This document provides information about performing and interpreting a normal semen analysis according to WHO guidelines. It discusses that a semen analysis examines the sperm count, morphology, and motility in the ejaculate. For a normal analysis, the sperm count should be between 35-200 million sperm per mL, most sperm should have a normal oval shape, and at least 50% of sperm should be actively motile. The document provides details on the procedures for examining each of these parameters under a microscope and calculating the sperm concentration in the ejaculate.

1. SEMEN ANALYSIS
Pandian M
Dept of Physiology
DYPMCKOP

2. PY.9.5 Interpret a normal semen analysis report
as per WHO guidelines & discuss the
results
1.Sperm count
2.Sperm morphology
3.Sperm motility

3. INTRODUCTION
•Semen is a white fluid that contains sperms.
•It is the collection of fluids from testes, seminal
vesicles, prostate gland and bulbourethral glands.
•Semen is discharged during sexual act and the process
of discharge of semen is called ejaculation.
•Testes contribute sperms.
•Prostate secretion gives milky appearance to the semen.
•Secretions from seminal vesicles and bulbourethral
glands provide mucoid consistency to semen.

4. Characteristic features
Volume.
• The average volume of semen per ejaculation is 2.5–3.5 mL after
an abstinence of 2 days.
• Volume of semen decreases with repeated ejaculations.
Appearance of semen is milky due to prostatic secretions.
Specific gravity is about 1.028.
Reaction is alkaline with a pH of 7.5.
The alkalinity is due to the prostatic secretions.
The alkaline semen brings the vaginal pH from 3.5–4 to 6–
6.5, the pH at which sperms show optimum motility.

5. Cont……..
Nature of the semen when ejaculated is liquid but soon it
coagulates in vitro or in the vagina, and finally
undergoes secondary liquefaction after about 15–30
min.
The clotting of semen soon after ejaculation helps
to retain it in the vagina for sometimes.
Lysis (fibrinolysin) later on would release the
sperms for their free movement into the uterine cavity
for fertilization.

6. Components of semen and their
characteristics
1. Spermatozoa. The normal sperm count varies
from 35 to 200 million/mL of semen with an
average of 100 million/mL.

7. 2. Secretions of seminal vesicles.
• Secretions of seminal vesicles contribute 60% of
the semen volume:
• The secretion from seminal vesicles is mucoid and
viscous fluid.
• It is neutral or slightly alkaline in nature.
• It contains fructose, phosphorylcholine,
ergothioneine, ascorbic acid, flavins and
prostaglandins.

8. Cont……
Functions subserved by the seminal vesicle secretions are:
• Nutrition to sperms after being ejaculated into the female
genital tract is provided by the fructose and other nutritive
substances from the seminal vesicle secretions.
• Clotting of semen soon after ejaculation into the female
genital tract occurs due to fibrinogen present in the
seminal vesicle secretions.
• Fertilization of ovum may be enhanced by the
prostaglandins present in the seminal vesicle secretion.

9. 3. Secretion of prostate gland.
• Secretion of prostate gland forms about 10% of the total semen
bulk:
• It contributes milky and alkaline fluid part of the semen.
Functions subserved by prostatic fluid component of semen are:
• Maintenance of optimum pH for fertilization (6–6.5) is the function
of alkaline prostate fluid, which neutralizes the acidity of vaginal
secretion.
• At this pH, the sperms become motile and the chances of fertilization
are enhanced.
Clotting of semen by converting fibrinogen (from seminal vesicles)
into a coagulum is caused by the clotting enzymes present in the
prostatic fluid.

10. 4. Secretion of bulbourethral gland.
•Secretion of bulbourethral gland and other
mucous glands provide mucoid consistency to
the semen after puberty.

11. Sperm morphology
•(a) Diagram of a human
mature sperm.
•(b) A close-up of the head
drawn from a different
angle.
•The acrosome contains
enzymes required for
fertilization of the ovum.

13. Sperm motility
•The definition of motility is the ability of an
organism or fluid to move.
•Sperm motility refers to the movement and
swimming of sperm.
•Poor sperm motility means that the sperm do
not swim properly, which can lead to male
infertility

15. Sperm count

16. Semen analysis

17. Apparatus
•Fresh sample of semen; 4% sodium bicarbonate
in 1% phenol; apparatus required as for WBC
counting
•Method – hanging drop preparation method

18. HAEMOCYTOMETER
NEUBAUER CHAMBER
RBC PIPETTE
WBC PIPETTE

20. Side view of Neubauer chamber

22. Procedure for Mobility
1. Obtain fresh sample of semen collected in a small
bottle.
2. Wait for 15 to 20 mins. So, that it may undergo
secondary liquefaction.
3. To assess the sperm motility, place a drop of semen
on the cover slip and invert it on a rim of plasticine on
the slide
4. Examine under high power objective and find out the
percentage of immobile to mobile sperms.

24. Determination of sperm count
i. Gently shake the sample to assure uniformity
ii. Draw semen in the WBC pipette up to 0.5 mark
iii. Draw in 4% sodium bicarbonate in 1% phenol sol.
Upto the mark of 11.
iv. Making a dilution of 1 in 20
v. Thoroughly shake the mixture, discard the first few
drops
vi. Then charge the counting chamber.
vii.Ensure uniform distribution

25. METHOD OF DILUTION
• Take blood up to the 0.5 mark without any air bubble
• Take the dilution fluid upto the 11 or 101 mark
• Keep the pipette between the palms and mix the fluid
• The fluid in the bulb gets mixed completely

26. Different between over and under charge

27. •Count the sperm under high power in the four
WBC squares and Follow rules for counting.

30. Calculation
•Number of sperms in 1 µL of sample
= N x 10/4 X 20
•Number of sperm in 1 µL (i.e. 1 cumm) of
sample
= N x 50 x 1000 (as 1 cc = 1000 µL)
N x 50,000

32. Reference
•Textbook of Practical Physiology
•A.K. Jain
•C.L. Ghai
•G.K.Pal
•Textbook of Medical Physiology
By Guyton & Hall, A.K. Jain

33. THANK YOU . . .