Examination of Semen

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Examination of Semen:


    -Semen is formed by testes present only in male.
    -Semen consists of spermatozoa and seminal plasma.
    -Semen examination is done in patient of sterility.
    -The cause of male sterility may be defective spermatozoa or number number of spermatozoa is quite less.
    -Semen examination is also done in person who are operated upon vasectomy, to check that spermatozoa are present in the semen.



Collection of Semen:
  • Patient should be asked not to pass semen for about 2-5 days before test.
  • Patient is asked to discharge semen using his hand (masturbstion).
  • Specimen of semen may be collected in clean and dry tube.
  • Specimen should reach laboratory for examination with 15 minutes.
  • Note the date, time of collection, and time of receiving specimen.
  • Examine semen only after liquefication (30-60 minutes).
Physical Examination:

Volume:
   - Measure the volume of semen in graduated cylinder.
   - Normal volume is 2-5 ml.

Color:
   - Normal Semen is opaque, white or grayish in color.
   - Normal Semen does not contain blood, pus or mucus.

Consistency:
  - Note down the consistency of specimen at the time of receiving it.
  - Find out the time taken for the semen to become liquid.
  - Fresh semen when passed out is viscid and clot quickly. It liquefies with in 20 - 60 minutes.

PH:
    - Dip a strip of Ph indicator paper (Ph between 6 and 8) in the semen and read the Ph given for the color most closely matching the test paper.
    - Normal Ph ranges from 7-7.7.

Microscopic Examination:

Sperm Motitility:
  • Sperm motility is seen at room temperature.
  • Place a drop of semen on a clean glass slide.
  • Cove a drop with a coverslip.  
  • Apply vaseline at the edge of coverslip to prevent drying.
  • Examine the slide under microscope under 10X objective.
  • When sperms are seen, shift to 40X objective.
  • Remember that is reduced by adjusting iris diapphram.
  • Count number of actievely motile (80%), Sluggishly motile or non-motile  (20%) sperm among 100 sperms. 
Sperm Count:
  • Pipette 0.95 ml of diluting fluid into a small bottle using 1 ml of graduated pipette.
  • Draw well mixed semen with WBC pipette up to 0.05 ml mark.
  • Blow 0.05 ml of semen into bottle containing 0.95 ml of diluting fluid (1:20 dilution).
  • Mix the contents well.
  • Keep coverslip properly properly over Neubauer chamber.
  • Add diluted semen with the help of pipette, by the side of coverslip. 
  • Leave this Chamber for 3 minutes on the bench. By this time, sperm will settele down .
  • Count the number of sperm in one large corner square (Area of one mm.sq). Use 10X objective and 6X eye piece.
  • While counting sperm, consider only complete sperms with intact heads and tails.
Calculation.

    Number of sperms in one large square = A
    Number of sperms per ml = A x 200000 millions.



Example:
    Number of sperms counted = 400
    Sperms count = 400 x 200000 = 80 millions / ml
    Normal sperm count = 60 - 150 millions / ml

Sperm Morphology:
  • Allow liquefication of semen in half to 1 hour.
  • Draw a smear of semen on a clear glass slide ( like thin blood smear ).
  • Dry the smear in air.
  • Fix the smear by passing it quickly over a flame once or twice.
  • Cool the slide and cover the smear with 25% aqueous basic fuchsin. keep it up for 5 minutes.
  • Wash with tap water and dry the smeared slide.
  • Examine the slide under oil immersion.
  • Count all sperm in the field.
  • Count defective sperms.
  • Change field and count a total of 100 sperms and report :
            Normal sperms = %
            Defective sperms = %
            (Defective sperms, should not be more than 20% in healthy man).



 Other Cells:
Also study other cells in semen like:
    - RBC
    -WBC
    - Epithelial cells.





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