Practical Guide to Sperm Analysis: Basic Andrology in Reproductive Medicine


PrefaceThe Usefulness of Sperm Viability Testing in Reproductive Technology: The Hypo-Osmotic Swelling Test, Laser and Motility StimulantsThe Scientific and Biological Bases of Testing Sperm ViabilityAnalytical Techniques to Test Sperm ViabilityClinical Options with Nonmotile SpermatozoaClinical Implications of Using Immotile Sperm for ICSIClinical Implications of Selecting Viable Immotile Sperm for Intracytoplasmic Sperm InjectionClinical Implications of Making Immotile Sperm Motile for Intracytoplasmic Sperm InjectionPentoxifylline and TheophyllineKartagener's SyndromeClinical Availability of Current Tests and the FutureREFERENCESHistory and Physical Examination: Male InfertilityIntroductionHistoryMale Sexual and Reproductive HistoryDevelopmental HistorySystemic IllnessPast Surgical HistoryFamily HistorySocial HistoryExposuresFemale Sexual and Reproductive HistoryTiming and Frequency of IntercourseCoital PracticesPhysicalConclusionREFERENCESThe Role of Hormonal Profiles to Forecast Male Fertility ChancesIntroductionHypothalamic-Pituitary-Gonadal AxisDiagnosis and Physiology of HypoandrogenismAssociation between Hormonal Dysfunction and Fertility PotentialUse of Hormonal Profiles to Predict Surgical and Medical Treatment OutcomesConclusionREFERENCESThe Application of Genetic Tests in an Assisted Reproduction Unit: KaryotypeIntroductionCytogenetic AnalysisKaryotype AnalysisFluorescent In Situ Hybridization AnalysisYq Microdeletion AnalysisSequencingConclusionsREFERENCESGenetic Testing of Y-Chromosome MicrodeletionHistory of the Y ChromosomeY Chromosome StructureY-Chromosome MicrodeletionsGenetic Testing of Y-ChromosomeMicrodeletionAZFa DeletionsAZFb and AZFbc DeletionsAZFc DeletionsThe Effect of Complete vs. Partial DeletionsHistological Phenotypes in YCMDClinical Presentation and AssessmentYCMD TestingIVF Success and Transgenerational Impacts of YCMDFuture of YCMDConclusionREFERENCESThe Application of Genetic Tests in an Assisted Reproduction Unit: Sperm FISHThe Scientific and Biological Bases of Sperm FISHAnalytical Techniques in Order to Measure Sperm FISHSperm FixationNucleus DecondensationDenaturation and HybridizationDetectionSignal Visualization and EvaluationClinical EvidenceClinical UseResultsSperm FISH in Normal Karyotype MenSperm FISH in Carriers of Chromosome AbnormalitiesMethods to Improve the ResultsTest AvailabilityREFERENCESThe Application of Genetic Tests in an Assisted Reproduction Unit: Cystic Fibrosis Carrier ScreeningIntroductionMale Infertility and CFTR GeneCFTR and Its Role in SpermatogenesisCFTR and Its Role in SpermiogenesisCFTR and Its Role in Sperm CapacitationConclusionREFERENCESThe Application of Genetic Tests in an Assisted Reproduction Unit: mRNA MicroarraysIntroductionAnalytical Techniques: Microarray Technology as a Diagnostic ToolThe Scientific and Biological Bases of Microarray Test: Involvement in Reproductive FunctionmRNA Content in Spermatozoa: EvidencesSperm Molecular Feature Required in ARTsmRNA Profile Donors versus PatientsmRNA Profile in IUIDifferentially Expressed TranscriptsExclusive Expressed TranscriptsGene Ontology AnalysisGenes Previously Described in Reproduction-Related Biological ProcessesmRNA Profile in IVFmRNA Profile in ICSIResultsREFERENCESThe Application of Genetic Tests in an Assisted Reproduction Unit: DNA FragmentationScientific Background of Sperm DNA DamageDNA Fragmentation TechniquesFlow CytometryFluorescence MicroscopyUse of Flow Cytometry in Sperm DNA Fragmentation TestsTerminal Transferase dUTP Nick-End LabelingSperm Chromatin Structure AssaySperm Chromatin DispersionAlkaline Comet AssayClinical Evidence of Relationships between Sperm DNA Damage and ART OutcomesClinical Interpretation of DNA Fragmentation Tests: Need for Agreed ThresholdsCosts and Clinical UseMethods to Improve Sperm DNA Fragmentation Testing Comparison of DNA Fragmentation TestsCustomized Sperm SelectionTest Availability and CostConclusionAcknowledgmentsConflict of InterestREFERENCESThe Application of Genetic Tests in an Assisted Reproduction Unit: DNA Methylation DefectsEpigenetics: DNA MethylationTest to Analyze Sperm DNA Methylation at Single Base-Pair ResolutionGeneral Considerations Sperm Cell IsolationSperm DNA IsolationSodium Bisulfite TreatmentSingle-Locus AnalysisLarge-Scale AnalysisCharacteristics of the Sperm MethylomeOrigin and Consequences of Sperm DNA Methylation DefectsOrigin of Sperm Methylation DefectsConsequences of Sperm DNA Methylation Defects on FertilityFinal RemarksREFERENCESThe Use of Sperm Ultra-Morphology Assessment in Assisted