Book contents
- Manual of Sperm Function Testing in Human Assisted Reproduction
- Cambridge Laboratory Manuals in Assisted Reproductive Technology
- Manual of Sperm Function Testing in Human Assisted Reproduction
- Copyright page
- Dedication
- Contents
- Contributors
- Short Biography
- Foreword
- Preface
- Introduction
- Chapter 1 Standard Semen Examination: Manual Semen Analysis
- Chapter 2 Standard Semen Analysis: Computer-Assisted Semen Analysis
- Chapter 3 Standard Semen Analysis: Home Sperm Testing
- Chapter 4 Standard Semen Analysis: Leukocytospermia
- Chapter 5 Standard Semen Analysis: Morphology
- Chapter 6 Sperm Vitality: Eosin-Nigrosin Dye Exclusion
- Chapter 7 Sperm Vitality: Hypo-Osmotic Swelling Test
- Chapter 8 Determination of Mitochondrial Membrane Potential by Flow Cytometry in Human Sperm Cells
- Chapter 9 Capacitation and Acrosome Reaction: Fluorescence Techniques to Determine Acrosome Reaction
- Chapter 10 Capacitation and Acrosome Reaction: Histochemical Techniques to Determine Acrosome Reaction
- Chapter 11 Zona Binding: Competitive Sperm-Binding Assay
- Chapter 12 Zona Binding: Hemizona Assay
- Chapter 13 Oolemma Binding: Sperm Penetration Assay
- Chapter 14 Oxidative Stress Testing: Direct Tests
- Chapter 15 Oxidative Stress Testing: Indirect Tests
- Chapter 16 Chromatin Condensation: Aniline Blue Stain
- Chapter 17 Chromatin Condensation: Chromomycin A3 (CMA3) Stain
- Chapter 18 Sperm Chromatin Structure: Toluidine Blue Staining
- Chapter 19 DNA Damage: TdT-Mediated dUTP Nick-End-Labelling Assay
- Chapter 20 DNA Damage: Sperm Chromatin Structure Assay
- Chapter 21 DNA Damage: COMET Assay
- Chapter 22 DNA Damage: Halo Sperm Test
- Chapter 23 DNA Damage: Fluorescent In-Situ Hybridization
- Chapter 24 Clinical Value of Sperm Function Tests
- Chapter 25 Future Developments: Sperm Proteomics
- Conclusion
- Index
- References
Chapter 23 - DNA Damage: Fluorescent In-Situ Hybridization
Published online by Cambridge University Press: 05 April 2021
- Manual of Sperm Function Testing in Human Assisted Reproduction
- Cambridge Laboratory Manuals in Assisted Reproductive Technology
- Manual of Sperm Function Testing in Human Assisted Reproduction
- Copyright page
- Dedication
- Contents
- Contributors
- Short Biography
- Foreword
- Preface
- Introduction
- Chapter 1 Standard Semen Examination: Manual Semen Analysis
- Chapter 2 Standard Semen Analysis: Computer-Assisted Semen Analysis
- Chapter 3 Standard Semen Analysis: Home Sperm Testing
- Chapter 4 Standard Semen Analysis: Leukocytospermia
- Chapter 5 Standard Semen Analysis: Morphology
- Chapter 6 Sperm Vitality: Eosin-Nigrosin Dye Exclusion
- Chapter 7 Sperm Vitality: Hypo-Osmotic Swelling Test
- Chapter 8 Determination of Mitochondrial Membrane Potential by Flow Cytometry in Human Sperm Cells
- Chapter 9 Capacitation and Acrosome Reaction: Fluorescence Techniques to Determine Acrosome Reaction
- Chapter 10 Capacitation and Acrosome Reaction: Histochemical Techniques to Determine Acrosome Reaction
- Chapter 11 Zona Binding: Competitive Sperm-Binding Assay
- Chapter 12 Zona Binding: Hemizona Assay
- Chapter 13 Oolemma Binding: Sperm Penetration Assay
- Chapter 14 Oxidative Stress Testing: Direct Tests
- Chapter 15 Oxidative Stress Testing: Indirect Tests
- Chapter 16 Chromatin Condensation: Aniline Blue Stain
- Chapter 17 Chromatin Condensation: Chromomycin A3 (CMA3) Stain
- Chapter 18 Sperm Chromatin Structure: Toluidine Blue Staining
- Chapter 19 DNA Damage: TdT-Mediated dUTP Nick-End-Labelling Assay
- Chapter 20 DNA Damage: Sperm Chromatin Structure Assay
- Chapter 21 DNA Damage: COMET Assay
- Chapter 22 DNA Damage: Halo Sperm Test
- Chapter 23 DNA Damage: Fluorescent In-Situ Hybridization
- Chapter 24 Clinical Value of Sperm Function Tests
- Chapter 25 Future Developments: Sperm Proteomics
- Conclusion
- Index
- References
Summary
Several sperm DNA integrity and chromatin quality assays have been developed and used for the detection of sperm DNA fragmentation and chromosomal aberrations [1, 2]. The Comet assay is one of the most commonly used single cell gel electrophoresis methods first developed by Östling and Johanson [3]. Damaged or fragmented sperm DNA is separated from undamaged DNA by electrophoresis. Fluorescent in situ hybridization (FISH) is a hybridization method that provides specific identification of selected DNA sequences and is frequently used for cytogenetic analysis of spermatozoa. These two methods were combined as the Comet-FISH technique first by Santos and colleagues in 1997 [4]. The Comet-FISH technique is a combination of two well-known methods, the Comet assay and FISH, and is a useful detection tool for screening of whole and region-specific DNA damage [5, 6]. In addition, the analysis of labeled DNA sequences and whole chromosomes of interest and identification of region-specific DNA and overall damage and repair in single cells is possible [7, 8]. Therefore, the modification of these assays interpolates hybridization with specific fluorescent-labeled probes to selected DNA sequences of interest after unwinding and electrophoresis [6] and developed a standardized Comet assay for human sperm with additional information for telomeres or special DNA sequences of interest. Thus, specific gene sequences can be detected using the Comet-FISH technique [9].
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- Manual of Sperm Function Testing in Human Assisted Reproduction , pp. 228 - 233Publisher: Cambridge University PressPrint publication year: 2021