Download or read book Self Learning Microfluidic Platform for Single Cell Imaging and Classification in Flow written by Iordania Constantinou and published by . This book was released on 2019 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Single-cell analysis commonly requires the confinement of cell suspensions in an analysis chamber or the precise positioning of single cells in small channels. Hydrodynamic flow focusing has been broadly utilized to achieve stream confinement in microchannels for such applications. As imaging flow cytometry gains popularity, the need for imaging-compatible microfluidic devices that allow for precise confinement of single cells in small volumes becomes increasingly important. At the same time, high-throughput single-cell imaging of cell populations produces vast amounts of complex data, which gives rise to the need for versatile algorithms for image analysis. In this work, we present a microfluidics-based platform for single-cell imaging in-flow and subsequent image analysis using variational autoencoders for unsupervised characterization of cellular mixtures. We use simple and robust Y-shaped microfluidic devices and demonstrate precise 3D particle confinement towards the microscope slide for high-resolution imaging. To demonstrate applicability, we use these devices to confine heterogeneous mixtures of yeast species, brightfield-image them in-flow and demonstrate fully unsupervised, as well as few-shot classification of single-cell images with 88% accuracy.
Download or read book Microfluidics for Cells and Other Organisms written by Danny van Noort and published by MDPI. This book was released on 2019-10-23 with total page 200 pages. Available in PDF, EPUB and Kindle. Book excerpt: Microfluidics-based devices play an important role in creating realistic microenvironments in which cell cultures can thrive. They can, for example, be used to monitor drug toxicity and perform medical diagnostics, and be in a static-, perfusion- or droplet-based device. They can also be used to study cell-cell, cell-matrix or cell-surface interactions. Cells can be either single cells, 3D cell cultures or co-cultures. Other organisms could include bacteria, zebra fish embryo, C. elegans, to name a few.
Download or read book Single Cell Analysis written by Tuhin Subhra Santra and published by MDPI. This book was released on 2021-06-02 with total page 254 pages. Available in PDF, EPUB and Kindle. Book excerpt: Cells are the most fundamental building block of all living organisms. The investigation of any type of disease mechanism and its progression still remains challenging due to cellular heterogeneity characteristics and physiological state of cells in a given population. The bulk measurement of millions of cells together can provide some general information on cells, but it cannot evolve the cellular heterogeneity and molecular dynamics in a certain cell population. Compared to this bulk or the average measurement of a large number of cells together, single-cell analysis can provide detailed information on each cell, which could assist in developing an understanding of the specific biological context of cells, such as tumor progression or issues around stem cells. Single-cell omics can provide valuable information about functional mutation and a copy number of variations of cells. Information from single-cell investigations can help to produce a better understanding of intracellular interactions and environmental responses of cellular organelles, which can be beneficial for therapeutics development and diagnostics purposes. This Special Issue is inviting articles related to single-cell analysis and its advantages, limitations, and future prospects regarding health benefits.
Download or read book Development of an Automated 3D Flow focusing Microfluidic Platform for Single Cell Sorting written by Yingkai Lyu and published by . This book was released on 2020 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Microfluidics for Single Cell Analysis written by Jin-Ming Lin and published by Springer Nature. This book was released on 2019-08-28 with total page 263 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book summarizes the various microfluidic-based approaches for single-cell capture, isolation, manipulation, culture and observation, lysis, and analysis. Single-cell analysis reveals the heterogeneities in morphology, functions, composition, and genetic performance of seemingly identical cells, and advances in single-cell analysis can overcome the difficulties arising due to cell heterogeneity in the diagnostics for a targeted model of disease. This book provides a detailed review of the state-of-the-art techniques presenting the pros and cons of each of these methods. It also offers lessons learned and tips from front-line investigators to help researchers overcome bottlenecks in their own studies. Highlighting a number of techniques, such as microfluidic droplet techniques, combined microfluidics-mass-spectrometry systems, and nanochannel sampling, it describes in detail a new microfluidic chip-based live single-cell extractor (LSCE) developed in the editor’s laboratory, which opens up new avenues to use open microfluidics in single-cell extraction, single-cell mass spectrometric analysis, single-cell adhesion analysis and subcellular operations. Serving as both an elementary introduction and advanced guidebook, this book interests and inspires scholars and students who are currently studying or wish to study microfluidics-based cell analysis methods.
