Parasite Inoculation and Quantification

Parasites were inoculated into 6 well plates of HFFs maintained in DMEM supplemented with 10% IFS and 10 μg/mL gentamicin, and allowed to grow undisturbed for 14 days. Plates were washed with PBS, and fixed for 10 min at room temperature with 100% ethanol. Staining was performed for 5 min at room temperature with 2 mL crystal violet solution per well (12.5 g crystal violet, 125 mL 100% ethanol, 500 mL 1% ammonium oxalate), followed by two washes with water and drying overnight.

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Waldman B.S., Schwarz D., Wadsworth MH I.I., Saeij J.P., Shalek A.K, & Lourido S. (2020). Identification of a Master Regulator of Differentiation in Toxoplasma. Cell, 180(2), 359-372.e16.

Publication 2020

Ammonium oxalate Crystal violet Ethanol Gentamicin Parasites

Corresponding Organization : Massachusetts Institute of Technology

Other organizations : Heidelberg University, Ragon Institute of MGH, MIT and Harvard, Broad Institute, University of California, Davis

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Variable analysis

independent variables

Parasite inoculation

dependent variables

Parasite growth

control variables

HFF cell culture in DMEM supplemented with 10% IFS and 10 μg/mL gentamicin
Incubation duration of 14 days
PBS washing
Fixation with 100% ethanol for 10 min at room temperature
Staining with crystal violet solution for 5 min at room temperature
Washing with water and drying overnight

controls

No positive or negative controls were explicitly mentioned in the provided information.

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