Semen Analysis Workshop Notes

SEMEN ANALYSIS WORKSHOP

To calculate the percent difference between two sperm counts, subtract the smaller count from the larger count and divide the resulting number by the average of the two counts.
If larger count = a, and smaller count = b, then percent difference is 2(a-b)/ (a+b) \times 100.
- Example:
  1. count a = 80.8, count b = 74.5
  2. 80.8 - 74.5 = 6.3
  3. 80.8 + 74.5 = 155.3; 155.3 / 2 = 77.7
  4. 6.3 / 77.7 = .081
  5. .081 \times 100 = 8.1 \%$ difference

Duration of abstinence: 3 days
Interval between ejaculation and start of analysis: 25 minutes
Appearance: Normal (Off-white-Grayish opalescent fluid). Abnormal: Any color/opacity/consistency other than normal.
Liquefaction: Normal (Liquefied by 60 minutes at room temperature). Abnormal: Not liquefied by 60 minutes at room temperature.
Viscosity: 4 = Normal, 1 = will not pipette. Abnormal viscosity can interfere with determination of sperm motility and concentration.

Volume (mL): Measurement = 2.0, Reference Limits = \geq 1.5
pH: Measurement = 8.0, Reference Limits = \geq 7.2
Sperm Concentration (x 10^6 /mL): Count 1 = 6.6, Reference Limits = \geq 15
White Blood Cell Conc. (x 10^6 /mL): <1, Reference Limits = <1
% Motile: a: motile = 50, Reference Limits = \geq 40\%. b: immotile = 50
Modal Progression: 2= definite forward progression, 1= weak/sluggish forward progression, 0= absence of forward progression
Total Motile Count (x 10^8 /Ejaculate): Calculated value = 6.6
Sperm Morphology (WHO Fifth Edition): 4\%-Lower Reference Limit
Morphology Comments: Results may be skewed due to low sperm concentration

WHO recommends distinguishing between progressive and non-progressive motile sperm.
2= definite forward progression
1= weak/sluggish forward progression
0= absence of forward progression
PROGRESSION RATING OF 2 @ > 32\%$$ MOTILITY
Total Motile Count (Volume x Concentration x Percent Motile)

Sperm must display an oval head, measuring 5-6 µm in length and 2.5 -3.5 µm in width
The acrosome proportion must comprise 40-70% of the head.
1-2 vacuoles of 1 µm or smaller is/are acceptable but larger or more vacuoles would place the sperm in the abnormal category.
There should be no obvious defects in the neck, midpiece or other parts of the flagellum.
The length of midpiece should be around 6-10 mm (6-7 µm also have been noted) and that of the flagellum be around 45-50 mm.
Any minor variations in shape or size which deviate from the normal parameters would place a sperm into the abnormal category.
Sperm with perfectly normal heads but with abnormal tails should be reported as a separate category in the comments section of the report for morphology.

Head defects: Tapered, Pyriform, Round, Amorphous, Vacuolated, Small acrosomal area, No acrosome
Neck and midpiece defects: Bent neck, Asymmetrical insertion, Thick insertion
Tail defects: Thin, Short, Bent, Coiled
Excess residual cytoplasm: > one third head

Sperm viability refers to a spermatozoa being alive and having a functional membrane despite the fact that it may be immotile.
Motility is an indication of viability but immotile sperm cannot be called dead (or non-viable) unless a differential (or vital) staining or another method of determining viability has been performed and determined to be non viable.
You should not call immotile sperm as “dead” or “non-viable” unless the vital staining has been performed.

A dye to determine the viability of sperm can be used in conjunction with semen analysis.
Using a dye to assess the viability of sperm in conjunction with IVF/ICSI is not allowed because of the possibility that we may not be able to wash the dye fully off sperm and the dye residues may be toxic to the embryos.

In the event that there is no sperm observed on the counting chamber, the sample must concentrated by centrifugation and the pellet thoroughly searched.

Reference slide
Links to pictures
Link to WHO 5th edition and assigned reading: Section 2.15, 2.16, 2.17- morphology, Section 2.6-Vitality
Be prepared to look at a completed semen analysis report and discuss what is wrong with the report and/or what the report tells us clinically. This will be conducted in the same manner that you would communicate with your physician or patient.