Search results for: techniques-in-cell-cycle-analysis

Techniques in Cell Cycle Analysis

Author : Gray, J.W., ed
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Techniques in Cell Cycle Analysis

Author : Joe W. Gray
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Quantification of the proliferative characteristics of normal and malignant cells has been of interest to oncolo gists and cancer biologists for almost three decades. This interest stems from (a) the fact that cancer is a disease of uncontrolled proliferation, (b) the finding that many of the commonly used anticancer agents are preferentially toxic to cells that are actively proliferating, and (c) the observa tion that significant differences in proliferation characteristics exist between normal and malignant cells. Initially, cell cycle analysis was pursued enthusiastically in the hope of gener ating information useful for the development of rational cancer therapy strategies; for example, by allowing identi fication of rapidly proliferating tumors against which cell cycle-specific agents could be used with maximum effec tiveness and by allowing rational scheduling of cell cyc- specific therapeutic agents to maximize the therapeutic ratio. Unfortunately, several difficulties have prevented realiza tion of the early promise of cell cycle analysis: Proliferative patterns of the normal and malignant tissues have been found to be substantially more complex than originally an ticipated, and synchronization of human tumors has proved remarkably difficult. Human tumors of the same type have proved highly variable, and the cytokinetic tools available for cell cycle analysis have been labor intensive, as well as somewhat subjective and in many cases inapplicable to humans. However, the potential for substantially improved cancer therapy remains if more accurate cytokinetic infor mation about human malignancies and normal tissues can be obtained in a timely fashion.

Techniques in Cell Cycle Analysis

Author : Joe W. Gray
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Cell Cycle Materials and Methods

Author : Michele Pagano
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During their lifetime, especially when growing and dividing, cells go through various steps of the cell cycle. Knowledge of the individual steps of the cell cycle will help us understand the development of a variety of diseases better, including cancer, and also to design new drugs against it. New techniques for studying the molecular basis of these processes have recently been developed and are described in detail in this manual. A glossary helps the reader to cope with the complex cell cycle terminology.

Techniques for Cytogenetic Investigation of the Malignant Cells and Cell Cycle Analysis in B cell Chronic Lymphocytic Leukaemia

Author : Christine Felicidad Stephenson
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Techniques for Cytogenetic Investigation of Malignant Cell Cycle Analysis in B cell Chronic Lymphocytic Leukaemia

Author : Christine Felicidad Stephenson
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Flow Cytometry

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Flow Cytometry, Second Edition provides a complete and comprehensive two volume laboratory guide and reference for the use of the most current methods in flow cytometry sample preparation and analysis. These essential techniques are described in a step-by-step format, supplemented by explanatory sections and trouble-shooting tips. The methods are accessible to all researchers and students in biomedical science and biology who must use flow cytometry to separate and analyze cells. Key Features * Completely revised and greatly expanded since the publication of the First Edition in 1990 * Methods cover cell death and cell cycle analyses Practical, handbook-style presentation works in lab or classroom * Unique comprehensive methodological coverage * Color plates illustrate techniques * In-depth treatment of procedures, including a description of each procedure: * Theoretical foundations * Critical aspects * Possible pitfalls * Written by authors with extensive experience who: * Developed or modified the techniques * Describe their experience with different instruments and applications to different cell systems * Are the Who's Who in Flow Cytometry

Cell Cycle Control and Dysregulation Protocols

Author : Antonio Giordano
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This volume focuces on emerging methodologies for studying the cell cycle, kinases and kinases inhibitors. Important issues are addressed including gene expression in vivo and in vitro, the analysis of cylin-dependent kinase inhibitors, protein degradation mediated by the proteosome, the analysis of the transformed cell phenotype, and innovative techniques to detect apoptosis. these highly sophisticated techniques for in vivo and in vitro studies comprise molecular biology, biochemistry, and various types of immunoassays. Describes protocols based on sophisticated techniques for in vivo and in vitro studies, including molecular biology, biochemistry, and immunoassays.

Cell Cycle Checkpoint Control Protocols

Author : Howard B. Lieberman
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A collection of basic cutting-edge techniques for studying the mechanisms underlying cell cycle regulation and checkpoint control. Using mammalian, yeast, and frog systems, these readily reproducible methods can be used to induce cell cycle checkpoints, detect changes in cell cycle progression, identify and analyze genes and proteins that regulate the process, and characterize chromosomal status as a function of cell cycle phase and progression. Each fully tested technique includes step-by-step instructions written by an investigator who routinely performs it, an introduction explaining the principle behind the method, equipment and reagent lists, and tips on troubleshooting and avoiding known pitfalls.

