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1st May 2012 @ 06:45
Data from Sandra Duffy, Griffith University Received April 25th 2012 Four compounds from the arylpyrrole series tested in a late stage anti-gametocyte imaging assay. Relevant compounds: [data]2349[/data] Note - mistake in picture ZHY 6 was sent, which is OSM-S-38, not OSM-S-37 as shown in the figure. Method: The assay involves the use of a Pfs16-GFP transgenic NF54 Plasmodium falciparum strain kindly provided by Dr David Fidock (Columbia University). Compound is added to 384 well imaging plates containing 20,000 highly synchronous late stage IV gametocytes. The plates are incubated for 72 hours in standard incubation conditions (5%CO2, 5%O2, 37oC, 5% humidity). After incubation the viability marker MitoTracker Red CM-H 2 XRos is added to the wells. The plates are then imaged on the OPERA confocal high-throughput imaging system. The images obtained are then analysed using an algorithm for determining the number of viable gametocytes still remaining in the compound treated wells based on viability marker fluorescent intensity and the morphology of the GFP expressing parasite. The number of gametocytes per well in this assay is the same number of parasites used in the asexual imaging assay for 3D7 and K1. Results: PMY10-2 has no significant activity whilst PMY14-1 has activity comparable to that in the asexual assay. (MHT: Others?) Raw data:[data]2305[/data] Caveat: It has to be noted that the compounds have been in storage for a while and the plates defrosted several times for screening. The inactive compound could have lost asexual activity at the same time due to some instability. This was not tested.
Attached Files
Gametocyte Assay
Gametocyte Compounds