ReproductionIntroductionLight and Electron MicroscopyMotile Sperm Organelle Morphology ExaminationFactors That Might Affect the Occurrence of Sperm Nuclear VacuolesCharacteristics and Origin of Sperm Head VacuolesSperm Vacuoles and Sperm DNA DamageFrom MSOME to Intracytoplasmic Morphologically Sperm InjectionREFERENCESThe Usefulness of Antisperm Antibodies TestingIntroductionScientific and Biological Bases of Evaluation of ASAEtiology of ASA Development in Men and WomenASA Effects on FertilityAnalytical Techniques to Measure the Presence of ASAMethods to Measure ASA in Spermatozoa and FluidsASA Incidence in Men and WomenASA Evaluation in Men Attending an Andrology Laboratory: Our ExperienceAssessment of ASA Incidence and Relationship with Routine Semen Parameters and Sperm Kinematics in a Large Population of PatientsLaboratory Guidelines for ASA Assessment in Sperm and Biological FluidsThe Direct MAR Test Reagents NeededThe Direct Immunobead Binding Test Reagents NeededThe Indirect IB TestConclusionsAcknowledgmentsREFERENCESThe Measurement of Oxidative Stress in Semen and Use in Assisted ReproductionSummaryIntroductionReactive Oxygen SpeciesCounteracting Reactive Oxygen SpeciesAn Inadequate BalanceOxidative Stress in the ART SettingMeasuring Oxidative StressMethods to Measure Oxidative StressChemiluminescenceMeasurement of ROS Using ChemiluminescenceROS Measurement by LuminometerCalculating and Analyzing The ResultsPrecautionary MeasuresNitroblue Tetrazolium TestMeasurement of ROS Using NBT AssayPrecautionary MeasuresFlow CytometryOxidation-Reduction PotentialMeasurement of ORP Using RedoxSYS SystemAbstract FindingsImmunocytochemistry and Western BlottingConclusionREFERENCESThe Importance of Sperm Surface Markers in Reproductive Success: Sperm Hyaluronan BindingIntroductionStructure of the Mature SpermatozoaSperm Surface MarkersHyaluronan Receptor Is a Marker of Sperm DevelopmentRelationship between Sperm Shape and HA BindingHA Binding and DNA IntegrityClinical Importance of HA BindingConclusionREFERENCESImportance of Sperm Surface Markers in Reproductive Success: Surface ChargeIntroductionSperm Surface Charge as a BiomarkerBiological Importance of Sperm Negative Surface ChargeCharge-Based Sperm Selection MethodsElectrophoretic Sperm SeparationMethodological Aspects of Electrophoretic Sperm SelectionSperm Quality Following Electrophoretic Sperm SeparationComparison between Electrophoretic Sperm Separation and Other MethodsClinical Importance of Sperm Preparation by ElectrophoreticConclusionZeta TestMethodology of Zeta TestQuality of Sperm Separated by Zeta TestComparison between the Zeta Test and Other MethodsClinical Importance of Sperm Preparation by Zeta TestConclusionMicro-Electrophoresis Sperm Selection MethodMethodological Aspects of Micro-ElectrophoresisQuality of Sperm Separated by MicroelectrophoresisDiversity of Charged Sperm in Raw Semen and DGC Prepared PopulationCharacteristics of Charged Sperm Population Analyzed by MicroelectrophoresisConclusionConclusion and Future ImplicationsREFERENCESThe Relevance in Reproductive Success of Sperm Head Polarization (Birefringence)IntroductionEvaluation of Sperm BirefringenceClinical SignificanceReproductive OutcomeConclusionsREFERENCESThe Applicability of Raman Spectroscopy in Sperm Diagnosis and Selection for Assisted ReproductionIntroductionRaman SpectroscopyUse of Raman Spectroscopy in Analysis of Male Reproductive Tissues, Fluids, and CellsConclusionsREFERENCESThe Use of Sperm Proteomics in the Assisted Reproduction LaboratoryThe Scientific and Biological Bases of ProteomicsProteomic TechniquesScientific EvidencePotential Clinical UseImprovement in the Reproductive CounselingImprovement in the Counseling of Azoospermic Patients Facing Invasive ProceduresImprovement of Clinical ProcessesDesign of Potential Fertility TherapiesDesign of Potential Anticonceptive StrategiesTest AvailabilityAcknowledgmentsREFERENCESEvaluation of Surgically Retrieved Spermatozoa and Its Usefulness in Assisted ReproductionIntroductionSperm Processing and SelectionMethods for Processing Epididymal and Testicular Sperm Samples Recovery of Epididymal SpermRecovery of Testicular SpermMechanical MethodsShredding MethodSqueezing MethodCell StrainerTissue Grinder MethodErythrocyte-Lysing Buffer MethodEnzymatic Digestion MethodSperm Selection MethodsSTFT/Mechanical Touch TechniqueThe Hypoosmotic Swelling TestChemical Motility EnhancersLaser-Assisted Immotile Sperm SelectionCryopreservation of Surgically Retrieved Testicular SpermThe Use of Surgically Retrieved Sperm in Assisted ReproductionConclusionREFERENCES
 
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