2. SeminalFluid
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Semen is body fluid that is ejaculated at the time
of orgasm, contain sperm & secretion of seminal
vesicle, prostate, Cowper's gland & urethral
gland.
3. Source Volume Characteristics
Urethral and
bulbourethral glands
0.1-0.2cc Viscous, clear
Testes, epididymides,
vas deferentia
0.1-0.2cc Sperm present
Prostate 0.5-1.0cc Acidic,watery
Seminal vesicles 1.0-3.0cc
Gelatinous, fructose
positive
Complete ejaculate 1.5-5.0cc Liquefies in 20-25min
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Semen
4. • Semen is viscous, neutral or slightly
alkaline & whitish opaque.
• 60 % semen volume is derived from seminal vesicle
which is also a major source of high FRUCTOSE
content of semen.
• Seminal vesicle secretion also provide
the substrate for the coagulation of the
semen following ejaculation.
• About 20 % of the volume of
semen is contributed by the prostate
gland.
⚬ It is milky in appearance & also rich in proteolytic enzymes are
responsible
for the liquefaction of semen.
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5. • About 10 -15 % of semen volume is also contributed by
epididymis, vasdeferens, cowper’s gland & uretheral
gland.
• Less than 5 % of semen volume is contributed by
Spermatozoa.
• The process of ejaculation result in the mixing of these
distinct fraction of semen.
• These enter the urethra individually in the rapid
succession.
• The third & final fraction consist of mucoid secretion
resulting from emptying of seminal vesicle.
• The semen specimen collected for routine examination
should contain all above mention fraction.
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7. Steps:SemenAnalysis
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In the first 5 minutes:
• Placing the specimen container on
the benchor in an
incubator (37 °C) for liquefaction.
Between 30 and 60 minutes:
• Assessing liquefaction and appearance of the
semen.
• Measuring semen volume.
• Measuring semen pH (if required).
Preparing a wet preparation for assessing
microscopic appearance,sperm motility
assessing spermand the dilution required for
number.
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8. StepsOfSemenAnalysis
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Between 30 and 60 minutes:
• Assessing sperm vitality (if the
percentageof motile cells is low).
• Making semen smears for assessing sperm
morphology.
• Making semen dilutions for assessing sperm
concentration.
• Assessing sperm number.
• Performing the mixed antiglobulinreaction
(MAR) test (if required).
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9. Steps
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Between 30 and 60 minutes:
• Assessing peroxidase-positive cells (if round cells
are present).
• Preparing spermatozoa for the immunobead test (if
required).
• Centrifuging semen (if biochemical markers are to be
assayed).
Within 3 hours:
• Sending samples to the microbiology laboratory (if
required).
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10. Steps
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After 4 hours:
• Fixing, staining and assessing
smears for sperm morphology.
Later on the same day (or on
a subsequent day if samples are
frozen):
• Assaying accessory gland markers (if
required).
• Performing the indirect
immunobead test (if required).
SAKSHI ALEX
11. Sample
Collection:
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Preparation:
• The sample should be collected in a private
room near the laboratory, in order to limit the
exposure of the semen to fluctuations in
temperature and to control the time between
collection and analysis.
• The sample should be collected after a
minimum of 2 days and a maximum of 7 days
of sexual abstinence. If additional samples
are required, the number of days of sexual
abstinence should be as constant as possible
at each visit.
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12. Sample
Collection:
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Preparation:
• The man should be given clear written and spoken
instructions concerning the collection of the semen
sample. These should emphasize that the semen
sample needs to be complete and that the man
should report any loss of any fraction of the sample.
• The following information should be recorded on
the report form: the man’s name, birth date and
personal code number, the period of abstinence, the
date and time of collection, the completeness of the
sample, any difficulties in producing the sample, and
the interval between collection and the start of the
semen analysis.
SAKSHI ALEX
13. SampleCollectionforDiagnostic/Researchpurpose:
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• The sample should be obtained by masturbation and
ejaculated into a clean, wide-mouthed container made of
glass or plastic, from a batch that has been confirmed to
be non-toxic for spermatozoa.
