Semen Analysis PDF

6/26/2024 ◦Semen consists of four components ◦ Contributed separately by the ◦ Testes and epididymis Physiology...

6/26/2024 ◦Semen consists of four components ◦ Contributed separately by the ◦ Testes and epididymis Physiology ◦ Spermatozoa produce in seminiferous tubules and mature in epididymis ◦ 5% of semen volume ◦ Seminal vessels ◦ Majority of fluid of semen, 60% ◦ Contains fructose – causes sperm motility ◦ Prostate ◦ Acidic fluid (acid phosphatase, citric acid) ◦ 20-30% of volume of semen ◦ Responsible for coagulation and liquefaction ◦ Bulbourethral glands ◦ 5% of volume of semen ◦ Alkaline fluid ◦ Normal semen specimen must have all four components Specimen Collection ◦ Reasons for testing – 1. fertility 2. to evaluate the success of vasectomy ◦ Proper collection is crucial for evaluation of fertility ◦First portion of the ejaculate is missing, then ◦ Sperm count will be decreased ◦ pH is falsely increased ◦ Specimen will not liquefy ◦Last portion of ejaculate is missing, then ◦ Semen volume is decreased ◦ Sperm count is falsely increased ◦ pH is falsely decreased ◦ Specimen will not clot 2  6/26/2024 Specimen Collection ◦ Requirements ◦ Abstinence at least for 2 days but not more than 7 days ◦ WHO – World Health Organization recommends 3 specimens between 7 days and 3 weeks (2 abnormal specimens = SIGNIFICANT) ◦ Detailed instructions should be given – written preferably ◦ Collection at the testing facility is preferred ◦ Delivery within 1 hour of collection at room temperature ◦ Positive identification ( name, birth date and date and time of collection) ◦ Collection should not have any nonspermicidial, condoms, or lubrication ◦ Should uses Standard Precautions Semen Analysis ◦ Macroscopic (TEST WITHIN 1 HOUR od collection) ◦ Appearance – Normal gray-white, translucent ◦ Abnormal = clear (sperm concentration low) = white (increased WBCs) = red (blood) = yellow (urine – which is toxic to sperm ◦ Liquification – fresh (clotted) should liquefy in 30 – 60 minutes. Clot >60 minutes = decreased prostatic enzymes ◦ Volume – Normal 2 – 5 mL – decreased volume could reflect improper collection ◦ Viscosity – consistent of fluid- draw up into pipette – report as 0 (watery) to 4 (gel-like) ◦ pH – Normal pH 7.2 – 8.0 - Increased pH = infection - Decreased pH = obstruction 3  6/26/2024 Semen Analysis ◦ Only one spermatozoon is needed to fertilize ◦ Various factors can affect the sperm ◦ Quantity – Normal >20 -250 M/mL, border-line 10 -20 M /mL ◦ Morphology of the head and tail ◦ Speed - Motility ◦ Microscopic ◦ Count on Neubauer chamber with a 1:20 dilution ◦ Diluting fluid is sodium bicarbonate & formalin ◦ Count in the four corners plus the center ◦ Side must match within 10% ◦ Automated instruments –Computer- assisted semen analysis ◦ Sperm Class analyzer ◦ CEROS CASA system ◦ Automated Sperm Quality Analyzer ◦ Motility Semen Analysis Microscopic continued ◦ Microscopic Motility – ◦ Need to have sperm with forward, progressive movement ◦ Well-mixed, liquefied semen specimen ◦ Examine within 1 hour; evaluate undiluted on glass slide with cover slip ◦ Estimate percentage with progressive, forward motion in 20 HP fields or, examine 200 sperm per slide and count the percentages of the different motile categories using a manual cell counter ◦ Grading can be done using a scale of 0 to 4, with 4 indicating rapid, straight-line movement and 0 indicating no movement ◦ A minimum motility of 50% with a rating of 2.0 after 1 hour is considered normal 4  6/26/2024 Semen Analysis Microscopic continued Additional Semen Analysis Sperm viability – mix with eosin-nigrosine stain to stain the dead sperm and count the number of dead per 100 sperm. Seminal fluid fructose energy for the sperm Normal ≥13 umol/ejaculate Anti-sperm antibodies – antibodies can be produced by male and female Microbial – Chlamydia, Mycoplasma & Ureaplasma Chemical – α-glucosidase, free L-carnitine, zinc, citric acid, glutamyl transpeptidase and prostatic acid phosphatase Post-vasectomy semen analysis Monthly testing beginning at 2 months Continues until no viable sperm are seen on a wet preparation Good Lab practice = Quality Control 5