Semen Analysis being one of the basic test and gold standard for male infertility evaluation. Guidelines by World Health Organization published in 2010 remain the most discussed and currently worldwide accepted in interpretation of semen examination.

1. SEMEN ANALYSIS & W.H.O CRITERIA
RAHUL KUMAR SEN M.Sc., Ph.D.
Vasundhara Hospital & Fertility Research Centre

2. INTRODUCTION
• Semen Analysis is the most important tool in
male infertility investigation.
• In the last two decades, researchers and clinicians
have relied on the World Health Organization
(WHO) criteria for the interpretation of basic
semen analysis.

3. Why Do Semen Analysis…?
In Truth, it is not really about determining the total
number or fastest sperm but rather:
• Does the man produce’s sperm…???
• Are the sperm’s alive…???
• Will the sperm make an embryo after
fertilization…???

4. • In order to establish consistency, the
WHO first published a manual for the
examination and preparation of
human semen in 1980.
• The manual has been regularly
updated (1980, 1987, 1992, 1999,
2010)
STANDARDIZATION

5. COMPOSITION OF SEMEN
a normal semen is mixture
of secretions from
•Testes,
•Epididymis,
•Seminal Vesicles,
•Prostrate Gland,
•Bulbourethral Glands
It has the presence of
•Spermatozoa,
•Sperm precursor cell,
other cells from genital
tract
•Leucocytes
•Some complex organic,
inorganic constituents

6. SEMEN COLLECTION
CLEAR & SIMPLE INSTRUCTIONS
IMPORTANCE OF
PERIOD OF ABSTINENCE
COLLECTED IN WARM,
NON-TOXIC, WIDE MOUTH
CONTAINER
USE OF LUBRICANTS, REGULAR
CONDOMS TO BE AVOIDED
USE OF NON-SPERMICIDAL
SOME PRELIM. INFO TO BE NOTED:
DATE, TIME & PLACE OF COLLECTION
IF ANY SPILLAGE OCCURED

7. SEMEN ANALYSIS
• To begin with simple inspection (wet
preparation) soon after liquefaction
(preferably after 15-45min) but no longer than
1 hr. to avoid dehydration and affects of
temperature on semen quality.

8. SEMEN EXAMINATION
Macroscopic parameters
• Volume
• Appearance
• Color
• Liquefaction
• Viscosity
• pH
Microscopic parameters
• Sperm concentration
• Sperm motility
• Morphology
• Leucocyte concentration
• agglutination

9. VOLUME
• 1.5 to 6 ml (WHO, 2010)
• The volume of an ejaculate is an indicator of various abnormalities.
HYPERSPERMIA (MORE VOLUME) HYPOSPERMIA (LESS VOLUME)
• Long period of abstinence
• In some cases of retrograde
ejaculation
• Loss of portion of ejaculate during
collection
• Short abstinence period
• Intake of certain drugs
• Following surgical procedures that
damage the sympathetic nervous
plexus
• resection of the prostate gland.

10. • Appearence – generally Opaque.
• Colour- Homogenous grey to opalescent.
• Odour- Distinctive musty.
Colour may also be different, i.e. red-brown red blood cells are present
(haemospermia)
Yellow: Jaundice or taking certain vitamins or drugs.
PALE /STRAW YELLOW: PYOSPERMIA

11. • Viscosity- a normal sample leaves the pipette in small
discrete drops. If viscosity is abnormal, the drop will
form a thread more than 2 cms long.
• pH- 7.2 to 8.0, reflects balance between pH of different
accessory gland secretions, to be measured after
liquefication.

12. Microscopic parameters
• Sperm concentration
• Sperm motility
• Morphology
• Leucocyte concentration
• Immature/ Round cell concentration
• Agglutination

13. Sperm Concentration
• the number of sperm found in an ejaculate.
• To be analyzed with either wet prepration/
makler’s chamber.
• Lower Limit to be at least 15 mill. per cc.
(WHO,2010)

14. Analysing multiple fields
Under 40x objective
As many fields can
be analysed
SPERM COUNTING

15. WET PREPRATION MAKLERS
CHAMBER

16. Sperm Motility
• to be assessed as soon after liquefaction,
preferably at 30 minutes, but in any case
within 1 hour, following ejaculation.
• Limit the deleterious effects of dehydration,
pH or changes in temperature on motility.
TOTAL MOTILITY = TOTAL NUMBER OF MOTILE SPERMS
/ TOTAL NUMBER OF SPERMS ANALYSED * 100

17. SPERM MOTILITY
Sperm motility
classified into 4
grades
• Group A (%)=
RLP (Rapid
Linear
Progressive)
• Group B (%)
Slow Linear,
Sluggish, Round
twitching
movements
• Group C (%)=
Immotile
A
B
C

18. Sperm Morphology
• Closely examination of a sperm, checking to see approximately what
percentage have a normal shape. The head, mid-section and tail are
evaluated.
• Different methods for staining for evaluation of sperm morphology have
been described (Ombelet et al. 1995).

19. A normal sperm
Head should be smooth and
generally oval shaped. There
should be a well defined
acrosomal region comprising 40-
60% of head area, it should not
contain any vacuole.
Mid piece/ neck should be
slender, regular and about the
same length as the sperm head
Tail is thinner than midpeice and
10times of head shape

24. Types of morphological defects
HEAD
• LARGE
• SMALL
• TAPERED
• PYRIFORM
• ROUND
• AMORPHOUS
• VACUOLATED
• DOUBLE
HEADED
NECK
• THICK
• IRREGULAR
• SHARPLY BENT
• THIN
• SHORT
TAIL
• SMOOTH
ANGULATED
BENDS
• SHORT
• MULTIPLE TAILS
• BROKEN

25. Leucocyte concentration
• Leukocytes are normally present in the seminal
fluid; however, a concentration of >1 × 106/mL is
considered abnormal (WHO1999).
• A positive correlation was observed between
leukocyte count and the total count of
microorganisms in semen sample.
• The excessive presence of leukocytes may be
detrimental to spermatozoa due to their
excessive production of reactive oxygen species
(ROS) and cytotoxic cytokines.

26. Agglutination
• Agglutination specifically refers to
motile spermatozoa sticking to each
other that causes limited motion

27. Cut off reference values for sperm parameters in
consecutive WHO manuals

28. (CASA)
Computer Assisted Semen Analysis
• Manual SA may lacks the ability to measure kinetics
& proper morphology of sperm
• Much attention can be given via Computer Assisted
Semen Analysis bcoz of its potential benefits

29. Inference
• It is performed as a part of the investigation of
the infertile couple and its interpretation plays
a major role in charting the treatment plan(s).
• It should be performed as on multiple
ejaculates (Keel 2006).

30. • Importance of the investigation in local language.
• Explanation of the procedure
• The abstinence period
• about the method of collection of the semen
• With interpretation of report and Interpreting treatment
modalities
Patient Counseling

31. • Medicines, such as cimetidine (Tagamet), male and female
hormones (testosterone, estrogen), undergoing chemotherapy
for cancer .
• Consuming higher Caffeine, alcohol, cocaine, marijuana, and
smoking tobacco.
• Temperature.
• Exposure to radiation, some chemicals (such as certain
pesticides or spermicides).
• An incomplete semen sample.
Factors affecting Semen Analysis

32. THANK YOU