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. 2017 Aug 16;7:8454. doi: 10.1038/s41598-017-09085-5

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Parasite detection in the brain of BALB/c mice infected with Leishmania donovani. (A) Kinetics of the parasite load in the brain detected by bioluminescence (upper panel) and in the cortex and base detected by RT-qPCR (bottom panel). The grey area corresponds to background signals. Data are represented as median and the interquartile range. (B) Representative images illustrating the 2D-bioluminescence analysis of the parasite detection in the brain at specific time points post infection. The regions of interest (ROIs) corresponding to the brain are represented as red circles. (C) In vivo tridimensional micro-computed tomography to localize the bioluminescence signals within the brain. Representative tridimensional reconstruction of an infected mouse skeleton at 120 days post infection exhibiting the x-axis (blue), y-axis (green) and z-axis (red), and two bioluminescent foci (red cubes). The cross-sectional images exhibit the coronal (z-axis), transaxial (y-axis) and sagittal (x-axis) views and allow the localization of both bioluminescent foci within the cranial cavity (red squares). See also Movie S1. (D,E) Parasite isolation from brain tissue. Percentage of positive parasite cultures from the cortex (D) and from the base (E) of infected BALB/c mice brains, according to different dilutions and time points (n = 3/time point). (F) Representative image of a parasite culture from a cortex at day 7 post infection, where it is possible to observe Leishmania donovani promastigotes with different morphologies (arrowhead) surrounded by cellular debris (*).