High-resolution Imaging of P. falciparum Egress

P. falciparum egress was imaged as previously described (Collins et al., 2013 (link); Das et al., 2015 (link)), using 1 μM (4-[7-[(dimethylamino)methyl]−2-(4-fluorphenyl)imidazo[1,2-α]pyridine-3-yl]pyrimidin-2-amine (compound 2) to tightly synchronise egress. Following removal of compound 2 by washing, parasites were suspended in fresh pre-warmed medium and introduced into a pre-warmed microscopy chamber on a temperature controlled microscope stage at 37°C. Beginning 6 min after washing off the compound 2, DIC images were collected at 5 s intervals for 30 min using a Nikon Eclipse Ni Microscope fitted with a Hamamatsu C11440 digital camera and converted to QuickTime movies using Nikon NIS-Elements software.

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Sherling E.S., Knuepfer E., Brzostowski J.A., Miller L.H., Blackman M.J, & van Ooij C. (2017). The Plasmodium falciparum rhoptry protein RhopH3 plays essential roles in host cell invasion and nutrient uptake. eLife, 6, e23239.

Publication 2017

Eclipse Ni Microscope NIS-Elements software

Amine Microscope Parasites Pyridine

Corresponding Organization :

Other organizations : The Francis Crick Institute, National Institute of Allergy and Infectious Diseases, National Institutes of Health, London School of Hygiene & Tropical Medicine

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Variable analysis

independent variables

1 μM (4-[7-[(dimethylamino)methyl]-2-(4-fluorphenyl)imidazo[1,2-α]pyridine-3-yl]pyrimidin-2-amine (compound 2)

dependent variables

P. falciparum egress

control variables

Temperature (37°C)
Time (6 min after washing off compound 2)

controls

Positive control: Compound 2 used to tightly synchronize egress
Negative control: Not explicitly mentioned

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