegans, both as an integrated organism and as a model system for research inquiries in cell, developmental, and molecular biology, as well as in genetics and pharmacology. Four primary sections--Genetic and Culture Methods, Neurobiology, Cell and Molecular Biology, and Genomics and Informatics--reflect the cross-disciplinary nature of C. elegans research. Because C. elegans is a simple and malleable organism with a small genome and few cell types, it provides an elegant demonstr.
Imaging Of Caenorhabditis Elegans With Near Infrared Spectrally Encoded Confocal Microscopy
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Author : Sadaf Rashtchian
language : en
Publisher:
Release Date : 2021
Imaging Of Caenorhabditis Elegans With Near Infrared Spectrally Encoded Confocal Microscopy written by Sadaf Rashtchian and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2021 with categories.
Caenorhabditis elegans is an optically transparent nematode that shares many gene orthologs and homologs with humans. C. elegans is widely used in large populations for genetic studies relevant to human biology and disease. Success of such studies frequently relies on the ability to image C. elegans structure at high-resolution and high-speed. This thesis reports on the feasibility of a high-speed variant of reflectance confocal microscopy, known as spectrally encoded confocal microscopy (SECM), for label-free imaging of C. elegans. The developed system utilizes near-infrared illumination in conjunction with refractive and diffractive optics to instantaneously image a confocal image line at a speed of up to 147 kHz with lateral and axial resolutions of 2m and 10m, respectively. Our imaging results from wild-type C. elegans and four mutant strains demonstrate the ability of SECM in revealing the overall geometry, key internal organs, and mutation-induced structural variations.
Expansion Microscopy For Cell Biology
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Author :
language : en
Publisher: Academic Press
Release Date : 2021-01-19
Expansion Microscopy For Cell Biology written by and has been published by Academic Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2021-01-19 with Science categories.
Expansion Microscopy for Cell Biology, Volume 161 in the Methods in Cell Biology series, compiles recent developments in expansion microscopy techniques (Pro-ExM, U-ExM, Ex-STED, X10, Ex-dSTORM, etc.) and their applications in cell biology, ranging from mitosis, centrioles or nuclear pore complex to plant cell, bacteria, Drosophila or neurons. Chapters in this new release include Protein-retention Expansion Microscopy: Improved Sub-cellular Imaging Resolution through Physical Specimen Expansion, Ultrastructure Expansion Microscopy (U-ExM), Expansion STED microscopy (ExSTED), Simple multi-color super-resolution by X10 microscopy, Expansion microscopy imaging of various neuronal structures, Mapping the neuronal cytoskeleton using expansion microscopy, Mechanical expansion microscopy, and much more. Provides the authority and expertise of leading contributors from an international board of authors Represents the latest release in the Methods in Cell Biology series Includes the latest information on Expansion Microscopy for Cell Biology
The Nematode Caenorhabditis Elegans
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Author : William Barry Wood
language : en
Publisher: Firefly Books
Release Date : 1988
The Nematode Caenorhabditis Elegans written by William Barry Wood and has been published by Firefly Books this book supported file pdf, txt, epub, kindle and other format this book has been release on 1988 with Caenorhabditis categories.
In 1965 Sydney Brenner chose the free-living nematode Caenorhabditis elegans as a promising model system for a concerted genetic, ultrastructural, and behavioral attack on the development and function of a simple nervous system. Since then, with the help of a growing number of investigators, knowledge about the biology of "the worm" has accumulated at a steadily accelerating pace to the extent that C. elegans is now probably the most completely understood metazoan in terms of anatomy, genetics, development, and behavior. The past few years have seen the completion of two major long-term projects that provide new insights into C. elegans development and lay important groundwork for future investigation: completion of the cell lineages of both sexes, from zygote to adult, and description of the complete anatomy at the level of electron microscope resolution, providing a complete "wiring diagram" of cell contacts in the animal. Recent years have also brought the first successes in molecularly cloning genes of developmental interest defined only by mutation, using transposon tagging as a generally applicable method for identification of the desired DNA sequences. Reintroduction of cloned DNA sequences into the genome has recently been accomplished. A physical map of the genome has been assembled with a combination of cosmid and YAC genes. This "Book of the Worm" serves as a reference source for C. elegans investigators as well as an introductory monograph for other biologists.
Optofluidic Add On Devices For Light Sheet Fluorescence Microscopy Of Caenorhabditis Elegans With Conventional Wide Field Microscopes
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Author : Mehran Behrouzi
language : en
Publisher:
Release Date : 2022
Optofluidic Add On Devices For Light Sheet Fluorescence Microscopy Of Caenorhabditis Elegans With Conventional Wide Field Microscopes written by Mehran Behrouzi and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2022 with categories.
This research reports on the development of low-cost and compact optofluidic add-on devices for converting a conventional wide-field microscope to a light sheet fluorescence microscope for immobilization-free, rapid and continuous imaging of the C. elegans model organism. The developed Polydimethylsiloxane (PDMS)-based optofluidic devices consist of an integrated PDMS cylindrical lens for on-chip light-sheet generation and a microfluidics channel for flow-based translation of samples through the light sheet. Validation experiments on several strains of C. elegans demonstrated the ability of the developed imaging platform in volumetric imaging of the fluorescence expressions of entire worms in a few seconds, at the single-neuron resolution, and with high contrast. Also, results of the primary neurodegeneration study demonstrated the ability of the platform on quantifying the fluorescent expressions of the worms and detecting different neurotoxin-induced phenotypes.
Electron Microscopy Of Caenorhabditis Elegans Muscle Mutants
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Author : Lorraine Santy
language : en
Publisher:
Release Date : 1990
Electron Microscopy Of Caenorhabditis Elegans Muscle Mutants written by Lorraine Santy and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 1990 with Caenorhabditis elegans categories.
In Vivo Investigation Of Protein Interactions In C Elegans By F Rster Resonance Energy Transfer Microscopy
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Author : Sophia Yannick Breusegem
language : en
Publisher:
Release Date : 2002
In Vivo Investigation Of Protein Interactions In C Elegans By F Rster Resonance Energy Transfer Microscopy written by Sophia Yannick Breusegem and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2002 with categories.