Sperm Function Tests are the keystones of evaluating functional condition of sperms. The fertility potential of a sperm will be decided not only with the number & motility but with the functional competence which is of utmost importance.
Ms. Doel Bose Pande discusses various semen preparation methods and principles. Common techniques include simple wash, swim-up, and density gradient centrifugation. The choice of method depends on factors like semen quality, intended use, and practical considerations. Density gradient is best for separating motile sperm from debris but is more time-consuming than direct swim-up. The goal is to recover high-quality sperm with minimal processing time and damage. Practical issues like sample volume and number of patients may also influence the choice of preparation technique.
This document discusses various sperm preparation methods used for intrauterine insemination (IUI). It describes the simple wash method, swim-up method, and density gradient centrifugation method. For each method, it provides the steps, including centrifugation speeds and times. It also covers preparing samples for retrograde ejaculation and samples from HIV-infected patients. The goal of sperm preparation is to select motile sperm and remove other seminal constituents like debris to maximize the chances of fertilization during IUI.
ICSI as it is presently performed is far from an ideal solution because the selection of sperm is based on the judgement of an embryologist, who is looking for the most normal appearing sperm available.
This document discusses various sperm preparation techniques used prior to assisted reproductive technologies. It begins by explaining the reasons for processing sperm samples, such as removing components that could prevent pregnancy and selecting morphologically normal, motile sperm free of debris. Several migration-based techniques are described, including swim-up, density gradient centrifugation, and magnetic activated cell sorting. Glass wool filtration and zeta potential techniques are also covered. The document discusses preparation of epididymal and testicular sperm samples. It concludes by outlining methods for preparing sperm from retrograde ejaculation samples.
This document summarizes key aspects of human oogenesis and oocyte quality assessment. It describes the process of oogenesis, including formation of follicles in the ovaries and maturation of oocytes. It also discusses morphological features used to assess oocyte quality, such as the cumulus-oocyte complex, nuclear maturity, size and shape, the zona pellucida, polar body morphology, and perivitelline space. While some studies found correlations between certain morphological features and development outcomes, others found no significant correlations. Overall assessment of oocyte quality relies on evaluating multiple morphological characteristics.
This document discusses sperm sorting techniques for men with high sperm DNA fragmentation index. It begins by defining sperm DNA damage and fragmentation, then discusses causes and indications for testing. It describes different tests to measure sperm DNA fragmentation and diagnostic cut-off points. The objectives and limitations of sperm sorting are outlined. Various sperm sorting techniques are explained, including swim-up, density gradient, magnetic activated cell sorting (MACS), motile sperm organelle morphology examination (MSOME), and surface charge-based sorting using hyaluronan binding. Advanced techniques like MSOME and MACS aim to select sperm with intact DNA and normal morphology to improve fertility outcomes.
The document outlines the key procedures involved in ICSI lab work for a gynecologist, as presented by Aboubakr Elnashar from Benha University in Egypt. It discusses 8 main procedures: 1) semen preparation, 2) oocyte identification, 3) oocyte denudation, 4) oocyte assessment, 5) oocyte injection, 6) embryo selection, 7) embryo transfer, and 8) cryopreservation. For each procedure, it provides details on the aim, methods, and considerations. The document serves as a reference for best practices in ICSI lab work to optimize outcomes.
Sperm retrieval techniques - nuts and boltsSandro Esteves
This document summarizes a training program in assisted reproductive technology focusing on azoospermia. It discusses the differences between obstructive and non-obstructive azoospermia and various sperm retrieval techniques such as PESA, MESA, TESA, and TESE. Success rates of sperm retrieval are provided for different causes of azoospermia. Predictive markers for sperm retrieval success are also examined, such as hormone levels, testicular volume, histopathology, and microdeletions. Micro-TESE is highlighted as a promising surgical method for non-obstructive azoospermia.
Ms. Doel Bose Pande discusses various semen preparation methods and principles. Common techniques include simple wash, swim-up, and density gradient centrifugation. The choice of method depends on factors like semen quality, intended use, and practical considerations. Density gradient is best for separating motile sperm from debris but is more time-consuming than direct swim-up. The goal is to recover high-quality sperm with minimal processing time and damage. Practical issues like sample volume and number of patients may also influence the choice of preparation technique.
This document discusses various sperm preparation methods used for intrauterine insemination (IUI). It describes the simple wash method, swim-up method, and density gradient centrifugation method. For each method, it provides the steps, including centrifugation speeds and times. It also covers preparing samples for retrograde ejaculation and samples from HIV-infected patients. The goal of sperm preparation is to select motile sperm and remove other seminal constituents like debris to maximize the chances of fertilization during IUI.
ICSI as it is presently performed is far from an ideal solution because the selection of sperm is based on the judgement of an embryologist, who is looking for the most normal appearing sperm available.
This document discusses various sperm preparation techniques used prior to assisted reproductive technologies. It begins by explaining the reasons for processing sperm samples, such as removing components that could prevent pregnancy and selecting morphologically normal, motile sperm free of debris. Several migration-based techniques are described, including swim-up, density gradient centrifugation, and magnetic activated cell sorting. Glass wool filtration and zeta potential techniques are also covered. The document discusses preparation of epididymal and testicular sperm samples. It concludes by outlining methods for preparing sperm from retrograde ejaculation samples.
This document summarizes key aspects of human oogenesis and oocyte quality assessment. It describes the process of oogenesis, including formation of follicles in the ovaries and maturation of oocytes. It also discusses morphological features used to assess oocyte quality, such as the cumulus-oocyte complex, nuclear maturity, size and shape, the zona pellucida, polar body morphology, and perivitelline space. While some studies found correlations between certain morphological features and development outcomes, others found no significant correlations. Overall assessment of oocyte quality relies on evaluating multiple morphological characteristics.
This document discusses sperm sorting techniques for men with high sperm DNA fragmentation index. It begins by defining sperm DNA damage and fragmentation, then discusses causes and indications for testing. It describes different tests to measure sperm DNA fragmentation and diagnostic cut-off points. The objectives and limitations of sperm sorting are outlined. Various sperm sorting techniques are explained, including swim-up, density gradient, magnetic activated cell sorting (MACS), motile sperm organelle morphology examination (MSOME), and surface charge-based sorting using hyaluronan binding. Advanced techniques like MSOME and MACS aim to select sperm with intact DNA and normal morphology to improve fertility outcomes.