Handbook of Plant Ecophysiology Techniques

Author : M. J. Reigosa Roger
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The Handbook of Plant Ecophysiology Techniques you have now in your hands is the result of several combined events and efforts. The birth of this handbook can be traced as far as 1997, when our Plant Ecophysiology lab at the University of Vigo hosted a practical course on Plant Ecophysiology Techniques. That course showed us how much useful a handbook presenting a bunch of techniques would be for the scientists beginning to work on Plant Ecophysiology. In fact, we wrote a short handbook explaining the basics of the techniques taught in that 1997 course: Flow cytometry to measure ploidy levels, Use of a Steady-State porometer to measure transpiration, In vivo measure of fluorescence, HPLC analysis of low molecular weight phenolics, Spectrophotometric determinations of free proline and soluble proteins, TLC polyamines contents measures, Isoenzymatic electrophoresis, Use of IRGA and oxygen electrode. That modest handbook, written in Spanish, was very helpful, both for the people who attended the course and for other who have used it for beginning to work in Plant Ecophysiology. The present Handbook is much more ambitious, and it includes more techniques. But we have also had in mind the young scientists beginning to work on Plant Ecophysiology. In 1999 François Pellissier leaded a proposal presented to the European Commission in the Fifth Framework Program in the High Level * Scientific Conferences, including three EuroLab Courses about lab and field techniques useful to improve allelopathic research.

Hydroxyurea

Author : David H. Worthington
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Hydroxyurea (10mM) blocks the exponential growth of Tetrahymena pyriformis (GL-I) populations by arresting progress through the cell cycle once the cells enter S-phase. Autoradiographic analysis reveals that HU reduces the uptake of tritiated thymidine (3H-TdR) into macronuclear DNA in S-phase cells to about 1/5 of that in untreated S-phase cells. These slowly synthesizing cells do not divide (even after 10 days) until the HU is removed. The HU-blocked cells are also rescued from S-phase arrest by adding 0. 5mM each of all four DNAprecursor deoxyribonucleosides (XdRs) thus indicating the primary effect of HU is the inhibition of the reduction of ribonucleosides. When the HU-block is removed by either washing the HU from the medium or by rescue with XdRs an excessive delay in recovering the capacity to divide is specifically induced in cells which were in S-phase at time of addition of HU. This delay may be due to the time needed to repair damage to the DNA which was induced specifically during S-phase. Fluoromicroscopic examination of euchrysine-stained cells exposed for a minimum of two hours starting in S-phase show a radical morphological change in the macronucleus. This phenomenon, termed the "halo effect," is characterized by the formation of apparently membrane-associated chromatin aggregates surrounding a constricted chromatin mass. Halo-induction by HU is S-phase specific and upon removal of the HU-block the halo remains until the first recovery division. The halo effect is interpreted as being a morphological indication of the cell's repair response to the damaging effects of HU upon macronuclear DNA. Observations of division of individual cells in microdrops, plus autoradiographic studies using 3H-TdR and standard cell cycle analysis techniques, reveal that HU blocks all cells in the initial 92% of S-phase but does not affect cells in the remaining 8 % of S-phase or in G2 and division. Thus the fraction of the population of cells that is in G2 can be approximately determined (within 6 minutes) by the fraction of the population able to divide in the presence of HU. This fraction can be related to the approximate duration of G2 after mathematically compensating for the age gradient. Thus G2 analysis of T. pyriformis (GL-I) is accomplished easily, quickly, and inexpensively with the use of HU. The possibilities of utilizing Sphase specific recovery delay and halo induction for simple means of determining the durations of G1 and S-phase respectively are also discussed. The addition of all four XdRs (0. 5mM) to exponentially growing cells in rich organic complete medium without HU results in the generation time of these cells being decreased by 20%. Cell cycle analysis with phosphorus-33 (33P) and experiments involving cell phase sensitivity to the presence of excess XdRs show that the excess XdRs are taken up and incorporated or pooled entirely during the initial 50% of S-phase. This acceleration of early S-phase completely accounts for the abbreviated generation time seen under these conditions. When HU is removed after 10 hours of treatment the cells undergo a period of recovery characterized by an increase in the duration of DNA synthesis and in the amount of DNA synthesized before they divide and enter a cell cycle abbreviated at the expense of G1. These results are related to a model for control of the initiation of DNA replication and cell division based upon the ratio of macronuclear DNA content to cytoplasmic mass.

