Download or read book Advanced Multiplexing Techniques in Single Molecule Localisation Microscopy written by Nazar Oleksiievets and published by . This book was released on 2022 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: DNA point accumulation for imaging in nanoscale topography (DNA-PAINT) is a powerful super-resolution technique highly suitable for multi-target (multiplexing) bio-imaging applications. However, multiplexed imaging of cells is still challenging due to the dense and sticky environment inside a cell. This thesis presents several novel techniques for multiplexed imaging, with the main focus on the DNA-PAINT super-resolution technique. At first, I demonstrate multiplexed imaging by using three different single-domain antibodies (nanobodies). They have a three times smaller size than conventiona...
Download or read book Development of Multiplexing Strategies for Electron and Super resolution Optical Microscopy written by Paul W. Tillberg and published by . This book was released on 2013 with total page 31 pages. Available in PDF, EPUB and Kindle. Book excerpt: The aim of this work is to increase the multiplexing capabilities of electron and super resolution optical microscopy. This will be done through the development of molecular-scale barcodes that can be resolved in one of the two high resolution imaging modes. In the optical domain, the number of colors available in stochastic optical reconstruction microscopy (STORM) will be increased by taking advantage of not only the spectral differences between STORM fluorophores but their kinetic properties as well. In the electron microscopy domain, the recently developed electron contrast-generating protein miniSOG will be concatenated to produce fully genetically encoded barcodes that can be resolved using standard transmission electron microscopy techniques. At the time of writing, the hardware for a STORM microscope has been assembled. Single molecule fluorescence blinking has been observed, though the imaging buffer still needs to be optimized for imaging. Concatamers of miniSOG have been generated and can be expressed in HEK cells and photo-oxidized.
Download or read book Advanced Single Molecule Localization Microscopy for Imaging Cellular Nuclei written by Shih-Ya Chen and published by . This book was released on 2020* with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: In this PhD research, single molecule localization microscopy (SMLM) was used to image nuclear structures with a resolution down to several nanometers.The scope of this PhD research is to develop a 3D SMLM microscope which can overcome several principle limitations in imaging nuclei in 3D. The advanced improvements during this PhD research include a broad range of research subjects associated to SMLM techniques. Firstly, one of the most common problems of a super-resolution microscope is sample drift, because a small sample drift may result in artefacts and can hamper the resolution. A speckle-based method was developed to correct sample drift without changing the standard design of the SMLM setup. This drift correction method can achieve a resolution of several nanometers. Secondly, another principle problem is that commonly used organic fluorophores are restricted in their photon budget. It is often observed that the chemical structure of fluorophores change after high laser irradiance resulting in photobleaching. A patterned illumination technique was developed which allows the user to define arbitrary regions of interest for illumination with a flat-top intensity profile. Thirdly, for SMLM in particular, a carefully adjusted chemical environment in the sample is recommended to induce sufficiently blinking signals of the organic fluorophores in combination with an appropriate laser irradiance. However, such an imaging buffer can degrade over time and may not be suitable for long time imaging. Nanographene was presented as a new class of fluorophores which have blinking properties without an imaging buffer. Therefore, the nanographenes facilitate a wide range of SMLM applications including bio-imaging and material science. These advanced developments are not only for imaging nuclei, but also applicable to applications in other biological researches and in material science.
Download or read book Single Molecule Fluorescence Spectroscopy and Imaging Advanced Methods and Applications in Life Sciences written by Arindam Ghosh and published by . This book was released on 2021 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The visualization of biological structures down to the molecular length scale has been recently made possible by the development of super-resolution fluorescence microscopy. These techniques now routinely resolve biological structures down to a few nanometers. Various super-resolution techniques have been developed, the most successful being Stimulated Depletion Emission (STED) microscopy and Single Molecule Localization Microscopy (SMLM). In what follows, I will focus on the latter class of techniques which is based on the fact that a single molecule image allows for localizing the molecul...
Download or read book Development of Advanced Single molecule Fluorescence Microscopy Techniques and Their Application for the Study of Protein protein Interactions in the Context of Immunological Signaling written by and published by . This book was released on 2023 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This thesis titled ``Development of Advanced Single-Molecule Fluorescence Microscopy Techniques and their Application for the Study of Protein-Protein Interactions in the Context of Immunological Signaling'' aims to make single-molecule imaging and the study of protein-protein interactions on membranes more accessible to the research community by providing new single-molecule fluorescence microscopy tools and techniques, and sharing them following an open-science approach. Besides the design, assembly, and characterization of a custom single-molecule TIRF microscope specifically tailored for live-cell and single-molecule imaging applications, the author presents an experimental platform designed for single-molecule analysis of ligand-induced binding, which was employed to investigate purified interleukin-2 and -15 receptors reconstituted on supported lipid bilayers. Furthermore, the author presents a novel approach called DNA-PAINT single-particle tracking (DNA-PAINT-SPT), an innovative method designed to extend single-molecule trajectory lengths of membrane proteins in live cells and reconstitution experiments. In the thesis, this method is applied for extensive quantitative studies of the binding kinetics of a membrane-anchored protein homodimer. Overall, the developments and applications presented in this thesis contribute to the advancement of single-molecule fluorescence microscopy techniques and open up new possibilities for investigating protein-protein interactions and immunological signaling in greater detail.
