Detection of P. falciparum-specific subpopulations of B-cells is important for studies of immunity in malaria. This protocol relies on the photostability and protein loading capacity of carboxylated quantum dots to detect a broad range of different P. falciparum-specific B-cells. Infected red blood cell ghosts, obtained by permeabilization of infected cells with Streptolysin O, are coupled with carboxyl quantum dots using N-ethyl-N-dimethylaminopropyl-carbodiimide condensation. Immunophenotyping of P. falciparum-specific B-cells is performed by flow cytometry using Fc-receptor block, quantum dot-infected red blood cell ghost conjugates, and fluorochrome-conjugated anti-human CD19 mouse monoclonal antibodies.
Keywords: B-cells; Flow cytometry; Ghost infected red blood cells; Malaria; Plasmodium falciparum; Quantum dots.
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