sperm dna fragmentation by dr.renukadevi
TRANSCRIPT
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DR.RENUKADEVI SENIOR EMBRYOLOGIST
ARC International Fertility & Research Centre
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SCD test • In brief, a tube containing agarose was first heated at 100 C
for 5 minutes to allow the agarose to melt. •After stabilization at 37 C, 25μL semen aliquots were added
to the tube, and a 15μL aliquot of the mixture was placed onto a pretreated microscope slide. •A coverslip was placed and the slide was kept in the
refrigerator for 5 minutes in order for the agarose to solidify.•The slide was then taken from the refrigerator and the coverslip removed.•Thereafter, the slide was immersed in the denaturation solution and incubated for 7 minutes.
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• The slide was then transferred to the lysis solution and incubated for 25 minutes. • Finally, the slide was washed by incubation in a Coupling jar
containing distilled water for 5 minutes, followed by incubation in ethanol solutions of 70%, 90%, and 100%, each for 2 minutes. • After air drying at room temperature, slides were stained with
Wright's stain, and analysis was carried out using bright-field microscopy.• Sperm containing non-damaged DNA were scored as the sperm
showing large or medium-sized haloes of dispersed chromatin surrounding a compact and well-defined core.
INTERPRETATION & RESULT:SDF%= 100 X No. of sperm with fragmented DNA
No. of sperm counted A. ≤30% SDF =EXCELLENT TO GOODB. >30% TO < 40% SDF =EXCELLENT TO GOODC. >40% TO <50% SDF =FAIR TO POORD. ≥50% SDF =VERY POOR
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