Semen is a “ thick, viscous, creamy, slightly yellowish or grayish” substance made up of  spermatozoa — commonly known as sperm — and a fluid called seminal plasma, secret from the male reproductive organs. The function of seminal plasma are: To provide motility to sperm To provide nutrition to spermatozoa

1. Ms Ankita R Bhatiya
Assistant Professor
SHREE P.M.PATEL COLLEGE OF
PARAMEDICAL SCIENCE N
TECHNOLOGY

2. It include:
1.What is Semen?
2.Production of Semen.
3.Indication of Semen
4.Collection of Semen.
5. Examination of Semen.

3. 1.What is Semen?
 Semen is a “ thick, viscous, creamy, slightly yellowish or
grayish” substance made up of spermatozoa —
commonly known as sperm — and a fluid called seminal
plasma, secret from the male reproductive organs.
 The function of seminal plasma are:
 To provide motility to sperm
 To provide nutrition to spermatozoa

4. 2.Semen production:
 The male reproductive system includes
the penis, scrotum, testes, epididymis,
vas deferens, prostate, and seminal vesicles.
 The penis and the urethra are part of
the urinary and reproductive systems.
 The scrotum, testes (testicles), epididymis,
vas deferens, seminal vesicles, and prostate
comprise the rest of the reproductive system.

5.  Overview of the Male Reproductive System
 The penis consists of the root (which is attached to the
lower abdominal structures and pelvic bones), the visible
part of the shaft, and the glans penis (the cone-shaped
end). The opening of the urethra (the channel that
transports semen and urine) is located at the tip of the
glans penis. The base of the glans penis is called the
corona. In uncircumcised males, the foreskin (prepuce)
extends from the corona to cover the glans penis.

6.  The scrotum is the thick-skinned sac that surrounds and protects
the testes. The scrotum also acts as a climate-control system for
the testes because they need to be slightly cooler than body
temperature for normal sperm development. The cremaster
muscles in the wall of the scrotum relax to allow the testes to hang
farther from the body to cool or contract to pull the testes closer to
the body for warmth or protection.
 The testes are oval bodies that average about 1.5 to 3 inches (4
to 7 centimeters) in length and 2 to 3 teaspoons (20 to 25
milliliters) in volume. Usually the left testis hangs slightly lower
than the right one. The testes have two primary functions:
 The Testes and Ovaries

7.  The epididymis consists of a single coiled microscopic
tube that measures almost 20 feet (6 meters) in length.
The epididymis collects sperm from the testis and
provides the environment for sperm to mature and
acquire the ability to move through the female
reproductive system and fertilize an ovum. One
epididymis lies against each testis.
 The vas deferens is a firm tube (the size of a strand of
spaghetti) that transports sperm from the epididymis.
One such duct travels from each epididymis to the back
of the prostate and joins with one of the two seminal
vesicles. In the scrotum, other structures, such as muscle
fibers, blood vessels, and nerves, also travel along with
each vas deferens and together form an intertwined
structure, the spermatic cord.
 The urethra serves a dual function in males. This

8.  The prostate lies just under the bladder and surrounds the
urethra. Walnut-sized in young men, the prostate enlarges
with age. When the prostate enlarges too much, it can block
urine flow through the urethra and cause bothersome urinary
symptoms.
 The seminal vesicles, located above the prostate, join with
the vas deferens to form the ejaculatory ducts, which travel
through the prostate. The prostate and the seminal vesicles
produce fluid that nourishes the sperm. This fluid provides
most of the volume of semen, the fluid in which the sperm is

9.  3.Indications for semen analysis:
1. Investigation of infertility
2. Post - vasectomy by confirming absence of sperm.
3. To support or disprove a denial of paternity on the
grounds of sterility.
4. To examine vaginal secretions or clothing stains for the
presence of semen in medico legal cases.
5. For selection of donors for artificial insemination.
6. For selection of assisted reproductive technology, e.g. in
vitro fertilization, gamete
intrafallopian transfer technique.

10.  4.Sample collection - Preparation
 Sexual abstinence – 2- 7 days
 Two separate samples at least 7 days apart
should be analyzed
 The duration of abstinence should be constant
 Sample collection - private room in the same
laboratory
 Recommended procedure – Masturbation
 Pre warmed (21oC), sterile, non-toxic, a clean,
dry,wide

11.  5. Precaution:
 The sample should not be collected in condoms,
since the powder or lubricant applied to the condoms
may be spermicidal.
 The container in which semen sample is collected
should be free from detergents.
 The specimen collected by masturbation in clinical
pathology laboratory. This allow a complete
examination of the semen particularly the
liquefaction time.

12.  6.Storage:
The semen specimen should be examined
immediately after collection.
It is necessary to store then do not store in
refrigerator, store at room tem.

13.  Clinical significance:
Low sperm count are observed when there
is suppression of gonad tropic hormone
estrogen or androgen.
Hyper & Hypothyroidism,trauma,infection,
damage of testis.
Oligozoospermia & azoospermia.

14.  Examination of Stool:
1. Physical examination:
2. Chemical examination:
3. Microscopic examination:

15.  Physical examination:
1.Volume:
Normally: b/w 2.5 to 5ml.
Low semen volume causes:
 – Obstruction of the ejaculatory duct
 – Congenital bilateral absence of the vas deferens
(CBAVD), a condition in which the seminal
 vesicles are also poorly developed.
 – Partial retrograde ejaculation
 – Androgen deficiency
 – Inadequate erection & improper mood at collection
 – Incomplete collection
High semen volume
 – may reflect active exudation in cases of active
inflammation of the accessory organs

16. 2. Color: Whitish or grayish
Slightly yellow – prolonged abstinence
Deep yellow – pyospermia, jaundice or
taking certain vitamins or drugs
Red-brown - red blood cells are present
(haemospermia)
Trauma to the genital tract
Inflammation
Tumor of the genital tract.

