Full Description
Selecting good-quality sperm for use in in-vitro fertilization is a key step in assisted reproduction. For many years purely morphological attributes have been used to assess suitability, but increasingly biochemical and molecular biological techniques are now identifying sperm with the best chances of producing viable and healthy embryos. Focusing on modern sperm function testing, this manual provides technical details of commonly used tests and gives an overview of the laboratory techniques used to evaluate sperm samples. Covering a variety of testing methods in detail, from manual and computer-assisted semen analysis to zona pellucida binding assays, and tests assessing sperm DNA damage such as the TUNEL assay. Describing the underlying science, practical advice for performing the tests is given, including tips for optimizing outcomes and trouble-shooting. This is an essential guide for reproductive medicine specialists, clinical andrologists, urologists and gynecologists working with sub-fertile men.
Contents
Introduction R. John Aitken; 1. Standard semen analysis: manual semen analysis Lars Björndahl; 2. Standard semen analysis: computer-assisted semen analysis Liana Maree; 3. Standard semen analysis: home sperm testing Manesh Panner Selvam Kumar, Christopher Douglas and Ashok Agarwal; 4. Standard semen analysis: leukocytospermia Renata Finelli and Ralf Henkel; 5. Standard semen analysis: morphology Roelof Menkveld; 6. Sperm vitality: eosin-nigrosin dye exclusion Rakesh Sharma and Ashok Agarwal; 7. Sperm vitality: hypo-osmotic swelling test Alexandre Rouen and Rajasingam S. Jeyendran; 8. Determination of mitochondrial membrane potential by flow cytometry in human sperm cells Nicolas Germain, Nathalie Jouy, Carole Marchetti and Philippe Marchetti; 9. Capacitation and acrosome reaction: fluorescence techniques to determine acrosome reaction Raúl S. Sánchez Gutiérrez, Fabiola Zambrano and Pamela Uribe; 10. Capacitation and acrosome reaction: histochemical techniques to determine acrosome reaction Brett Nixon, Shenae Café, Elizabeth Bromfield, Geoffry De Iuliis and Matthew Dun; 11. Zona binding: competitive sperm-binding assay De Yi Liu, Ying Zhong and Zi-Na Wen; 12. Zona binding: hemizona assay Shubhadeep Roychoudhury and Daniel Franken; 13. Oolemma binding: sperm penetration assay Anup Shah and Kathleen Hwang; 14. Oxidative stress testing: direct tests Renata Finelli, Manesh Kumar, Panner Selvam and Ashok Agarwal; 15. Oxidative stress testing: indirect tests Rakesh Sharma, Kathy Robert and Ashok Agarwal; 16. Chromatin condensation: aniline blue stain Jesse JP, Vidhu Dhawan and Rima Dada; 17. Chromatin condensation: chromomycin A3 (CMA3) stain Sara Marchiani, Lara Tamburrino, Francesca Benini, Sandra Pellegrini and Elisabetta Baldi; 18. Sperm chromatin structure: Toluidine Blue Staining Mohammed Nasr-Esfahani and Marziyeh Tavalaee; 19. DNA damage: TdT-mediated dUTP nick-end-labelling (TUNEL) assay Rakesh Sharma, Concetta Iovine and Ashok Agarwal; 20. DNA damage: sperm chromatin structure assay (SCSA) Don Evenson; 21. DNA damage: COMET assay Luke Simon, Benjamin R. Emery and Douglas T. Carrell; 22. DNA damage: halo sperm test Jaime Gosálvez and José Luís Fernández; 23. DNA damage: fluorescent in-situ hybridization (FISH) Sezgin Güneş; 24. Clinical value of sperm function tests Ahmad Majzoub; 25. Future developments: sperm proteomics Manesh Panner Selvam Kumar, Saradha Baskaran and Ashok Agarwal; 26. Conclusion Ahmad Majzoub, Ralf Henkel and Ashok Agarwal.
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