"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)
DNA Damage Detection In Situ, Ex Vivo and In Vivo: Methods and Protocols
Vladimir V. Didenko, Editor
Table of Contents
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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 Immunohistochemistry 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 Immunohistochemistry. 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 -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
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.