The document outlines the key procedures involved in ICSI lab work for a gynecologist, as presented by Aboubakr Elnashar from Benha University in Egypt. It discusses 8 main procedures: 1) semen preparation, 2) oocyte identification, 3) oocyte denudation, 4) oocyte assessment, 5) oocyte injection, 6) embryo selection, 7) embryo transfer, and 8) cryopreservation. For each procedure, it provides details on the aim, methods, and considerations. The document serves as a reference for best practices in ICSI lab work to optimize outcomes.
Sperm retrieval techniques - nuts and boltsSandro Esteves
This document summarizes a training program in assisted reproductive technology focusing on azoospermia. It discusses the differences between obstructive and non-obstructive azoospermia and various sperm retrieval techniques such as PESA, MESA, TESA, and TESE. Success rates of sperm retrieval are provided for different causes of azoospermia. Predictive markers for sperm retrieval success are also examined, such as hormone levels, testicular volume, histopathology, and microdeletions. Micro-TESE is highlighted as a promising surgical method for non-obstructive azoospermia.
Oocyte retrieval involves three key steps:
1) Anesthetizing the patient using either conscious sedation with a paracervical block or general anesthesia. 2) Guiding an ultrasound-monitored needle into each follicle to aspirate the follicular fluid and oocyte. 3) Using a suction pump set to 90-120 mmHg for mature follicles and 40-60 mmHg for immature follicles to aspirate the fluid and oocyte without damaging the cumulus-oocyte complex. Precautions like antibiotics and monitoring for bleeding are important to minimize risks of complications.
The document provides information about andrology laboratory services for male infertility evaluation and treatment. It discusses:
- Tests offered including semen analysis, specialized tests of sperm function and morphology, sperm processing for infertility treatments, and cryopreservation.
- Procedures for semen sample collection, transport, and analysis following WHO standards, including macroscopic examination of volume, pH, and microscopic examination of motility, concentration, vitality, and morphology.
- Uses of semen analysis to diagnose infertility issues, identify treatment options, and assess effectiveness of treatments like vasectomy reversal. Computer-assisted semen analysis is also discussed.
Sperm DNA Fragmentation in Male InfertilitySandro Esteves
This document summarizes a presentation on sperm DNA fragmentation (SDF) and male infertility. It discusses how SDF provides different information than routine semen analysis and is a better prognostic indicator. Elevated SDF is associated with infertility, poor assisted reproductive technology outcomes, and miscarriage. Several methods can assess SDF but differ in their ability to directly or indirectly measure damage. Lifestyle changes like reducing stress and smoking, treating underlying conditions, and using oral antioxidants can help lower SDF. Varicocele repair is also effective at reducing SDF levels in men with the condition.
Sperm DNA Fragmentation (Oxidative stress, DNA damage and apoptosis, Test, Techniques, Relation to other semen parameters, Relationship to leucocytes, Relation to ICSI outcomes, Clinical applications, significance and limitations)
in this lecture i tried to summarize the most important normal morphological features of oocyte \ Follicle( including process of oogenesis and female mammalian meiosis) then i tried to summarize abnormal oocyte morphology
This document discusses various surgical sperm retrieval techniques for assisted reproduction, including:
1) Percutaneous epididymal sperm aspiration (PESA) and microsurgical epididymal sperm aspiration (MESA) are used to retrieve sperm from the epididymis in cases of obstructive azoospermia.
2) Testicular sperm aspiration (TESA) and testicular sperm extraction (TESE) are used to retrieve sperm directly from the testes in cases of non-obstructive azoospermia or previous failed epididymal sperm retrieval.
3) Microsurgical testicular sperm extraction (Micro-TESE) uses an operating microscope to identify and extract semin
This document discusses methods for scoring embryos at different stages of development during in vitro fertilization (IVF). It describes parameters evaluated for pronuclear zygotes, cleaved embryos on days 2-3 of development, and blastocysts on days 4-5. Parameters include pronuclear morphology, blastomere number and symmetry, fragmentation, compaction and expansion. Scoring systems assign quality grades based on these parameters to select the most developmentally advanced embryo for transfer. Precise embryo examination and selection of the highest quality embryo can reduce multiple pregnancies resulting from IVF.
This document discusses sperm cryopreservation, including the aims, techniques, factors affecting results, and future issues. The key points are:
- Sperm cryopreservation preserves sperm cells at sub-zero temperatures for future use, such as for fertility treatments. Slow freezing and rapid freezing are two common techniques.
- Factors like cryoprotectants, cooling/thawing rates, and semen quality can impact sperm survival after thawing. Semen preparation before freezing may improve outcomes.
- While some studies found cryopreservation does not affect reproductive success rates with ICSI, its effects on sperm DNA integrity are still unclear and require more research. Proper cryopreservation protocols aim to minimize DNA damage
Human spermatozoa can tolerate a range of temperature. They are not very sensitive to damage caused by cooling possibly because of high membrane fluidity which is used as a technique to preserve spermatozoa in adverse conditions. cryopreservation technology has been a boon in every aspect of infertility & ART practice.
Recent advances in assisted reproductive technology include:
1. The 1978 birth of Louise Brown, the first "test-tube baby", using in vitro fertilization without ovarian stimulation.
2. Developments like intracytoplasmic sperm injection (ICSI) and preimplantation genetic diagnosis (PGD) that have improved treatment options for male factor infertility and genetic disorders.
3. Continued research on techniques such as cryopreservation of eggs/embryos, stem cell therapy, and cloning that could further advance reproductive medicine if proven successful and safe.
PPT-Embryo grading and ART Summary.pptxKajal530634
Embryo grading is important in IVF to select good quality embryos for transfer based on developmental rate and morphology. The most followed grading systems are Gardner and Istanbul consensus, which assess embryos daily from fertilization to blastocyst stage based on criteria like cell number, size, and fragmentation. Good quality embryos with early cleavage and cell number on day 2 often develop into good blastocysts. Donor oocyte and sperm criteria and screening are also outlined to follow regulations. Oocyte donors can donate up to 7 oocytes only once in their lifetime from age 23-35.
