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In Situ Detection of DNA Damage

Methods and Protocols

  • Book
  • © 2002

Overview

Editors:
  1. Vladimir V. Didenko

    1. Baylor College of Medicine, Houston

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Part of the book series: Methods in Molecular Biology (MIMB, volume 203)

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Table of contents (22 protocols)

  1. Front Matter

    Pages i-xvii
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  2. Labeling DNA Breaks Using Terminal Transferase (TUNEL Assay)

    1. Front Matter

      Pages 1-1
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    2. Labeling DNA Damage with Terminal Transferase

      • P. Roy Walker, Christine Carson, Julie Leblanc, Marianna Sikorska
      Pages 3-19

    3. TUNEL Assay

      • Deryk T. Loo
      Pages 21-30

    4. Electron Microscopic Detection of DNA Damage Labeled by TUNEL

      • Antonio Migheli
      Pages 31-39

    5. Quantitative Differentiation of Both Free 3′ OH and 5′ OH DNA Ends Using Terminal Transferase-Based Labeling Combined with Transmission Electron Microscopy

      • Yoshinori Otsuki, Yuko Ito
      Pages 41-54

    6. Determination of Three-Dimensional Distribution of Apoptotic DNA Damage by Combination of TUNEL and Quick-Freezing and Deep-Etching Techniques

      • Shinichi Ohno, Takeshi Baba, Nobuo Terada, Yasuhisa Fuji
      Pages 55-68

    7. In Situ Detection of DNA Strand Breaks in Analysis of Apoptosis by Flow- and Laser-Scanning Cytometry

      • Zbigniew Darzynkiewicz, Elzbieta Bedner, Piotr Smolewski
      Pages 69-75

  3. Labeling DNA Breaks Using DNA Polymerase I or its Klenow Fragment

    1. Front Matter

      Pages 79-79
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    2. DNA Damage Detection Using DNA Polymerase I or its Klenow Fragment

      • Jan Hein van Dierendonck
      Pages 81-108

    3. Labeling DNA Breaks In Situ by Klenow Enzyme

      • Katherine A. Wood
      Pages 109-119

    4. In Situ Nick Translation at the Electron Microscopic Level

      • Marc Thiry
      Pages 121-130

  4. Labeling DNA Breaks Using Ligase

    1. Front Matter

      Pages 131-131
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    2. In Situ DNA Ligation as a Method for Labeling Apoptotic Cells in Tissue Sections

      • Peter J. Hornsby, Vladimir V. Didenko
      Pages 133-141

    3. Detection of Specific Double-Strand DNA Breaks and Apoptosis In Situ Using T4 DNA Ligase

      • Vladimir V. Didenko
      Pages 143-151

    4. In Situ Detection of Double-Strand DNA Breaks with Terminal 5′OH Groups

      • Vladimir V. Didenko, Hop Ngo, David S. Baskin
      Pages 153-159

  5. Detection of DNA Breaks in Agarose Trapped Cells: Comet Assay and Related Techniques

    1. Front Matter

      Pages 161-161
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    2. The Comet Assay

      • Andrew R. Collins
      Pages 163-177

    3. The Comet Assay

      • Peggy L. Olive
      Pages 179-194

    4. Ultrasensitive Detection of DNA Damage by the Combination of the Comet and TUNEL Assays

      • Andrei L. Kindzelskii, Howard R. Petty
      Pages 195-201
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Keywords

About this book

Detection and analysis of DNA damage is of critical importance in a variety of biological disciplines studying apoptosis, cell cycle and cell di- sion, carcinogenesis, tumor growth, embryogenesis and aging, neu- degenerative and heart diseases, anticancer drug development, environmental and radiobiological research, and others. Individual cells within the same tissue or in cell culture may vary in the extent of their DNA damage and, consequently, can display different re- tions to it. These differences between individual cells in the same cell popu- tion are detected using in situ approaches. In situ is a Latin term meaning “on site” or “in place.” It is used to denote the processes occurring or detected in their place of origin. In mole- lar and cell biology this usually refers to undisrupted mounted cells or tissue sections. In that meaning “in situ” is used as part of the terms “in situ PCR,” “in situ transcription,” “in situ hybridization,” “in situ end labeling,” and “in situ ligation.” Sometimes the “in situ” term is applied at the subcellular level to cells disrupted in the process of analysis, for example, in the detection of specific sequences in chromosomes using fluorescent in situ hybridization (FISH). Historically, the term was used primarily in methods dealing with nucleic acids.

Reviews

"Investigators working in the general area of DNA damage, including apoptosis, will find this book of use. The protocols are sufficiently detailed so as to allow experienced laboratory workers access to the described methods. . .the broad spectrum of methods described allows for individual investigators to exercise options."-Doody's Health Sciences Book Review Journal

"Here, a panel of experts describe all the major in situ techniques for studying DNA damage and apoptosis, and how approaches originally designed to label apoptotic cells can be used for DNA damage analysis (and vice versa)." - European Journal of Histochemistry

Editors and Affiliations

  • Baylor College of Medicine, Houston

    Vladimir V. Didenko

Bibliographic Information

  • Book Title: In Situ Detection of DNA Damage

  • Book Subtitle: Methods and Protocols

  • Editors: Vladimir V. Didenko

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1385/1592591795

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media New York 2002

  • Hardcover ISBN: 978-0-89603-952-0Published: 30 April 2002

  • Softcover ISBN: 978-1-61737-272-8Published: 10 November 2010

  • eBook ISBN: 978-1-59259-179-4Published: 05 February 2008

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XVII, 313

  • Number of Illustrations: 133 b/w illustrations, 31 illustrations in colour

  • Topics: Human Genetics, Cell Biology

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