Urinalysis and other Body Fluids Lab - 06 Seminalysis

ANALYSIS OF URINE AND OTHER BODY FLUIDS LABORATORY SEMINALYSIS MACROSCOPIC • SEMINALYSIS INTRODUCTION • • • • Advances in the field of andrology and assisted reproductive technology (ART), and increased concern over fertility, particularly by couples choosing to have children later in life, have resulted in increased emphasis on semen analysis. Patients with abnormal results on the routine semen analysis performed in the clinical laboratory often are referred to specialized andrology laboratories for further testing to determine the need for in vitro fertilization (IVF). In addition to fertility testing, the clinical laboratory performs postvasectomy semen analysis and forensic analyses to determine the presence of semen. Can be used in forensics Around 35°C is conducive to produce sperm • • MICROSCOPIC • PRINCIPLE • • Semen specimen Counting chambers (improved Neubauer) Sodium bicarbonate Formalin (neutral) Distilled Water 25% aqueous solution of basic fuchsin Microscope PROCEDURE SAMPLE COLLECTION • Motility 1. Place a large drop of mixed, liquefied specimen on a clear slide, cover with a cover glass. 2. Count the sperm cells showing progressive forward movement and nonmotile sperm cell in 20 HPF. C 3. Calculate the % of motile sperm cells % motility = total sperm cells - non-motile sperm cells x 100 total sperm cells Microscopy MATERIALS • • • • • • • Gross Appearance o Freshly ejaculated specimen consists of an opaque grayish to yellowish semigelatinous mass. ▪ Pearly white Volume o Measure the volume in a small graduated cylinder o The amount varies between few drops and 10mL o The average is 2-5mL. Samples less than 1.5 are considered below normal. Viscosity o Freshly ejaculated semen has a high degree of viscosity Preparation: o The sample should be collected in a private room near the laboratory, in order to limit the exposure of the semen to fluctuations in temperature and to control the time between collection and analysis. o The sample should be collected after a minimum of 2 days and a maximum of 7 days of sexual abstinence. If additional samples are required, the number of days of sexual abstinence should be as constant as possible at each visit. o The man should be given clear written and spoken instructions concerning the collection of the semen sample. These should emphasize that the semen sample needs to be complete and that the man should report any loss of any fraction of the sample. o The following information should be recorded on the report form: the man’s name, birth date and personal code number, the period of abstinence, the date and time of collection, the completeness of the sample, any difficulties in producing the sample, and the interval between collection and the start of the semen analysis. o Liquefaction time of sperm - 20-30 mins at room temp ▪ Included in the report • Sperm Cell Count 1. Mix the specimen thoroughly, shaking gently. 2. Use the leukocyte counting pipette and draw the semen up to the 0.5 mark, then dilute to 11 mark with the following diluting fluid: ▪ Sodium Bicarbonate - 5gms ▪ Formalin (Neutral) - 1cc ▪ Distilled Water - 100cc 3. Charge the mixture on a hemocytometer counting chamber. Allow the cells to settle for 2 minutes and count cells either in 5 RBC counting squares or 2 WBC counting squares Morphology 1. After liquefaction of the semen, prepare a thin smear on a clean glass slide. The smear should be made within 2 hours. 2. Dry in the air and fix gently with heat by passing the slide through the flame. 3. Cover the slide under with 25% aqueous solution of basic fucshin and stain for 5 minutes. 4. Wash with tap water, dry and examine under the oil immersion objective of the microscope. 5. The heads of the sperms are dark red color with the tail and the middle piece pink. 6. Determine the number of spermatozoa in microscopic field disregarding their morphology. 7. Examine 100 to 200 sperms for abnormalities ▪ Head – too small, too large, pointed ragged edges and a typical distribution of chromatic vacuoles or double heads. ▪ Middle Piece – absent, bifurcated or swollen ▪ Tail – double, rudimentary or absent 8. Report percentage of abnormal and normal forms. RESULTS OF THE TEST/S • Normal Values for Semen Analysis o Volume - 2 - 5Ml o Viscosity - Pours in droplets o pH - 7.2 – 8.0 o Sperm Concentration - > 20 million / mL o Sperm Count - > 40 million / ejaculate o Motility - > 50 % within one hour o Morphology - > 14 % normal forms (strict criteria) > 30 % normal forms (routine criteria) o Liquefaction time - within 20 minutes OTHER NOTES • • • • Chilled/cooled water - alternative for diluting fluid in sperm cell count Any smear can be used o Smear is used for observing morphology Video Testicle o