Manual of Sperm Function Testing in Human Assisted Reproduction

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.

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.

• Written by an international team of experts, bringing diverse experience and knowledge bases to the book
• Covers a variety of different techniques, suitable for a range of infertility causes, and includes descriptions of the underlying science to aid readers in their understanding
• Includes practical advice for performing the tests as well as tips for optimizing the outcomes and trouble-shooting any problems that may arise

Editors

Ashok Agarwal, The Cleveland Clinic Foundation, Cleveland, OH
Ashok Agarwal is the Director of the Clinical Andrology Laboratory and Reproductive Tissue Bank and the Director of Research at the American Center for Reproductive Medicine. He holds these positions at The Cleveland Clinic Foundation, where he is a Professor of Surgery (Urology) at the Lerner College of Medicine of Case Western Reserve University, Cleveland, Ohio, USA.

Ralf Henkel, University of the Western Cape, South Africa
Ralf Henkel is the Chief Scientific Advisor for LogixX Pharma Ltd and Editor-in-Chief of Andrologia. He is also an Extraordinary Professor at the University of the Western Cape, Bellville, South Africa and an Honorary Professor at the Universidad Peruana Cayetano Heredia, Lima, Peru, American Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH.

Ahmad Majzoub, Hamad Medical Corporation, Doha
Ahmad Majzoub is a Consultant in the Department of Urology and Program Director of the Andrology Fellowship at the Hamad Medical Corporation, Doha and an Assistant Professor of Clinical Urology at Weill Cornell Medicine, Doha, Qatar.

Contributors

R. John Aitken, Lars Björndahl, Liana Maree, Manesh Panner Selvam Kumar, Christopher Douglas, Ashok Agarwal, Renata Finelli, Ralf Henkel, Roelof Menkveld, Rakesh Sharma, Alexandre Rouen, Rajasingam S. Jeyendran, Nicolas Germain, Nathalie Jouy, Carole Marchetti, Philippe Marchetti, Raúl S. Sánchez Gutiérrez, Fabiola Zambrano, Pamela Uribe, Brett Nixon, Shenae Café, Elizabeth Bromfield, Geoffry De Iuliis, Matthew Dun, De Yi Liu, Ying Zhong, Zi-Na Wen, Shubhadeep Roychoudhury, Daniel Franken, Anup Shah, Kathleen Hwang, Kathy Robert, Jesse JP, Vidhu Dhawan, Rima Dada, Sara Marchiani, Lara Tamburrino, Francesca Benini, Sandra Pellegrini, Elisabetta Baldi, Mohammed Nasr-Esfahani, Marziyeh Tavalaee, Concetta Iovine, Don Evenson, Luke Simon, Benjamin R. Emery, Douglas T. Carrell, Jaime Gosálvez, José Luís Fernández, Sezgin Güneş, Ahmad Majzoub, Saradha Baskaran