Identifying good-quality sperm for use in in-vitro fertilization is a key step in assisted reproduction. Focusing on modern sperm function testing, a variety of testing methods are covered in detail, from manual and computer-assisted semen analysis to zona pellucida binding assays, and tests assessing DNA damage.
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.
Arvustused
'This is truly a compendium of human sperm function assays. The chapters include background information, some detailed protocols, interesting and insightful drawings, and photographs and insights from the authors/creators of the assays ... it should have a spot on the shelves of every biologist with an interest in human sperm function.' Karla Saner Amigh, Doody's Notes
Muu info
Focusing on modern sperm function testing, this guide is essential in selecting sperm that will produce viable and healthy embryos.
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ix | |
Short Biography |
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xii | |
Foreword |
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xiii | |
Denny Sakkas Preface |
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xv | |
Introduction |
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1 | (5) |
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1 Standard Semen Examination Analysis: Manual Semen Analysis |
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6 | (5) |
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2 Standard Semen Analysis: Computer-Assisted Semen Analysis |
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11 | (12) |
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3 Standard Semen Analysis: Home Sperm Testing |
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23 | (8) |
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Manesh Kumar Panner Selvam |
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4 Standard Semen Analysis: Leukocytospermia |
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31 | (8) |
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5 Standard Semen Analysis: Morphology |
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39 | (8) |
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6 Sperm Vitality: Eosin-Nigrosin Dye Exclusion |
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47 | (5) |
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7 Sperm Vitality: Hypo-Osmotic Swelling Test |
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52 | (6) |
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8 Determination of Mitochondrial Membrane Potential by Flow Cytometry in Human Sperm Cells |
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58 | (14) |
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9 Capacitation and Acrosome Reaction: Fluorescence Techniques to Determine Acrosome Reaction |
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72 | (9) |
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10 Capacitation and Acrosome Reaction: Histochemical Techniques to Determine Acrosome Reaction |
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81 | (12) |
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11 Zona Binding: Competitive Sperm-Binding Assay |
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93 | (7) |
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12 Zona Binding: Hemizona Assay |
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100 | (6) |
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13 Oolemma Binding: Sperm Penetration Assay |
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106 | (5) |
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14 Oxidative Stress Testing: Direct Tests |
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111 | (12) |
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Manesh Kumar Panner Selvam |
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15 Oxidative Stress Testing: Indirect Tests |
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123 | (19) |
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16 Chromatin Condensation: Aniline Blue Stain |
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142 | (9) |
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17 Chromatin Condensation: Chromomycin A3 (CMA3) Stain |
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151 | (5) |
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18 Sperm Chromatin Structure: Toluidine Blue Staining |
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156 | (7) |
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19 DNA Damage: TdT-Mediated dUTP Nick-End-Labelling Assay |
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163 | (29) |
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20 DNA Damage: Sperm Chromatin Structure Assay |
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192 | (10) |
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21 DNA Damage: COMET Assay |
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202 | (11) |
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22 DNA Damage: Halo Sperm Test |
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213 | (15) |
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23 DNA Damage: Fluorescent In-Situ Hybridization |
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228 | (6) |
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24 Clinical Value of Sperm Function Tests |
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234 | (11) |
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25 Future Developments: Sperm Proteomics |
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245 | (11) |
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Conclusion |
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256 | (3) |
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Index |
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259 | |
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 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 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.