Media is used in IVF to keep cells wet, feed them, and control the environment. There are different types of media for gametes, fertilization, cleavage, and blastocyst stages. While studies have compared various media formulations, no clear treatment effect has been found on clinical outcomes like live birth or ongoing pregnancy rates. Optimal media aims to mimic the natural embryo environment with constant temperature, pH, and avoidance of contaminants.
This document provides an overview of IVF and ICSI procedures. It discusses that IVF involves fertilizing eggs with sperm in a lab dish, then transferring embryos into the uterus. ICSI is used for severe male factor infertility and involves injecting a single sperm into each egg. Both aim to increase the chances of fertilization. The document outlines the steps of ovarian stimulation, egg retrieval, sperm preparation, fertilization, embryo culture, and embryo transfer.
Sperm DNA Fragmentation from the Male Infertility Specialist's PerspectiveSandro Esteves
This document summarizes a presentation on sperm DNA fragmentation from the perspective of a male infertility specialist. It discusses the relationship between sperm DNA fragmentation and infertility, methods for assessing sperm DNA fragmentation, and strategies for managing elevated sperm DNA fragmentation levels. The key points are that sperm DNA fragmentation provides different and more prognostically useful information than a semen analysis, is mainly caused by oxidative stress during sperm transit through the epididymis, and is associated with infertility, poor assisted reproductive technology outcomes, and miscarriage.
Clinical Utility of Sperm DNA Fragmentation Testing in Male Infertility Treat...Sandro Esteves
1) Sperm DNA fragmentation (SDF) testing provides important information about male fertility beyond conventional semen analysis.
2) Higher levels of SDF are associated with lower rates of pregnancy and live birth with infertility treatments like IUI and IVF/ICSI. It can also increase the risk of miscarriage.
3) Validated tests for SDF include the sperm chromatin dispersion test and TUNEL assay. Research is ongoing to improve testing methods and interpretation of results to help guide patient care.
This document discusses azoospermia, which is defined as the absence of sperm in the ejaculate. It identifies the main categories for the etiology of azoospermia as pre-testicular, testicular, and post-testicular. For patients presenting with azoospermia, the evaluation aims to determine the cause and includes medical history, physical exam, hormone levels, imaging, and potentially genetic testing. The diagnosis and treatment approach differs depending on factors like ejaculate volume, hormone levels, and results of further testing.
Antisperm antibody, presentation task in Infertility class. Our study program is Andrology, Medical Faculty, Airlangga University.
Visit us in:
Andrologi FK UNAIR: http://spesialis1.andrologi.fk.unair.ac.id/
FK UNAIR: http://fk.unair.ac.id/
UNAIR: http://unair.ac.id/
This document discusses poor ovarian responders in assisted reproductive technology treatment. It defines poor ovarian response based on the Bologna criteria of fewer than 3 oocytes retrieved with conventional stimulation. Poor responders make up 9-24% of IVF patients. The document outlines limitations of the Bologna criteria and introduces the POSEIDON classification system, which categorizes patients based on age, ovarian reserve markers, and ovarian response. It discusses markers for predicting poor response and reviews strategies for managing poor responders, including use of recombinant FSH, increasing FSH dosage, adding recombinant LH, and DHEA supplementation.
This document provides information about azoospermia, including definitions, types, causes, and sperm retrieval techniques. It defines azoospermia as the absence of sperm in the ejaculate. There are two main types - obstructive azoospermia, which is due to blockages, and non-obstructive azoospermia, which is due to testicular failure. Various sperm retrieval techniques are described that can be used depending on the type of azoospermia, including microsurgical epididymal sperm aspiration, percutaneous epididymal sperm aspiration, and testicular sperm extraction. Complications of the procedures include hematoma, infection, and testicular fibrosis.
The document outlines the key considerations for setting up an ART lab, including location, equipment, consumables, procedures, and quality control. The lab must be in a safe, pathogen-free environment with stable temperature and air quality. Critical equipment includes incubators, laminar flow hoods, micromanipulators, medical refrigerators, and liquid nitrogen tanks. Consumables must be established brands and not expired. Procedures must be documented and strictly followed. Quality control such as temperature monitoring is crucial.
This document discusses semen analysis, which plays a key role in evaluating male infertility. It describes the normal parameters for semen volume, pH, liquefaction time, viscosity, sperm concentration, motility, morphology, and viability. The roles of the prostate, seminal vesicles, and other glands in contributing to the semen are outlined. Standard procedures for semen collection and analysis are provided, along with causes of abnormal results and limitations of semen analysis. Advanced computer-assisted techniques for further evaluating sperm motility and function are also mentioned.
This document provides an overview of semen analysis. It discusses the indications, gross examination, microscopic examination including motility, morphology, viability and count. It also covers chemical examinations like fructose and acid phosphatase tests. Immunological assays for antisperm antibodies and microbiological assays are summarized. Key sperm function tests like sperm penetration and hypoosmotic swelling tests are mentioned. The document concludes with an overview of semen cryopreservation.
Oocyte retrieval involves three key steps:
1) Anesthetizing the patient using either conscious sedation with a paracervical block or general anesthesia. 2) Guiding an ultrasound-monitored needle into each follicle to aspirate the follicular fluid and oocyte. 3) Using a suction pump set to 90-120 mmHg for mature follicles and 40-60 mmHg for immature follicles to aspirate the fluid and oocyte without damaging the cumulus-oocyte complex. Precautions like antibiotics and monitoring for bleeding are important to minimize risks of complications.
The document provides information about andrology laboratory services for male infertility evaluation and treatment. It discusses:
- Tests offered including semen analysis, specialized tests of sperm function and morphology, sperm processing for infertility treatments, and cryopreservation.
- Procedures for semen sample collection, transport, and analysis following WHO standards, including macroscopic examination of volume, pH, and microscopic examination of motility, concentration, vitality, and morphology.
- Uses of semen analysis to diagnose infertility issues, identify treatment options, and assess effectiveness of treatments like vasectomy reversal. Computer-assisted semen analysis is also discussed.
Sperm DNA Fragmentation in Male InfertilitySandro Esteves
This document summarizes a presentation on sperm DNA fragmentation (SDF) and male infertility. It discusses how SDF provides different information than routine semen analysis and is a better prognostic indicator. Elevated SDF is associated with infertility, poor assisted reproductive technology outcomes, and miscarriage. Several methods can assess SDF but differ in their ability to directly or indirectly measure damage. Lifestyle changes like reducing stress and smoking, treating underlying conditions, and using oral antioxidants can help lower SDF. Varicocele repair is also effective at reducing SDF levels in men with the condition.
