Semen Fluid Lecture Notes PDF

SEMINAL FLUID Dr. RANIA SAAD SULIMAN.Associate Professor of Medical Laboratory Sciences.Consultant in Histopathology and Cytology 1  PHYSIOLOGY  Semen is composed of four fractions that are contributed by the testes, epididymis, seminal ve...

SEMINAL FLUID Dr. RANIA SAAD SULIMAN.Associate Professor of Medical Laboratory Sciences.Consultant in Histopathology and Cytology 1  PHYSIOLOGY  Semen is composed of four fractions that are contributed by the testes, epididymis, seminal vessels, prostate, and bulbourethral glands  the mixing of all four fractions during ejaculation is essential for the production of a normal semen 2 specimen 3  ANATOMY, COMPOSITION AND FORMATION  Testes: source of sperm  Seminal vesicles:  produce the majority of the fluid present in semen (60% to 70%).  The fluid contains a high concentration of fructose & nutrients (spermatozoa metabolize the fructose for the energy needed for the flagella to propel them through the female reproductive tract) 4  SEMEN COMPOSITION AND FORMATION  Prostate gland:  The muscular prostate gland, located just below the bladder  Surrounds the upper urethra and aids in propelling the sperm through the urethra by contractions during ejaculation  Approximately 20% to 30% of the semen volume is acidic fluid produced by the prostate gland  The acidic fluid contains high concentrations of acid phosphatase, citric acid, zinc, and proteolytic enzymes responsible for both the coagulation and liquefaction of the semen following ejaculation 5  2nd source of fluid  SEMEN COMPOSITION AND FORMATION  Bulbourethral glands:  Contribute about 5% of the seminal fluid volume  Thick alkaline mucous-like fluid that neutralizes acids (prostate secretions and the vagina)  located below the prostate. 6  SEMINAL FLUIDS  Spermatozoa: produced in the testes, mature in the epididymis. 7  SEMINAL FLUIDS (REASONS FOR TESTING)  Infertility issues: more often a problem with the woman, but easy to rule-out the male.  Post-vasectomy: frequent reason for testing  Test at one month intervals until 2 consecutive months are negative for sperm  Forensic analysis of fluid as being semen:  chemical test for enzyme: acid phosphatase. 8  SPECIMEN COLLECTION  Sterile container should be used  No lubricants, spermicides, condoms, etc.  2-3 days of sexual abstinence required (but not more than 5 days that have higher volumes and decreased motility)  Ideal collection site is at laboratory  If other, specimen must be kept warm and delivered to lab within 1 hour  Time of collection important (& must be9 recorded)  SEMINAL FLUIDS  When performing fertility testing, two or three samples are usually tested at 2-week intervals (if 2 samples are abnormal this considered significant). 10  SEMEN ANALYSIS  The semen analysis for fertility evaluation consists of both macroscopic and microscopic examination.  Parameters reported include appearance, volume, viscosity, pH, sperm concentration and count, motility, and morphology. 11  SEMINAL FLUIDS APPEARANCE  Grey-white, translucent, characteristic musty odor  Increased white turbidity indicates increased WBC and likely infection (leukocyte esterase test part of UA dipstick, may be useful)  Red coloration indicates RBCs  Yellow coloration can indicate urine contamination, medications, prolonged abstinence 12  VOLUME & VISCOSITY& PH  Volume:  Normal semen volume ranges between 2 and 5 mL.  Viscosity:  From a pipette tip, specimen should pour in droplets, and not be too stringy  Rating 0 = water-like through 4 = gel-like  Increased viscosity & / incomplete liquefaction will reduce sperm motility  pH:  NV = 7.2 –8.0  Alkaline to offset the vaginal environment  Sperm count: 13  NV = 20-160 million / mL  PHYSICAL CHARACTERISTICS  Liquefaction:  fresh specimen will clot, then liquefy within 30 minutes  Persistence of clot is abnormal  All further evaluation must wait until liquefaction is complete. 14  MICROSCOPIC EXAMINATION  Done 30 - 60 min after collection after liquidification has occurred  Motility:  Motility is a very necessary quality of sperm.  Using a hemocytometer, examine a drop of undiluted specimen using high dry objective 15  EXAMINATION MOTILITY  Subjective evaluation  Rated from “0” to 4.0”  4.0 : rapid and straight line movement  3.0 : slower, and some lateral movement  2.0 : slow forward progression, noticeable lateral movement  1.0 : no forward progression  0 : no movement  NV = 50% showing 2.0 or greater motility at 1 hour after collection  New technology –CASA (computer assisted semen analysis) provides objective evaluation, but only found in specialized 16 labs.  EXAMINATION MORPHOLOGY  Oval/egg shaped head (3x5µm) which is half covered with an enzyme laden acrosomal cap  Middle piece which provides energy  Tail piece of 45 µm long 17  EXAMINATION MORPHOLOGY  At least 200 cells evaluated on smear (Wright’s, Giemsa or Papanicolaou) stained.  Usually evaluated by pathologist, or cytologist  Looking for double heads, pin heads, giant heads, or amorphous heads, double, coiled, or missing tails, etc.  Normal = < 30% abnormal forms  WBC, RBC, bacteria presence should be noted (infection)  Round cells (neutrophils and immature sperm) should 18 be noted as well. 19 MICROSCOPIC EXAMINATION  Sperm count NV= 20 –160 million/mL  Make 1 to 20 dilution with the traditional diluting fluid which contains sodium bicarbonate and formalin (saline and distilled water can be used), which immobilize and preserve the cells  Count 5 small squares (within the center large square that used for RBCs) of the Neubauer 20 hemacytometer. 21  MICROSCOPIC EXAMINATION  Example: 52 cells (mature sperm) x 20 5 (squares) x 0.004  This provides results as ___ cells / uL;  Normal values are reported as ___ cells / mL  Must multiply X 1000 to convert uL to mL  = 52.0 x 106 / mL 22  OTHER TESTS (INFERTILITY)  Sperm viability:  Smear made from specimen mixed with eosin- nigrosin stain and evaluated under microscope.  Living cells appear blue-white  Dead cells appear red against blue background  At least 50-75 % should be live cells 23  SEMINAL FLUIDS  Examination of fluid as to being semen (forensic)  Acid phosphatase–highly sensitive, as no other body fluid contains as high level (2500 units compared to 5 units)  ABO, HLA typing  DNA analysis  UV light scan, semen fluoresces green/white 24