Arif, Muhammad, Rajpoot, Nasir M. (Nasir Mahmood), Nattkemper, Tim W., Technow, Ulrike, Chakraborty, Trinad, Fisch, Nicole, Jensen, Nickels A. and Niehaus, Karsten. (2011) Quantification of cell infection caused by Listeria monocytogenes invasion. Journal of Biotechnology, Vol.154 (No.1). pp. 76-83. ISSN 0168-1656

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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.

Item Type:	Journal Article
Subjects:	Q Science > QA Mathematics > QA76 Electronic computers. Computer science. Computer software
Divisions:	Faculty of Science > Computer Science
Journal or Publication Title:	Journal of Biotechnology
Publisher:	Elsevier Science BV
ISSN:	0168-1656
Date:	10 June 2011
Volume:	Vol.154
Number:	No.1
Page Range:	pp. 76-83
Identification Number:	10.1016/j.jbiotec.2011.03.008
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Restricted or Subscription Access
URI:	http://wrap.warwick.ac.uk/id/eprint/42382

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