Automated Microfluidic Platforms To Facilitate Nerve Degeneration Studies With C Elegans

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Automated Microfluidic Platforms To Facilitate Nerve Degeneration Studies With C Elegans

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Author : Navid Ghorashian
language : en
Publisher:
Release Date : 2013

Automated Microfluidic Platforms To Facilitate Nerve Degeneration Studies With C Elegans written by Navid Ghorashian and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2013 with categories.

With its well-characterized genome, simple anatomy, and vast array of uses in molecular biology, the roundworm, Caenorhabditis elegans (C. elegans) is a well-established model organism in neurobiology. Concurrently, neurodegenerative diseases are some the most devastating and least understood ailments in modern medicine, making high-throughput approaches to understand their fundamental mechanisms imperative to developing new therapies. The worm's physical length-scales and simple genetics make it an ideal in vivo tool for high-throughput screening platforms. Concurrently, microfluidic technology has been used to make devices that manipulate these animals in a multitude of fashions to study various biological phenomena. With these considerations in mind, we have developed microfluidic platforms to facilitate optical interrogation of neurodegenerative and neuroregenerative phenomena in C. elegans for large-scale screens. First we developed a multiwell format device with 16 on-chip reservoirs to house and quickly deliver distinct worm populations to any liquid-format imaging platform. The system achieved unprecedented delivery speeds, avoided any population cross-contamination, and maintained animal viability. We then expanded this platform into a 64-well device that acted as a modular plug and play system for simple manipulation by conventional high-throughput liquid handling systems. The chip could be manipulated in the same fashion as a multiwell plate and interfaced with a novel pneumatic gasket system to achieve delivery speeds that were two-fold faster than those attained on the 16-well device. In addition, we worked to develop potential optical interrogation platform that could be fed populations of worms by the aforementioned delivery systems. This microfluidic chip consisted of an array of parallel traps to house individual worms over long durations for time-lapse studies of nerve regeneration after cuts to single axons mediated by a femtosecond pulse laser. Specifically, the platform was designed for regeneration studies in the C. elegans PQR neuron.

Automated Parallel Immobilization Microfluidic Platforms For High Throughput Neuronal Degeneration Studies With C Elegans

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Author : Evan Marley Hegarty
language : en
Publisher:
Release Date : 2014

Automated Parallel Immobilization Microfluidic Platforms For High Throughput Neuronal Degeneration Studies With C Elegans written by Evan Marley Hegarty and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2014 with categories.

C. elegans has emerged as an invaluable model organism for in vivo neurobiology research to understand disease mechanisms and pathology relevant in humans. Simple anatomy, short lifecycle, fully characterized genome, and miniature body scale make these nematodes an ideal model organism for phenotyping and bio-molecular studies using microfluidic platforms. Advancements in soft-lithography have improved the functionality of microfluidic technology for C. elegans, leading to whole organism studies in high-throughput manner that were not otherwise possible. In order to study phenomena that require large amounts of data such as drug screens for neurological disorders and phenotyping, high-throughput imaging platforms with high-speed, high-resolution image acquisition become essential. With this in mind, we have developed and tested microfluidic immobilization devices to enable high-throughput optical interrogation of C. elegans for neurodegenerative diseases and large scale drug screens. Initially, we designed, developed, and tested single-layer and double-layer SU8 mold PDMS chips with parallel tapered channels to immobilize 40 adult C. elegans for high-resolution fluorescence imaging of their neurons in a parallel manner. vi We achieved over 90% immobilization efficiency using these initial devices, but could achieve only ~50% of the trapped worms with proper orientation to allow scoring of the VC neurons of interest. To improve worm orientation, we developed a three-layer microfluidic chip that can immobilize and orient the adult worms for optical interrogation of these VC neurons with 90% efficiency. Finally, we scaled the platform to accommodate a large scale platform with standard multi-well format on-chip wells where each well leads to the optimized trapping channels. The final optimized multi-well platform provides comprehensive easy to use 96-well microfluidic system to orient, immobilize, and image adult C. elegans in high-throughput manner. The novel gasket system can pressurize the multi-well device pre-loaded with 96 individual worm populations. Using a sequence of on-off applied gasket pressure, we can orient and immobilize worms in all 96 devices simultaneously in less than 5 minutes. Custom designed software can capture 12 z-stack images per worm from all 96-well in less than 12 minutes. With 95% trapping efficiency, approximately 90% of the worms can be scored successfully for neuronal phenotyping of VC neurons. This 96-well platform and the automated imaging system enable high-throughput optical interrogation of adult C. elegans for large-scale drug screens relating to ageing and various neurodegenerative diseases.

