Semen Analysis

Semen Analysis

Semen Analysis looks at the volume and quality of a man’s sperm. It is one of the first steps to detecting male fertility issues. The test also shows whether a vasectomy was successful. Semen analysis involves collecting a semen sample and evaluating it in a lab. The Semen test is the primary test in the diagnosis of quinsy. The following information is noted in this test:

  • Volume
  • pH
  • Sperm Count
  • Morphology of Sperm
  • Percentage of Motile Spermatozoa

Sample of Semen:

  1. Sample for semen test is obtained in clean dry and leak proof bottle/container. The patient is instructed to deliver the sample to the laboratory within 15-30 minutes. It is better to get the sample in the laboratory itself.
  2. Sexual intercourse should be abstained from for three days before giving the sample for semen test.

Special Precautions:

The laboratory worker should process the semen sample with extreme care because it may contain the following pathogens:

Volume of Semen:

Semen is in a condensed semi-solid state after it is excreted from the body. Its state changes to liquid form in about an hour. When the semen turns into a liquid state, the quantity can be determined in any marked small cylinder.

Normal Volume: More than 2 ml.

Semen Analysis

Determining the pH:

pH paper is used to determine pH. Add a drop of semen to the pH paper with a pipette.

Normal pH (within one hour) 7.2

Count of Motile Sperm:


  • Place a drop of liquid cement on a clean glass slide and cover with a cover slip.
  • Focus the sample in the microscope, first at 10 power and then at ×40.
  • Take a sample of 100 sperm by focusing under a microscope and see how many are motile and how many are not.

Normal movement (within one hour) in more than 50% movement

Live or Dead Sperm:

  • Put one drop of liquid semen (10-15 µl) on a glass slide and add one drop of Eosin (Eosin) to ceramic and place the slide.
  • After two minutes, focus the slide in the microscope. Look first at 10 and then at ×40.
  • Live sperm do not stain while dead sperm do.
  • Find the number of live and dead sperm out of 100 sperm.

The normal amount of live dead (more than 75 sperm are alive within an hour).

Total Count of Sperm:


  • Pour one milliliter of semen into a clean dry marked small cylinder with a pipette.
  • Then add 20 ml of sodium bicarbonate formalin and mix well. This will be the ratio of 1:20 to the total sample of Simmons. Keep this mixture for five minutes.
  • Focus the Neubauer Ruled Chamber in the microscope and fill it with the above cement mixture.
  • Wait five minutes after filling the chamber and focus the sample at 10 and count the amount of sperm in the two large square boxes.
  • The volume in 2 large boxes is sq mm 2.

Sperm Morphology:


  • Make a thin layer of cement on the glass, and slide. Allow it to air dry and mix in 7/7 95% ethane and allow to air dry.
  • Then wash the fixed layer with sodium bicarbonate formalin solution, then dip the slide in clean water 4-5 times.
  • Apply Carbon Fuchsine (1:20) to the slide and leave for 3 minutes, then wash it off.
  • Add Loeffler Methylene (Blue) as a counter stain to the slide and wash after 2 minutes.
  • Focus the slide under the microscope and see the shapes of the sperm.
  1. Nucleus of Head – (Dark Blue)
  2. The cytoplasm of Head – (Pale Blue)
  3. Middle Piece & Tail – (Pink Red)

Count the different shapes in one hundred sperm, and separate normal and abnormal among them.

Semen Analysis

Normal Sperm Morphology:

Normal sperm is 50-70 micrometers in length. The head is oval with an acrosome cap on top. The head is followed by the middle part and then the long tail.

Normal Values: 50% or more sperm should look normal.

Abnormal Sperm Morphology:

The following forms may be seen.

  • Sperm size may be too small or too large.
  • A change in the size or shape of the head.
  • Acres should not be covered.
  • Vacuoles or chromatin in the nucleus.
  • The middle part (Middle Piece) is missing or the size is very large.
  • The middle part is divided into two parts.
  • Crooked midsection.
  • The disappearance of the tail, splitting into 2 parts or becoming crooked.

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