1. Semen Semen is made up of fluid and of sperm.
The Fluid component is produced in various male reproductive organs;
- The seminal vesicles produces a slightly alkaline fluid which makes up 50% of the semen fluid (it contains citric acid , flavins , fructose and potassium , these substances provide nutritional support for the spermatozoa).
- The prostate gland contributes an acidic fluid containing acid phosphates , citric acid and proteolitic enzymes (including a clotting enzyme) , which makes up 20% of the semen fluid.
The rest is contributed by other male reproductive organs, such as the bulb urethral and urethral glands , which produce a fluid important in lubricating the male reproductive system.

2. Semen The Spermatozoa is formed in the testis .
The spermatozoa take over 70 days to develop and are produced solely in the testicles from a cell called a spermatogonium.
The spermatogonium divides to produce spermatocytes, which then develop into spermatids.
The spermatid develops its familiar tail and the cell gradually acquires the ability to move by beating its tail.
The spermatid eventually develops into a mature spermatozoan.
The fluid produced by the testicles contains several chemicals, but is particularly rich in testosterone
The process of spermatozoa formation is under hormonal control including Testosterone ,Leuteinizing hormone LH , and Follicle- stimulating hormone FSH.
Upon ejaculation the fluids from all these sources form the mixture semen.

3. Normal Semen Analysis Range
Normal Values ;
Volume ; 2.0 – 5.0 ml.
pH ;
Colour ; White , grayish white.
Viscosity; Viscous.
Liquefaction ; after 30 min.
Sperm concentration ; 20 – 250 million / ml.
Agglutination; None.
Motility ; > 60% progressively motile.
Penetration; > 30 mm.
Viability ; > 75% .
Normal Morphology ; > 70%.
Immature forms ; < 2%.
Leukocytes ; None – Occasional.

4. Normal Semen Analysis Range
Red Blood Cells ; None.
Epithelial Cells ; None – few.
Bacteria ; None.
Fructose ; 1+ to ( 300mg /dl)
Specimen Collection ;
It should be collected in a plastic or glass container , or in a special type of condom known as a collection condom. It should be performed after a h absence of sexual activity so it contains an accurate account of sperm count and viability.
The specimen should arrive at the laboratory as soon after collection as possible so that an accurate Liquefaction time can be reported.
During transportation it should be kept near body temperature.
The patient should be asked if any of the sample is lost since the highest concentration of sperm is in the first part of ejaculation.

5. Semen Analysis Semen Analysis involves; 1-Macroscopical analysis ;
A semen analysis evaluates certain characteristics of a male's semen and the sperm contained in the semen. It may be done while investigating a couple's infertility or after a vasectomy to verify that the procedure was successful.
Semen Analysis involves;
1-Macroscopical analysis ;
Aa) Liquefaction; As soon as the sample arrives it is observed for liquefaction
time beyond 60min is abnormal .
The semen is initially 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 minutes. Abnormal liquefaction may be caused by prostatic abnormalities, e.g. prostatitis. Increased viscosity may affect sperm motility.
b)Appearance; Semen normal can range from white , grey to light yellow.
The higher the flavin concentration in semen the darker the yellow colour.
The colour of semen might be affected by certain drugs.

6. Semen Analysis Brown or red coloured semen indicates blood.
A highly turbid semen usually contains leukocytes in large amounts indicating
infection of the reproductive tract infection.
c) Volume ; normal volume is 2-5ml , Low volume may indicate partial or complete blockage of the seminal vesicals, or that the man was born without seminal vesicles .
d)Viscosity; Normal semen is highly viscose and can be observed when measuring the volume . The increased viscosity is demonstrated by the formation of a string of fluid as the specimen is dispensed from the pipette.
2- Microscopic Analysis;
Sperm count (sperm concentration);
It can be done by automated
methods or manually by a hemocytometer.
Normal Count million / ml
Oligospermia ; sperm count less than
20 million / ml.

7. Semen Analysis Azospermia; complete absence of sperm.
Low count can be due to chromosomal disorders, ductal obstruction , drugs , radiation …… .
b) Motility; Fertilization of an ova is dependent on the ability of the sperm to reach and unite with it.
A small drop of liquefied semen is placed on a pre-warm slide and cover slipped , movement of sperms is observed under microscope.
It should be evaluated within 1h of specimen collection. At least 80% of the sperms should show forward progress in a normal semen sample.
C)Agglutination; It can be observed during motility observation. A true agglutination is when sperms clump head to head or tail to tail , which indicate anti-sperm Ab in semen which might cause infertility , and must be confirmed by specific immunological tests.
d)Viability; Determining whether the non-motile sperm is viable or not is important in establishing the cause of infwertility.
The membranes of dead sperms are damaged and can easily take up eosin stain.
The viable sperms do not allow the stain to penetrate leaving a colorless sperm.

8. Semen Analysis
d)Viability; Determining whether the non-motile sperm is viable or not is important in establishing the cause of infwertility.
The membranes of dead sperms are damaged and can easily take up eosin stain.
The viable sperms do not allow the stain to penetrate leaving a colorless sperm.

9. Semen Analysis Normally > 75% of sperms are viable.
e) Penetration; Even though the sperm might be viable and motile the male can still face infertility problems if the sperms are incapable of penetrating through the cervical mucus.
The semen sample penetration through bovine cervical mucus is observed and evaluated .
f) Morphology; Refers to the shape and size of the sperm. A normal sperm has an oval head and a tail times longer than the head. Abnormal sperm morphology can be a contributing factor to infertility. Abnormal sperm morphology can include defects in head or tail such as a double headed sperm , a kinked , a twisted tail, these abnormalities can impair the sperms ability to reach and fertilize the egg.
Causes of abnormal morphology include;
Congenital testicle abnormalities.
Fever
Illicit drug use.
If any abnormal morphology is seen the semen analysis should be repeated in four to two weeks to check if the changes are temporary or permenant. .

10. Morphology
Normal Morphology
Abnormal Morphology

11. Semen Analysis
3-Immunological Analysis; Once an Agglutination test is positive more specific tests should be performed to confirm the finding and to determine the Ab responsible.
The anti-sperm Ab can be present in both the serum and semen of the individual and some times women can develop Ab against partners sperms.Kibrick method is one of the common immunological methods used .It involves incubating fresh liquified semen with serum from the male involved or his female partner. Agglutination is observed macroscopically and titre recorded.
4-Chemical Analysis (such as); The purpose of the Fructose Test is to measure the amount of Fructose in human semen . Fructose in semen reflects the secretory function of seminal vesicles. Lower levels could indicate problems with the seminal vesicles.
Fructose + indole => heat/acid => complex which absorbs at wavelength of nmFructose level, a normal level of fructose is at least 3mg/ml

12. Semen Analysis
pH; a normal range of Acidic ejaculate (lower pH value) may indicate one or both of the seminal vesicles are blocked. A basic ejaculate (higher pH value) may indicate an infection .A pH value outside of the normal range is harmful to sperm.
5-Microbiology;
Urigenital infections caused by microrganisms are responsibl;e for 15% of male infertility cases ( some can cause anti-sperm antibody production).
Thus when suspected a semen sample is cultured and examined for microorganisms.