When the iRBCs are thawed, are they visualized under the microscope to check if healthy infected red blood cells are added to co-culture?

protocol Protocol: Purification, freezing, and thawing of 3D7 P. falciparum-infected red blood cells and antibody-dependent cellular cytotoxicity (ADCC ) assay


  1. How did you ensure that you had Schizonts after thawing before they were used in the assay?


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Savannah N. Lewis Author Answered Mar 16, 2023

Hi, to answer your questions:

  1. We don't add additional healthy red blood cells after counting - we use what we thaw from the frozen stocks, keeping in mind that the live yield will be about ~50% of the number frozen. The best way to do this is via light microscopy and counting live cells only (they have a "halo").
  2. We verify Pf cultures are schizonts visually by taking a blood smear immediately before magnet purifying and freezing them. We also ensure the cultures are highly synchronous via sorbitol synchronization before purification.


Hope this helps! Let us know if there is anything else we can clarify.

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Grace Turyasingura Answered Mar 16, 2023

Indiana University School of Medicine

Thank you , this is helpful insight.

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