Approximately 200 μL of
supernatant was left with particle fraction on the bottom of the tubes to avoid the uptake of
nanoparticle clusters in the pipette. Tubes with spermatozoa were placed in a 200 ml water bath
and cooled to 5 °C, over next 1 ½ h. Post-sort sperm concentration was re-assessed by
hemocytometer and adjusted to 40 x 106 spermatozoa/mL. Following nanopurification, an equal
volume of egg-yolk citrate extender containing 14% glycerol were added in 3 aliquots of 20%,
30%, and 50% of the ½ volume amount. Color-coded straws were filled with 10 (10M CON,
10M UBI, 10M PNA) or 20 (20M CON) million control or nanopurified spermatozoa and
Approximately 200 μL ofsupernatant was left with particle fraction on the bottom of the tubes to avoid the uptake ofnanoparticle clusters in the pipette. Tubes with spermatozoa were placed in a 200 ml water bathand cooled to 5 °C, over next 1 ½ h. Post-sort sperm concentration was re-assessed byhemocytometer and adjusted to 40 x 106 spermatozoa/mL. Following nanopurification, an equalvolume of egg-yolk citrate extender containing 14% glycerol were added in 3 aliquots of 20%,30%, and 50% of the ½ volume amount. Color-coded straws were filled with 10 (10M CON,10M UBI, 10M PNA) or 20 (20M CON) million control or nanopurified spermatozoa and
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