Sperm DNA Fragmentation (Oxidative stress, DNA damage and apoptosis, Test, Techniques, Relation to other semen parameters, Relationship to leucocytes, Relation to ICSI outcomes, Clinical applications, significance and limitations)
in this lecture i tried to summarize the most important normal morphological features of oocyte \ Follicle( including process of oogenesis and female mammalian meiosis) then i tried to summarize abnormal oocyte morphology
This document discusses various surgical sperm retrieval techniques for assisted reproduction, including:
1) Percutaneous epididymal sperm aspiration (PESA) and microsurgical epididymal sperm aspiration (MESA) are used to retrieve sperm from the epididymis in cases of obstructive azoospermia.
2) Testicular sperm aspiration (TESA) and testicular sperm extraction (TESE) are used to retrieve sperm directly from the testes in cases of non-obstructive azoospermia or previous failed epididymal sperm retrieval.
3) Microsurgical testicular sperm extraction (Micro-TESE) uses an operating microscope to identify and extract semin
This document discusses methods for scoring embryos at different stages of development during in vitro fertilization (IVF). It describes parameters evaluated for pronuclear zygotes, cleaved embryos on days 2-3 of development, and blastocysts on days 4-5. Parameters include pronuclear morphology, blastomere number and symmetry, fragmentation, compaction and expansion. Scoring systems assign quality grades based on these parameters to select the most developmentally advanced embryo for transfer. Precise embryo examination and selection of the highest quality embryo can reduce multiple pregnancies resulting from IVF.
This document discusses sperm cryopreservation, including the aims, techniques, factors affecting results, and future issues. The key points are:
- Sperm cryopreservation preserves sperm cells at sub-zero temperatures for future use, such as for fertility treatments. Slow freezing and rapid freezing are two common techniques.
- Factors like cryoprotectants, cooling/thawing rates, and semen quality can impact sperm survival after thawing. Semen preparation before freezing may improve outcomes.
- While some studies found cryopreservation does not affect reproductive success rates with ICSI, its effects on sperm DNA integrity are still unclear and require more research. Proper cryopreservation protocols aim to minimize DNA damage
Human spermatozoa can tolerate a range of temperature. They are not very sensitive to damage caused by cooling possibly because of high membrane fluidity which is used as a technique to preserve spermatozoa in adverse conditions. cryopreservation technology has been a boon in every aspect of infertility & ART practice.
Recent advances in assisted reproductive technology include:
1. The 1978 birth of Louise Brown, the first "test-tube baby", using in vitro fertilization without ovarian stimulation.
2. Developments like intracytoplasmic sperm injection (ICSI) and preimplantation genetic diagnosis (PGD) that have improved treatment options for male factor infertility and genetic disorders.
3. Continued research on techniques such as cryopreservation of eggs/embryos, stem cell therapy, and cloning that could further advance reproductive medicine if proven successful and safe.
PPT-Embryo grading and ART Summary.pptxKajal530634
Embryo grading is important in IVF to select good quality embryos for transfer based on developmental rate and morphology. The most followed grading systems are Gardner and Istanbul consensus, which assess embryos daily from fertilization to blastocyst stage based on criteria like cell number, size, and fragmentation. Good quality embryos with early cleavage and cell number on day 2 often develop into good blastocysts. Donor oocyte and sperm criteria and screening are also outlined to follow regulations. Oocyte donors can donate up to 7 oocytes only once in their lifetime from age 23-35.
Media is used in IVF to keep cells wet, feed them, and control the environment. There are different types of media for gametes, fertilization, cleavage, and blastocyst stages. While studies have compared various media formulations, no clear treatment effect has been found on clinical outcomes like live birth or ongoing pregnancy rates. Optimal media aims to mimic the natural embryo environment with constant temperature, pH, and avoidance of contaminants.
This document provides an overview of IVF and ICSI procedures. It discusses that IVF involves fertilizing eggs with sperm in a lab dish, then transferring embryos into the uterus. ICSI is used for severe male factor infertility and involves injecting a single sperm into each egg. Both aim to increase the chances of fertilization. The document outlines the steps of ovarian stimulation, egg retrieval, sperm preparation, fertilization, embryo culture, and embryo transfer.
Sperm DNA Fragmentation from the Male Infertility Specialist's PerspectiveSandro Esteves
This document summarizes a presentation on sperm DNA fragmentation from the perspective of a male infertility specialist. It discusses the relationship between sperm DNA fragmentation and infertility, methods for assessing sperm DNA fragmentation, and strategies for managing elevated sperm DNA fragmentation levels. The key points are that sperm DNA fragmentation provides different and more prognostically useful information than a semen analysis, is mainly caused by oxidative stress during sperm transit through the epididymis, and is associated with infertility, poor assisted reproductive technology outcomes, and miscarriage.
Clinical Utility of Sperm DNA Fragmentation Testing in Male Infertility Treat...Sandro Esteves
1) Sperm DNA fragmentation (SDF) testing provides important information about male fertility beyond conventional semen analysis.
2) Higher levels of SDF are associated with lower rates of pregnancy and live birth with infertility treatments like IUI and IVF/ICSI. It can also increase the risk of miscarriage.
3) Validated tests for SDF include the sperm chromatin dispersion test and TUNEL assay. Research is ongoing to improve testing methods and interpretation of results to help guide patient care.
This document discusses azoospermia, which is defined as the absence of sperm in the ejaculate. It identifies the main categories for the etiology of azoospermia as pre-testicular, testicular, and post-testicular. For patients presenting with azoospermia, the evaluation aims to determine the cause and includes medical history, physical exam, hormone levels, imaging, and potentially genetic testing. The diagnosis and treatment approach differs depending on factors like ejaculate volume, hormone levels, and results of further testing.
Antisperm antibody, presentation task in Infertility class. Our study program is Andrology, Medical Faculty, Airlangga University.
