Cytotoxic T-lymphocytes are capable of killing tumor cells. However, when tumor growth rates exceed the T-lymphocyte killing rate, tumor cells are able to spread nonetheless. Compound screening experiments were conducted to identify substances capable of modulating T-lymphocyte killing efficiency, quantifiable by the ratio between healthy and apoptotic tumor cells. High-throughput fluorescence microscopy was used to identify tumor cells in either state.
For the quantification of cell numbers, an automated segmentation pipeline was implemented as a macro in ImageJ. In addition to the macro, a plug-in was constructed to aid the analysis of the imaging and segmentation data by automatically generating scatter plots with a gating option. By selection gating, the generated scatter plots give access to additional functionality such as sub-population cell counting, cell identification, contour finding and the generation of (sub-image) morphology galleries.
High-throughput, T-Lymphocyte, Tumor, Gating, Contour finding
Louis Wolf, MSc obtained his masters degree in biology in 2007. Since then he has been working as a JAVA programmer for a number of employees. In 2010 Louis started working for the Microscopic Imaging Centre (MIC) and Nijmegen Centre for Molecular Life Sciences (NCMLS) within the Radboud University Nijmegen Medical Centre (RUNMC). At the RUNMC, he aides researchers by automating image analysis tasks. The main focus of his work is on quantification of the (collective) migration of cancer cells. Interests include microbiology, 3D reconstruction and web techniques. Besides his work for the RUNMC, Louis is a self employed computer programmer and part-time student in computing sciences.
Presenting author: Louis Wolf
Organisation: Radboud University Nijmegen Medical Centre
co-authors: Cindy Dieteren
Jeroen van der Laak