Introduction of Semen Analysis

Semen analysis is the branch of laboratory medicine that deals with the examination of semen which reflects the condition of male fertility. It is the most common test suggested by a gynecologist or general physician. Sample collection is a little bit hard than urine collection since it is collected either during sexual intercourse or masturbation. A sterile container same as urine culture is provided for collection but there is prior to collection some laboratory precautions are mandatory like 3 days free from sexual intercourse or coitus, the technique of sample collection, time and place of sample collection, collection, and examination time is also necessary for semen analysis and semen should examine within 30 minutes.

List of Tests in Semen Analysis

Semen analysis completes into subheadings like physical examination, chemical examination, and microscopic examination.

Physical Examination

• Colour and appearance
• Viscosity
• Volume
• Liquefaction time

Chemical Examination

• PH
• Fructose

Microscopic Examination

• Sperm count
• Sperms Motility after 2,3 and 6 Hours
• Abnormal forms

Other Findings

• Pus cells
• Epithelial cells
• RBCs
• Trichomonas or other microbes

Unit and Normal Range of Semen Analysis

	Semen Analysis
	Physical Examination
Test	Value	Normal Finding
Color and appearance		Greyish-white, Viscid, opaque
Viscosity		The specimen of normal viscosity can be poured drop by drop.
Volume		2-5 mL
Liquefaction time		20-30 minutes
	Chemical Examination
PH		7-2-7.8
Fructose		150-300 mg/dl
	Microscopic Examination
Sperm count		40-300 million/mL
Motility after 2,3 and 6 Hours		60%
Abnormal forms		0-20%
	Other Findings
Pus cells		1-2/HPF
Epithelial cells		1-2/HPF
RBCs		Absent
Trichomonas and other microbes		Absent
Note-

Suggestions on Semen Analysis

Always try to know who is testing your specimen because he or she is playing with body samples and the proper finding never ignores good track of your life in case of diagnosis, normal activities, and so on. To give this sense, your report should have the complete name of the examiner ( laboratory person), his/her signature, council number, and logo of the institution (hospital/ diagnostic center).

Procedure of Sperm Count

The procedure for sperm count, also known as a semen analysis, involves several steps to evaluate the health and viability of a man’s sperm. This test is commonly used to investigate male fertility and includes the assessment of sperm concentration, motility, morphology, and other parameters. Here is a detailed procedure for conducting a sperm count:

1. Sample Collection

• Preparation: The patient should abstain from ejaculation for 2-5 days before the test to ensure an optimal sperm count.
• Collection Method: The semen sample is typically collected through masturbation into a sterile container. It is important to collect the entire ejaculate, as the first part of the ejaculate contains the highest concentration of sperm.
• Alternative Methods: If masturbation is not possible, semen can be collected during intercourse using a special condom (without spermicides) or via other medical procedures.
• Timing: The sample should be delivered to the laboratory within 1 hour of collection to ensure accurate results. It should be kept at body temperature during transport.

2. Macroscopic Examination

• Volume: Measure the volume of the semen sample. Normal volume ranges from 1.5 to 5 milliliters.
• Color and Appearance: Assess the color and appearance of the sample. Normal semen is typically whitish-gray and homogeneous.
• Viscosity: Evaluate the viscosity by observing how the semen forms threads when allowed to drop from a pipette. Normal semen liquefies within 15-30 minutes after ejaculation.
• pH: Measure the pH of the semen, which should be between 7.2 and 8.0.

3. Microscopic Examination

• Sperm Concentration (Count):
  • Preparation: Mix the semen sample well to ensure homogeneity.
  • Dilution: Dilute the semen sample with a fixative solution (e.g., sodium bicarbonate-formalin) to immobilize the sperm.
  • Counting Chamber: Load the diluted sample onto a counting chamber (e.g., Neubauer hemocytometer).
  • Counting: Count the number of sperm in specific grids of the counting chamber under a microscope. Calculate the concentration using the formula:

• Normal Range: A normal sperm concentration is typically above 15 million sperm per milliliter.
• Motility:
  • Assessment: Evaluate the motility of sperm in a fresh sample by observing their movement under a microscope.
  • Classification: Classify sperm motility into progressive motility, non-progressive motility, and immotility.
  • Calculation: Calculate the percentage of sperm with progressive motility (PR) and total motility (PR + NP).
  • Normal Range: A normal motility rate is typically above 40% total motile sperm and 32% progressively motile sperm.
• Morphology:
  • Staining: Stain a smear of the semen sample with a staining solution (e.g., Papanicolaou stain) to visualize sperm morphology.
  • Assessment: Examine the stained smear under a microscope and evaluate the shape and structure of the sperm.
  • Classification: Classify sperm as normal or abnormal based on their head, midpiece, and tail morphology.
  • Normal Range: A normal morphology rate is typically above 4% normal forms according to strict criteria (e.g., Kruger criteria).

4. Additional Tests (if needed)

• Vitality Test: Evaluate the percentage of live sperm using a staining method (e.g., eosin-nigrosin stain).
• Sperm Agglutination: Check for the presence of sperm agglutination, which may indicate an immunological cause of infertility.
• Leukocyte Count: Assess the presence of white blood cells in the semen, which may indicate infection or inflammation.
• Biochemical Tests: Measure the levels of fructose, zinc, and other biochemical markers in the semen.

5. Interpretation and Reporting

• Analysis: Compare the results with the reference values provided by the World Health Organization (WHO) or other guidelines.
• Report: Provide a comprehensive report detailing the findings of the semen analysis, including sperm count, motility, morphology, and any additional tests performed.

Reference Values (WHO 2010)

• Volume: ≥1.5 ml
• pH: ≥7.2
• Sperm Concentration: ≥15 million/ml
• Total Sperm Count: ≥39 million/ejaculate
• Motility: ≥40% total motility (PR + NP) or ≥32% progressive motility (PR)
• Morphology: ≥4% normal forms (strict criteria)

Gallery of Semen Analysis

Direct Semen Microscopy

Sperms at 100X magnification

Spermatozoa at 400X magnification

Human Sperms at 1600X Magnification

Human Sperms at 4000X Magnification

Diluted Semen Ready for Observation

Charging diluted semen in Neubauer Chamber

Human Sperms in Neubauer Chamber at 400X Magnification

An eosin-nigrosin stained smear of semen showing sperms

Human sperms in Ziehl-Neelsen stained smear of Semen

Sperms in Leishman stained smear of semen at a magnification of 2000X

Human Sperms in Giemsa Stained smear of Semen

Human spermatozoa in Gram-Stained Smear of Semen

Human live sperms in semen Microscopy