Download and read online Live Cell Imaging in PDF and EPUB 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 and read online Imaging and Spectroscopic Analysis of Living Cells in PDF and EPUB This volume of Methods in Enzymology is the second of three parts looking at current methodology for the imaging and spectroscopic analysis of live cells. The chapters provide hints and tricks not available in primary research publications. It is an invaluable resource for academics, researchers and students alike. Expert authors who are leaders in the field Extensively referenced and useful figures and tables Provides hints and tricks to facilitate reproduction of methods
Download and read online Imaging and Spectroscopic Analysis of Living Cells in PDF and EPUB This volume is the second of 3 parts looking at current methodology for the imaging and spectroscopic analysis of live cells. The chapters provide hints and tricks not available in primary research publications. It is a useful resource for academics, researchers and students alike.
Download and read online Live Cell Imaging in PDF and EPUB Now a routine tool in biomedical and life science research, live cell imaging has made major progress enabling this core biochemical, cell, and molecular biology technique to become even more powerful, versatile, and affordable. In Live Cell Imaging: Methods and Protocols, a panel of expert contributors provide a comprehensive compendium of experimental approaches to live cell imaging in the form of several overview chapters followed by representative examples and case studies covering different aspects of the most current methodology. By examining a range of state-of-the-art protocols extensively validated in complex biological studies, this volume highlights new experimental and instrumental opportunities and helps researchers to select appropriate imaging methods for their specific biological questions and measurement tasks. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Live Cell Imaging: Methods and Protocols promises to contribute greatly to the further development and dissemination of this fundamentally important technology which spans across many disciplines including molecular and cell biology, chemistry, physics, optics, engineering, cell physiology, and medicine.
Download and read online Live Cell Imaging Cell Tracking and Lineage Analysis as a Tool to Investigate Dynamic Culture Processes in Heterogeneous Cell Systems in PDF and EPUB Live cell imaging can be used to study dynamic cellular systems at single cell resolution. In heterogeneous cell populations, analyzing cell properties at the single cell level reduces the generalization of results caused by population-based assays. This thesis details the implementation of live cell imaging and single cell tracking to characterize heterogeneous cell systems undergoing dynamic processes over multiple generations. This approach enables the consideration of both spatial and temporal variables as well as the mapping of cell phenotype trajectories along their generational lineages. Cell-, lineage-, and colony-level properties are used as descriptors of the underlying molecular mechanisms that they are produced by. These may be unexpected, emergent properties that can not be predicted or completely characterized at the molecular level. Analysis of these properties can reveal and characterize the properties and processes of dynamic, heterogeneous cell systems. Live cell imaging culture strategies were developed to enable characterization of both two- and three-dimensional cell systems. Computational modeling was performed to evaluate the conditions imposed by a confined imaging chamber that enables single cell resolution imaging of monolayer and multilayer cell systems. Imaging chamber dimensions and cell colony/aggregate sizes were calculated that would prevent the introduction of metabolite transport limitations and allow for stable, long term imaging. Methods for single cell tracking and analysis were also developed, which produces a database detailing the tracked, observed and extracted properties of every cell and colony, while maintaining the lineage structure of the data. Visualizations such as lineages, histograms and scatter plots were implemented to enable interactive data analysis and querying. These methods were used to characterize heterogeneity in two separate cell systems: human islet of Langerhans-derived progenitor cells, and human embryonic stem cells. Islet-derived progenitors are an expandable source of cells with potential for treatment of diabetes. Here, it was shown that there is an unequal contribution of islets to the progenitor derivation process. Islet-derived progenitors consist of two distinct sub-populations of cells that were distinguished by morphological identification during live cell image analysis. These sub-populations possess unique proliferation profiles and appear to exist in a dynamic state with each other. Three-dimensional tracking of islet progenitor derivation was implemented, but suffered from a lack of resolution to capture the dynamic nature of the transformation process. However, entire islets were imaged and tracked successfully under maintenance conditions, suggesting that this system may be useful for other cell types. These results highlight that live cell imaging and cell tracking may not be suitable for all cell systems and that inclusion of other analytical information, such as immunocytochemistry, would improve the power of cell tracking analysis. Human embryonic stem cell cultures were studied using live cell imaging to identify the mechanisms by which they differentiate to produce supportive niche cells. Cell tracking, morphology scoring and lineage analysis revealed a previously unappreciated level of heterogeneity within human embryonic stem cell colonies. The results show that a sub-population of human embryonic stem cells exist that are precursors to niche cell differentiation. However, these cells exist in a dynamic equilibrium with self-renewing stem cells, which is dependant on the presence of existing local niche cells. Sub-optimal niche conditions leads to the production of niche differentiation-competent cells and, significantly, considerable cell death. The effect of cell death is the clonal selection of self-renewing cells that contribute to colony expansion. Overall, these results highlight the importance of the co-transfer of existing niche cells and the dynamic balance that regulates human embryonic stem cell self-renewal and differentiation. This thesis displays the utility of live cell imaging, cell tracking and cell, colony and lineage analysis for studying dynamic heterogeneous systems. Furthermore, it highlight the fact that cell-, lineage- and colony-level analysis can uncover previously unappreciated heterogeneity and unknown sub-populations of cells. The system does not rely on characterization at the molecular level, but uses higher order measures to generalize them. However, future incorporation of cell, lineage and colony information with molecular-level information may results in analytical power not possible from either level alone. Such systems will be valuable tools in the growing fields of stem cell biology and systems biology.
