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 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 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 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 Live Cell Imaging Technology Development for Cancer Research in PDF and EPUB The results show that the timing of depolyploidization in MC cells correlates with increased multipolar divisions, up-regulation of meiosis-specific genes, and the production of mononucleated cell progeny. It was confirmed that mononucleated cells are produced from multipolar divisions and these cells are capable of resuming normal divisions [Ianzini et al., 2009]. The implications for the induction of meiosis as a mechanism of survival after radiation treatment are discussed. In the second project, the effects of long-term fluorescence excitation light exposure are examined through measurements of cell division and cell death. In the field of live cell imaging, probably the most modern and most widely utilized technique is fluorescence detection for intracellular organelles, proteins, and molecules. While the technologies required to label and detect fluorescent molecules in a cell are well developed, they are not idealized for long term measurements as both the probes and excitation light are toxic to the cells [Wang and Nixon, 1978; Bradley and Sharkey, 1977]. From the event analysis data it was determined that fluorescence excitation light is toxic to multiple cell lines observed as the reduction of normal cell division, induction of cell death, and apparent morphological aberrations.
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 Celebrating 50 Years of Live Cell Imaging in PDF and EPUB
Download and read online Live cell imaging of transcription activation of drosophila heat shock genes and characterization of water soluble quantum dots in PDF and EPUB
Download and read online International Review of Cell and Molecular Biology in PDF and EPUB International Review of Cell & Molecular Biology presents current advances and comprehensive reviews in cell biology—both plant and animal. Articles address structure and control of gene expression, nucleocytoplasmic interactions, control of cell development and differentiation, and cell transformation and growth. * Authored by some of the foremost scientists in the field * Provides up-to-date information and directions for future research * Valuable reference material for advanced undergraduates, graduate students and professional scientists
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 Neurons Methods and Applications for the Cell Biologist in PDF and EPUB Neurons: Methods and Applications for the Cell Biologist lays out numerous simple techniques for growing and carrying out experiments with many varieties of neurons. Subjects include peripheral and central neurons from vertebrate and invertebrate sources, as well as neuron-like cell lines. It also explains recent advances in our ability to introduce exogenous proteins and genes to neurons in culture. Procedures for successful protein infiltration, biolistic transfection, electroporation, and viral transgenic methods in neurons are also presented. Contains culture methodology for more than a dozen types of CNS and PNS neurons Includes most recent and reliable techniques from expert practitioners for specific experimental applications Addresses the latest strategies for transfecting neurons
Download and read online Aptamer Sensors for Live cell Imaging of Pol II Promoter Activity in PDF and EPUB
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 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