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DNA Damage Detection In Situ, Ex Vivo, and In Vivo

Methods and Protocols

  • Book
  • © 2011

Overview

Editors:
  1. Vladimir V. Didenko

    1. Baylor College of Medicine, Depts. Neurosurgery and Molecular & Cell, University of Houston, Houston, USA

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  • The first publication of its kind combines in situ, ex vivo and in vivo techniques
  • Comprehensive evaluation of various types of DNA damage
  • Detection in live tissues, blood, urine, sperm, cell cultures and tissue sections
  • Detection range from subcellular to the level of a whole live organism
  • Methods for noninvasive detection and monitoring of DNA damage
  • Includes supplementary material: sn.pub/extras

Part of the book series: Methods in Molecular Biology (MIMB, volume 682)

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Table of contents (22 protocols)

  1. Front Matter

    Pages i-xiii
    Download chapter PDF

  2. Detection in Tissue Sections

    1. Front Matter

      Pages 1-1
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    2. In Situ Detection of Apoptosis by the TUNEL Assay: An Overview of Techniques

      • Deryk T. Loo
      Pages 3-13

    3. Combination of TUNEL Assay with Immunohistochemistry for Simultaneous Detection of DNA Fragmentation and Oxidative Cell Damage

      • Alexander E. Kalyuzhny
      Pages 15-27

    4. EM-ISEL: A Useful Tool to Visualize DNA Damage at the Ultrastructural Level

      • Antonio Migheli
      Pages 29-36

    5. In Situ Labeling of DNA Breaks and Apoptosis by T7 DNA Polymerase

      • Vladimir V. Didenko
      Pages 37-48

    6. In Situ Ligation: A Decade and a Half of Experience

      • Peter J. Hornsby, Vladimir V. Didenko
      Pages 49-63

    7. In Situ Ligation Simplified: Using PCR Fragments for Detection of Double-Strand DNA Breaks in Tissue Sections

      • Vladimir V. Didenko
      Pages 65-75

    8. 5′OH DNA Breaks in Apoptosis and Their Labeling by Topoisomerase-Based Approach

      • Vladimir V. Didenko
      Pages 77-87

  3. Detection in Cell Cultures

    1. Front Matter

      Pages 89-89
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    2. Detection of DNA Strand Breaks in Apoptotic Cells by Flow- and Image-Cytometry

      • Zbigniew Darzynkiewicz, Hong Zhao
      Pages 91-101

    3. Fluorochrome-Labeled Inhibitors of Caspases: Convenient In Vitro and In Vivo Markers of Apoptotic Cells for Cytometric Analysis

      • Zbigniew Darzynkiewicz, Piotr Pozarowski, Brian W. Lee, Gary L. Johnson
      Pages 103-114

    4. Combining Fluorescent In Situ Hybridization with the Comet Assay for Targeted Examination of DNA Damage and Repair

      • Sergey Shaposhnikov, Preben D. Thomsen, Andrew R. Collins
      Pages 115-132

    5. Simultaneous Labeling of Single- and Double-Strand DNA Breaks by DNA Breakage Detection-FISH (DBD-FISH)

      • José Luis Fernández, Dioleyda Cajigal, Jaime Gosálvez
      Pages 133-147

    6. Co-localization of DNA Repair Proteins with UV-Induced DNA Damage in Locally Irradiated Cells

      • Jennifer Guerrero-Santoro, Arthur S. Levine, Vesna Rapić-Otrin
      Pages 149-161

  4. Detection in Live Tissues, Blood, Urine, Sperm

    1. Front Matter

      Pages 163-163
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    2. Ultrasound Imaging of Apoptosis: Spectroscopic Detection of DNA-Damage Effects at High and Low Frequencies

      • Roxana M. Vlad, Michael C. Kolios, Gregory J. Czarnota
      Pages 165-187

    3. Quantifying Etheno–DNA Adducts in Human Tissues, White Blood Cells, and Urine by Ultrasensitive 32P-Postlabeling and Immunohistochemistry

      • Jagadeesan Nair, Urmila J. Nair, Xin Sun, Ying Wang, Khelifa Arab, Helmut Bartsch
      Pages 189-205

    4. ELISpot Assay as a Tool to Study Oxidative Stress in Peripheral Blood Mononuclear Cells

      • Jodi Hagen, Jeffrey P. Houchins, Alexander E. Kalyuzhny
      Pages 207-215

    5. Cytokinesis-Block Micronucleus Cytome Assay in Lymphocytes

      • Philip Thomas, Michael Fenech
      Pages 217-234
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About this book

Recent advances in organic chemistry, fluorescent microscopy, and materials science have created an entirely new range of techniques and probes for imaging DNA damage in molecular and cellular biology. In DNA Damage Detection In Situ, Ex Vivo, and In Vivo: Methods and Protocols, expert researchers explore the latest advances in the area, covering both recent and established techniques to detect and quantify DNA damage at scales ranging from subcellular to the level of a whole live organism. Chapters present all major assays used in molecular and cellular biology for the labeling of DNA damage in situ, ex vivo, and in vivo. Composed in the highly successful Methods in Molecular Biology™ series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls. Comprehensive and current, DNA Damage Detection In Situ, Ex Vivo, and In Vivo: Methods and Protocols is an essential handbook for novice and experienced researchers in a variety of fields, including molecular and cellular biology, experimental and clinical pathology, toxicology, radiobiology, oncology, embryology, experimental pharmacology, drug design, and environmental science.

Reviews

From the reviews:

“This volume is presenting the major essays used nowadays in cell and molecular biology laboratories to detect in situ, ex vivo and in vivo DNA damages. Enjoyable and interesting enough is the long explanation devoted by Prof. Didenko … to the use of the terms in vivo, ex vivo and in situ … . Well done! … . Those of us interested in whatsoever way to DNA damage studies will surely find this book very useful.” (Carlo Alberto Redi, European Journal of Histochemistry, Vol. 55, 2011)

Editors and Affiliations

  • Baylor College of Medicine, Depts. Neurosurgery and Molecular & Cell, University of Houston, Houston, USA

    Vladimir V. Didenko

Bibliographic Information

  • Book Title: DNA Damage Detection In Situ, Ex Vivo, and In Vivo

  • Book Subtitle: Methods and Protocols

  • Editors: Vladimir V. Didenko

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-60327-409-8

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC 2011

  • Hardcover ISBN: 978-1-60327-408-1Published: 08 November 2010

  • Softcover ISBN: 978-1-4939-6198-6Published: 23 August 2016

  • eBook ISBN: 978-1-60327-409-8Published: 08 November 2010

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XIII, 313

  • Topics: Human Genetics, Gene Function

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