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Yomna Mazid El-Hamd Assist. Lecturer

Conventional Vs Computerized Semen Analysis. By. Yomna Mazid El-Hamd Assist. Lecturer. Conventional Vs Computerized Semen Analysis. Semen Analysis. Semen analysis is the examination of freshly ejaculated seminal fluid.

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Yomna Mazid El-Hamd Assist. Lecturer

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  1. Conventional Vs Computerized Semen Analysis By Yomna Mazid El-Hamd Assist. Lecturer

  2. Conventional Vs Computerized Semen Analysis

  3. Semen Analysis • Semen analysis is the examination of freshly ejaculated seminal fluid. • It is a keystone in the clinical evaluation of the infertile male patient.

  4. Semen Analysis It includes the examination of : • Spermatozoa(concentration, count, motility, viability, and morphology). • Seminal fluid(volume, PH, viscosity, liquifaction, fructose, neutral a-glucosidase, zinc, citric acid, acid phosphatase) • Other cells present in semen (immature germ cells, WBCs, RBCs).

  5. Indications • Assessment of male fertility . • After a vasectomy to confirm the success of the procedure & absence of sperm. • In suspected case of rape, a microscopic exam for sperm is performed on vaginal swabs and clothes. (together with tests for acid phosphatase and prostate specific antigen to determine the presence of seminal fluid).

  6. Relation to fertility • The characteristics measured by semen analysis are only some of the factors insemen quality. • It is found that 30% of men with a normal semen analysis actually have abnormal sperm function. • Conversely, men with poor semen analysis results may go on to father children.

  7. Source of semen • Testes, epididymis, vas deference:0.1-0.2cc, contain sperms & represent 1% of volume. • Prostate:0.5-1.0cc, Acidic, watery & represent 15-30% of volume . • Seminal vesicles:1.0-3.0 cc, Gelatinous, fructose positive & represent 45-80% of volume . • Urethral and bulbourethral glands:0.1-0.2cc, Viscous & clear. • Complete ejaculate:2.0-5.0-cc & Liquefies in 20-25min.

  8. Semen Sample Collection Methods • The most common way to collect a semen sample is throughmasturbation, directing the sample into a clean cup. • A sample may also be collected during intercourse in a special type ofcondomknown as acollection condom.Collection condoms are made from silicone or polyurethane, aslatexis somewhat harmful to sperm. • A third option for collecting a sample is through(coitus interruptus)withdrawal. With this technique, the man removes his penis from his partner near the end of intercourse and ejaculates into a cup.

  9. Semen Sample Collection Methods • Finally, if a blockage in thevas deferensis suspected, semen can be taken directly from theepididymis.Such a collection is calledpercutaneous epididymal sperm aspiration)PESA). • Alternatively, the testicular tissueitself, instead of the sperm produced can be investigated. Then, the collecting method is called TESE.

  10. Semen Sample Collection Methods • The sample should be collected after a minimum of 48 hours and no longer than 7 days of sexual abstinence. • When the duration of abstinence is more than 7 days, sperm motility, i.e. the proportion of spermatozoa with rapid progressive motility, may decline. • If the duration of abstinence is <48h, sperm concentration may be reduced, but motility will probably not be affected.

  11. Semen Sample Collection Methods • Typically two to three semen analyses should obtained over a 3 month period prior making any final conclusion regarding sperm quality & quantity. • Recent febrile illness or exposure gonadotoxic agent may affect spermatogenesis for up to 3 months, therefore semen analysis has to be postponed.

  12. Semen Sample Collection Methods • The sample must kept at room temperature. • Examinations should be performed within one hour of sample collection. • Examinations should be performed after waiting about 30 minutes after ejaculation, to allow the semen to liquefy.

  13. WHO guidlines of semen analysis • In response to a growing need for the standardization of human semen, the WHO first published a Laboratory Manual For Examination of Human Semen in 1980. • The second,third&fourth editions, of the manual followed in 1987,1992 & 1999 respectively. • Over the past 30 years, the manual has been translated into a number of languages & recognized as providing global standards and used extensively by research and clinical laboratories throughout the world.

