Activity #5

SEMINAL FUID EXAMINATION

OBJECTIVES: At the end of this activity, the student shall:

1. Explain the significance of seminal fluid analysis
2. Discuss methods of collection, transport, handling, processing, preservation and proper disposal
3. Characterize an acceptable specimen for routine testing
4. Describe a normal seminal fluid
5. Perform the following parameters in a routine semen analysis:
a. Physical examination ( volume, color, pH, liquefaction time, viscosity)
b. Sperm motility
c. Sperm count
d. Sperm morphology
6. Arrive accurate calculation of sperm concentration and sperm count
7. Discuss principle involved in each test performed
8. Correlate test results with pathologic conditions
9. Discuss variables that affect test results
10. Apply concepts of quality assurance and quality control
11. Report test results based on the standard format

Materials:
Per group: per class:
Hemocytometer & tally counter safranin stain
Cold water petroleum jelly
2 slides & 1 cover slip
Dropper, applicator stick
Macroscopic Examination:
Take note of the appearance, volume, pH, viscosity of the semen and the liquefaction time.

Microscopic examination:
1. Motility
a. Hanging drop Method
1. Place 1 drop of semen on a pre-warmed slide
2. Cover with coverslip which has been ringed with petroleum jelly
3. Scan several fields until a total of at least 200 sperm cells have been observed
4. Take note of the percentage of sperm cells showing actual progressive motion
Sperm Motility Grading
4.0 a Rapid, straight line motility
3.0 b slower speed, some lateral movement
2.0 b slow forward progression, noticeable lateral movement
1.0 c No forward progression
0 d No movement
Normal = > 50% motile sperm cells w/in 1 hr.

2. Morphology
Procedure:
1. Prepare a smear (same as in preparation of blood films)
2. Dip once into either 95% ethanol or 50% ethanol ether
3. Air dry
4. Stain with safranin (3-5 dips)
5. Rinse with water
6. Air dry

Examine at least 200 cells under oil immersion objective and take note the percentage of normal and abnormal forms.
Observe appearance of both head and tail.

Normal sperm morphology

Head oval shaped 0.5 um long and 3 um wide acrosomal cap ½ of the head for ovum penetration
Middle piece contains mitochondria- provides energy for flagellar tail motion
Tail 45 um long flagellar tail
Abnormal head double, tapered, giant, amorphous, pinhead, constricted
Abnormal tail double, coiled, bent

3. Sperm cell count and sperm concentration

Normal value - sperm concentration: >20million/ml
Sperm count: >40 million/ejaculation
Diluting fluids- Na bicarbonate, formalin, cold water

Procedure:
a. Suck liquefied semen up to 0.5 mark of WBC pipet
b. Suck diluting fluid up to 11 mark
c. Mix, discard 3-5 drops and charge in a counting chamber
d. Stand for 2 minutes ( for the immobilized sperm to settle)
e. Count sperm cells in 2 sq. mm (2 WBC squares)
Formula: Sperm cells/ml= sperm cells counted x 100,000