Semen analysis Spermogram

1. Semen analysis(Spermogram)

2. Collected in : - a glass or plastic jar free of any preservative -warm ( at least to room T. ) Evaluated : within 30 min after collection (no examination after 2 hrs)

3. Record : 1- The name of the man 2- The period of abstinence 3- The date & time of collection 4- Completeness of collection 5- Difficulties in producing the sample 6- The interval between collection & analysis

4. Two samples should be collected for initial evaluation( interval :7 days � 3 weeks )

5. Macroscopic analysis ; 1- PH (measured with PH paper) 2-Volume (measured by weight or using graduated cylinder) 3-Appearance: abnormal smell ,color or viscosity( thread shorter than 2 cm) 4-Liquefaction time (above 60 min ) bromelain ? 5-Seeding for bacterial analysis

6. Microscopic analysis : 1-Sperm concentration : (total number , viability & sp. agglutination) - Centrifuge if indicated - Examine at least 200 sp. For motility 2- Presence of other cellular elements

7. Centrifugation properties : 1- sp. <1-2 / field 1 ml at 600 g for 15 min 2-no sp. 1 ml at 3000 g for 15 min

8. Preparation for routine analysis: A fixed volume of 10 ul semen onto a clean glass slide covered with a 22mm / 22mm coverslip Evaluation after one minute ( for stabilization ) Assessment at 37 C ( or a constant temperature between 20 � 40 C )

9. Grading system for sperm motility a. Rapid progressive motility (>25 um/s at 37 C and >20 um/s at 20 C) b. Slow or sluggish progressive motility c. Nonprogressive motility (<5 um/s ) d. Immotility

10. Preliminary estimation of sperm concentraton: This is used to decide the dilution for determining the sperm concentration by haemacytometer. - Counting spermatozoa per 400+ field - Preparing appropriate diluent (distilled water + NaHCO3 + formalin + gentian violet or trypan blue ) - Dilute the semen based on estimated sperm concentration

11. Assessment of sperm concentration : 1 - It should be determined using the haemocytometer method - 10 ul of the thoroughly mixed diluted specimen is transferred to the counting chamber - Counting spermatozoa in the central square of the grid (25 large squares ) , the number of large squares are determined based on the number of sp. per each square ( to 200 sp. ) - Calculate total number by using the formula.

12. Assessment of sperm concentration: 2 � alternative method : - 1 ml well-mixed sample + 19 ml cold water - Fill a hematology counting chamber - Count number of sperm seen in a 1 mm2 area ( 16 small squares ) . This number is then multiplied by 200,000 ) � Double figure and add 5 zeros �!!

13. Limitations of the porocedure : Delayed examination of the specimen Collection in improper container Exposure of the specimen to temperature extremes during transport Abnormally low sperm count allowing for evaluation of less than 200 spermatozoa Use of dirty or contaminated supplies

14. Semen culture : Abstinence for 5-7 days Passing urine before obtaining the sample C leang the genitalia with soap Collect the specimen in a sterile container Examine the specmen within 3 hrs after collection Culture in a G+ and G- medium