Visit us in:
Andrologi FK UNAIR: http://spesialis1.andrologi.fk.unair.ac.id/
FK UNAIR: http://fk.unair.ac.id/
UNAIR: http://unair.ac.id/
This document discusses poor ovarian responders in assisted reproductive technology treatment. It defines poor ovarian response based on the Bologna criteria of fewer than 3 oocytes retrieved with conventional stimulation. Poor responders make up 9-24% of IVF patients. The document outlines limitations of the Bologna criteria and introduces the POSEIDON classification system, which categorizes patients based on age, ovarian reserve markers, and ovarian response. It discusses markers for predicting poor response and reviews strategies for managing poor responders, including use of recombinant FSH, increasing FSH dosage, adding recombinant LH, and DHEA supplementation.
This document provides information about azoospermia, including definitions, types, causes, and sperm retrieval techniques. It defines azoospermia as the absence of sperm in the ejaculate. There are two main types - obstructive azoospermia, which is due to blockages, and non-obstructive azoospermia, which is due to testicular failure. Various sperm retrieval techniques are described that can be used depending on the type of azoospermia, including microsurgical epididymal sperm aspiration, percutaneous epididymal sperm aspiration, and testicular sperm extraction. Complications of the procedures include hematoma, infection, and testicular fibrosis.
The document outlines the key considerations for setting up an ART lab, including location, equipment, consumables, procedures, and quality control. The lab must be in a safe, pathogen-free environment with stable temperature and air quality. Critical equipment includes incubators, laminar flow hoods, micromanipulators, medical refrigerators, and liquid nitrogen tanks. Consumables must be established brands and not expired. Procedures must be documented and strictly followed. Quality control such as temperature monitoring is crucial.
This document discusses semen analysis, which plays a key role in evaluating male infertility. It describes the normal parameters for semen volume, pH, liquefaction time, viscosity, sperm concentration, motility, morphology, and viability. The roles of the prostate, seminal vesicles, and other glands in contributing to the semen are outlined. Standard procedures for semen collection and analysis are provided, along with causes of abnormal results and limitations of semen analysis. Advanced computer-assisted techniques for further evaluating sperm motility and function are also mentioned.
This document provides an overview of semen analysis. It discusses the indications, gross examination, microscopic examination including motility, morphology, viability and count. It also covers chemical examinations like fructose and acid phosphatase tests. Immunological assays for antisperm antibodies and microbiological assays are summarized. Key sperm function tests like sperm penetration and hypoosmotic swelling tests are mentioned. The document concludes with an overview of semen cryopreservation.
Semen examination for B.Sc. MLT studentsVamsi kumar
This document provides an overview of semen analysis. It discusses the indications, gross examination, microscopic examination including motility, morphology, viability and count. It also covers chemical examinations like fructose and acid phosphatase tests. Immunological assays for antisperm antibodies are described. Microbiological assays to check for infections are mentioned. Sperm function tests like penetration and swelling tests are listed. Finally, the document touches on semen cryopreservation and its indications.
This document discusses procedures for semen analysis, which is the first test performed to investigate male infertility. It describes how semen is examined physically, microscopically, chemically, and through immunological and microbiological assays. Tests evaluate semen volume, pH, motility, count, morphology, and the presence of fructose or acid phosphatase. Additional sperm function tests and cryopreservation are discussed. Semen donation is also summarized as a procedure to help individuals conceive.
Semen is produced in the male reproductive tract and contains sperm. A semen analysis evaluates several parameters of semen including volume, pH, sperm count, motility, viability, and morphology. It is used to assess male fertility and can help with infertility diagnosis. A semen sample is collected through masturbation after 2-7 days of abstinence and sent for analysis within an hour. The analysis involves examining the sample under a microscope to evaluate the number and health of sperm present. Abnormal results may indicate issues that could impact a man's fertility.
This document provides information about semen analysis, including its history, components, and examination process. It discusses key points such as:
- Sperm were first observed microscopically in 1677 by Antonie van Leeuwenhoek. Semen contains sperm cells and secretions from the testes, prostate, and seminal vesicles.
- A normal semen analysis involves examining samples macroscopically and microscopically to evaluate volume, pH, viscosity, sperm concentration, motility, morphology, and the presence of any abnormalities.
- Automated systems like the SQA-V Gold can analyze semen samples in about 75 seconds by measuring optical density, motility, and translating these
The correct answer is A. Harassment of the woman.
Section 498A of IPC defines cruelty as including any willful conduct which is of such a nature as is likely to drive the woman to commit suicide or to cause grave injury or danger to life, limb or health (whether mental or physical) of the woman; or harassment of the woman where such harassment is with a view to coercing her or any person related to her to meet any unlawful demand for any property or valuable security or is on account of failure by her or any person related to her to meet such demand.
Newer Modalities for Semen Testing | Male Infertility | Seeds Of InnocenceSOI Delhi
Male Infertility is a inability that causes pregnancy in a female fertile. Male infertility is commonly due to Low sperm Count. Soi provides best male infertility treatment in delhi, ghaziabad - India. For more information call us 9810350512
This document provides information about performing a semen analysis. It discusses the structures involved in semen production, contributions to semen volume, indications for a semen analysis, how to collect a semen sample, and the various tests that can be done on a semen sample including physical examination, microscopic examination of sperm count, motility, viability and morphology, immunologic analysis for antisperm antibodies, biochemical analysis of fructose, and sperm function tests. A normal semen analysis provides important information for investigating male factor infertility while abnormal results can help identify potential causes of reduced fertility.
The document discusses semen analysis, including:
1. It describes the different fractions that make up semen and their functions, such as nourishing sperm.
2. Semen analysis provides information on sperm production, male duct patency, accessory gland function, and ejaculation. It is used to evaluate male infertility, vasectomy effectiveness, and artificial insemination suitability.
3. Parameters examined in semen analysis include volume, pH, sperm count, motility, morphology, and presence of round cells. Tests are performed within a few hours of collection to assess these parameters.
4. Multiple factors can affect semen analysis results, so repeated testing is often needed for an accurate assessment
SELECTION OF SEMEN EJACULATES FOR FREEZINGAswiniSivan
Semen evaluation involves microscopic and biochemical analysis to assess sperm quality and fertility potential. Microscopic evaluation includes examining sperm motility, concentration, morphology, viability. Biochemical tests evaluate metabolic activity and membrane integrity. Modern techniques assess sperm function through penetration and DNA integrity assays. Together, these analyses provide a comprehensive view of semen quality and identify potential infertility issues.
