<i>Leishmania amazonensis</i> lodge and multiply within giant acidic vacuoles.

<p>Six days-old macrophages were seeded into 384-well microplates and incubated with <i>L. amazonensis Ds</i>Red2 expressing amastigotes for 48 h. (<b>A</b>) Phase contrast image demonstrating the lodging of the amastigotes within giant vacuoles. Images of living cells were acquired with a 100× immersion objective on a Zeiss Axiovert 200 M microscope. Scale bar corresponds to 5 µm. (<b>B–D</b>) Images of living infected macrophage cultures incubated 60 min with fluorescent reporters were acquired on a confocal plate reader (Opera QEHS) after 3 days of incubation with the compounds. 20× confocal images acquired from samples treated with (<b>B</b>) 1% DMSO (C-), (<b>C</b>) 0.5 µM Amphotericin B (C+) and (<b>D</b>) 180 µM Cycloheximide (<b>C†</b>) are depicted. Scale bars represent 20 µm. Macrophage nuclei and PV were stained with Hoechst 33342 (Blue) and LysoTracker DND-26 (Green), respectively. Amastigotes expressing <i>Ds</i>Red2 are represented in red.