Semen Analysis Examination – Overview, Procedure, and Interpretation

Semen Analysis Examination – Overview, Procedure, and Interpretation

This presentation provides a detailed overview of Semen Analysis, an essential diagnostic test in the evaluation of male fertility and reproductive health. It covers the purpose, collection methods, macroscopic and microscopic parameters, and interpretation of results according to WHO guidelines.

Key Highlights:

Definition and importance of semen analysis

Steps for semen sample collection and precautions

Physical characteristics: volume, color, pH, viscosity, liquefaction time

Microscopic findings: sperm count, motility, morphology, vitality, and presence of abnormal cells

Factors affecting semen quality

Normal vs. abnormal findings and clinical significance


This slide deck is designed for medical, nursing, and laboratory science students, as well as healthcare professionals involved in reproductive diagnostics and laboratory analysis.

1. . Presented by Dr.Nayan Mehar MD 1 st year Department of RNVV ( Roga Nidan Evum Vikriti Vigyan ) SEMEN EXAMINATION

2. General Consideration Analysis of semen is one of the important parameters for gonodal function. A normal semen is practically a guarantee of normal androgenicity . Inadequacies on the part of male contribute to a significant percentage of infertility problems it is often requested before more complicated and expensive examination of female. In addition to infertility studies, as a part of forensic studies, semen examination may be requested to examine vaginal secretions or clothing stains for the presence of semen in alleged or suspected rape case. 2

3. Purpose of semen analysis Evaluation of infertility Routine follow up of patients undergone vasectomy Artificial insemination Examination of stored semen specimen ( May be in the case of a husband away from home for a long period and when the wife is undergoing complicated infertility therapy ) For men whose future fertility is threatened may be by the need of Radiotherapy or Chemotherapy in the treatment of cancer 3

4. Specimen collection Length of Abstinence 2-3 days is an adequate length of abstinence for the proper assessment of semen quality. Site of production of semen specimen Patient should be allowed to choose where he wishes to produce specimen. It may be at home or in a room of lab. or hospital which is quiet, secluded and which is guarantee total privacy. Time of production of semen specimen A semen specimen is best produced in the morning for an assessment of motility 6 hrs after production. The presence of any “deadline” for specimen production should be avoided. 4

5. Specimen collection (Precautions) It is very important to relax the patient and allow the production of the specimen to take place in surroundings which are the most comfortable both emotionally and physically for patient. The production of semen for analysis in a situation of stress can lead to inadequate ejaculation. This may lead to misdiagnosis towards the evaluation of infertility problem of a couple. The collection of semen specimen must be organized by experience staff. 5

6. Method of production of semen Specimen The production of semen by masturbation usually results in a specimen that is complete and uncontaminated For those patients who are reluctant to produce semen by masturbation, specimens may be collected by coitus interruptus . (Ordinarily condoms must never be used to collect semen specimens. Most condoms contain spermicidal powder which swiftly obliterates all semen motility) 6

7. Containers used for semen Specimen All patients should be provided with clean and dry wide mouth glass bottle or plastic container (50 ml) Detergents are highly toxic to sperm. Hence there should not be even trace of soap or detergent in the container Water is equally lethal to sperm (hence the container should be dry) There must not be a rubber lining to the lid as contact between the rubber lid and the semen may result in sperm death The container should be correctly labeled 7

8. Storage and transport of the Specimen Sperms are easily damaged by either excessive heat or excessive cold Semen specimen should not be left for any length of time in direct sunlight The semen specimen should be brought to the laboratory at close to body temperature It is best but not essential that the semen specimen is delivered to the laboratory within two hours of production 8

9. Number of Specimen for analysis A reasonable accurate evaluation of only the fertility status of the patient can usually be achieved by assessing at least three samples of semen collected on different days Great care must be taken both by the clinician and by the lab to ensure that semen specimens are an accurate reflection of the patient's fertility and not rendered invalid by stress, abnormal or incomplete ejaculation or by the action of physical or chemical damage during the transport to lab Crystals of spermicidal powder in condom (if it is used by the patient without caring for instructions) and that of talcum powder may be seen in semen as artifact. 9

10. Physical examination of semen Volume of semen and presence of spermatozoa Can be measured by using a 10 ml graduated measuring cylinder or a 10 ml graduated centrifuge tube Normal semen volume: 2-6 ml Volume as low as 1 ml and as high as 10 ml may be regarded as normal 10

11. Physical examination of semen Terms used to express Volume and presence of spermatozoa Aspermia : Means total absence of ejaculate (a rare phenomenon) Azoospermia : Absence of spermatozoa in semen Oligospermia ( Hypospermia ): A reduction in the volume of the ejaculate Hyperspermia : Increase in semen volume (a rare phenomenon) 11

12. Physical examination of semen Colour of the seminal fluid Normally pale grey-yellow opalescent fluid. The colour may change due to the following reason Presence of urine: may occur with bladder neck function disturbances and ejaculation. (it can be detected by consistency of semen and by uniferous odor of semen and by high urea content) Urine has a lethal effect on sperm and there may be poor motility or completely immotile sperm Presence of blood: may appear pink if in traces may appear bright red if amount is large Infection in seminal vesicle or prostate Trauma or malignancy of testis Prostate carcinoma in older 12

13. Physical examination of semen pH of semen Normal fresh semen is 7.9-8.1. May slowly fall on storage. Can be measured by using pH paper Liquefaction of semen: Semen is ejaculated in liquid form and it forms a gel like clot within 5-20 min. Determination of liquefaction ( Usually assessed visually) Unliquefied semen: forms a gel like coagulum Partially liquefied : May contain many small gel like clots Fully liquefied: No such clots are seen and semen appears completely fluid (Alfa-amylase is used to liquefy unliquefied semen) 13

