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In Fluorescent Protein-Based Biosensors: Methods and Protocols, experts in the field have assembled a series of protocols describing several methods in which fluorescent protein-based reporters can be used to gain unique insights into the regulation of cellular signal transduction. Genetically encodable fluorescent biosensors have allowed researchers to observe biochemical processes within the endogenous cellular environment with unprecedented spatiotemporal resolution. As the number and diversity of available biosensors grows, it is increasingly important to equip researchers with an understanding of the key concepts underlying the design and application of genetically encodable fluorescent biosensors to live cell imaging. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Fluorescent Protein-Based Biosensors: Methods and Protocols promises to be a valuable resource for researchers interested in applying current biosensors to the study of biochemical processes in living cells as well as those interested in developing novel biosensors to visualize other cellular phenomena.
Cellular Senescence and Aging, Volume 181 in the Methods in Cell Biology series, highlights new advances in the field, with this new volume presenting interesting chapters on topics such as assessing polyglutamine tract aggregation in the nematode Caenorhabditis elegans, Generation of glial cell-derived neurotrophic factor (gdnf) morphants in zebrafish larvae by cerebroventricular microinjection of vivo morpholino, Methods for detection of mitochondrial reactive oxygen species in senescent cells, Assessment of cell cycle progression and mitotic slippage by videomicroscopy, The original colorimetric method to detect cellular senescence, and more.Additional sections cover Assessing microbiota composition in the context of aging, Assessing chronological aging in Saccharomyces cerevisiae, Image processing and supervised machine learning for retinal microglia characterization in senescence, Measuring telomerase activity using TRAP assays, High throughput assessment of cellular senescence, Detection of radiation-induced senescence by the Debacq-Chainiaux protocol: Improvements and upgrade in the detection of positive events, Dynamic and scalable assessment of the senescence-associated secretory phenotype (SASP), Flow cytometry-assisted quantification of cell cycle arrest in cancer cells treated with CDK4/6 inhibitors, and more. - Provides the authority and expertise of leading contributors from an international board of authors - Presents the latest release in the Methods in Cell Biology series - Updated release includes the latest information on Cellular Senescence and Aging
Monitoring brain function with light in vivo has become a reality. The technology 33 of detecting and interpreting patterns of reflected light has reached a degree of 34 maturity that now permits high spatial and temporal resolution visualization at both 35 the systems and cellular levels. There now exist several optical imaging methodolo- 36 gies, based on either hemodynamic changes in nervous tissue or neurally induced 37 light scattering changes, that can be used to measure ongoing activity in the brain. 38 These include the techniques of intrinsic signal optical imaging, near-infrared optical 39 imaging, fast optical imaging based on scattered light, optical imaging with voltage 40 sensitive dyes, and two-photon imaging of hemodynamic signals. The purpose of 41 this volume is to capture some of the latest applications of these methodologies to 42 the study of cerebral cortical function. 43 This volume begins with an overview and history of optical imaging and its use 44 in the study of brain function. Several chapters are devoted to the method of intrin- 45 sic signal optical imaging, a method used to record the minute changes in optical 46 absorption due to hemodynamic changes that accompanies cortical activity. Since the 47 detected hemodynamic changes are highly localized, this method has excellent 48 spatial resolution (50–100 μm ), a resolution sufficient for visualization of fundamen- 49 tal modules of cerebral cortical function.
This open access book describes marked advances in imaging technology that have enabled the visualization of phenomena in ways formerly believed to be completelyimpossible. These technologies have made major contributions to the elucidation of the pathology of diseases as well as to their diagnosis and therapy. The volume presents various studies from molecular imaging to clinical imaging. It also focuses on innovative, creative, advanced research that gives full play to imaging technology inthe broad sense, while exploring cross-disciplinary areas in which individual research fields interact and pursuing the development of new techniques where they fuse together. The book is separated into three parts, the first of which addresses the topic of visualizing and controlling molecules for life. Th e second part is devoted to imaging of disease mechanisms, while the final part comprises studies on the application of imaging technologies to diagnosis and therapy. Th e book contains the proceedings of the 12th Uehara International Symposium 2017, “Make Life Visible” sponsored by the Uehara Memorial Foundation and held from June 12 to 14, 2017. It is written by leading scientists in the field and is an open access publication under a CC BY 4.0 license.
From the Lab to Clinical Settings-Advances in Quantitative, Noninvasive Optical DiagnosticsNoninvasive fluorescence imaging techniques, novel fluorescent labels, and natural biomarkers are revolutionizing our knowledge of cellular processes, signaling and metabolic pathways, the underlying mechanisms for health problems, and the identification of n
New Methods and Sensors for Membrane and Cell Volume Research, Volume 88 provides an overview of novel experimental approaches to study both the cell membrane and the under-membrane space – the cytosol, which have lately began drawing renewed attention. The book's overall emphasis is on fluorescent and FRET-based sensors, however, other optical (such as variants of transmission microscopy) and non-optical methods (neutron scattering and mass spectrometry) also have dedicated chapters. This volume provides a rare review of experimental approaches to study intracellular phase transitions, as well as anion channels, membrane tension and dynamics, and other topics of intense current interest. - Describes novel FRET-based membrane sensors - Reviews selected non-optical approaches to membrane structure and dynamics - Describes traditional and modern aspects of cell volume research, such as phase transitions and macromolecular crowding
Fluorescent proteins are intimately connected to research in the life sciences. Tagging of gene products with fluorescent proteins has revolutionized all areas of biosciences, ranging from fundamental biochemistry to clinical oncology, to environmental research. The discovery of the Green Fluorescent Protein, its first, seminal application and the ingenious development of a broad palette of fluorescence proteins of other colours, was consequently recognised with the Nobel Prize for Chemistry in 2008. Fluorescent Proteins II highlights the physicochemical and biophysical aspects of fluorescent protein technology beyond imaging. It is tailored to meet the needs of physicists, chemists and biologists who are interested in the fundamental properties of fluorescent proteins, while also focussing on specific applications. The implementations described are cutting-edge studies and exemplify how the physical and chemical properties of fluorescent proteins can stimulate novel findings in life sciences.
This book provides an update for the rapidly developing technology known as “optogenetics”, which is the use of genetically encoded light-sensitive molecular elements (usually derived from lower organisms) to control or report various physiological and biochemical processes within the cell. Two ongoing clinical trials use optogenetic tools for vision restoration, and optogenetic strategies have been suggested as novel therapies for several neurological, psychiatric and cardiac disorders. This Special Issue comprises two reviews and seven experimental papers on different types of light-sensitive modules widely used in optogenetic studies. These papers demonstrate the efficiency and versatility of optogenetics and are expected to be equally relevant for advanced users and beginners considering using optogenetic tools in their research.