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Maximize sperm survival during density gradient centrifugation.
PureSperm Buffer is designed specifically for two functions; first as a balanced salt solution for diluting PureSperm 100 to make up two layers of different densities for the gradient, and secondly, to reduce viscosity in the semen sample. The formula of PureSperm Buffer is optimized for maximum sperm survival and greatest yield during density gradient centrifugation. Watch video tutorial
PureSperm Buffer is a sterile, isotonic, salt solution. It is optimised for the dilution of PureSperm 100 in the preparation of density gradients for the separation and purification of human sperm in ART. This gradient system effectively separates normal sperm from lymphocytes, epithelial cells, abnormal, immature and senescent sperm, cell debris, bacteria and seminal fluid.
Prepare two PureSperm gradients for each semen sample. This reduces the risk of overloading a single gradient, provides security when handling tubes or recovering sperm pellets, and provides two tubes to balance the centrifuge rotor.
It can be extremely difficult to obtain a high yield from motile sperm from highly viscous semen samples. Nidacon has developed a method that could be helpful to you in the lab. You simply add PureSperm Buffer to the ejaculate, incubate at 37°C for 15-30 minutes and the sample is ready to use. This will give you a much higher proportion of motile sperm.
Protocol:
For sperm preparation from a viscous semen sample:
1. Bring all solutions to room temperature (17 to 27°C).
2. Measure the volume of the semen sample.
3. Dilute 1+3 , 1 part PureSperm Buffer and 3 parts semen sample
(e.g. 0.5 ml PureSperm Buffer + 1.5 ml semen sample).
4. Incubate at 37°C for 15-30 minutes.
5. Mix using a pipette.
6. Ready for sperm preparation on a density gradient.
Article No. / Name / Volume
PSB-100 / PureSperm Buffer / 100 mL