Contextual automated 3D analysis of subcellular organelles adapted to high-content screening.

Journal of Biomolecular Screening
Thierry DorvalRegis Grailhe

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 comp...Continue Reading

References

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Jan 9, 2004·Proceedings of the National Academy of Sciences of the United States of America·Rose E Goodchild, William T Dauer
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Aug 17, 2005·Journal of Biomolecular Screening·Dmytro Kevorkov, Vladimir Makarenkov

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Citations

Jul 28, 2013·Methods : a Companion to Methods in Enzymology·Charilaos MousoulisCorey P Neu
Jan 29, 2013·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·Anthony P LeonardYuri K Peterson
Aug 15, 2014·Neurotherapeutics : the Journal of the American Society for Experimental NeuroTherapeutics·Howard J Worman, William T Dauer
Jun 1, 2012·Expert Opinion on Drug Discovery·Martin Oheim
Dec 8, 2010·Neurobiology of Disease·D Cristopher BraggNutan Sharma

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Methods Mentioned

BETA
fluorescence
confocal microscopy
flow cytometry
fluorescence microscopy
fluorescence-activated cell sorting

Software Mentioned

VTK
FACSDiva
Windows [UNK]

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