Intended Use; General Information - COOK Medical IVF Media Instructions For Use Manual

DIRECTIONS FOR USE
• Aseptic technique should be used.
• Semen should be processed within one hour of collection.
• Spermient must be diluted with Gamete Buffer (K-SIGB) to an appropriate concentration for
sperm preparation (e.g. 40% and 80% gradients).
• Gradients should be prepared immediately prior to use.
• Raw semen should not be centrifuged at any time.
• Allow the semen to liquefy at 37°C for 30 minutes.
• Warm the Spermient to 37°C for a minimum of 4 hours prior to use.
• Prepare two gradients by adding 1.5 mL of 40% under laid with 1.5 mL of 80% in conical
bottom tubes.
• Load 60% or up to 2.0 mL of the ejaculate onto one gradient and 40% or up to 1.2 mL onto
the other.
• Centrifuge 20 minutes at 300 g then carefully remove the seminal plasma, the upper
interface, the 40% layer and the lower interface. Leave the remainder of the 80% intact.
• Recover the sperm pellet using a clean pasteur pipette and re-suspend in 3 mL of Gamete
Buffer (K-SIGB).
• Centrifuge for 10 minutes at 600 g.
• Repeat the washing step in a further 3 mL of Gamete Buffer (K-SIGB).
• Remove the supernatant and re-suspend the pellet in a small volume (approximately 200 µL)
of Sperm Medium (K-SISM) or Fertilization Medium (K-SIFM).
• Count sperm and calculate the concentration. Adjust as required.
• Store in a 6% CO incubator at 37°C until required.
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SYDNEY IVF SPERM MEDIUM K SISM 20, K SISM 50 & K SISM 100

INTENDED USE

The Sperm medium intended for use during in vitro fertilization procedures to process sperm.

GENERAL INFORMATION

Sperm Medium is supplemented with Human serum albumin (10 mg/mL) and gentamicin
(0.01 mg/mL).
Ready to use after equilibration to 37°C and 6% CO
STORAGE AND STABILITY
Sperm Medium must be stored in original unopened container, refrigerated at 2-8°C. Do not
freeze.
DIRECTIONS FOR USE
• Aseptic technique should be used.
• Semen should be processed within one hour of collection.
• Warm the Sperm Medium to 37°C and equilibrate in a 6% CO
4 hours prior to use.
• Allow the semen to liquefy at 37°C for 30 minutes.
• Gently underlay aliquots (100-300 µL) of fully liquefied semen under 0.5 mL aliquots of
equilibrated Sperm Medium in round bottom 5 mL tubes.
• Place tubes in a test tube rack so that the tubes are at 60º to the horizontal. Place the rack in
the CO incubator.
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.
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incubator for a minimum of
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