Download or read book Microfluidic Methods for Molecular Biology written by Chang Lu and published by Springer. This book was released on 2016-05-14 with total page 382 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book covers the state-of-the-art research on molecular biology assays and molecular techniques enabled or enhanced by microfluidic platforms. Topics covered include microfluidic methods for cellular separations and single cell studies, droplet-based approaches to study protein expression and forensics, and microfluidic in situ hybridization for RNA analysis. Key molecular biology studies using model organisms are reviewed in detail. This is an ideal book for students and researchers in the microfluidics and molecular biology fields as well as engineers working in the biotechnology industry. This book also: Reviews exhaustively the latest techniques for single-cell genetic, epigenetic, metabolomic, and proteomic analysis Illustrates microfluidic approaches for inverse metabolic engineering, as well as analysis of circulating exosomes Broadens readers’ understanding of microfluidics convection-based PCR technology, microfluidic RNA-seq, and microfluidics for robust mobile diagnostics
Download or read book Microtechnology for Cell Manipulation and Sorting written by Wonhee Lee and published by Springer. This book was released on 2016-10-05 with total page 287 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book delves into the recent developments in the microscale and microfluidic technologies that allow manipulation at the single and cell aggregate level. Expert authors review the dominant mechanisms that manipulate and sort biological structures, making this a state-of-the-art overview of conventional cell sorting techniques, the principles of microfluidics, and of microfluidic devices. All chapters highlight the benefits and drawbacks of each technique they discuss, which include magnetic, electrical, optical, acoustic, gravity/sedimentation, inertial, deformability, and aqueous two-phase systems as the dominant mechanisms utilized by microfluidic devices to handle biological samples. Each chapter explains the physics of the mechanism at work, and reviews common geometries and devices to help readers decide the type of style of device required for various applications. This book is appropriate for graduate-level biomedical engineering and analytical chemistry students, as well as engineers and scientists working in the biotechnology industry.
Download or read book Cell Analysis on Microfluidics written by Jin-Ming Lin and published by Springer. This book was released on 2017-10-25 with total page 435 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book presents a detailed overview of the design, formatting, application, and development of microfluidic chips in the context of cell biology research, enumerating each element involved in microfluidics-based cell analysis, discussing its history, status quo, and future prospects, It also offers an extensive review of the research completed in the past decade, including numerous color figures. The individual chapters are based on the respective authors' studies and experiences, providing tips from the frontline to help researchers overcome bottlenecks in their own work. It highlights a number of cutting-edge techniques, such as 3D cell culture, microfluidic droplet technique, and microfluidic chip-mass spectrometry interfaces, offering a first-hand impression of the latest trends in the field and suggesting new research directions. Serving as both an elementary introduction and advanced guidebook, the book interests and inspires scholars and students who are currently studying microfluidics-based cell analysis methods as well as those who wish to do so.
Download or read book Development of Multi purpose Microfluidic Platform for Single cell and Pairwise Cell Interaction Assay written by Young-Ji Kim and published by . This book was released on 2009 with total page 95 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Imaging Flow Cytometry written by Natasha S. Barteneva and published by Humana. This book was released on 2015-11-23 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This detailed volume for the first time explores techniques and protocols involving quantitative imaging flow cytometry (IFC), which has revolutionized our ability to analyze cells, cellular clusters, and populations in a remarkable fashion. Beginning with an introduction to technology, the book continues with sections addressing protocols for studies on the cell nucleus, nucleic acids, and FISH techniques using an IFC instrument, immune response analysis and drug screening, IFC protocols for apoptosis and cell death analysis, as well as morphological analysis and the identification of rare cells. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Imaging Flow Cytometry: Methods and Protocols will be a critical source for all laboratories seeking to implement IFC in their research studies.
