Precision Matters: How Gradient Preparation Determines Your Results

19 Feb 2025 | News, Tips & Tricks

All solutions at room temperature

  • Prepare the gradient using 2 ml layers.
  • Prepare the gradient no more than 30 minutes before use to prevent the layers from mixing.
  • Avoid overloading—do not exceed the recommended 1.5 ml of sample
  • Follow the recommended centrifugation settings: 300g for 20 minutes.
  • Retrieve the pellet immediately after centrifugation stops.
  • Wash the pellet using a hSA containing medium: 500g for 10 minutes.

Using smaller volumes, uneven layers, or overloading the gradient can lead to blockage, spermatozoa aggregation, and rafting (the accumulation of sperm and other components in the gradient layers). These issues can significantly reduce yield and number of motile sperm after preparation.

This film will show the different layers after a centrifugation

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