Cellebiologi (cytologi)

Plant microtubules are key elements of cell growth, division and morphogenesis. In addition to their role in plant development and architecture, they have emerged as regulatory elements of signalling and important targets of evolution.Since the publication of the first edition of Plant Microtubules in 2000, our understanding of microtubules and their manifold functions have advanced substantially. Consisting of the following three parts, this book highlights the morphogenetic potential of plant microtubules from three general viewpoints: Microtubules and Morphogenesis: control of cell axis during division and expansion, cross-talk with actin filaments, mechanical properties of the cell wall. Microtubules and Environment: the role of microtubules during the sensing or response of environmental factors such as pathogens or abiotic stresses. Microtubules and Evolution: complexity and specialization of plant microtubules in the context of plant evolution. The book is an invaluable source of information for researchers as well as for graduate and advanced students.

Mammalian and Avian Transgenesis presents a collection of novel methods for the production of a wide range of transgenic animals. The manual focuses largely on mice, but also contains protocols for successful transgenesis in rats, cows, pigs and birds. The manual provides detailed, step-by-step protocols covering all aspects of the production of transgenic animals, including the use of lentiviral vectors in gene transfer, intracytoplasmic sperm injection, nuclear transfer, large insert transgenesis, conditional gene expression systems, the use of reporter genes in transgenesis and transgenesis in large animals and birds. The text is supplemented by superb color photos. While the focus is on newly established techniques, the fundamental methods of transgenesis are also covered for those new to the field. Thus this manual is perfectly suited for those wishing to adopt new technologies in transgenesis.

This new and completely revised edition brings several new elements to the reader. Each chapter begins with aseries of Major Study Topics and con- cludes with some Questions for Reviewand Discussion. I also have added a glossary to assist students with unfamiliar terms. This edition offers a greater emphasis on molecular biology and genetics than was present in either of the previous editions. The sequence of topics has also changed so that basic regulation and recombination are introduced early to provide a basis for subsequent discussion. I have preserved the pre- sentation of basic virology and genetic transfer processes while expanding coverage of plasmid molecular biology. All of what I would consider to be essential material occurs within the first 13chapters. The final four chapters are shorter, optional material and are not interdependent; they can be used in any order or omitted at an instructor's discretion. It is a pleasure to acknowledge the able assistance of the editorial and production staff of Springer-Verlag. I am also grateful to my colleagues who were so patient with my questions and to Rene Allard for his helpful com- ments on the first six chapters.

The human body contains many specialized tissues that are capable of fulfilling an incredible variety of functions necessary for our survival. This volume in the Human Cell Culture Series focuses on mesenchymal tissues and cells. The in vitro study of mesenchymal cells is perhaps the oldest form of human cell culture, beginning with the culturing of fibroblasts. Fibroblasts have long been generically described in the literature, arising from many tissue types upon in vitro cell culture. However, recent studies, many enabled by new molecular biology techniques, have shown considerable diversity in fibroblast type and function, as described within this volume. Mesenchymal tissue types that are described within include bone, cartilage, tendons and ligaments, muscle, adipose tissue, and skin (dermis). The proper function of these tissues is predominantly dependent upon the proper proliferation, differentiation, and function of the mesenchymal cells which make up the tissue. Recent advancements in primary human mesenchymal cell culture have led to remarkable progress in the study of these tissues. Landmark experiments have now demonstrated a stem cell basis for many of these tissues, and, furthermore, significant plasticity and inter-conversion of stem cells between these tissues, resulting in a great deal of contemporary excitement and controversy. Newly-developed mesenchymal cell culture techniques have even lead to novel clinical practices for the treatment of disease.

In the early 1980s, a few scientists started working on a Xenopus transcription factor, TFIIIA. They soon discovered a novel domain associated with zinc, and named this domain "e;zinc finger. "e; Th e number of proteins with similar zinc fingers grew quickly and these proteins are now called C2H2, Cys2His2 or classical zinc finger proteins. To date, about 24,000 C2H2 zinc finger proteins have been recognized. Approximately 700 human genes, or more than 2% of the genome, have been estimated to encode C2H2 finger proteins. From the beginning these proteins were thought to be numerous, but no one could have predicted such a huge number. Perhaps thousands of scientists are now working on C2H2 zinc finger proteins fi-om variou s viewpoints. This field is a good example of how a new science begins with the insight of a few scientists and how it develops by efforts of numerous independent scientists, in contrast to a policy-driven scientific project, such as the Human Genome Project, with goals clearly set at its inception and with work performed by a huge collaboration throughout the world. As more zinc finger proteins were discovered, several subfamilies, such as C2C2, CCHC, CCCH, LIM, RING, TAZ, and FYVE emerged, increasing our understanding of zinc fingers. The knowledge was overwhelming. Moreover, scientists began defining the term "e;zinc finger"e; differently and using various names for identical zinc fingers. These complications may explain why no single comprehensive resource of zinc finger proteins was available before this publication.

Cell culture is cell cloning technology that simulates in vivo environment conditions such as asepsis, appropriate temperature, and pH as well as certain nutritional conditions to enable cells to survive, grow, reproduce, and maintain their structure and function. Cell culture can be used to grow human, animal, plant, and microbial cells. Each type of cell culture has its own characteristics and essential conditions. This book focuses on the advanced technology and applications of cell culture in the research and practice of medical and life sciences. Chapters address such topics as primary cancer cell cultures, 2D and 3D cell cultures, stem cells, nanotechnology, and more.