Parallel And Serial Microfluidic Platforms For Femtosecond Laser Axotomy In Caenorhabditis Elegans For Nerve Regeneration Studies

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Author : Sertan Kutal Gokce
language : en
Publisher:
Release Date : 2016

Parallel And Serial Microfluidic Platforms For Femtosecond Laser Axotomy In Caenorhabditis Elegans For Nerve Regeneration Studies written by Sertan Kutal Gokce and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2016 with categories.

Understanding the molecular basis of nerve regeneration can potentiate the development of novel and efficient treatments for neurodegenerative diseases. Severing axons in the small nematode Caenorhabditis elegans (C. elegans) with femtosecond laser surgery and then observing the subsequent axonal regrowth is a promising approach to understand the molecular mechanisms of nerve regeneration in vivo. Effective and reversible immobilization of the nematodes is necessary for both axotomy and follow-up imaging. However, conventional worm handling techniques such as using anesthetics or polystyrene beads are labor-intensive and time-consuming processes, hindering high-throughput. Microfluidic devices enable the manipulation of the nematodes on a single chip with unprecedented throughput and integrity. This dissertation introduces two comprehensive microfluidic systems for femtosecond laser axotomy in C. elegans, offering several advantages over conventional techniques in terms of speed and the ability for automation of the tedious axotomy experiments. The first microfluidic system is an automated serial microfluidic platform for performing femtosecond laser axotomy in C. elegans. The microfluidic platform along with a custom-developed automation program isolates a single nematode from a pre-loaded population, immobilizes the nematode, and performs femtosecond laser axotomy. The full automation of the axotomy process is achieved by combining efficient image analysis methodologies with synchronized valve and flow progression in the microfluidic chip to perform multiple surgeries in a serial and automated manner. The serial automated microfluidic platform reduces the time required to perform axotomies within individual worms to ~ 17 s/worm. The second microfluidic system is a parallelized multitrap microfluidic platform, “worm hospital”, that allows on-chip axotomy, post-surgery housing for recovery, and imaging of nerve regeneration on a single chip. The microfluidic platform features 20 trapping channels for laser axotomy and subsequent post-surgery imaging, and a perfusion area to house the worms after laser axotomy. This microfluidic platform is a single-flow Polydimethylsiloxane (PDMS) layer device and has no active control PDMS layer, which reduces the fabrication and operation complexity of the chip, especially for non-expert users. The roles of neurodevelopmental genes in the Wnt/Frizzled pathway on the nerve regeneration was investigated using the “worm hospital”. In summary, the microfluidic platforms presented in this dissertation enabled performing femtosecond laser axotomy in C. elegans in a fast and repeatable manner with a controllable microenvironment. Both microfluidic platforms offer promising methodologies for prospective large-scale screening of genes involved in nerve regeneration with a high throughput in an automated manner.

Parallelized Microfluidic Devices For High Throughput Nerve Regeneration Studies In Caenorhabditis Elegans

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Author : Navid Ghorashian
language : en
Publisher:
Release Date : 2010

Parallelized Microfluidic Devices For High Throughput Nerve Regeneration Studies In Caenorhabditis Elegans written by Navid Ghorashian and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2010 with categories.

The nexus of engineering and molecular biology has given birth to high-throughput technologies that allow biologists and medical scientists to produce previously unattainable amounts of data to better understand the molecular basis of many biological phenomena. Here, we describe the development of an enabling biotechnology, commonly known as microfluidics in the fabrication of high-throughput systems to study nerve degeneration and regeneration in the well-defined model nematode, Caenorhabditis elegans (C. elegans). Our lab previously demonstrated how femtosecond (fs) laser pulses could precisely cut nerve axons in C. elegans, and we observed axonal regeneration in vivo in single worms that were immobilized on anesthetic treated agar pads. We then developed a microfluidic device capable of immobilizing one worm at a time with a deformable membrane to perform these experiments without agar pads or anesthetics. Here, we describe the development of improved microfluidic devices that can trap and immobilize up to 24 individual worms in parallel chambers for high-throughput axotomy and subsequent imaging of nerve regeneration in a single platform. We tested different micro-channel designs and geometries to optimize specific parameters: (1) the initial trapping of a single worm in each immobilization chamber, simultaneously, (2) immobilization of single worms for imaging and fs-laser axotomy, and (3) long term storage of worms on-chip for imaging of regeneration at different time points after the initial axon cut.

Micro Nano Devices For Chemical Analysis

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Author : Manabu Tokeshi
language : en
Publisher: MDPI
Release Date : 2018-07-03

Micro Nano Devices For Chemical Analysis written by Manabu Tokeshi and has been published by MDPI this book supported file pdf, txt, epub, kindle and other format this book has been release on 2018-07-03 with Electronic book categories.

This book is a printed edition of the Special Issue "Micro/Nano Devices for Chemical Analysis" that was published in Micromachines

C Elegans Atlas

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Author : David H. Hall
language : en
Publisher: CSHL Press
Release Date : 2008

C Elegans Atlas written by David H. Hall and has been published by CSHL Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008 with Caenorhabditis elegans categories.

