Semen analysis by Pandian M, Tutor Dept of Physiology DYPMCKOP, this ppt for MBBS,BPTH, BDS and all pre,para medicos

Interpret a normal semen analysis report as per WHO guidelines & discuss the results
1.Sperm count
2.Sperm morphology
3.Sperm motility

1. SEMEN ANALYSIS Pandian M Dept of Physiology DYPMCKOP

2. PY.9.5 Interpret a normal semen analysis report as per WHO guidelines & discuss the results   1.Sperm count   2.Sperm morphology   3.Sperm motility

3. INTRODUCTION Semen is a white fluid that contains sperms. It is the collection of fluids from testes, seminal vesicles, prostate gland and bulbourethral glands. Semen is discharged during sexual act and the process of discharge of semen is called ejaculation. Testes contribute sperms. Prostate secretion gives milky appearance to the semen. Secretions from seminal vesicles and bulbourethral glands provide mucoid consistency to semen.

4. Characteristic features Volume . The average volume of semen per ejaculation is 2.5–3.5 mL after an abstinence of 2 days . Volume of semen decreases with repeated ejaculations. Appearance of semen is milky due to prostatic secretions. Specific gravity is about 1.028. Reaction is alkaline with a pH of 7.5. The alkalinity is due to the prostatic secretions. The alkaline semen brings the vaginal pH from 3.5–4 to 6–6.5, the pH at which sperms show optimum motility .

5. Cont …….. Nature of the semen when ejaculated is liquid but soon it coagulates in vitro or in the vagina, and finally undergoes secondary liquefaction after about 15–30 min. The clotting of semen soon after ejaculation helps to retain it in the vagina for sometimes. Lysis ( fibrinolysin ) later on would release the sperms for their free movement into the uterine cavity for fertilization.

6. Components of semen and their characteristics 1. Spermatozoa. The normal sperm count varies from 35 to 200 million/mL of semen with an average of 100 million/ mL.

7. 2. Secretions of seminal vesicles . Secretions of seminal vesicles contribute 60% of the semen volume: The secretion from seminal vesicles is mucoid and viscous fluid . It is neutral or slightly alkaline in nature. It contains fructose, phosphorylcholine , ergothioneine , ascorbic acid, flavins and prostaglandins .

8. Cont …… Functions subserved by the seminal vesicle secretions are: Nutrition to sperms after being ejaculated into the female genital tract is provided by the fructose and other nutritive substances from the seminal vesicle secretions. Clotting of semen soon after ejaculation into the female genital tract occurs due to fibrinogen present in the seminal vesicle secretions. Fertilization of ovum may be enhanced by the prostaglandins present in the seminal vesicle secretion.

9. 3. Secretion of prostate gland . Secretion of prostate gland forms about 10% of the total semen bulk: It contributes milky and alkaline fluid part of the semen. Functions subserved by prostatic fluid component of semen are: Maintenance of optimum pH for fertilization (6–6.5) is the function of alkaline prostate fluid, which neutralizes the acidity of vaginal secretion. At this pH, the sperms become motile and the chances of fertilization are enhanced. Clotting of semen by converting fibrinogen (from seminal vesicles ) into a coagulum is caused by the clotting enzymes present in the prostatic fluid.

10. 4. Secretion of bulbourethral gland. Secretion of bulbourethral gland and other mucous glands provide mucoid consistency to the semen after puberty.

11. Sperm morphology (a) Diagram of a human mature sperm. ( b) A close-up of the head drawn from a different angle. The acrosome contains enzymes required for fertilization of the ovum.

12. Sperm motility The definition of  motility  is the ability of an organism or fluid to move.  Sperm motility  refers to the movement and swimming of  sperm . Poor   sperm motility  means that the  sperm  do not swim properly, which can lead to male infertility

13. Sperm count

14. Semen analysis

15. Apparatus Fresh sample of semen; 4% sodium bicarbonate in 1% phenol; apparatus required as for WBC counting Method – hanging drop preparation method

16. HAEMOCYTOMETER NEUBAUER CHAMBER RBC PIPETTE WBC PIPETTE

17. Side view of Neubauer chamber

18. Procedure for Mobility 1. Obtain fresh sample of semen collected in a small bottle. 2. Wait for 15 to 20 mins . So, that it may undergo secondary liquefaction. 3. To assess the sperm motility, place a drop of semen on the cover slip and invert it on a rim of plasticine on the slide 4. Examine under high power objective and find out the percentage of immobile to mobile sperms.

19. Determination of sperm count Gently shake the sample to assure uniformity Draw semen in the WBC pipette up to 0.5 mark Draw in 4% sodium bicarbonate in 1% phenol sol. Upto the mark of 11. Making a dilution of 1 in 20 Thoroughly shake the mixture, discard the first few drops Then charge the counting chamber. Ensure uniform distribution

20. METHOD OF DILUTION Take blood up to the 0.5 mark without any air bubble Take the dilution fluid upto the 11 or 101 mark Keep the pipette between the palms and mix the fluid The fluid in the bulb gets mixed completely

21. Different between over and under charge

22. Count the sperm under high power in the four WBC squares and Follow rules for counting.

23. Calculation Number of sperms in 1 µL of sample = N x 10/4 X 20 Number of sperm in 1 µL (i.e. 1 cumm ) of sample = N x 50 x 1000 (as 1 cc = 1000 µL ) N x 50,000

24. Reference Textbook of Practical Physiology A.K. Jain C.L. Ghai G.K.Pal Textbook of Medical Physiology By Guyton & Hall, A.K. Jain

25. THANK YOU . . .