title: An image-based algorithm for precise and accurate high throughput assessment of drug activity against the human parasite Trypanosoma cruzi.
abstract: We present a customized high content (image-based) and high throughput screening algorithm for the quantification of Trypanosoma cruzi infection in host cells. Based solely on DNA staining and single-channel images, the algorithm precisely segments and identifies the nuclei and cytoplasm of mammalian host cells as well as the intracellular parasites infecting the cells. The algorithm outputs statistical parameters including the total number of cells, number of infected cells and the total number of parasites per image, the average number of parasites per infected cell, and the infection ratio (defined as the number of infected cells divided by the total number of cells). Accurate and precise estimation of these parameters allow for both quantification of compound activity against parasites, as well as the compound cytotoxicity, thus eliminating the need for an additional toxicity-assay, hereby reducing screening costs significantly. We validate the performance of the algorithm using two known drugs against T.cruzi: Benznidazole and Nifurtimox. Also, we have checked the performance of the cell detection with manual inspection of the images. Finally, from the titration of the two compounds, we confirm that the algorithm provides the expected half maximal effective concentration (EC50) of the anti-T. cruzi activity.
authors: Image Mining (IM) Group, Institut Pasteur Korea, Seongnam-si, Gyeonggi-do, South Korea ; Department of Mathematics, Seoul National University (SNU), Gwanak-Gu, Seoul, South Korea.
authors: Seunghyun
authors: S
authors: Moon
authors: Center for Neglected Diseases (CND3), Institut Pasteur Korea, Seongnam-si, Gyeonggi-do, South Korea.
authors: Jair L
authors: JL
authors: Siqueira-Neto
authors: Center for Neglected Diseases (CND3), Institut Pasteur Korea, Seongnam-si, Gyeonggi-do, South Korea.
authors: Carolina Borsoi
authors: CB
authors: Moraes
authors: Chemical Biology of Pathogen (CBP) Group, Institut Pasteur Korea, Seongnam-si, Gyeonggi-do, South Korea.
authors: Gyongseon
authors: G
authors: Yang
authors: Department of Mathematics, Seoul National University (SNU), Gwanak-Gu, Seoul, South Korea.
authors: Myungjoo
authors: M
authors: Kang
authors: Center for Neglected Diseases (CND3), Institut Pasteur Korea, Seongnam-si, Gyeonggi-do, South Korea.
authors: Lucio H
authors: LH
authors: Freitas-Junior
authors: Image Mining (IM) Group, Institut Pasteur Korea, Seongnam-si, Gyeonggi-do, South Korea.
authors: Michael A E
authors: MA
authors: Hansen
mesh_terms: D000465:Algorithms; D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D004353:Drug Evaluation, Preclinical; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D010271:Parasites / Q000166:cytology / Q000187:drug effects*; D021261:Parasitic Sensitivity Tests; D015203:Reproducibility of Results; D014344:Trypanocidal Agents / Q000032:analysis* / Q000494:pharmacology*; D014349:Trypanosoma cruzi / Q000166:cytology / Q000187:drug effects*
chemical_list: D014344:Trypanocidal Agents