COOK Medical IVF Media Instructions For Use Manual page 5

SYDNEY IVF SPERM GRADIENT K SISG 20 & K SISG 50
INTENDED USE
Sperm Gradient Kit is used to separate sperm based on density, using density gradient
solutions.
GENERAL INFORMATION
The solutions are supplemented with Human serum albumin (10 mg/mL) and gentamicin
(0.01 mg/mL).
Ready to use after equilibration to 37°C.
STORAGE AND STABILITY
Sperm Gradient Kit 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.
• 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 Sperm Gradient Kit to 37°C for a minimum of four 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 SPERMIENT K SISP 20 & K SISP 100
INTENDED USE
Spermient is used to separate sperm based on density, using density gradient solutions.
GENERAL INFORMATION
Spermient is supplemented with Human serum albumin (10 mg/mL) and gentamicin (0.01 mg/mL).
Ready to use after equilibration to 37°C.
STORAGE AND STABILITY
Spermient must be stored in original unopened container, refrigerated at 2-8°C. Do not freeze.
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