title: A novel organelle map framework for high-content cell morphology analysis in high throughput.
abstract: A screening procedure was developed that takes advantage of the cellular normalization by micropatterning and a novel quantitative organelle mapping approach that allows unbiased and automated cell morphology comparison using black-box statistical testing. Micropatterns of extracellular matrix proteins force cells to adopt a reproducible shape and distribution of intracellular compartments avoiding strong cell-to-cell variation that is a major limitation of classical culture conditions. To detect changes in cell morphology induced by compound treatment, fluorescently labeled intracellular structures from several tens of micropatterned cells were transformed into probabilistic density maps. Then, the similarity or difference between two given density maps was quantified using statistical testing that evaluates differences directly from the data without additional analysis or any subjective decision. The versatility of this organelle mapping approach for different magnifications and its performance for different cell shapes has been assessed. Density-based analysis detected changes in cell morphology due to compound treatment in a small-scale proof-of-principle screen demonstrating its compatibility with high-throughput screening. This novel tool for high-content and high-throughput cellular phenotyping can potentially be used for a wide range of applications from drug screening to careful characterization of cellular processes.
authors: 1Molecular Mechanisms of Intracellular Transport, Unité Mixte de Recherche144 Centre National de la Recherche Scientifique/Institut Curie, Paris, France.
authors: Kristine
authors: K
authors: Schauer
authors: Jean-Philippe
authors: JP
authors: Grossier
authors: Tarn
authors: T
authors: Duong
authors: Violaine
authors: V
authors: Chapuis
authors: Sébastien
authors: S
authors: Degot
authors: Aurianne
authors: A
authors: Lescure
authors: Elaine
authors: E
authors: Del Nery
authors: Bruno
authors: B
authors: Goud
mesh_terms: D048430:Cell Shape; D003599:Cytoskeleton / Q000378:metabolism; D057166:High-Throughput Screening Assays; D006801:Humans; D007091:Image Processing, Computer-Assisted; D015388:Organelles
keywords: Rab7; cell biology; density mapping; lysosomes