1 × 106 Hepa 1-6 hepatocyte cells resuspended in DMEM (Sigma, St. Louis, MO, USA) with 10% v/v FCS (Sigma) were plated in 40-mm glass-bottom culture dishes (Greiner bio-one, Monroe, NC, USA, at 5 × 105 cells/well) for at least 4 h before inoculation with 1 × 104 freshly isolated or cryopreserved P. berghei sporozoites. The infected cultures were washed twice with PBS and imaged at 36 h post-infection (p.i.) (Applied Precision DeltaVision Microscope System, GE Healthcare, Issaquah, WA, USA) [27 (link)]. Cryopreserved Pb-Luc-GFP parasites were visualized using the GFP channel to assess conservation of transgenic properties following cryopreservation.
For in vivo infections, 1 × 103 freshly isolated or cryopreserved P. berghei or P. yoelli sporozoites were enumerated and resuspended in 200 µL complete normal saline before inoculating into B6 mice intravenously (i.v.). The parasite burdens or immune responses were assessed at 24 h, 44 h, or 7–10 days p.i.
For in vivo infections, 1 × 103 freshly isolated or cryopreserved P. berghei or P. yoelli sporozoites were enumerated and resuspended in 200 µL complete normal saline before inoculating into B6 mice intravenously (i.v.). The parasite burdens or immune responses were assessed at 24 h, 44 h, or 7–10 days p.i.
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