  14. WHO guidlines of semen analysis • Despite this success, it has become apparent that some recommendations from previous editions of the manual needed to be revised in light of new evidence. • Prompted by these considerations, WHO established the fifth edition in 2010.

  15. WHO criteria of semen analysis

  16. Gradation of sperm motility (WHO)-2010 • Grade 4 (A):Sperm with progressive motility. These are the strongest and swim fast in a straight line. • Grade 3(B): Spermwith non-linear forward motility. These also move forward but tend to travel in a curved or crooked motion. • Grade 2 (C):These have non-progressive motility because they do not move forward despite they move their tails. • Grade 1(D):These are immotile and fail to moveat all.

  17. WHO normal ranges of semen analysis

  18. Nomenclature for semen variable (WHO) • Normal semen quality Normospermic • No ejaculate Aspermia • Low volume Hypospermia • High volume Hyperspermia • No spermatozoa Azoospermia

  19. Nomenclature for semen variable (WHO) • Low spermatozoa conc. Oligozoospermia • Low spermatozoal motility Asthenozoospermia • Low normal morphology Teratozoospermia • Excessively high sperm conc., Polyzoospermia • WBCs in semen Pyospermia • RBCs in semen Heamatospermia • Non viable sperms (dead). Necrozospermia

  20. Globozoospermia • Globozoospermia (sperm with round heads), the Golgi apparatus is not transformed into the acrosome that is needed for fertilization.

  21. Cryptozoospermia: low sperm numbers: • If no spermatozoa are observed in the wet preparations, azoospermia can be suspected. • But repeated analysis & centrifugation may reveal apparently absent or hidden sperms. • Therefore, the definition of azoospermia should change and only be used if no spermatozoa are found in the sediment of a centrifuged sample.

  22. Distinction between leucocytes and spermatogenesis cells • The distinction between spermatogenesis cells and leucocytes is important particularly in cases of azoospermia. • In azoospermia, the presence of spermatogenesis cells indicates a testicular malfunction, while the presence of leucocytes in the absence of any spermatogenesis cells suggests that the azoospemia might be due to an obstuction problem. • Spermatogenesis cells present in semen are usually degenerating, and can sometimes be confused with leucocytes. In particular, multinucleated spermatids can be confused with polymorphonuclear leucocytes (PMN). • Staining of endogenous peroxidases present in PMN can help to distinguish between these two cell types.

  23. Semen analysis : Aspermia Aspermia: • Neuropathic failure of emission • Complete retrograde ejaculation

  24. Semen analysis : appearance Very clear: • Low sperm count. Brown: • Hematospermia.

  25. Semen analysis : Volume Low volume: • Incomplete collection. • Seminal vesicle agenesis. • Ejaculatory ducts obstruction. • Retrograde ejaculation. High volume: • Long periods of abstinence. • Varicocele.

  26. Semen analysis: Liquefaction & Viscosity The semen is ejaculated in liquefied state but quickly coagulate by the action of protein kinase secreted by the seminal vesicles. Proteolytic enzymes from the prostate liquefy coagulum in 20-25 minutes. ↑Liquefaction time ( > 30 min ): • Prostate dysfunction ↑Viscosity: • Prostate dysfunction

  27. Semen analysis : PH ↑PH • Prostate dysfunction. • Infection. ↓PH • Seminal vesicle dysfunction or agenesis. • Vas deferens agenesis. • Ejaculatory ducts obstruction. • Incomplete collection.

  28. Semen analysis and prostate dysfunction • Delayed liquefaction • Increased viscosity • Increased pH • Decreased Zinc concentration

  29. Semen analysis and seminal vesicles dysfunction • Low volume • Decreased fructose concentration • Decreased PH

  30. Semen Analysis Advantages: • It's noninvasive and inexpensive. • It's a brief test that produces fast and reliable results. Disadvantages: • The patient may be too embarrassed to masturbate and ejaculate in the doctor's office, and a sample from home may not be suitable for analysis because of exposure to heat or light.