Download or read book Controlled Synthesis of Nanoparticles in Microheterogeneous Systems written by Vincenzo Turco Liveri and published by Springer Science & Business Media. This book was released on 2006-01-04 with total page 202 pages. Available in PDF, EPUB and Kindle. Book excerpt: The first step in developing nanoscience and nanotechnology is the production of nanoparticles. Controlled Synthesis of Nanoparticles in Microheterogeneous Systems contains descriptions of one of the most powerful bottom-up methods of synthesizing size controlled and stable nanoparticles. This method is based on the use of surfactant-containing microheterogeneous systems: liquid crystals, monolayers and multilayers, solutions of direct and reversed micelles, direct and reversed vesicles, and water-in-oil and oil-in-water microemulsions. The author is prominent in the field of physico-chemical characterization of microheterogeneous systems and their use as ideal solvent and reaction media for the production and long-term storage of nanomaterials. This is the first book that attempts to unify the knowledge necessary for judicious manipulation of surfactant-based systems and a fine tuning of geometric and physico-chemical properties of nanoparticles of a wide variety of substances. Prof. Turco Liveri has chosen to write an easy-to-read book aiming to be evocative rather than exhaustive. Because of the intense interest in nanoscience and nanomaterials, this book is an important fundamental work that fits ideally into the series Nanostructure Science and Technology and will be useful for a wide range of students and young researchers involved in the study and manipulation of matter at the atomic level.
Download or read book Advanced Fluorescence Microscopy written by Peter J. Verveer and published by Humana. This book was released on 2016-08-23 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume provides an overview of advanced fluorescence microscopy, covering a broad range of methods. Each chapter focuses on a different method and provides a practical guide for application in biological systems. Written in the highly successful Methods in Molecular Biology series format, 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 cutting-edge, Advanced Fluorescence Microscopy: Methods and Protocols seeks to provide scientists with methods for biological systems that are of interest.
Download or read book Development of Advanced Methods for Super resolution Microscopy Data Analysis and Segmentation written by Leonid Andronov and published by . This book was released on 2018 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Among the super-resolution methods single-molecule localization microscopy (SMLM) is remarkable not only for best practically achievable resolution but also for the direct access to properties of individual molecules. The primary data of SMLM are the coordinates of individual fluorophores, which is a relatively rare data type in fluorescence microscopy. Therefore, specially adapted methods for processing of these data have to be developed. I developed the software SharpViSu and ClusterViSu that allow for most important data processing steps, namely for correction of drift and chromatic aberrations, selection of localization events, reconstruction of data in 2D images or 3D volumes using different visualization techniques, estimation of resolution with Fourier ring correlation, and segmentation using K- and L-Ripley functions. Additionally, I developed a method for segmentation of 2D and 3D localization data based on Voronoi diagrams, which allows for automatic and unambiguous cluster analysis thanks to noise modeling with Monte-Carlo simulations. Using advanced data processing methods, I demonstrated clustering of CENP-A in the centromeric regions of the cell nucleus and structural transitions of these clusters upon the CENP-A deposition in early G1 phase of the cell cycle.
Download or read book Studying Large Multi Protein Complexes Using Single Molecule Localization Microscopy written by David Virant and published by . This book was released on 2019 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Automated Highly Multiplexed Super resolution Imaging of Protein Nano architecture in Cells and Tissues written by Maja Klevanski and published by . This book was released on 2019 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Selective Autophagy written by Ioannis P. Nezis and published by Springer Nature. This book was released on with total page 250 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Live Cell Imaging written by Robert D. Goldman and published by CSHL Press. This book was released on 2005 with total page 658 pages. Available in PDF, EPUB and Kindle. Book excerpt: Recent advances in imaging technology reveal, in real time and great detail, critical changes in living cells and organisms. This manual is a compendium of emerging techniques, organized into two parts: specific methods such as fluorescent labeling, and delivery and detection of labeled molecules in cells; and experimental approaches ranging from the detection of single molecules to the study of dynamic processes in organelles, organs, and whole animals. Although presented primarily as a laboratory manual, the book includes introductory and background material and could be used as a textbook in advanced courses. It also includes a DVD containing movies of living cells in action, created by investigators using the imaging techniques discussed in the book. The editors, David Spector and Robert Goldman, whose previous book was Cells: A Laboratory Manual,are highly respected investigators who have taught microscopy courses at Cold Spring Harbor Laboratory, the Marine Biology Laboratory at Woods Hole, and Northwestern University.