This new edition explores lab protocols describing new techniques to study cytotoxic T-cells (CTLs), as well as chapters of a more general discursive nature, all with an emphasis on the use of systems biology in immunology. Beginning with phenotypical characterization of CTL populations, the volume continues with in vitro and in vivo cytotoxicity assays, methods to detect senescent T cells, in vivo and in vitro models to understand immune and bone cells cross-talk, microscopy and in vivo imaging, as well as ¿Omics¿ approaches and molecular methods, concluding with chapters on CTL involvement in transplantation and link microbiota-immunity. Written for the highly successful Methods in Molecular Biology series, chapters feature the kind of detail and key implementation advice for best results in the lab. Authoritative and up-to-date, Cytotoxic T-Cells: Methods and Protocols, Second Edition serves as an ideal guidefor researchers working with these vital cells.

This detailed book highlights recent advances in molecular imaging techniques and protocols, designed to be immediately applicable in global bio-laboratories. The chapters are categorized into seven major groups according to the reporter materials, such as imaging with passive optical readouts, activatable bioluminescent probes, functional substrates and luciferases, organic fluorescent probes, BRET probes, FRET probes, as well as with advanced instrumentation. 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, Live Cell Imaging: Methods and Protocols aims to direct and inspire researchers into creating smarter, next-generation imaging techniques that are truly quantitative, highly sensitive, and readily comprehended, in the effort to engender deeper understanding of biological systems and break new ground in the research fields of life science.

Transcriptome Profiling: Progress and Prospects assists readers in assessing and interpreting a large number of genes, up to and including an entire genome. It provides key insights into the latest tools and techniques used in transcriptomics and its relevant topics which can reveal a global snapshot of the complete RNA component of a cell at a given time. This snapshot, in turn, enables the distinction between different cell types, different disease states, and different time points during development. Transcriptome analysis has been a key area of biological inquiry for decades. The next-generation sequencing technologies have revolutionized transcriptomics by providing opportunities for multidimensional examinations of cellular transcriptomes in which high-throughput expression data are obtained at a single-base resolution. Transcriptome analysis has evolved from the detection of single RNA molecules to large-scale gene expression profiling and genome annotation initiatives. Written by a team of global experts, key topics in Transcriptome Profiling include transcriptome characterization, expression analysis of transcripts, transcriptome and gene regulation, transcriptome profiling and human health, medicinal plants transcriptomics, transcriptomics and genetic engineering, transcriptomics in agriculture, and phylotranscriptomics.

This volume provides a wide range of imaging protocols that can be tailored to specific organisms or cell-types. Chapters guide readers through fixed-cell, live-cell, phenotype screening, super-resolution, intravital imaging techniques, and fluorescence life-time imaging microscopy (FLIM). 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, Confocal Microscopy: Methods and Protocols aims to ensure successful results in the further study of this vital field.

This detailed book explores the fundamentals of myofibroblast biology in tissue repair, fibrosis, and tumors as well as providing cutting-edge laboratory methods used to investigate myofibroblast functions in physiological and pathological settings in vitro and in vivo, as written by leading academic scientists. Section I of this volume focuses on fundamental methods to study myofibroblast biology and covers topics such as methods for detecting myofibroblasts and senescent myofibroblasts in cell culture and histology, single cell RNA sequencing to identify myofibroblast subsets in fibrotic tissues, and functional assays to assess TGF-¿ activation, myofibroblast apoptosis, or matrix deposition and crosslinking. Section II discusses methods to investigate the mechanobiology of myofibroblasts in vitro, including the fabrication of functional hydrogels with tunable stiffness, the use of atomic force microscopy to characterize matrix and cellular stiffness, as well as molecular assays to assess fibroblast mechanotransduction pathways and durotaxis. Section III describes multiple animal models to investigate myofibroblast functions across organs in vivo as well as human organoid systems, precision tissue slices and decellularized 3D tissue scaffolds to assess myofibroblast functions in relevant human ex vivo models. 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 extensive, Myofibroblasts: Methods and Protocols is an essential collection for researchers delving into the processes and effects of these important cells.

This first book on high-speed atomic force microscopy (HS-AFM) is intended for students and biologists who want to use HS-AFM in their research. It provides straightforward explanations of the principle and techniques of AFM and HS-AFM. Numerous examples of HS-AFM studies on proteins demonstrate how to apply this new form of microscopy to specific biological problems. Several precautions for successful imaging and the preparation of cantilever tips and substrate surfaces will greatly benefit first-time users of HS-AFM. In turn, the instrumentation techniques detailed in Chapter 4 can be skipped, but will be useful for engineers and scientists who want to develop the next generation of high-speed scanning probe microscopes for biology.The book is intended to facilitate the first-time use of this new technique, and to inspire students and researchers to tackle their own specific biological problems by directly observing dynamic events occurring in the nanoscopic world. Microscopy in biology has recently entered a new era with the advent of high-speed atomic force microscopy (HS-AFM). Unlike optical microscopy, electron microscopy, and conventional slow AFM, it allows us to directly observe biological molecules in physiological environments. Molecular "e;movies"e; created using HS-AFM can directly reveal how molecules behave and operate, without the need for subsequent complex analyses and roundabout interpretations. It also allows us to directly monitor morphological change in live cells, and dynamic molecular events occurring on the surfaces of living bacteria and intracellular organelles. As HS-AFM instruments were recently commercialized, in the near future HS-AFM is expected to become a common tool in biology, and will enhance and accelerate our understanding of biological phenomena.