To determine antibody specificity for the parasite stages of interest, we generated formalin-fixed and paraffin-embedded mock tissue samples containing cultured RBCs infected with asexual and sexual P. falciparum parasite stages. Briefly, 100 μl of packed RBCs, derived either from asexual cultures or from individual time points of gametocyte development time courses, were mixed with thrombin 2 and fetal bovine plasma, wrapped in lens paper, and fixed with formalin. Cell clots were then placed into a paraffin wax block with a composite of surgical tissue specimens obtained from the Brigham and Women’s Hospital (Boston) for BMs that were analyzed in the autopsy study and sliced at a thickness of 3 μm. Using these samples, parasite antibodies and host antibodies were titrated for specificity and against host tissue background labeling.

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