• The specimen container should be kept at ambient
temperature, between 20 °C and 37 °C, to avoid large
changes in temperature that may affect the spermatozoa
after they are ejaculated into it. It must be labelled with
the man’s name and identification number, and the date
and time of collection.
• The specimen container is placed on the bench or in
an incubator (37 °C) while the semen liquefies.
• Note in the report if the sample is incomplete,
especially if the first, sperm-rich fraction may be missing.
If the sample is incomplete, a second sample should be
collected, again after an abstinence period of 2–7 days.
SAKSHI ALEX
15. Collectionofsemenathome:
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• A sample may be collected at home in exceptional circumstances,
such as a demonstrated inability to produce a sample by
masturbation in the clinic or the lack of adequate facilities near the
laboratory.
• The man should be given clear written and spoken instructions
concerning the collection and transport of the semen sample. These
should emphasize that the semen sample needs to be complete,
i.e. all the ejaculate is collected, including the first, sperm-rich
portion, and that the man should report any loss of any fraction of
the sample. It should be noted in the report if the sample is
incomplete.
• The man should be given a pre-weighed container, labelled with his
name and
identification number.
• The man should record the time of semen production and
deliver the sample to the laboratory within 1 hour of collection.
• During transport to the lab., the sample should be kept between 20
°C - 37 °C.
• The report should note that the sample was collected at home
or another location outside the laboratory.
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16. Safehandlingofspecimens:
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• Semen samples may contain dangerous infectious
agents (e.g. human immunodeficiency virus (HIV),
hepatitis viruses or herpes simplex virus) and should
therefore be handled as a biohazard. If the sample is
to be processed for bioassay, intra-uterine
insemination (IUI), in-vitro fertilization (IVF) or
intracytoplasmic sperm injection (ICSI), or if semen
culture is to be performed, sterile materials and
techniques must be used. Safety guidelines as
outlined in Appendix 2 should be strictly followed;
good laboratory practice is fundamental to laboratory
safety (WHO, 2004).
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17. Precautions
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• Specimen should not be collected in ordinary
condoms since the powder or lubricant
applied to the condom may be spermicidal.
• The container in which the semen sample is
collected should be free from detergent.
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18. Storage
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• The semen specimen should be examined
immediately after collection.
• It should be kept at room temperature.
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19. • History To Be Noted:
• Name
• Date and time of collection
• Length of abstinence
• Interval between collection and analysis
• History of fever
• Drug intake
• Alcohol abuse
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21. • Optionaltests:
• Alpha galactosidase (neutral): 20mU
or more.
• Zinc (total): 2.4 micromol or more
• Citric acid(total): 52 micromol or more
• Acid phosphatase: 200U or more
• Fructose: 13 micromol or more
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24. Viscosity:
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substrate
coagulum
produced by seminal
vesicles. liquefies
spontaneously to
form
• Freshly ejaculated semen is highly
viscous due to
The
a
translucent, viscous fluid in a three stage process.
• Action of a prostatic clotting enzyme.
• Liquefaction is initiated by enzymes of prostatic
origin.
• Protein fragments are further degraded into free
amino acids and ammonia.
• Failure to liquefy indicates inadequate prostatic
secretion. To liquefy, add bromeline, plasmin or
chymotrypsin.
SAKSHI ALEX
25. • Normal liquefaction time: 20-60min
• Viscosity of liquefied semen can be estimated by: Gentle
aspiration into a 5ml pipette, then allowing the semen to drop
by gravity. Observe the length of the thread.
• Normal semen leaves as small discrete drops.
• Increased viscosity is associated with poor invasion of
cervical mucus in post-coital studies as well as decreased
ability to fertilize ovum.
• Absence of viscosity points to reduced cell content.
The semen from males with b/l congenital absence of vas
deferentia & seminal vesicles fails to coagulate due to absence
of substrate.
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