DNA Damage Detection In Situ, Ex Vivo, and In Vivo: Methods and Protocols / Edition 1

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Overview

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 Prools, 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 Prools 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.

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Editorial Reviews

Doody's Review Service
Reviewer: Omer Iqbal, MD, FACC, FESC (Loyola University Medical Center)
Description: This is an excellent, beautifully illustrated resource on recent and established techniques to detect and quantify DNA damage ranging from the subcellular to the level of a whole live organism.
Purpose: The book is useful for newcomers to the field and experienced researchers. It is also intended to serve as a guide for novice molecular biology scientists to master the required techniques by providing detailed chapters with simple to follow protocols and describing possible technical pitfalls and limitations.
Audience: Novice molecular biology scientists, established researchers, and scientists from other fields, such as clinical scientists and nanobiotechnology specialists, are the intended audience. Researchers in many fields including molecular and cell biology, experimental and clinical pathology, toxicology, radiobiology, oncology, embryology, experimental pharmacology, drug design, and environmental science also will find this book quite useful. It also will be useful for graduate students interested in this field.
Features: The first of the book's three parts has seven chapters covering fixed tissue sections. It contains enzymatic approaches such as in situ detection of apoptosis by TUNEL assay and combination of TUNEL assay with immunohistochemistry for simultaneous detection of DNA fragmentation and cell damage. It also covers EM_ISEL, a useful tool to visualize DNA damage. The five chapters in the second part of the book covers detection of DNA strand damage and apoptosis in cultured cells. Nine chapters in the final part deal with methods developed either for in vivo detection, directly in a living organism, or for samples taken from the body, ex vivo — in blood, urine, and sperm. The assays often permit monitoring levels of DNA damage. The book is well illustrated with both color and black-and-white figures.
Assessment: This is an excellent resource for all major assays used in molecular and cell biology for labeling of DNA damage in situ, ex vivo, and in vivo. Graduate students, novice molecular biology scientists, and established investigators in this field will find this book of great help.
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Product Details

  • ISBN-13: 9781603274081
  • Publisher: Springer-Verlag New York, LLC
  • Publication date: 11/8/2010
  • Series: Methods in Molecular Biology Series , #682
  • Edition description: 2011
  • Edition number: 1
  • Pages: 314
  • Product dimensions: 7.00 (w) x 10.10 (h) x 0.90 (d)

Table of Contents

DNA Damage Detection In Situ, Ex Vivo and In Vivo: Methods and Prools
Vladimir V. Didenko, Editor
Table of Contents
________________________________________________________________

Table of Contents

Preface

Contributors

Part 1. DETECTION IN TISSUE SECTIONS
1 In Situ Detection of Apoptosis by the TUNEL Assay: An Overview of Techniques. Deryk T. Loo
2 Combination of TUNEL Assay with Immunohishemistry for Simultaneous Detection of DNA Fragmentation and Oxidative Cell Damage. Alexander E. Kalyuzhny
3 EM-ISEL: A Useful Tool to Visualize DNA Damage at the Ultrastructural Level. Antonio Migheli
4 In Situ Labeling of DNA Breaks and Apoptosis by T7 DNA Polymerase. Vladimir V. Didenko
5 In Situ Ligation: a Decade and a Half of Experience. Peter J. Hornsby and Vladimir V. Didenko
6 In Situ Ligation Simplified: Using PCR Fragments for Detection of Double-Strand DNA Breaks in Tissue Sections. Vladimir V. Didenko
7 5’OH DNA Breaks in Apoptosis and their Labeling by Topoisomerase-Based Approach. Vladimir V. Didenko

Part 2. DETECTION IN CELL CULTURES
8 Detection of DNA Strand Breaks in Apoptotic Cells by Flow- and Image-Cytometry. Zbigniew Darzynkiewicz and Hong Zhao
9 Fluorochrome-Labeled Inhibitors of Caspases (FLICA): Convenient In Vitro and In Vivo Markers of Apoptotic Cells for Cytometric Analysis. Zbigniew Darzynkiewicz, Piotr Pozarowski, Brian W. Lee and Gary L. Johnson
10 Combining Fluorescent In Situ Hybridization with the Comet Assay for Targeted Examination of DNA Damage and Repair. Sergey Shaposhnikov, Preben D. Thomsen, and Andrew R. Collins
11 Simultaneous Labeling of Single- and Double-Strand DNA Breaks by DNA Breakage Detection-FISH (DBD-FISH). José Luis Fernández, Dioleyda Cajigal, Jaime Gosálvez
12 Co-Localization of DNA Repair Proteins with UV-induced DNA Damage in Locally Irradiated Cells. Jennifer Guerrero-Santoro, Arthur S. Levine, and Vesna Rapić-Otrin

Part 3. DETECTION IN LIVE TISSUES, BLOOD, URINE, SPERM
13 Ultrasound Imaging of Apoptosis: Spectroscopic Detection of DNA-Damage Effects at High and Low-Frequencies. Roxana M. Vlad, Michael C. Kolios and Gregory J. Czarnota

14 Quantifying Etheno-DNA Adducts in Human Tissues, White Blood Cells and Urine by Ultrasensitive 32P-postlabeling and Immunohishemistry. Jagadeesan Nair, Urmila J. Nair, Xin Sun, Ying Wang, Khelifa Arab, and Helmut Bartsch

15 ELISpot Assay as a Tool to Study Oxidative Stress in Peripheral Blood Mononuclear Cells. Jodi Hagen, J.P. Houchins and Alexander E. Kalyuzhny

16 Cytokinesis-Block Micronucleus Cytome assay in Lymphocytes. Philip Thomas and Michael Fenech

17 Buccal Micronucleus Cytome Assay. Philip Thomas and Michael Fenech

18 γ-H2AX Detection in Peripheral Blood Lymphocytes, Splenocytes, Bone Marrow, Xenografts and Skin.
Christophe E. Redon, Asako Nakamura, Olivier Sordet, Jennifer S. Dickey, Ksenia Gouliaeva, Brian Tabb, Scott Lawrence, Robert J. Kinders, William M. Bonner, Olga A. Sedelnikova

19 Immunologic Detection of Benzo(a)pyrene-DNA Adducts. Regina M. Santella and Yu-Jing Zhang

20 Non-Invasive Assessment of Oxidatively Damaged DNA: Liquid Chromatography-Tandem Mass Spectrometry Analysis of Urinary 8-Oxo-7,8-dihydro-2’-deoxyguanosine.Vilas Mistry, Friederike Teichert, Jatinderpal.K.Sandhu, Rajinder Singh, Mark D. Evans, , Peter B. Farmer, and Marcus S. Cooke

21 Assessing sperm DNA fragmentation with the Sperm Chromatin Dispersion (SCD) test. José Luis Fernández, Dioleyda Cajigal, Carmen López-Fernández, Jaime Gosálvez

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