Download or read book Microfluidic Platform for Multi parameter Analysis of Live Single Cells written by Meryl Rodrigues and published by . This book was released on 2014 with total page 54 pages. Available in PDF, EPUB and Kindle. Book excerpt: Cellular heterogeneity is a key factor in various cellular processes as well as in disease development, especially associated with immune response and cancer progression. Cell-to-cell variability is considered to be one of the major obstacles in early detection and successful treatment of cancer. Most present technologies are based on bulk cell analysis, which results in averaging out the results acquired from a group of cells and hence missing important information about individual cells and their behavior. Understanding the cellular behavior at the single-cell level can help in obtaining a complete profile of the cell and to get a more in-depth knowledge of cellular processes. For example, measuring transmembrane fluxes oxygen can provide a direct readout of the cell metabolism. The goal of this thesis is to design, optimize and implement a device that can measure the oxygen consumption rate (OCR) of live single cells. A microfluidic device has been designed with the ability to rapidly seal and unseal microchambers containing individual cells and an extracellular optical oxygen sensor for measuring the OCR of live single cells. The device consists of two parts, one with the sensor in microwells (top half) and the other with channels and cells trapped in Pachinko-type micro-traps (bottom half). When the two parts of the device are placed together the wells enclose each cell. Oil is flown in through the channels of the device to produce isolated and sealed microchamber around each cell. Different fluids can be flowed in and out of the device, alternating with oil, to rapidly switch between sealed and unsealed microenvironment around each cell. A fluorescent ratiometric dual pH and oxygen sensor is placed in each well. The thesis focuses on measuring changes in the oxygen consumption rate of each cell within a well. Live and dead cells are identified using a fluorescent live/dead cell assay. Finally, the technology is designed to be scalable for high-throughput applications by controlling the flow rate of the system and increasing the cell array density.
Download or read book Microfluidic Tools for Connecting Single Cell Optical and Gene Expression Phenotype written by Jesse Qiuxu Zhang and published by . This book was released on 2020 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: The single cell is the fundamental unit of biology. Understanding how the identity of individual cells in multicellular organisms contribute to their function remains a key question in Biology. Traditionally, most observations of cells were made through imaging-based techniques. Today, advances in Next Generation Sequencing have led to the widespread adoption of sequencing-based techniques for investigating the genotype and phenotype at single-cell resolution. Microfluidics, including droplet-based microfluidics, have been instrumental in the most successful commercial single-cell genomics platforms.Integrating sequencing and imaging techniques will provide additional information than either of the techniques alone. Both single-cell imaging and genomics techniques measure orthogonal targets, and when combined reveal additional insights into cellular function. However, when performing sequential single-cell assays, there currently exists a tradeoff between throughput and information content. This dissertation will describe progress made towards reducing that gap. I will describe novel microfluidic platforms and techniques and applications involving integrating single-cell sequencing and optical measurements at high throughput. The microfluidics tools that will be discussed in this Dissertation aim to be a platform for performing single-cell multi-parameter and multi-omics techniques that will help further our understanding of cellular identity and how genotype informs phenotype at the single-cell level.
Download or read book A Microfluidic Platform Enables Large Scale Single Cell Screening to Identify Genes Involved in Bacterial Persistence written by Amanda Verpoorte and published by . This book was released on 2017 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Mots-clés de l'autrice: microfluidics ; screening platform ; high-content screening ; time-lapse microscopy ; bacterial persistence ; isoniazid ; phenotypic heterogeneity ; Mycobacterium smegmatis ; single-cell analysis.