Derived from the acclaimed online “WormAtlas,†C. elegansAtlas is a large-format, full-color atlas of the hermaphroditic form of the model organism C. elegans, known affectionately as “the worm†by workers in the field. Prepared by the editors of the WormAtlas Consortium, David H. Hall and Zeynep F. Altun, this book combines explanatory text with copious, labeled, color illustrations and electron micrographs of the major body systems of C. elegans. Also included are electron microscopy cross sections of the worm. This laboratory reference is essential for the working worm biologist, at the bench and at the microscope, and provides a superb companion to the C. elegansII monograph. It is also a valuable tool for investigators in the fields of developmental biology, neurobiology, reproductive biology, gene expression, and molecular biology.

Micro And Nano Systems For Biophysical Studies Of Cells And Small Organisms

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Author : Xinyu Liu
language : en
Publisher: Academic Press
Release Date : 2021-08-14

Micro And Nano Systems For Biophysical Studies Of Cells And Small Organisms written by Xinyu Liu and has been published by Academic Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2021-08-14 with Business & Economics categories.

Micro and Nano Systems for Biophysical Studies of Cells and Small Organisms provides a comprehensive introduction to the state-of-the-art micro and nano systems that have recently been developed and applied to biophysical studies of cells and small organisms. These micro and nano systems span from microelectromechanical systems (MEMS) and microfluidic devices to robotic micro-nanomanipulation systems. These biophysical studies range from cell mechanics to the neural science of worms and Drosophila. This book will help readers understand the fundamentals surrounding the development of these tools and teach them the most recent advances in cellular and organismal biophysics enabled by these technologies. Comprehensive coverage of micro and nano-system technology and application to biophysical studies of cells and small organisms. Highlights the most recent advances in cellular and organismal biophysics enabled by micro and nano systems. Insightful outlook on future directions and trends in each chapter covering a sub-area of the book topic.

Fly Pushing

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Author : Ralph J. Greenspan
language : en
Publisher: CSHL Press
Release Date : 2004

Fly Pushing written by Ralph J. Greenspan and has been published by CSHL Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2004 with Science categories.

A second edition of the classic handbook has become a standard in the Drosophila field. This edition is expanded to include topics in which classical genetic strategies have been augmented with new molecular tools. Included are such new techniques as homologous recombination, RNAi, new mapping techniques, and new mosaic marking techniques.

Applications Of Microfluidic Systems In Biology And Medicine

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Author : Manabu Tokeshi
language : en
Publisher: Springer
Release Date : 2024-10-30

Applications Of Microfluidic Systems In Biology And Medicine written by Manabu Tokeshi and has been published by Springer this book supported file pdf, txt, epub, kindle and other format this book has been release on 2024-10-30 with Science categories.

This book is the second edition of the one originally published in 2016, which focused on state-of-the-art microfluidic research in medical and biological applications. Similar to the first edition, beginners in the field —undergraduates, engineers, biologists, medical and pharmaceutical researchers—will easily learn to understand microfluidic-based medical and biological applications. Because a wide range of topics is summarized here, it also helps experts to learn more about fields outside their own specialties. In this second edition, significant revisions have been made to chapters covering technologies that have seen major advancements, such as acoustofluidics, protein crystallography, organ-on-a-chip systems, nanopore sensing, and paper-based microfluidics. In addition, the chapters on cancer diagnosis using exosomes and single-cell sequencing using droplet microfluidics, which are attracting attention as new technologies, have been newly added. Readers will be convinced that microfluidic devices have great potential for medical and biological applications.

Micrornas

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Author : Krishnarao Appasani
language : en
Publisher: Cambridge University Press
Release Date : 2009-08-20

Micrornas written by Krishnarao Appasani and has been published by Cambridge University Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2009-08-20 with Science categories.

MicroRNAs (miRNAs) are RNA molecules, conserved by evolution, that regulate gene expressions and their recent discovery is revolutionising both basic biomedical research and drug discovery. Expression levels of MiRNAs have been found to vary between tissues and with developmental stages and hence evaluation of the global expression of miRNAs potentially provides opportunities to identify regulatory points for many different biological processes. This wide-ranging reference work, written by leading experts from both academia and industry, will be an invaluable resource for all those wishing to use miRNA techniques in their own research, from graduate students, post-docs and researchers in academia to those working in R&D in biotechnology and pharmaceutical companies who need to understand this emerging technology. From the discovery of miRNAs and their functions to their detection and role in disease biology, this volume uniquely integrates the basic science with industry application towards drug validation, diagnostic and therapeutic development. Forewords by: Sidney Altman, Yale University, Winner of the Nobel Prize in Chemistry, 1989 and Victor R. Ambros, Dartmouth Medical School, Co-discoverer of MicroRNAs