Download and read online Cell Imaging Techniques in PDF and EPUB Cell imaging methodologies have now become essential research tools for a variety of disciplines that traditionally had not relied on them. In Cell Imaging Techniques: Methods and Protocols, distinguished international researchers describe in detail their state-of-the-art methods for the microscopic imaging of cells and molecules. The authors cover a wide spectrum of complementary techniques, including such methods as fluorescence microscopy, electron microscopy, atomic force microscopy, and laser scanning cytometry. Additional protocols on confocal scanning laser microscopy, quantitative computer-assisted image analysis, laser-capture microdissection, microarray image scanning, near-field scanning optical microscopy, and reflection contrast microscopy round out this eclectic collection of cutting-edge imaging techniques now available. The authors also discuss preparative methods for particles and cells by transmission electron microscopy. The protocols follow the successful Methods in Molecular Biology series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Timely and highly practical, Cell Imaging Techniques: Methods and Protocols provides researchers and clinicians with a richly useful guide to selecting and performing the best imaging method from a bewildering variety of microscopy-based techniques.
Download and read online Handbook of Single Molecule Biophysics in PDF and EPUB This handbook describes experimental techniques to monitor and manipulate individual biomolecules, including fluorescence detection, atomic force microscopy, and optical and magnetic trapping. It includes single-molecule studies of physical properties of biomolecules such as folding, polymer physics of protein and DNA, enzymology and biochemistry, single molecules in the membrane, and single-molecule techniques in living cells.
Download and read online Celebrating 50 Years of Live Cell Imaging in PDF and EPUB
Download and read online Cell Imaging in PDF and EPUB This book covers the key techniques that can be employed in any lab with access to cell imaging equipment, even if they do not currently specialize in imaging. It focuses on live cell imaging and light microscopy applications, but is equally relevant to the imaging of fixed specimens.
Download and read online Confocal Microscopy and Multiphoton Excitation Microscopy in PDF and EPUB This text guides you through the principles and practical techniques of confocal and multiphoton microscopy. It also describes the historical connections and parallel inventions that resulted in modern techniques of live cell imaging and their use in biology and medicine. You will find comparisons of different types of confocal and multiphoton microscopes, solutions to the problems one would encounter when using various microscopic techniques, tips on selecting equipment, and an extensive annotated bibliography of additional resources.
Download and read online Trafficking Dynamics of ERQC Proteins Through Live Cell Imaging in PDF and EPUB
Download and read online Quantitative Imaging in Cell Biology in PDF and EPUB This new volume, number 123, of Methods in Cell Biology looks at methods for quantitative imaging in cell biology. It covers both theoretical and practical aspects of using optical fluorescence microscopy and image analysis techniques for quantitative applications. The introductory chapters cover fundamental concepts and techniques important for obtaining accurate and precise quantitative data from imaging systems. These chapters address how choice of microscope, fluorophores, and digital detector impact the quality of quantitative data, and include step-by-step protocols for capturing and analyzing quantitative images. Common quantitative applications, including co-localization, ratiometric imaging, and counting molecules, are covered in detail. Practical chapters cover topics critical to getting the most out of your imaging system, from microscope maintenance to creating standardized samples for measuring resolution. Later chapters cover recent advances in quantitative imaging techniques, including super-resolution and light sheet microscopy. With cutting-edge material, this comprehensive collection is intended to guide researchers for years to come. Covers sections on model systems and functional studies, imaging-based approaches and emerging studies Chapters are written by experts in the field Cutting-edge material
Download and read online Microbial Imaging in PDF and EPUB Recent advances in molecular technology have provided new microbial imaging tools, not only complementing more classical methods, but in many cases significantly enhancing the sensitivity and efficiency in which studies may be conducted. These technologies are applicable to a wide range of problems in contemporary microbiology, including strain selection, understanding microbial structure, function and pathophysiology, as well as in the development of anti-microbial agents and vaccines. This volume emphasizes detailed methodology, provides a theoretical background and lists potential applications of specific imaging tools. * Edited by two experts in the field * Applicable to a broad Microbiology readership * Highly illustrated * Provides in-depth accounts from scientists working with cutting edge technologies * Facilitates researchers who involve Microbial Imaging in their work
Download and read online Handbook of Biological Confocal Microscopy in PDF and EPUB Once the second edition was safely off to the printer, the 110 larger world of micro-CT and micro-MRI and the smaller world authors breathed a sigh of relief and relaxed, secure in the belief revealed by the scanning and transmission electron microscopes. that they would “never have to do that again. ” That lasted for 10 To round out the story we even have a chapter on what PowerPoint years. When we ?nally awoke, it seemed that a lot had happened. does to the results, and the annotated bibliography has been In particular, people were trying to use the Handbook as a text- updated and extended. book even though it lacked the practical chapters needed. There As with the previous editions, the editor enjoyed a tremendous had been tremendous progress in lasers and ?ber-optics and in our amount of good will and cooperation from the 124 authors understanding of the mechanisms underlying photobleaching and involved. Both I, and the light microscopy community in general, phototoxicity. It was time for a new book. I contacted “the usual owe them all a great debt of gratitude. On a more personal note, I suspects” and almost all agreed as long as the deadline was still a would like to thank Kathy Lyons and her associates at Springer for year away.
Download and read online Analysis of FGF Receptor Signalling and Trafficking by Live cell Imaging in PDF and EPUB