14. Physical examination of semen Viscosity of semen Normal viscosity is defined as that which allow semen to be poured drop by drop out of a container It can be quantified by measuring the time taken to drop 1 drop of semen to leave a standard pipette. (Capillary tube of 10 cm length containing 0.1 ml of semen) Hyperviscid semen: May fail to coat the cervix and may easily drain out of the vagina after intercourse Reduction of viscosity : May be achieved by mixing the viscid semen with sperm free seminal fluid from another man. 14

15. Sperm count Visual assessment By using improved neubauer counting chamber Sperm counting based on laser principle This involves the use of laser light. From the deflection of a beam of laser light, a computer can determine not only the sperm concentration but also the sperm motility as well as the presence or absence of forward progression.

16. Expression of motility of Sperm Asthenozoospermia : an absence or a marked reduction in sperm movement Oligoasthenozoospermia : abnormalities of sperm movement associated with a reduction in sperm numbers. If a sperm is showing any movement, the sperm is said to be motile for the purpose of semen analysis, sperm motility must be recorded as the percentage motility i.E percentage of the total sperm that are showing movement. The number of motile sperm per ml is calculated as motile sperm as below: [Sperm conc./ml x % motility x semen volume/ 100]

17. Expression of motility of Sperm The estimation of percentage motility must be performed soon after the production of the semen sample and such assessment should be repeated at 3 hrs and 6 hrs of production. Gradation of Sperm movement (0 - 4 scale) Definition Description 0 - None An absence of forward progression 1 - Poor Weak or sluggish forward progression 2 - Moderate Defining forward progression 3 - Good Good forward movement with progression 4 - Excellent Vigorous rapid forward progression

18. Assessment of Morphology of Sperm The terms commonly used in relation to abnormalities in the morphology of spermatozoa Teratozoospermia : means the presence of an increase in the numbers of morphologically abnormal sperm in a sample Teratoasthenozoospermia : Describes the abnormal sperm showing poor or absent motility 18 Head abnormalities Tail abnormalities Marked reduction in size Coiled tails Marked increase in size Eccentrically inserted tails Presence of vacuoles within the head Hairpin deformities Tapering or pyriform head Duplicate or triplicate tails

19. Other Microscopic Abnormalities White blood cells : Present in small numbers in normal semen but presence of large numbers indicates inflammation. Epithelial cells: Presence in large numbers does not have pathological significance. May be present due to urethritis . Red blood cells: Not usually present in semen. Presence indicates infection or possibility of malignant disease within genital tract. Protozoa & Bacteria: Presence indicates infection Most common protozoa is Trichomonas and visible bacteria may be Staphylococcus and streptococci. Particulate matter Crystals of endogenous substances such as spermine may be present. The exogenous substances may be spermicidal powder or talcum powder .   19

20. Important biochemical investigations Determination of semen fructose Important component of seminal vesicle provides energy source to sperm Absence of fructose results in completely immotile sperm. Determination of citric acid Also present in prostatic fluid and may also be used as marker of prostatic secretion. Determination of acid phosphatase Secreted by prostate gland and useful marker of prostatic function. Used by forensic lab as a test for presence of semen. Also as a test for presence of prostatic contribution in an ejaculate. 20

21. Interpretation of semen analysis Physical Examination Test Normal finding Abnormal finding & Clinical condition Colour Appearance Greyishwhite Viscid, opaque Increased turbidity may be associated with inflammatory process in some part of reproductive tract Viscosity Can be poured drop by drop Increased viscosity indicates poor invasion of cervical mucus after post coitus. Absence indiactes reduced cell contents Volume 2-5 ml Lesser amount may arouse suspicion of deficiency and premature weakening by vaginal acidity Liquefaction time 20-30 min Failure indicates infertility Congenital absence of seminal vesicle and vas deferens fails to coagulate

22. Interpretation of semen analysis Chemical examination Test Normal finding Abnormal finding & Clinical condition pH 7.2 – 7.8 Less than 7 are frequently associated with semen consisting largely of prostatic secretion due to congenital aplasia of the vas deferens and seminal vesicles. Fructose 150-300 mg/dl Disorders of seminal vesicle may lead to reduction in fructose concentration. Absence of fructose results in immobile sperm formation.

23. Interpretation of semen analysis Microscopic Examination Test Normal finding Abnormal finding & Clinical condition Sperm count 40-300 millions/ml Comp Absence: Azoospermia Reduced: Oligispermia Mumps, Orchitis , Prostatitis , Occlusion or absence of efferent ducts, Hypopituitarism , Oestrogen secreting tumor, Hypo-hyperthyroidism Motility after 2 hrs, 3 hrs, 6 hrs 60-95% Motile forms decrease by about 5%/hr after the 4 th hr following collection. Motility <60% may indicate infertility Abnormal forms 0-20% > 20% abnormal form indicate infertility Heads: Too small, Large, double, pointed Middle piece : absent, bifurcated or swollen Tail : Double, curved, rudimentary or absent

24. Interpretation Of Semen Analysis Other Findings Test Normal finding Abnormal finding & clinical condition Pus cells 1 – 2/ hpf Increased indicates inflammation due to infection in some part of reproductive system Infection of seminal vesicle Epithelial cells 1 – 2/ hpf Increased number: not significant Red blood cells Absent Tuberculosis of seminal vesicle, rupture of blood vessel, infection of prostate, Vit C deficiency Trichomonas Absent Trichomonas infection (motile flagellete with pus cells