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abstract. Parameters for the Amnis FlowSight imaging flow cytometer were optimized to quantitate the median
inhibitory concentration (IC50) of drugs against the malaria parasite Plasmodium falciparum in vitro. Cytotoxicity was
performed on 96-well plates as modified from Druilhe and Gentilini (1982), using acridine orange to measure DNA. Starting
from a non-treated parasite culture previously scored for parasitemia using a RAL kit (RAL Diagnostics, France), we tried
various gating strategies involving the brightfield and the FITC channels. A sequence of gatings consisting of FOCUSED ->
SINGLE -> INTERNALIZED -> BRIGHT DETAIL INTENSITY vs INTENSITY resulted in a scatter plot with at least 3 parasitized
erythrocyte populations.
The sum of these populations corresponded to parasitemia determined by microscopy. The gating algorithm was then
applied on drug treated parasites resulting in the consistent appearance of the scatter plot and its populations. Counts for the
R2 population resulted in reproducible dose response curves. The IC50 for chloroquine and artemisinin determined from these
sigmoid curves corresponded with literature values and historical data from the laboratory. Furthermore, a small but distinct
new population of parasitized erythrocytes tagged R4 appeared consistently. We are currently investigating the hypothesis
that this population might constitute a marker for a drug endpoint, perhaps as an indicator of apoptosis.
methods.