Preparation Protocol for Buffy Coat from Whole Blood

GUIDELINE

The buffy coat is a fraction of whole blood that is rich in white blood cells (WBCs) and is named after its "buff-like" color. Preparation of a buffy coat from whole blood samples precludes many downstream applications. During processing, whole blood is centrifuged to form distinct blood fractions.

METHODS

  • Add an equal volume of recommended medium to whole blood and mix gently.
  • Centrifuge at 800 x g for 10 minutes at room temperature (15-25°C) with the brake off.
  • Remove the concentrated leukocyte band (this is the buffy coat), plus a small portion of the plasma and concentrated red blood cells (RBCs). The target is to concentrate the leukocytes approximately 5-fold while maintaining an equivalent ratio of leukocytes to RBCs (e.g. collect 2 mL of buffy coat when starting with 10 mL of whole blood).

Creative Bioarray Relevant Recommendations

NOTES

  • Invert the media and whole blood to mix gently.
  • Remove the sample from the centrifuge carefully, so as not to disturb the distinct blood fractions.

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For research use only. Not for any other purpose.