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

by Vladimir V. Didenko

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.

Product Details

ISBN-13:	9781493961986
Publisher:	Springer-Verlag New York, LLC
Publication date:	08/23/2016
Series:	Methods in Molecular Biology , #682
Edition description:	Softcover reprint of the original 1st ed. 2011
Pages:	313
Product dimensions:	7.01(w) x 10.00(h) x (d)

Table of Contents

In Situ Detection of Apoptosis by the TUNEL Assay: An Overview of Techniques.-Combination of TUNEL Assay with Immunohishemistry for Simultaneous Detection of DNA Fragmentation and Oxidative Cell Damage.- EM-ISEL: A Useful Tool to Visualize DNA Damage at the Ultrastructural Level.- In Situ Labeling of DNA Breaks and Apoptosis by T7 DNA Polymerase.- In Situ Ligation: a Decade and a Half of Experience.-In Situ Ligation Simplified: Using PCR Fragments for Detection of Double-Strand DNA Breaks in Tissue Sections.- 5’OH DNA Breaks in Apoptosis and their Labeling by Topoisomerase-Based Approach.- Detection of DNA Strand Breaks in Apoptotic Cells by Flow- and Image-Cytometry.- Fluorochrome-Labeled Inhibitors of Caspases (FLICA): Convenient In Vitro and In Vivo Markers of Apoptotic Cells for Cytometric Analysis.- Combining Fluorescent In Situ Hybridization with the Comet Assay for Targeted Examination of DNA Damage and Repair. Simultaneous Labeling of Single- and Double-Strand DNA Breaks by DNA Breakage Detection-FISH (DBD-FISH).- Co-Localization of DNA Repair Proteins with UV-induced DNA Damage in Locally Irradiated Cells.- Ultrasound Imaging of Apoptosis: Spectroscopic Detection of DNA-Damage Effects at High and Low-Frequencies. - Quantifying Etheno-DNA Adducts in Human Tissues, White Blood Cells and Urine by Ultrasensitive 32P-postlabeling and Immunohishemistry.- ELISpot Assay as a Tool to Study Oxidative Stress in Peripheral Blood Mononuclear Cells.- Cytokinesis-Block Micronucleus Cytome assay in Lymphocytes.- Buccal Micronucleus Cytome Assay.- -H2AX Detection in Peripheral Blood Lymphocytes, Splenocytes, Bone Marrow, Xenografts and Skin.- Immunologic Detection of Benzo(a)pyrene-DNA Adducts.- Non-Invasive Assessment of Oxidatively Damaged DNA: Liquid Chromatography-Tandem Mass Spectrometry Analysis of Urinary 8-Oxo-7,8-dihydro-2’-deoxyguanosine.- Assessing sperm DNA fragmentation with the Sperm Chromatin Dispersion (SCD) test.

Editorial Reviews

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.

Doody's Review Service

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 Hishemistry, Vol. 55, 2011)

From the Publisher