The semen analysis test is the most important infertility test. Semen analysis is the first step in the medical evaluation of male infertility.  The semen analysis will help determine whether there is a male factor involved in the couple's sub-fertility. If so, an evaluation is usually indicated. Some findings of the semen analysis suggest certain specific potential problems. For example, an increased white blood cell count may indicate infection or inflammation. However, other abnormalities in many of the main parameters are non-specific. For example, there are a number of different causes for a decreased count or diminished movement. Some of these causes have other serious medical implications. A thorough evaluation helps determine the cause of an abnormal semen analysis and rules out medical problems.

Semen Collection Instruction

1. You should abstain from any sexual activity for at least 2-3 days (but not longer than 7 days) before producing a sample.
2. A semen sample should be collected after you have washed your hands and penis with soap and water , be sure to rinse away all of  the soap residue.  Dry thoroughly before collecting the sample.
3. The sample should be collected by masturbation directly into a sterile clean and dry container. If masturbation could not be possible , coitus interruptus can be accepted.
4. Condoms or lubricant must not be used because they contain agent that kill sperm.
5. Sterile, wide-mouthed containers ready for used can be obtained from our hospital.
6. In case the husband cannot produce a specimen at the hospital and the collection happen outside, the sample should be handed to the hospital within 1 or 2 hours.

Semen Tests

Macroscopic Examination

1. Volume This is a measurement of the volume of the ejaculate. Normal is 1.5 milliliters or greater. The volume may be low if a man is anxious when producing a specimen, if all of the specimen is not caught in the collection container, or if there are hormonal abnormalities or ductal blockages.

2. pH The pH of normal semen is slightly alkaline.  Normal is 7.2 or greater.

3. Viscosity Viscosity is measured after complete liquefaction has occured. Viscosity is considered "normal" if the liquefied specimen can be poured from a graduated beaker drop by drop with no attaching agglutinum between drops.

4. Liquefaction Semen is a thick gel at the time of ejaculation and normally becomes liquid  within 20 minutes after ejaculation. Liquefaction time is a measure of the time it takes for the semen to liquefy.

Microscopic Examination

1. Viability Viability of spermatozoa is a valuable quality parameter which is measured by permeability of the plasma membrane to specific stains. The most common staining procedure has been eosin/nigrosin being easy to use, requiring only a basic microscope. Its drawback is that assessment of viability can be extremely subjective due to the varying intensity of background coloration. Dead

sperm is red staining and  alive sperm is coloress (white). 58% or more of the sperm should be alive.

.2. MAR test Mixed agglutination reaction. This test is screening test. This test is done on the semen sample in order to check the presence or absence of antisperm antibodies and the type of these antibodies (IgA, IgG or IgM). Antibodies that bind to sperm may reduce fertility. If the test is negative, this is reassuring

3. White Blood Cells The semen may contain a high number of white blood cells, which may be an indication of either infection or inflammation. White blood cells are considered significant if more than one million are found in each milliliter of the ejaculate.

4. Red Blood Cells Red cell count should be less than 1 million per ml. If there are more cells, this may be an indication of an infection.

5. Round cell Round cells are cells that have round shape such as Epithelial cells, white blood cells, and immature germ cells; spermatid, spermatocytes, spermatogonia.

Computer Assisted Semen Analysis (CASA)

The use of computer asisted semen analysis has advanced the ability to study and understand sperm function as it relates to human infertility. The major advances have been in the ability to more accurately determine sperm concentration (counts) and motility (movement).

6. Sperm concentration (Sperm count) This is a measurement of how many million sperm there are in each milliliter of fluid. There are various techniques for obtaining this number - some prove to be more accurate than others. Normal sperm concentration is 15 or geater million per milliliter.

7. Motility This describes the percentage of sperm that are moving. Forty percent or more of the sperm should be moving.

• Motile sperm marked with green track

• Progressive sperm marked with blue track

• Slow sperm marked with purple track

• Static sperm marked with red dot

8. Morphology This describes the shape of the sperm. The sperm are stained by Diff-quick stain that compound Methanol, Eosin, and Methylene blue. After that the sperm are examined under a CASA and must meet specific sets of criteria for several sperm characteristics in order to be considered normal. Normal is four or more percent of the sperm should be normal by WHO criteria and fourteen or more percent by Strict criteria.

WHO Normal range for Semen Characteristics 4^th edition VS 5^th edition