Download or read book Advances in Medical Imaging Detection and Diagnosis written by Raj Bawa and published by CRC Press. This book was released on 2023-10-31 with total page 1541 pages. Available in PDF, EPUB and Kindle. Book excerpt: Medical care is the most critical issue of our time and will be so for the foreseeable future. In this regard, the pace and sophistication of advances in medicine in the past two decades have been truly breathtaking. This has necessitated a growing need for comprehensive reference resources that highlight current issues in specific sectors of medicine. Keeping this in mind, each volume in the Current Issues in Medicine series is a stand‐alone text that provides a broad survey of various important topics in a focused area of medicine—all accomplished in a user-friendly yet interconnected format. This volume addresses advances in medical imaging, detection, and diagnostic technologies. Technological innovations in these sectors of medicine continue to provide for safer, more accurate, and faster diagnosis for patients. This translates into superior prognosis and better patient compliance, while reducing morbidity and mortality. Hence, it is imperative that practitioners stay current with these latest advances to provide the best care for nursing and clinical practices. While recognizing how expansive and multifaceted these areas of medicine are, Advances in Medical Imaging, Detection, and Diagnosis addresses crucial recent progress, integrating the knowledge and experience of experts from academia and the clinic. The multidisciplinary approach reflected makes this volume a valuable reference resource for medical practitioners, medical students, nurses, fellows, residents, undergraduate and graduate students, educators, venture capitalists, policymakers, and biomedical researchers. A wide audience will benefit from having this volume on their bookshelf: health care systems, the pharmaceutical industry, academia, and government.
Download or read book Fundamentals of Advanced Omics Technologies From Genes to Metabolites written by and published by Newnes. This book was released on 2014-02-14 with total page 488 pages. Available in PDF, EPUB and Kindle. Book excerpt: Fundamentals of Advanced Omics Technologies: From Genes to Metabolites covers the fundamental aspects of the new instrumental and methodological developments in omics technologies, including those related to genomics, transcriptomics, epigenetics, proteomics and metabolomics, as well as other omics approaches such as glycomics, peptidomics and foodomics. The principal applications are presented in the following complementary volume. The chapters discuss in detail omics technologies, DNA microarray analysis, next-generation sequencing technologies, genome-wide analysis of methylation and histone modifications, emerging nanotechniques in proteomics, imaging mass spectrometry in proteomics, recent quantitative proteomics approaches, and advances in high-resolution NMR-based metabolomics, as well as MS-based non-targeted metabolomics and metabolome analysis by CE-MS, global glycomics analyses, foodomics, and high resolution analytical tools for quantitative peptidomics. Key aspects related to chemometrics, bioinformatics, data treatment, data integration and systems biology, deep-sequencing data analysis, statistical approaches for the analysis of microarray data, the integration of transcriptome and metabolome data and computational approaches for visualization and integration of omics data are also covered. Covers the latest advances in instrumentation, experimental design, sample preparation, and data analysis Provides thorough explanations and descriptions of specific omics technologies Describes advanced tools and methodologies for data pretreatment, storage, curation and analysis, as well as data integration
Download or read book Practical Bayesian Inference written by Coryn A. L. Bailer-Jones and published by Cambridge University Press. This book was released on 2017-04-27 with total page 306 pages. Available in PDF, EPUB and Kindle. Book excerpt: Science is fundamentally about learning from data, and doing so in the presence of uncertainty. This volume is an introduction to the major concepts of probability and statistics, and the computational tools for analysing and interpreting data. It describes the Bayesian approach, and explains how this can be used to fit and compare models in a range of problems. Topics covered include regression, parameter estimation, model assessment, and Monte Carlo methods, as well as widely used classical methods such as regularization and hypothesis testing. The emphasis throughout is on the principles, the unifying probabilistic approach, and showing how the methods can be implemented in practice. R code (with explanations) is included and is available online, so readers can reproduce the plots and results for themselves. Aimed primarily at undergraduate and graduate students, these techniques can be applied to a wide range of data analysis problems beyond the scope of this work.
Download or read book Single Molecule Image Analysis written by Christian Franke and published by Frontiers Media SA. This book was released on 2022-12-15 with total page 152 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Brain Derived Neurotrophic Factor BDNF written by Carlos B. Duarte and published by Humana. This book was released on 2019-07-10 with total page 258 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume discusses the latest techniques and methodologies used in the neurotrophin field to study the physiology of the Brain-Derived Neurotrophic Factor (BDNF). The book provides the tools essential to any researcher that intends to explore the peculiar regulation of this neurotrophin from gene expression to its release and the signaling by TrkB receptors, as well as to study the neuronal responses to BDNF or the role of neurotrophin in various neurological diseases. In Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory. Cutting edge and comprehensive, Brain-Derived Neurotrophic Factor(BDNF) is a valuable resource for researchers who want to further study the diversity of physiological roles of this important growth factor.
