title: Contextual automated 3D analysis of subcellular organelles adapted to high-content screening.
abstract: Advances in automated imaging microscopy allow fast acquisitions of multidimensional biological samples. Those microscopes open new possibilities for analyzing subcellular structures and spatial cellular arrangements. In this article, the authors describe a 3D image analysis framework adapted to medium-throughput screening. Upon adaptive and regularized segmentation, followed by precise 3D reconstruction, they achieve automatic quantification of numerous relevant 3D descriptors related to the shape, texture, and fluorescence intensity of multiple stained subcellular structures. A global analysis of the 3D reconstructed scene shows additional possibilities to quantify the relative position of organelles. Implementing this methodology, the authors analyzed the subcellular reorganization of the nucleus, the Golgi apparatus, and the centrioles occurring during the cell cycle. In addition, they quantified the effect of a genetic mutation associated with the early onset primary dystonia on the redistribution of torsinA from the bulk endoplasmic reticulum to the perinuclear space of the nuclear envelope. They show that their method enables the classification of various translocation levels of torsinA and opens the possibility for compound-based screening campaigns restoring the normal torsinA phenotype.
authors.affiliation: Cellular Differentiation, Institut Pasteur Korea, Seongnam-si, Gyeonggi-do, Korea. dorvalt@ip-korea.org
authors.forename: Thierry
authors.initials: T
authors.lastname: Dorval
authors.forename: Arnaud
authors.initials: A
authors.lastname: Ogier
authors.forename: Auguste
authors.initials: A
authors.lastname: Genovesio
authors.forename: Hye Kuyon
authors.initials: HK
authors.lastname: Lim
authors.forename: Do Yoon
authors.initials: DY
authors.lastname: Kwon
authors.forename: Joo-Hyun
authors.initials: JH
authors.lastname: Lee
authors.forename: Howard J
authors.initials: HJ
authors.lastname: Worman
authors.forename: William
authors.initials: W
authors.lastname: Dauer
authors.forename: Regis
authors.initials: R
authors.lastname: Grailhe
mesh_terms: D000818:Animals; D001331:Automation / Q000379:methods*; D002453:Cell Cycle; D002467:Cell Nucleus / Q000378:metabolism; D020821:Dystonic Disorders / Q000175:diagnosis; D006367:HeLa Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D051379:Mice; D008954:Models, Biological; D018832:Molecular Chaperones / Q000378:metabolism; D015388:Organelles / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D018832:Molecular Chaperones