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CM Pot 1 Week 4 - Semen Analysis Notes

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Jeeves Christian Josol
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7 views

CM Pot 1 Week 4 - Semen Analysis Notes

Uploaded by

Jeeves Christian Josol
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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SEMEN ANALYIS

3 purposes
- fertility testing
- postvasectomy semen analysis
-forensic analysis
PHYSIOLOGY

Composition of Semen
5% Spermatozoa Seminiferous tubule
- site of spermatogenesis
-_________________
Epididymis
___________________
60-70% Seminal Fluid Produced by Seminal Vesicles
(TERMINAL) - provides nutrients
- rich in fructose for motility
20-30% Prostate Fluid Acidic Fluid
Contains Zinc, ACP, Citric Acid, and other enzymes
5% Bulbourethral Fluid Thick alkaline mucus
Neutralizes acidity from the prostatic secretion and vagina
SPECIMEN COLLECTION
1. Abstinence of _______

2. In fertility testing WHO recommends two or three samples be collected not less than 7 days or more
than 3 weeks apart, with two abnormal samples considered significant.

3. Collect the entire ejaculate Methods: masturbation, coitus interruptus, condom method – use a non-
spermicidal, non-lubricant containing rubber or silastic condom

4. Specimen should be delivered to the laboratory within ____ of collection at room temp

5. Take note of the time of specimen collection, specimen receipt and liquefaction

6. Analysis should be done after liquefaction (usually _______ minutes)

7. If after 2 hours if the specimen has not liquefied, add Dulbecco’s phosphate-buffered saline, alpha
chymotrypsin, or bromelain to induce liquefaction

8. Specimen awaiting analysis should be kept at ____

9. Semen specimen are potential reservoir of HIV and Hepatitis

10. Jelly-like granules (gelatinous bodies) may be present in liquefied semen specimens and have no
clinical significance

1st portion ejaculate missing Last portion ejaculate missing


Sperm Count Decreased Increased
Ph increased Decreased
Specimen State Won’t liquefy Won’t Clot
SEMEN ANALYSIS

APPEARANCE
- gray-white color, translucent and musty odor
- INCREASED WHITE TURBIDITY = presence of WBC ad infxn

LIQUEFACTION
- jelly like granules = no significance
- mucus strands= may interfere with semen analysis

VOLUME
- 2-5 mL
- Decreased volume is more frequently associated with infertility and may indicate improper functioning
of one of the semenproducing organs, primarily the seminal vesicles

VISCOSITY
- droplets longer than 2 cm are considered highly viscous and considered abnormal
- Rating of 0 ( watery ) to 4 (gel-like) = used for reporting
- Increased viscosity and incomplete liquefaction impede testing for sperm motility, sperm
concentration, antisperm antibody detection, and measurement of biochemical markers.

pH
Normal = __ to __
Increase = ______________
Decrease = _______________
SPERM CONCENTRATION AND COUNT
Normal value = 20-250 million/ ml
➢ Borderline = between 10-20 million/ml
➢ Methods:
1. Improved Neubauer counting chamber
Dilution: 1:20 using a mechanical (positive displacement) pipette
Diluents: Formalin, Sodium bicarbonate, saline, distilled water
Purpose of diluents: To immobilize the sperm

2. Makler Counting chamber


- For undiluted specimen
-Uses heat to immobilize sperm cells
CALCULATING ROUND CELLS
-Peroxidase-positive granulocytes are the predominant form of leukocyte in semen and can be further
differentiated from spermatogenic cells and lymphocytes using a peroxidase stain

-
-This method can be used when counting cannot be performed during the hemocytometer count and to
verify counts performed by hemocytometer.
- > 1 million WBCs per ejaculate = an inflammatory condition associated with infection and poor sperm
quality and may impair sperm motility and DNA integrity

ADDITIONAL TESTING

Modified Bloom’s test


Reagent = ___________________
 ✓ Count the number of dead cells in a 100 sperm using brightfield or phase contrast microscope
 ✓ Living sperm = unstained, bluish white (at least 50%)
 ✓ Dead sperm = red with a purple background
(DONE)
 Many immobile cells  Defective flagellum
 Many immotile and NOnviable cells  Epididymal pathology
SEMINAL FLUID FRUCTOSE
- tested within 2 hrs to prevent fructolysis
- Screening test: Resorcinol test = (+) orange or orange-red color
- Low fructose levels are caused by abnormalities of the
 seminal vesicles,
 bilateral congenital absence of the vas deferens,
 of the ejaculatory duct, partial retrograde ejaculation, and
 androgen deficiency
- A normal quantitative level of fructose is equal to or greater than 13 µmol per ejaculate

AntiSperm Antibodies
- Tests to detect antibody-coate sperms are the mixed agglutination reaction (MAR) test and the
immunobead test.
- The MAR test is a screening procedure used primarily to detect the presence of immunoglobulin G
(IgG) antibodies
- A finding of less than 10% of the motile sperm attached to the particles is considered normal.

- Head-directed antibodies can interfere with penetration into the cervical mucosa or ovum, whereas
tail-directed antibodies affect movement through the cervical mucosa
Head-direct antibodies Penetration
Tail-directed movement
MICROBIAL AND CHEMICAL TESTING

ROUND CELLS
If greater than 1 million WBC/ ml =
If greater than 1 million spermatids/ml
Normal value
This is a test for chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum
-A more specific method for detection of sperm history is the detection of seminal glycoprotein p30
(prostatic specific antigen [PSA]

-
POST VASECTOMY SEMEN ANALYSIS
- Recommended testing includes examining a wet preparation using phase microscopy for the presence
of motile and nonmotile sperm. A negative wet preparation is followed by specimen centrifugation for
10 minutes and examination of the sediment
- A single “motile” sperm on a wet preparation is an indication of an unsuccessful vasectomy.

VARICOCELE
 Hardening of veins that drains the testes
 Most common cause of male infertility
 Sperm head = tapered head

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