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Dedication; Foreword; Preface; Acknowledgements; Contents; About the Authors; Abbreviations; 1: Semen Analysis: An Overview; 1.1 Semen Quality Evaluation; 1.2 Harvest of Spermatozoa; 1.3 Storage of Semen; 1.4 Evaluation of Semen Quality Parameters (SQP); 1.4.1 Sperm Assays; 1.4.1.1 The Classical Spermiogram; 1.4.1.2 Advancement in Sperm Assays; Literature Cited; Key Reference; 2: Selection of Spermatozoa; 2.1 Introduction; 2.2 Selecting Procedure for Sperm Separation; 2.3 Comparison of Sperm Selection Protocols; 2.4 Sperm Selection Protocols; 2.4.1 Simple Washing of Spermatozoa
2.4.2 Sperm Swim-Up Technique2.4.2.1 Direct Swim-Up I; 2.4.2.2 Direct Swim-Up II; 2.4.3 Discontinuous Density Gradients; 2.4.4 Glass Wool Filtration Column; 2.5 Preparing Testicular and Epididymal Spermatozoa; 2.6 Processing Sperm Suspensions for ICSI; 2.7 Background Information; Literature Cited; Key References; 3: Handling and Applications of Optical Devices; 3.1 Introduction; 3.2 General Principles of Light Microscopy; 3.2.1 The Formation of the Image; 3.2.1.1 Microscope Lenses and their Defects; 3.2.1.1.1 Axial Aberrations; 3.2.1.1.2 Off-Axial Aberrations; 3.3 Eye Pieces
3.4 Objectives3.5 Condensers; 3.6 Illumination; 3.7 Fluorescence Microscopy; 3.8 Phase Contrast Microscopy; 3.9 Bright-Field Microscopy; 4: Basic Semen Assays; 4.1 Introduction; 4.2 Basic Semen Assays; 4.2.1 Macroscopic Evaluation of Ejaculate; 4.2.1.1 Ejaculate Volume; 4.2.1.2 Colour; 4.2.1.3 Consistency; 4.2.1.4 Density; 4.2.1.5 Viscosity; 4.2.1.6 Hydrogen Ion Concentration (pH) Value; 4.2.1.7 Osmotic Pressure; 4.2.1.8 Odour; 4.2.2 Microscopic Evaluation of Ejaculate; 4.2.2.1 Microbial Load of Semen; 4.3 Optional Assays in Semen Evaluation
4.3.1 Evaluation of Aggregation of Spermatozoa4.3.2 Evaluation of Agglutination of Spermatozoa; 4.3.3 Examination of Cellular Elements Other Than Spermatozoa; 4.3.4 Leucocyte Measurement in Semen Sample; 4.3.4.1 Screening of Granulocytes; 4.3.4.2 Panleukocyte (CD45) Immunocytochemical Staining; 4.4 Background Information; Literature Cited; Key References; 5: Estimates of Sperm Motility; 5.1 Introduction; 5.2 Microscopy for Motility Evaluation; 5.3 Preparing a Wet Smear for Motility Estimates; 5.4 Estimates of Motility; 5.4.1 Mass Activity
5.4.2 Estimates of Sperm Motility by Direct Count5.4.3 Estimates of Sperm Motility Using Haemocytometer; 5.4.4 Post-thaw Motility; 5.4.5 Motility Index; 5.5 Computer-Aided Sperm Analysis (CASA); 5.5.1 Applications of CASA; 5.5.1.1 Sperm Concentration and Motility; 5.5.1.2 Computer-Aided Sperm Morphometric Assessment (CASMA); 5.5.1.2.1 Information; Literature Cited; Key References; 6: Evaluating Sperm Cell Viability and Membrane Integrity; 6.1 Introduction; 6.2 Comparison of Different Staining Procedures for Viability Evaluation; 6.3 Staining for Vitality Determination
2.4.2 Sperm Swim-Up Technique2.4.2.1 Direct Swim-Up I; 2.4.2.2 Direct Swim-Up II; 2.4.3 Discontinuous Density Gradients; 2.4.4 Glass Wool Filtration Column; 2.5 Preparing Testicular and Epididymal Spermatozoa; 2.6 Processing Sperm Suspensions for ICSI; 2.7 Background Information; Literature Cited; Key References; 3: Handling and Applications of Optical Devices; 3.1 Introduction; 3.2 General Principles of Light Microscopy; 3.2.1 The Formation of the Image; 3.2.1.1 Microscope Lenses and their Defects; 3.2.1.1.1 Axial Aberrations; 3.2.1.1.2 Off-Axial Aberrations; 3.3 Eye Pieces
3.4 Objectives3.5 Condensers; 3.6 Illumination; 3.7 Fluorescence Microscopy; 3.8 Phase Contrast Microscopy; 3.9 Bright-Field Microscopy; 4: Basic Semen Assays; 4.1 Introduction; 4.2 Basic Semen Assays; 4.2.1 Macroscopic Evaluation of Ejaculate; 4.2.1.1 Ejaculate Volume; 4.2.1.2 Colour; 4.2.1.3 Consistency; 4.2.1.4 Density; 4.2.1.5 Viscosity; 4.2.1.6 Hydrogen Ion Concentration (pH) Value; 4.2.1.7 Osmotic Pressure; 4.2.1.8 Odour; 4.2.2 Microscopic Evaluation of Ejaculate; 4.2.2.1 Microbial Load of Semen; 4.3 Optional Assays in Semen Evaluation
4.3.1 Evaluation of Aggregation of Spermatozoa4.3.2 Evaluation of Agglutination of Spermatozoa; 4.3.3 Examination of Cellular Elements Other Than Spermatozoa; 4.3.4 Leucocyte Measurement in Semen Sample; 4.3.4.1 Screening of Granulocytes; 4.3.4.2 Panleukocyte (CD45) Immunocytochemical Staining; 4.4 Background Information; Literature Cited; Key References; 5: Estimates of Sperm Motility; 5.1 Introduction; 5.2 Microscopy for Motility Evaluation; 5.3 Preparing a Wet Smear for Motility Estimates; 5.4 Estimates of Motility; 5.4.1 Mass Activity
5.4.2 Estimates of Sperm Motility by Direct Count5.4.3 Estimates of Sperm Motility Using Haemocytometer; 5.4.4 Post-thaw Motility; 5.4.5 Motility Index; 5.5 Computer-Aided Sperm Analysis (CASA); 5.5.1 Applications of CASA; 5.5.1.1 Sperm Concentration and Motility; 5.5.1.2 Computer-Aided Sperm Morphometric Assessment (CASMA); 5.5.1.2.1 Information; Literature Cited; Key References; 6: Evaluating Sperm Cell Viability and Membrane Integrity; 6.1 Introduction; 6.2 Comparison of Different Staining Procedures for Viability Evaluation; 6.3 Staining for Vitality Determination