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Semen Analysis UG

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21 views21 pages

Semen Analysis UG

Uploaded by

bijjamsaivardhan
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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SEMEN ANALYSIS

Infertility - Failure of a couple


to conceive/achieve a clinical
pregnancy after 12 months or
more of regular unprotected
coitus.
Causes of Infertility

• Male Infertility (30%)


• Female Infertility (30%)
• Both Male & Female
Infertility(30%)
• Unexplained (10%)
Approach to Male Infertility
Semen Analysis

• Collected after about 3 days of sexual abstinence.


• Obtained by masturbation.
• Collected in a clean, dry, sterile, and
leakproof wide-mouthed plastic container.

• Brought to laboratory within 1 hour of collection.

• During transport to the laboratory, the


specimen should be kept as close to body
temperature as possible.
• Two semen specimens should be examined that are
collected 2-3 weeks apart
Semen Analysis

Test Result
Physical examination Time to liquefaction, viscosity, volume, pH,
color.
Microscopic examination Sperm count, vitality, motility, morphology,
and proportion of white cells
Immunologic analysis Antisperm antibodies
SpermMAR test
Immunobead test
Bacteriologic analysis Detection of infection
Sperm function tests Postcoital test, cervical mucus penetration
test, Hamster egg penetration assay,
hypoosmotic swelling of flagella, and
computer-assisted semen analysis
Physical Examination
Appearance :
Normal - Gray-white color, translucent
White turbidity - Presence of WBCs (infection)
Red color - RBCs (infection,malignancy)
Yellow color - Urine contamination, specimen collection
following prolonged abstinence and
medications.
Physical Examination
Viscosity : Slightly viscous and easily drawn into a pipette.
Incompletely liquefied specimens will be clumped
and highly viscous.
Increased viscosity and incomplete liquefaction will
impede sperm motility.
Factors affecting semen viscosity
•Dehydration
•Infections of prostate/seminal vesicles
•Drugs
Semen Analysis
Physical Examination
Volume: N= 1.5-5 ml
Decreased volume frequently associated with infertility
indicates improper functioning of one of the semen-
producing organs.
pH : Normal pHis 7.2 to 8.0
Increased pH – infections within the reproductive tract.
decreased pH – increased prostatic fluid(acidic)
low semen volume.
Semen Analysis
Microscopic Examination
Sperm Motility
• A drop of semen is placed on a glass slide, covered with a coverslip that is
then ringed with petroleum jelly to prevent dehydration, and examined
under 40× objective.

• At least 200 spermatozoa are counted in several different microscopic


fields.
Progressive motility (PR): spermatozoa moving actively, either linearly
Non-progressive motility (NPR): all other patterns of motility with an absence
of progression
Immotility (IM): no movement
Sperm Viability
• Viable sperm cells will not take up the eosin Y stain, whereas non-viable cells
will be stained pink-red.

Method
1. Mix one drop of semen with 1 drop of eosin-nigrosin solution and incubate for
30 seconds.
2. A smear is made from a drop placed on a glass slide.

3. The smear is air-dried and examined under oil immersion objective


Sperm Count
1. Semen is diluted 1:20 with sodium bicaronate- formalin diluting fluid ( ml
liquified semen in graduated tube and fill with diluting fluid to 20ml mark &
mix well)
2 . Place a coverslip over Neubauer counting chamber filled with well mixed
diluted semen sample.
3. Spematozoa is counted in 4 large corner squares. N: >15mill/ml
Sperm Morphology (N:>4%)
Normal values of semen analysis (World Health Organization,2010)

Test Result
Volume ≥ 1.5 ml
pH ≥ 7.2
Sperm concentration ≥15 million/ml
Total sperm count per ejaculate 39 million
Morphology ≥ 4.0% sperms with normal morphology
Vitality ≥ 58% live forms
White blood cells ≤ 1 million/ml
Motility within 1 hour of ejaculation
Total motility (PR + NPR) ≥ 40% rapidly progressive
Progressive motility (PR) ≥32% progressive
Mixed antiglobulin reaction (MAR) test <50% motile sperms with adherent particles
Immunobead test <50% motile sperms with adherent particles
Biochemical variables of semen analysis (World Health Organization,2010)

Test Result
Total fructose (seminal vesicle marker) ≥13 μmol/ejaculate
Total zinc (Prostate marker) ≥2.4 μmol/ejaculate
Total acid phosphatase (Prostate marker) ≥200 U/ejaculate
Total citric acid (Prostate marker) ≥52 μmol/ejaculate
α-glucosidase (Epididymis marker) ≥20 mU/ejaculate
Carnitine (Epididymis marker) 0.8-2.9 μmol/ejaculate
Terminology Inference
Normozoospermia All semen parameters normal

Oligozoospermia Sperm concentration < 15 million/ml


mild to moderate: 5-15 million/ml
severe: < 5 million/ml)
Azoospermia Absence of motile/viable sperm in the
semen.
Aspermia Absence of ejaculate/semen

Asthenozoospermia Reduced sperm motility


<40% of sperms show total( progressive and
non-progressive) motility
or
<32% sperms showing progressive motility.

Teratozoospermia Spermatozoa with reduced proportion of


normal morphology (< 4% or increased
proportion of abnormal forms)

Leukocytospermia >1 million white blood cells/ml of semen

Oligoasthenoteratozoospermia All sperm variables are abnormal

Necrozoospermia All sperms are non-motile or non-viable


Sperm Function Tests
Hypoosmotic swelling of flagella
Sperm Function Tests
Computer assisted semen analysis

•Automated system measures various characteristics of the spermatozoa


•The semen specimen is placed on the stage of the microscope.
• This microscope has a high resolution videocamera attached.
• The videocamera feeds data into the computer where it undergoes analysis by software.
•A computerized semen analysis will give many more parameters that are useful to the
pathologist.
•More importantly, the results are accurate and reproducible.
Sperm Function Tests
Computer assisted semen analysis - sperm motility

Squiggly green lines =>individual sperm paths


Red dots => sperm that are not moving
Sperm Function Tests
Computer assisted sperm morphology

The software measures individual sperm cells “Strict Morphology”


and shapes A normal percentage under Kruger’scriteria is 14%
THANK YOU

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