Quantification of cell infection caused by Listeria monocytogenes invasion |
| |
Authors: | Muhammad Arif Nasir M Rajpoot Tim W NattkemperUlrike Technow Trinad ChakrabortyNicole Fisch Nickels A JensenKarsten Niehaus |
| |
Institution: | a Department of Electrical Engineering, Pakistan Institute of Engineering & Applied Sciences (PIEAS), P.O. Nilore, Islamabad 45650, Pakistan b Department of Computer Science, University of Warwick, Coventry CV4 7AL, United Kingdom c Biodata Mining Group, Faculty of Technology, Bielefeld University, Germany d Giessen University, Research Center for Infectious Disease, Germany e Faculty of Biology, Department for Proteome and Metabolome Research, Bielefeld University, Germany |
| |
Abstract: | Listeria monocytogenes causes a life-threatening food-borne disease known as Listeriosis. Elderly, immunocompromised, and pregnant women are primarily the victims of this facultative intracellular Gram-positive pathogen. Since the bacteria survive intracellularly within the human host cells they are protected against the immune system and poorly accessed by many antibiotics. In order to screen pharmaceutical substances for their ability to interfere with the infection, persistence and release of L. monocytogenes a high content assay is required. We established a high content screen (HCS) using the RAW 264.7 mouse macrophage cell line seeded into 96-well glass bottom microplates. Cells were infected with GFP-expressing L. monocytogenes and stained thereafter with Hoechst 33342. Automated image acquisition was carried out by the ScanR screening station. We have developed an algorithm that automatically grades cells in microscopy images of fluorescent-tagged Listeria for the severity of infection. The grading accuracy of this newly developed algorithm is 97.1% as compared to a 74.3% grading accuracy we obtained using the commercial Olympus ScanR software. |
| |
Keywords: | High content screen Listeria monocytogenes Bioimage informatics Bacterial infection Cell segmentation Pattern recognition |
本文献已被 ScienceDirect 等数据库收录! |
|