Guide to Flow Cytometry Methods

Author : W. McLean Grogan
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Discusses the methodology and procedures used in studies of the cell cycle, cell development and differentiation, ageing, immunology, membrane fluidity, and aneuploidy analysis of the 15 most common forms of cancer. Described techniques of analysis include preparation of single-cell suspensions, DNA

Flow Cytometry Protocols

Author : Mark J. Jaroszeski
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Features researchers and experimentalists who update the field of flow cytometry with detailed accounts of their improved, and well-tested protocols and applications. This title includes techniques that take advantage of the instrumentation and probes that have been developed to advance the field of flow cytometry and improve standard methods.

Cell Cycle Oscillators

Author : Amanda S. Coutts
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​ This volume brings together a unique collection of protocols that cover standard, novel, and specialized techniques. Cell Cycle Oscillators: Methods and Protocols guides readers through recent progress in the field from both holistic and reductionist perspectives, providing the latest developments in molecular biology techniques, biochemistry, and computational analysis used for studying oscillatory networks. 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, Cell Cycle Oscillators: Methods and Protocols will serve as an invaluable reference to gain further insight into the complex and incompletely understood processes that are involved in the cell cycle and its regulation by oscillatory networks.

Flow Cytometry Applications in Cell Culture

Author : Mohamed Al-Rubeai
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This work present practical, biotechnological applications of flow cytometry techniques for the study of animal, plant and microbial cells, explaining methodologies for sample preparation, staining and analysis. It discusses cell variability in cell culture processes and shows how the quantitative analysis of heterogeneous populations aids in the biotechnological exploitation of cells.

Cell Biology

Author : Julio E. Celis
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V. 1: cell and tissue culture and associated techniques; Primary cultures from embyonic and newborn tissues; Culture of specific cell types; Cell separation techniques; Model systems to study differentiation; cell cycle analysis; Assays of tumorigenicity, invasion, and others; Cytotoxic and cell growth assays; Senescence and apoptosis; Electrophysiological methods; Histocultures and organ cultures; Other cell types and organisms; Viruses; Appendices; v. 2: Organelles and cellular structures; Assays; Antibodies; Immunocytochemistry; Vital staining of cells; v. 3: Light microscopy and contrast generation; Electron microscopy; Intracellular measurments; Cytogenetics and in situ hybridization; transgenic and gene knockouts; v. 4: Transfer of macromelcules and small molecules; Expression systems; Differential gene expression; Proteins; Appendix; List of suppliers; Subject index.

Mitosis and Meiosis

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Mitosis and Meiosis details the wide variety of methods currently used to study how cells divide as yeast and insect spermatocytes, higher plants, and sea urchin zygotes. With chapters covering micromanipulation of chromosomes and making, expressing, and imaging GFP-fusion proteins, this volume contains state-of-the-art "how to" secrets that allow researchers to obtain novel information on the biology of centrosomes and kinetochores and how these organelles interact to form the spindle. Chapters Contain Information On: * How to generate, screen, and study mutants of mitosis in yeast, fungi, and flies * Techniques to best image fluorescent and nonfluorescent tagged dividing cells * The use and action of mitoclastic drugs * How to generate antibodies to mitotic components and inject them into cells * Methods that can also be used to obtain information on cellular processes in nondividing cells

Methods in Cell Biology

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Critically acclaimed for more than 25 years, the Methods in Cell Biology series provides an indispensable tool for the researcher. Each volume is carefully edited by experts to contain state-of-the-art reviews and step-by-step protocols. Techniques are described completely so that methods are made accessible to users.

Flow Cytometry with Plant Cells

Author : Jaroslav Dolezel
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Targeted at beginners as well as experienced users, this handy reference explains the benefits and uses of flow cytometery in the study of plants and their genomes. Following a brief introduction that highlights general considerations when analyzing plant cells by flow cytometric methods, the book goes on to discuss examples of application in plant genetics, genomic analysis, cell cycle analysis, marine organism analysis and breeding studies. With its list of general reading and a glossary of terms, this first reference on FCM in plants fills a real gap by providing first-hand practical hints for the growing community of plant geneticists.

Journal of Cell Science

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