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MP34:Kuhn:LabelFreeCytotoxicityScreeningByDHM

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 J. Kühn1, M. Chambon1, J. Mena1, Y. Emery2, P. Marquet3, P. Magistretti3 and G. Turcatti1


1EPFL, Biomolecular Screening Facililty (BSF), Lausanne, Switzerland
2Lyncée Tec SA, PSE-A, Lausanne, Switzerland
3EPFL, Laboratory of Neuroenergetics and Cellular Dynamics (LNDC), Lausanne, Switzerland


Abstract

Phenotypic screening by fluorescence imaging is widely used for investigating a variety of biological processes in academic screening platforms as well as in modern pharmacological research. Usually, the goal is to detect cellular phenotypes induced by compounds (i.e. chemicals and siRNAs). For putting into evidence the biomarkers characterizing the different phenotypes, multiple labels and labeling approaches are often required leading to multiple cell-associated parameters defining or characterizing a given phenotype. Those derived from morphological changes appear as highly relevant biomarkers for getting information about various biological processes within the cell.

Here we focus on cell viability, which is of paramount importance in the pharmacological context and in general, in the bioactive probes discovery field. Early toxicological profile of compounds is valuable for the selection of candidate molecules for hits validation, secondary screen and for the hits follow-up process. Moreover, the further characterization of the cell death type, apoptosis, necrosis, or autophagy, may provide additional relevant information about the compounds effects on a given cell type.

For overcoming most of the limitations associated with automated fluorescence microscopy screens, we present a novel non-invasive label-free technology called Digital Holographic Microscopy (DHM) for routine use to monitor cell viability for phenotypic toxicological profiling of chemicals and siRNAs. This DHM technology can not only avoid the use of multiple invasive and sometime costly dyes, but its data output is naturally quantitative (proportional to dry mass) and the technique is free of focusing issues thanks to a unique “digital refocusing” feature. Here we are showing an excellent agreement between DHM-based cytotoxicity screening assays and fluorescence microscopy “control” screens based on Hoechst and Propidium Iodide dyes. Moreover the DHM data post-processing appears as entirely compatible with common cell imaging software, like e.g. CellProfiler.

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