This document provides information about performing a semen analysis, including why it is requested, how to examine various parameters of a semen sample, and what the results may indicate. It discusses examining semen for volume, viscosity, pH, motility, concentration, and morphology. Abnormal results in these parameters can help locate problems in the male reproductive system, such as infections, obstructions, or issues with the testes, pituitary, or prostate. Precise techniques are described for tasks like measuring volume, assessing motility and concentration using a hemocytometer, and evaluating samples under a microscope.
The document discusses investigations for male infertility. It lists various tests that may be conducted to evaluate male fertility including semen analysis, hormone levels, imaging tests, and genetic testing. Semen analysis is described as the primary test, examining parameters like volume, pH, sperm concentration, motility, morphology, and vitality. Abnormal results on semen analysis may indicate issues like low sperm count, poor motility, or abnormal morphology which could require further testing and treatment.
This document provides information about evaluating semen quality through various tests and examinations. It describes how to collect and transport semen samples, the normal characteristics of semen, and different abnormalities that can be detected through physical, microscopic, biochemical and other analyses of semen samples. The goal of semen analysis is to assess male fertility and detect any issues that may be causing infertility.
The semen analysis is the most important test for male fertility. It examines semen volume, sperm count, motility, morphology, and other factors. Abnormal results may indicate infertility issues. Hormone tests of blood and urine examine reproductive hormone levels which are important for ovulation. Ultrasound can check the ovaries, uterus, and fallopian tubes for issues like endometriosis that impact fertility. Other tests include hysterosalpingography which uses dye and x-rays to examine the fallopian tubes for blockages. Together these tests provide information to diagnose and treat fertility problems.
This document provides information about evaluating abnormalities in a semen analysis panel, including:
- The indications, sample collection/transport procedures, and normal ranges for semen volume, pH, motility, concentration, morphology, and other tests.
- How to interpret abnormalities in these parameters, such as low/high volume, pH, motility, oligospermia/azoospermia, teratozoospermia and their potential causes.
- Quality control procedures like repeat testing, and transient defects that could affect initial semen analysis results.
Four clinical cases are then presented to demonstrate applying this evaluation and interpretation of semen analysis results.
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2. sperm is a male reproductive cell, which is motile.
It is 60µm long and 2-5µm wide and contains 23 set
of chromosomes
3. ATTRIBUTES OF NORMAL SPERMATOZOA
1-Plasma membrane
2-Outer acrosomal membrane
3-Acrosome
4-Inner acrosomal membrane
5-Nucleus
6-Proximal centriole
7-Rest of the distal centriole
8-Thick outer longitudinal
fibers
9-Mitochondrion
10-Axoneme
11-Anulus
12-Ring fibers
A-Head, B-Neck, C-Mid
piece, D-Principal piece, E-
End piece
• TO EXHIBIT A NORMAL STRUCTURE
• PERSISTS MOTILE MOVEMENTS TO PASS THROUGH REPRODUCTIVE TRACT BARRIERS
• PROPERLY PACKAGED AND STABLE DNA
• MAINTAIN FUNCTIONAL METABOLIC PATHWAYS (RECEPTOR PROTIENS,
ACROSOMAL ENZYMES, SURVIVAL PROTEINS TO MAINTAIN IN FEMALE TRACT )
4. SFTs are the keystones of male
infertility evaluation
“In vivo” and “in vitro” conditions
sperm functions are same
The fertility will be decided not
only with the number but also the
functional competence of sperms
5. SPERM FUNCTION TESTS
Semen analysis is an essential component when
assessing male reproductive capacity
Semen analysis incompletely provides a
comprehensive assessment of male fertility potential
Sperm function tests adds important data regarding
sperm ability for fertilization and impact in post
embryo development
Use of SFT in conjunction with semen analysis
contributes more efficient treatment strategy
6. SO, WHAT ARE SPERM FUNCTIONS?
TO BE PRESENT IN AN EJACULATE ?
TO BE IN A GOOD NUMBER?
TO BE MOTILE ?
HAVE A GOOD MOTILE SPEED?
NAAAAAAAAAHHHHHHHH…….
7. SPERM FUNCTIONS ARE
TO FERTILIZE AN OOCYTE
DELIVER INTACT DNA INTO OOCYTE
ACHIEVE HEALTHY PREGNANCY
8. Advent of sperm function tests
Patient demand derived innovation
Patient satisfactory treatment approaches
Detailed assessment in Sperm assessments to rule
out cause
Translation of benchtop to bedside
9. SPERM FUNCTIONS
BASIC FUNCTION IS TO FERTILIZE AN OVUM AND
CONTRIBUTES CHROMOSOMES
sperm functions including their potential for movement, cervical
mucus penetration,
capacitation, zona recognition, the acrosome reaction and
sperm-oocyte fusion
19. Motility
Microscopic examination
Computer assisted semen analyzers (CASA)
Advantages
High accuracy
Quantitative information on the kinematic parameters of
spermatozoa (forward progression and hyper activated
motility, characteristic of capacitated cells)
Utility
Productivity of testis
Patency of GT
Substantial motility reveals severe infection and
inflammatory reactions in RT
Abnormal sequence of ejaculation
Disorders of sex glands
Correlated with DNA fragmentation
20. Eosin and Nigrosin Procedure
1- Mix 50µl of semen with 2 drops of eosin
2- mix 3 drops of Nigrosin after 30 seconds
3- Prepare a slide after 30 seconds and leave it for dry
4- examine immediately after drying with oil immersion at 100x
Utility
Progressive motility < 40%
LRL-58%
21. Hypo-Osmotic Swelling (HOS) test
Procedure
1- Incubate 1ml of HOS solution for 5-10 minutes
2- mix 0.1 ml of semen with HOS Solution and incubate
again for 30 minutes
3- Make a wet preparation with 10-15µls of incubated
sample
4- Examine at 40x
Utility
Discover active sperms
The sample can be reused
22. BIOCHEMICAL TESTS
Poor-quality semen may result from testicular production
of abnormal spermatozoa, or from post-testicular
damage to spermatozoa in the epididymis or the
ejaculate from abnormal accessory gland secretions.
Secretions from accessory glands can be measured to
assess gland function
An infection can sometimes cause a decrease in the
secretion of these markers, but the total amount of
markers present may still be within the normal range.
23. BIOCHEMICAL TESTS
The amount of zinc, citric acid (Möllering & Gruber, 1966) or acid
phosphatase (Heite & Wetterauer, 1979) in semen gives a reliable
measure of prostate gland secretion, and there are good
correlations between these markers.