  31. Semen Analysis • Standard semen analysis is a rather subjective technique andassociated with large inter-laboratory variation,which makes it virtually impossible to comparesperm motility assessments performed by different laboratories. • In an attempt to make the assessments of semen qualitymoreobjective and detailed, tools for computer-assisted semen analysis(CASA) have been developed.

  32. Computer Assisted Semen Analysis (CASA) • A CASA system is an automated, objective and standardized equipment that allows to assess all the parameters of sperm in a semen sample. • CASA are most often used for the assessment of sperm concentration and motility following the WHO criteria. • By use of CASA several specificmotility parameters in a more detailed manner such as velocity can be obtained.

  33. SQA-V • The SQA-V ( Sperm Quality Analyzer), is a high performance sperm analysis instrument used to test male fertility. • It combines electro-optics, computer algorithms and video microscopy to provide a precise and accurate 75 second automated semen analysis.

  34. SQA-V • The main reason the SQA-V is a growing instrument among the semen analysis community is due to the speed, accuracy, and precision to run a semen sample. • In addition, the SQA-V semen analysis eliminates inter-operator variables from the manual method. • The SQA-V still provides 16 clinical parameters including: Sperm Concentration, Rapid Progressive motility, Slow Progressive motility, Non-progressive motility, Immotility, %Normal Morphology, and more.

  35. ISAS • Integrated Semen Analysis Systemis a CASA system based on image analysis • ISAScan be considered as the most complete and easiest-to-use system in market • ISASanalyzes motility and concentration in more than 17 sperm parameters • ISASanalyzes automatically stained morphometry, giving 15 parameters and DNA fragmentation analysis.

  36. ISAS • Motility and concentration analysis give to the customers more tan 17 sperm parameters. • AlsoISASanalyzes automatically stained morphometry, giving 15 parameters and DNA fragmentation analysis. • ISAShas been developed to be used in several species, from classical species like human, boar or bull to new research species like cod or some small rodents

  37. Sperm Class Analyzer • SCA provides fast, accurate and objectively repeatable results. This would be impossible to attain using traditional (subjective) methods. This CASA system has the following modules: • SCA Motility & Concentration: The system provides immediately and objectively detailed results of motility and concentration. • SCA Morphology: Following a manual or automatic image capture, a precise morphological analysis of each spermatozoa is provided. • SCA DNA Fragmentation: Analysis of samples prepared for study of the DNA fragmentation provides detailed numerical data for each cell. • SCA Vitality: Automatically analyze the vitality of a sperm sample.

  38. IVOS • The IVOS is an Integrated Visual Optical System for sperm analysis.

  39. IVOS • The (IVOS) is found in hospitals, universities, IVF clinics, pharmaceutical companies, contract labs, reproductive toxicology labs, veterinary clinics and animal breeding facilities around the world. • The standard IVOS software may be used to analyze sperm of multiple species, with a specific program geared toward analysis of rat sperm. • The IVOS is unique in that it is the only CASA system that integrates the optical system within the unit, so that an external microscope is not needed.

  40. IVOS • A field of sperm are analyzed in just 0.5 second with a level of accuracy unobtainable by visual assessment. • Results calculated include count, concentration, motility, progressive motility, curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN), straightness (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF).

  41. CEROS • Also built upon the same analysis algorithms and software interface as the IVOS, theCEROSoffers the same level of accuracy and reliability for sperm analysis. • The CEROS is also compatible with both the Dimensions and Metrix morphology options and can be used for to analyze sperm from all species except rat.

  42. Advantages of CASA • The subsequent introduction of CASA in the laboratory routine not only improved accuracy on data retrieval and interpretation, but reduced as well the time required far the exam performance and the possibility of errors due to subjective evaluation of technicians. • The major problems whit CASA is represented by the very high cost of the instruments that suggests its use only in laboratories that perform a high number of exam.

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