Download or read book Development and Application of a Microfluidic Platform for Single cell Cultivation of CHO Suspension Cell Lines written by Julian Schmitz and published by . This book was released on 2022 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Integrated Microfluidic Device for Single cell High Throughput Screening in Dynamic Gene Expression Analysis written by Lawrence Kwan Yeung Hui and published by . This book was released on 2008 with total page 60 pages. Available in PDF, EPUB and Kindle. Book excerpt: Over the past decade, interest in microfluidics has surged as applications have trended towards novel biological assays. Specifically, the ability of microfluidics to parallelize cellular studies through array-based chip designs has attracted researchers interested in investigating cellular function under a wide variety of environmental conditions. The capability of microfluidic devices to control microenvironment conditions and induce dynamic perturbation to cellular systems makes microfluidics (or "lab-on-a-chip") an attractive platform to study gene expression dynamics. In this project, the functionality of microfluidic technology is exploited to design and construct a device for isolation and observation of cells in high throughput. The integration of a concentration gradient with homogenous medium within each chamber was designed specifically to investigate gene regulation in Saccharomyces cerevisiae under various concentrations of chemical inducers. These devices were designed to sustain cells for extended periods of time with high temporal resolution to study dynamic gene expression in single cells. The device builds on previous studies by probing up to eight distinct cell cultures in parallel. The microfluidic platform was then used to study yeast cells at various levels of inducer perturbations. Further experimentation revealed the utility of a parallel gradient by producing an induction curve of the yeast response. Such high-throughput designs will prove essential to yeast systems biology research as it strives to understand the complex regulatory interactions that dictate cell function by probing vast regions of parameter space.
Download or read book Microfluidics for Single cell Analysis written by and published by . This book was released on 2019 with total page 263 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book summarizes the various microfluidic-based approaches for single-cell capture, isolation, manipulation, culture and observation, lysis, and analysis. Single-cell analysis reveals the heterogeneities in morphology, functions, composition, and genetic performance of seemingly identical cells, and advances in single-cell analysis can overcome the difficulties arising due to cell heterogeneity in the diagnostics for a targeted model of disease. This book provides a detailed review of the state-of-the-art techniques presenting the pros and cons of each of these methods. It also offers lessons learned and tips from front-line investigators to help researchers overcome bottlenecks in their own studies. Highlighting a number of techniques, such as microfluidic droplet techniques, combined microfluidics-mass-spectrometry systems, and nanochannel sampling, it describes in detail a new microfluidic chip-based live single-cell extractor (LSCE) developed in the editor's laboratory, which opens up new avenues to use open microfluidics in single-cell extraction, single-cell mass spectrometric analysis, single-cell adhesion analysis and subcellular operations. Serving as both an elementary introduction and advanced guidebook, this book interests and inspires scholars and students who are currently studying or wish to study microfluidics-based cell analysis methods.
Download or read book High Throughput Microfluidic Technologies for Cell Separation and Single cell Analysis written by Lidan Wu (Ph. D.) and published by . This book was released on 2016 with total page 102 pages. Available in PDF, EPUB and Kindle. Book excerpt: The heterogeneity of individual cellular behavior in response to physical and chemical stimuli has raised increasing attention in many biological processes. There is great incentive in developing techniques for high throughput single-cell measurements and manipulations. Particularly, cell size has been recognized as an important parameter in single cell study and pericellular protease activity plays a key role in regulating the microenvironment of individual cells. Therefore, this thesis focuses on establishing new methods to address the issues of cell size and single cell protease measurement. We first develop a size-based cell separation technique using Dean-coupled inertial microfluidic sorter. Separation of cells by size before downstream assays might be beneficial in simplifying the system and facilitating the discovery of rare subpopulations through enrichment of cells with certain size range or cell cycle phase. By investigating the particle focusing and separation mechanisms in curved microfluidic channel, we develop a novel design of inertial microfluidic sorter with higher separation resolution and then demonstrate its capacity in leukocyte isolation from blood. This novel cell sorter would be a promising alternative to many other cell separation problems. We then establish a microfluidic platform for functional measurement of single cell pericellular proteases, including both those secreted and expressed on cell surface. We apply the platform to studying the PMA-mediated protease response of HepG2 cells at single-cell level and reveal the diversity in the dynamic patterns of single-cell protease activity profile upon drug stimulation. We also present the preliminary exploration of single-cell protease activity behavior in anticancer drug resistance development. Lastly, we explore the applicability of our platform for single-cell shedding measurement. Protease-mediated molecular shedding is one of the key mechanisms through which individual cells actively regulate their own microenvironment. However, the amount of molecules being shed for individual cells is extremely low, posing significant challenges in detecting shedding quantitatively. By means of analytical analysis and numerical simulations, we investigate the intrinsic noise of low-abundance molecule detection. Experimental characterizations have also been performed to evaluate the impact of practical factors on actual readout variation.
