9. GUIDELINES FOR SEMEN COLLECTION NOT RECOMMENDED COLD/SUN EXPOSED SAMPLE 10 NOT RECOMMENDED SAMPLE DURING FEVER 9 NOT RECOMMENDED ORDINARY CONDOM 8 NOT RECOMMENDED COITUS INTERRUPTUS 7 ROOM TEMPERATURE TEMPERATURE 6 CLEAN,WIDE MOUTHED GLASS /PLASTICCONTAINER CONTAINER 5 MASTURBATION METHOD OF COLLECTION 4 2 – 4 WEEKS PERIOD BETWEEN 2 SAMPLES 3 TWO IF RESULT IS ABNORMAL NO. OF SAMPLES 2 > 2 AND < 7 DAYS(3-5 DAYS) ABSINENCE 1 RECOMMENDATIONS CRITERIA
43. DEFINITION OF NORMAL SPERM < 20 % of head area Up to 4 Ambiguous 4) VACUOLES 4.5-5 u long,2.5-3.5 u wide .length/width = 1.5-1.75 3-5 u long, 2-3 u wide ( width = ½ -2/3 of length 3-5 u long, 2-3 u wide 4-5 u long, 2-3 u wide 3)SIZE 40-70 % of head surface >1/3 of head surface 40-70 % of head surface Approx.1/2 to 2/3 of head surface 2) ACROSOME Oval Oval Perfect oval, smooth border Oval HEAD 1) SHAPE WHO 3 rd WHO 2nd STRICT ASCP
44. DEFINITION OF NORMAL SPERM < 1/3 of head area < ½ of head area CD (cytoplasmic droplet 7 – 8 u LONG 7 – 8 u LONG LENGTH 1.5 X HEAD 5 u LONG 2) SIZE : LENGTH Axially attached Axially attached Axially attached Straight regular outline Straight regular outline Slender straight regular outline - MID PIECE 1) WHO 3 rd WHO 2 nd STRICT ASCP
45. DEFINITION OF NORMAL SPERM > Or =45 u long > 45 u long 10 x head 50 -55 u long 3)LENGTH Thinner than mid piece 1 u at base, 0.1 u at end 2) WIDTH Slender uncoiled ,regular outline Slender uncoiled ,regular outline Uniform size , uncoiled TAIL PIECE 1) WHO 3 rd WHO 2 nd STRICT ASCP
46. CATEGORIES DEFECTS Large -Small -Tapering -Amorphous -Double -Pyriform -Pin -Round -Vacuolated -Large -Small -Tapering -Amorphous -Double -Pyriform -Pin -Round BORDERLINE 1)NORMAL - Slight deviation from oval 2)ABNORMAL -Acrosome <40 %or >70 % of head area -Round head -Small head -Tapered head -Double head -Large head -Acrosomal abnormality -Post Acrosomal abnormality -bicephalic -Paired -Large head -Small head -Vacuolated HEAD WHO 3 rd WHO 2 nd STRICT ASCP
66. SPERM COUNT 0.4 1 2 4 CONVERSION FACTORS (no. of LARGE squares counted = 4 ) DIL.SEMEN(SEMEN + DILUENT ) SPERMS PER HIGH POWER FIELD 1:50 (1 + 49 ) > 200 million 4 1:20 (1 + 19 ) 40 – 200 million 3 1:10 (1+9 ) 15 – 40 million 2 1:5 ( 1 + 4 ) < 15million 1
67.
68. NORMAL VALUES FOR HUMAN SEMEN 50 % OR MORE LIVE VIABILITY 7 < 1 MILLION / ML WBC’s 6 15 % OR MORE NORMAL FORMS MORPHOLOGY 5 50 % OR MORE MOTILE MOTILITY 4 4O MILLION OR MORE/ ML SPERM COUNT 3 7.2 OR MORE pH 2 2.0 ML OR MORE VOLUME 1
72. SPERM FUNCTIONS TESTS PRINCIPLE TEST ESTIMATES THE FULL INCIDENCE OF SPERMS HAVING FULL COMPLEMENT OF MITOCHONDRIA CONTAINING RESP. ENZYMES SPERM MITOCHONDRIAL ACTIVITY INDEX TESTING THE ABILITY OF SPERMATOZOA HEAD NUCLEUS TO DECONDENSE. SPERM NUCLEAR CHROMATIN DECONDENSATION TEST. ACROSOMAL ENZYMES DISSOLVE PROTEIN & ASSESS IT UNDER PHASE CONTRAST MICROSCOPE TEST FOR ACROSOME INTACTNESS ASSESSMENT OF MEMBRANE FUNCTION BY EXPOSING THE SPERM TO HYPO OSMOTIC SOLUTION HYPO OSMOTIC SWELLING TEST