Secretory capacity of the seminal vesicles. Fructose in semen reflects
the secretory function of the seminal vesicles.
Secretory capacity of the epididymis. L-Carnitine, GPC and neutral
–glucosidase are epididymal markers used clinically. Neutral -
glucosidase has been shown to be more specific and sensitive for
epididymal disorders than L-carnitine and GPC (Cooper et al.,
1990a).
There are two isoforms of –glucosidase in the seminal plasma: the
major, neutral form originates solely from the epididymis, and the
minor, acidic form, mainly from the prostate.
24. MEASUREMENT OF ZINC
Spectrophotometric Assay
The volumes of semen and reagents can be
proportionally adjusted for spectrophotometers
using 3-ml or 1-ml cuvettes.
The lower reference limit for zinc is 2.4 mol per
ejaculate (Cooper et al., 1991 and unpublished
data from TG Cooper).
25. MEASUREMENT OF FRUCTOSE
Fructose is an energy source for sperm motility,
Azoospermia negative fructose results may indicate
absence of SV/VD or any obstruction
Can be assessed via both ways qualitative &
quantitative
Quantitatively,
The lower reference limit for fructose is 13 mol per
ejaculate
26. MEASUREMENT OF FRUCTOSE
Fructose testing should be considered for patients
with azoospermia, low volumes
Determined by heating semen in a strong acid in the
presence of resorcinol. Fructose gives red color
upon heating.
Low fructose in semen is characteristic of ejaculatory
duct obstruction, bilateral congenital absence of the
vas deferens
27. MEASUREMENT OF GLUCOSIDASE
Seminal plasma contains both a neutral -glucosidase
isoenzyme, which originates in the epididymis, and
an acid isoenzyme contributed by the prostate.
The lower reference limit for neutral -glucosidase is
20 mU per ejaculate
28. CASA
(COMPUTER AIDED SEMEN ANALYSIS)
Until recently, it was not feasible to measure sperm
concentration by computer aided sperm analysis (CASA)
because of difficulties in distinguishing spermatozoa from
particulate debris (ESHRE, 1998).
Several manufacturers produce CASA systems. These
machines are capable of measuring sperm motility and
kinematics, and some can also be used to estimate sperm
concentration.
CASA, including assessment of motility, concentration and
morphology, has two advantages over manual methods: it
has high precision and it provides quantitative data on the
kinematic parameters of spermatozoa (forward progression,
hyperactive motility)
29. CASA
(COMPUTER AIDED SEMEN ANALYSIS)
Some studies have suggested that CASA estimates
of concentration and movement characteristics of
progressively motile spermatozoa are significantly
related to fertilization rates in vitro and in vivo, as
well as to time to conception
Many factors affect the performance of CASA
instruments, e.g. sample preparation, frame rate,
sperm concentration and counting-chamber depth
32. Sperm Antibodies
Because mature spermatozoa are formed after
puberty, they can be recognized as foreign protein by
the male immune system. If there is a breach in this so-
called “blood–testis barrier,”
These antibodies can then come into contact with the
sperm and may cause them to clump.
There are several tests currently employed for detecting
the presence of sperm antibodies.
The two most common are mixed agglutination
reaction (MAR) and the Immunobead binding test.
33. Mixed Agglutination Reaction (MAR)
This test is performed by mixing semen, IgG- or IgA-
coated latex beads or red blood cells, and IgG or IgA
antiserum on a microscope slide. The slides are
incubated and observed at 400× magnification. At
least 200 sperm are counted. If antibodies are present,
the sperm will form clumps with the coated latex beads
or coated red blood cells. If antibodies are absent, the
sperm will swim freely.
The WHO considers a level of binding of ≥50% to be
clinically significant. This test is used only for detection
of direct antibodies in men, and is not specific for the
location of bead attachment to the sperm.
34. The Immunobead Binding Test
This test is performed by combining IgG- or IgA-coated
latex beads and washed sperm on a slide. The sperm
must be removed from the seminal plasma by washing
the sample with media plus bovine serum albumin (BSA).
The presence of human protein on the surface of the
sperm interferes with the binding of the immunobeads
to the sperm, and thus may mask a positive result. After
washing, the sperm is placed on a slide with IgG- or
IgA-coated latex beads and is read at 200× or 400×
magnification. If antibodies are present, the small
beads will attach directly to the sperm.
The level of binding of ≥50% is commonly considered
to be clinically significant
35. SPERM ACROSOME
The acrosome is an intracellular organelle, similar to a
lysosome, which forms a cap-like structure over the
apical portion of the sperm nucleus (36). The acrosome
contains multiple hydrolytic enzymes, including
hyaluronidase, neuraminidase, proacrosin,
phospholipase, and acid phosphatase, which, when
released, are thought to facilitate sperm passage
through the cumulus mass, and possibly the zona
pellucida.
In fact, only acrosome-reacted sperm is capable of
penetrating the zona pellucida, binding to the
oolemma, and fusing with the oocyte
36. SPERM ACROSOME
This has led to the necessity for the development of
biochemical markers for the acrosome reaction.
Contemporary assays for the determination of
acrosomal status employ fluorescent plant lectins or
monoclonal antibodies, which can be detected much
more easily with fluorescence microscopy.
one of the predominant enzymes that is present in
the acrosome is proacrosin. The enzymatic action of
acrosin is not necessarily correlated to the presence
of an intact acrosome
37. HEMI-ZONA BINDING ASSAY
a significant correlation between tests of sperm–zona
pellucida binding and subsequent fertilization in ART
the hemizona assay (HZA) employs sperm and
nonviable oocytes in an in vitro assessment of
fertilization
This test assesses the ability of sperm to bind to the
zona itself. Although the HZA is relatively expensive,
labor intensive, and difficult to perform, there are some
data that suggest that the HZA may help to identify
individuals with a poor prognosis for success with ART
38. HEMIZONA BINDING ASSAY
oocytes that failed to fertilize during an ART
procedure are bisected, and then sperm from a
proven fertile donor (500,000/mL) is added to one
hemizona, while sperm from the subject male is
added to the other hemizona. Following a four-hour
incubation, each hemizona is removed and pipetted
in order to dislodge loosely attached sperm. A
comparison or hemizona index (HZI) is then
calculated
39. HEMI-ZONA BINDING ASSAY
Unlike several other tests of sperm function,
a cutoff value
(35%)
has been identified as a predictor of IVF success.
