Design of the RNA imaging P. falciparum growth assay. Cultures were incubated in a 96-well microplate with antimalarial compounds in complete medium for 72 hrs under a standard gas environment at 37°C. After the drug incubation period, cultures were diluted to a 0.025% hematrocrit to form a monolayer of erythrocytes in a 96-well microplate. RNA dyes were added and incubated at 37°C for 30 min to stain parasites in infected erythrocytes. An automated fluorescent microscope, the Pathway HT, was used to detect and obtain images of fluorescent stained parasites, as described in Materials and Methods. Transmitted light images were also obtained to verify erythrocyte infection. Fourteen dyes were found to have similar intensity to DAPI and SYBR Green I in the primary screen. 132A, 107E, and 107F dyes were found to have a higher affinity to RNA, displaying a diffused staining throughout the parasite, in the secondary screening.
Cervantes et al. BMC Cell Biology 2009 10:45 doi:10.1186/1471-2121-10-45