TY - JOUR
T1 - Contextual automated 3D analysis of subcellular organelles adapted to high-content screening
AU - Dorval, Thierry
AU - Ogier, Arnaud
AU - Genovesio, Auguste
AU - Hye Kuyon Lim, Kuyon Lim
AU - Do Yoon Kwon, Yoon Kwon
AU - Lee, Joo Hyun
AU - Worman, Howard J.
AU - Dauer, William
AU - Grailhe, Regis
N1 - Funding Information:
We gratefully acknowledge MEST Korea, Gyonggido, KISTI, and the Bachmann-Strauss Dystonia & Parkinson Foundation for funding support. We acknowledge the excellent technical assistance of Honggun Lee.
PY - 2010/8
Y1 - 2010/8
N2 - 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.
AB - 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.
KW - automation
KW - image analysis
KW - image-based screening
KW - phenotypic classification
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U2 - 10.1177/1087057110374993
DO - 10.1177/1087057110374993
M3 - Article
C2 - 20639502
AN - SCOPUS:77956028557
SN - 1087-0571
VL - 15
SP - 847
EP - 857
JO - Journal of Biomolecular Screening
JF - Journal of Biomolecular Screening
IS - 7
ER -