17.  3.Appreance: Viscid, Opaque
Less opaque : if sperm concentration is very low
 4.Viscosity:
when ejaculated, semen is fairly viscid
and it falls drop by drop
Viscosity of the sample estimated After liquefaction
Procedure:
 gently aspirate sample into a wide-bore 5ml plastic
disposable pipette allow the semen to drop by gravity.
 Observe the length of any thread.
A normal sample leaves the pipette in small discrete
drops.
If viscosity is abnormal , the drop will form a thread more

18. 5. Liquefaction time:
Normal – 15- 30 minutes after collection
The complete sample usually liquefies within 15 minutes
at room temperature
If the semen does not liquefy within 30 minutes, do not
proceed with semen analysis but wait for another 30
minutes.
Rarely it may take up to 60 minutes or more.
If complete liquefaction does not occur within 60 minutes,
this should be recorded.
Semen samples collected at home or by condom will

19.  2. Chemical examination:
1. FRUCTOSE TEST
 This test determines androgen deficiency or
ejaculatory obstruction of semen ; the level of
seminal fructose is low in both these conditions.
 Normal seminal fructose level is 150-300mg/dl
 Fructose is measured by qualitatively by
resorcinol test.

20.  Procedure:
 5 ml resorcinol reagent + 0.5 ml of semen in a
test tube
 Mix and place in a boiling water bath for 5min or
heat.
 Observations: Red colored ppt. in 30 seconds.
 In quantitative assays, this is compared with a
known fructose standard at 490nm.
 Normal level of fructose: 150-300mg/dl.
 Reduced levels: Seminal vesicle dysfunction,
High sperm count, obstructed vas deferens

21.  2.PH:
 Reference value - ≥ 7.2
 Measured after liquefaction , preferably after 30 minutes
 Should be measured within 1 hour of ejaculation since it is influenced by the
loss of CO2 that occurs after production.
 Procedure:
 For normal samples: pH paper in the range 6.0 to 10.0 should be used.
 For viscous samples, the pH of a small aliquot of the semen can be
measured using a pH meter designed for measurement of viscous solutions.
 pH < 7.0
 – With absence of sperm - ejaculatory duct obstruction or congenital bilateral
absence of the vas
 deferens, a condition in which seminal vesicles are also poorly developed.
 – contaminated with urine.
 Semen pH increases with time, as natural buffering decreases, so high pH
values may provide little clinically useful information.

22.  3. Microscopic examination:
 1.Sperm Motility:
 Procedure:
 Place a drop of liquefied semen on clean glass slide.
 Put a cover slip over it and examined it under the
microscope .
 Normally , within 2 hours of ejaculation , more than 60%
of spermatozoa are vigorously motile and in 6-8 hours
95-60% are still motile is.
 If motility is less than 50%, a stain for viability such as
methylene blue or eosin Y with nigrosin as counte rsatin
can be done.
 Heads of non-motile sperms are stained positive for the
dye.
 Sterile moles have <25% motility of sperms
 Motile forms decrease by about 5% per hr after the 4th hr

23.  Types of Motility:
 Darting motility: The sperm are moving on their self.(Grade-4)
 Very strong activity with forward progression- (Grade 3)
 Decent sperm activity with forward progression- (Grade 2)
 Sluggish motility: The sperm are active and are transferred to other
place very slowly.(grade-1)
 Necrozoospermia: Sperm are present but they are immotile they can
not move.(Grade-0)
 Oligozosopermia: Here few sperms are only motile.(Grade-2)
 Azoospermia: Sperm are present in high power.(Grade-0)

24. 2.Sperm count:
Sperm count is same as WBC count.
The calculation is similar to WBC formula expect that the reported
sperm count is per ml instead of per cumm. So result is
multiplied with thousand to convert cumm to ml.
 The composition of diluting fluid is as under:
 Sodium bicarbonate 5 gm
 Formalin (neutral) 1 ml
 Distilled water 100 ml
 This is done in Neubauer’s (hemacytometer) chamber
using a WBC pipette.
 Draw liquefied semen in WBC pipette up to marks 1 and
then the diluting fluid up to mark 11.
 After mixing it properly, charge the chamber.
 Allow the spermatozoa to settle down in 2 minutes.
 Examine under microscope and count the number of
spermatozoa in one large peripheral square.
 Normal Range: 40-300 millions/ml

25.  3.Sperm morphology:
 Procedure:
 Prepare a thin smear from liquefied semen on a glass slide and stain
with a Romanowsky stains ,
 Observe at least 200 sperms for any abnormality in their morphology
 Normally 80% of sperm are normal
 The abnormal forms of sperms are with double head , swollen and
pointed head , double tail and rudimentary forms.
 Observation:
1.Spermatozoa head: Light blue
2.Nuclear posterior: Dark blue
3.Bodies & tail: Red or pink
4.Spermatozoa size: 50-70 UM
 Observation for other abnormalities:
Abnormal:0-20%
More than 20% abnormal form may be associated with infertility.

26.  Following abnormalities are observed in
spermatozoa: