Upon fixation and immunostaining, cell culture plates were analyzed to determine the number and size of the parasites using a CellInsight High Content Screening platform equipped with the Studio HCS software (Thermo Fisher Scientific). Uninuclear hepatic parasite forms observed beyond D4 were considered to be putative hypnozoites, whereas multinucleate forms were classed as schizonts. The parasite size reduction was calculated on the average object area using the total surface area of each selected object (μm2). The high-content imaging approach has been described previously (Bosson-Vanga et al, 2018 (link)). To assess cell toxicity of infusion for hepatic cultures, fixed plates were further scanned for the DAPI signal representing host nuclei. The analysis was based on counting of total cell nuclei.
Ashraf K., Tajeri S., Arnold C.S., Amanzougaghene N., Franetich J.F., Vantaux A., Soulard V., Bordessoulles M., Cazals G., Bousema T., van Gemert G.J., Le Grand R., Dereuddre-Bosquet N., Barale J.C., Witkowski B., Snounou G., Duval R., Botté C.Y, & Mazier D. (2021). Artemisinin-independent inhibitory activity of Artemisia sp. infusions against different Plasmodium stages including relapse-causing hypnozoites. Life Science Alliance, 5(3), e202101237.
Cell Cell cultureDapi Nuclei of cell ParasiteSchizonts
Corresponding Organization : Institut de Recherche pour le Développement
Other organizations :
Institut Pasteur du Cambodge, Institut Pasteur, Institut des Biomolécules Max Mousseron, Université de Montpellier, Radboud University Nijmegen, Direction de la Recherche Fondamentale, Commissariat à l'Énergie Atomique et aux Énergies Alternatives, Université Paris-Sud, Infectious Disease Models and Innovative Therapies, CEA Paris-Saclay - Etablissement de Fontenay-aux-roses
Proportion of uninuclear hepatic parasite forms (putative hypnozoites)
Proportion of multinucleate forms (schizonts)
Total cell nuclei (host cells)
control variables
Cell culture plates
CellInsight High Content Screening platform
Studio HCS software
Fixation and immunostaining
DAPI signal for host nuclei
Annotations
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