40. MANNOSE BINDING ASSAY
Another test has been developed in order to assess
the ability of sperm to bind to the zona.
involves the recognition by a sperm surface receptor
of a specific complementary receptor on the surface
of the zona pellucida.
This zona receptor appears to be a glycoprotein,
the predominant sugar moiety of which is mannose
In vitro assays in which labeled probes of mannose
conjugated to albumin are co-incubated with semen
specimens allow for the differential staining of
sperm
invites further study
42. PICSI- Physiological ICSI
In the process of fertilization, the sperm bind
to the hyaluronic acid (HA) present in the
cumulus oophorus by HspA2 protein, with
hyaluronidase activity that induces acrosome
reaction.
The PICSI device, a dish similar to ICSI dish,
contains microdots of hyaluronan hydrogel
which needs to be hydrated by media before
ICSI.
Selectively binding of sperm to the culture
dish shows:
More viability,
Non DNA-fragmented, and
with lower rates of aneuploidies.
43. Selection of HA binding spermatozoa
Incubation, RT, 10
min
Add sperm to the hyaluronan
microdot
Gentle aspiration of a bound
spermatozoa
After 15 min, the proportion of hyaluronan-bound spermatozoa, that exhibited vigorous tail
beating, were assessed versus the unbound motile spermatozoa
45. ASSAYS FOR SPERM DNA INTEGRITY
The most current area of investigation into sperm
function involves the assessment of sperm DNA
integrity
Sperm chromatin has been demonstrated to be
packaged very differently from chromatin in
somatic cells.
As there are many ways in which this DNA
organization or the sperm chromatin itself can be
damaged,
47. ASSAYS FOR SPERM DNA INTEGRITY
SPERM KARYOTYPING
SPERM FISH ANALYSIS
SPERM DNA FRAGMENTATION TESTS
48. INDICATIONS FOR SPERM DNA ASSESSMENT
Unexplained infertility
Repeated ART Failures
Recurrent Pregnancy Losses
Prediction of ART outcomes
Assessment of DNA/Genetic Integrity
- Post Chemotherapy
- Advanced Paternal Age
49. DETECTION OF SPERM DNA DAMAGE
Strand breaks in DNA
Strand breaks in DNA
Susceptibility to
Denaturation
50. OTHER TESTS FOR
DETECTION OF SPERM DNA DAMAGE
Strand breaks in DNA (incorporation of probes at site of damage)
DNA breakage detection FISH (DBD-FISH) (potential DNA damage)
In situ nick translation (ISNT) (real DNA damage)
8-Hydroxydeoxy-guanosine (8-OHdG) (real DNA damage)
Susceptibility to denaturation in an acidic solution
Halosperm test (potential DNA damage)
Chromatin condensation
Aniline blue stain
Toluidine blue stain
ChromomycinA3 stain
51. Sperm Chromatin Dispersion Assay
sperm are immersed in an agarose matrix on a slide,
treated with an acid solution to denature DNA that contains
breaks, and
then treated with lysis buffer to remove membranes and
proteins.
The agarose matrix allows working with unfixed sperm on a
slide in a suspension-like environment.
Removal of nuclear proteins results in nucleoids with a
central core and a peripheral halo of dispersed DNA loops.
Sperm nuclei with elevated DNA fragmentation produce
very small or no halos of DNA dispersion, whereas those
sperm with low levels of DNA fragmentation release their
DNA loops forming large halos.
53. SPERM CHROMATIN STRUCTURE ASSAY
The SCSA measures the level of DNA
fragmentation after denaturation by using
acridine orange staining and flow cytometry.
Double-stranded (native) DNA stains green
and single stranded (denatured) DNA stains
red.
SCSA also detects immature sperm that have
increased histones rather than protamines.
54. SPERM CHROMATIN STRUCTURE ASSAY
• SCSA is an accurate and
reproducible test for
measuring sperm DNA
damage
• Test requires a highly skilled
staff, expensive equipment
which limits its widespread
use
55. TUNEL Assay
(Terminal deoxytransferase-mediated dUTP nick end-labeling)
TUNEL is a direct method to detect single and
double strand breaks in the DNA
Detects DNA strand breaks resulting from apoptotic
signaling cascades
TUNEL can be used in a clinical setting using
flowcytometer.
56. TUNEL Assay
(Terminal deoxytransferase-mediated dUTP nick end-labeling)
This single-step staining method labels
DNA breaks with fluorescein
isothiocyanate (FITC)-dUTP followed
by flow cytometric analysis.
TUNEL utilizes a template-
independent DNA polymerase called
TdT which non-preferentially adds
deoxyribonucleotides to 3´-hydroxyl
(OH) single- and double-stranded
DNA.
dUTP is the substrate that is added
by the TdT enzyme to the free 3´-OH
break-ends of DNA
57. COMET Assay
Comet assay (single cell gel electrophoresis) commonly
used to detect extent of DNA damage in spermatozoa
with exposure to an electric field under neutral or
alkaline conditions
Damage is measured by measuring displacement
between genetic material of nucleus „comet head‟ and
resulting tail
58. COMET Assay
Embedding of sperm in agarose gel
Lysing of cells and decondensation of DNA
Unwinding of DNA
Separation and staining of DNA fragments
Image and statistical analysis
Quantifies actual damage in individual sperm
COMET images showing
1. Damaged
2. Undamaged DNA
59. UTILITY OF SDF TESTS
SDF is mainly oxidative stress mediated
Elevated SDF is associated with infertility, Poor ART
outcomes and recurrent miscarriages
SDF gives different information than routine semen
analysis and have better prognostic value
≤ 15% DFI EXCELLENT TO GOOD SPERM DNA INTEGRITY
> 15% TO < 25% GOOD TO FAIR SPERM DNA INEGRITY
≥ 25% TO <50% FAIR TO POOR SPERM DNA INEGRITY
≥ 50% VERY POOR SPERM DNA INEGRITY
63. Utilities of SFTs
Diagnosis of infertility
Selection of preparation
Selection of insemination method
Selection of sperms for Micromanipulation
Reduce embryonic development abnormalities
Give an overview of overall outcome