Contextual automated 3D analysis of subcellular organelles adapted to high-content screening.
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.
Dorval
Thierry
T
Cellular Differentiation, Institut Pasteur Korea, Seongnam-si, Gyeonggi-do, Korea. dorvalt@ip-korea.org
Ogier
Arnaud
A
Genovesio
Auguste
A
Lim
Hye Kuyon
HK
Kwon
Do Yoon
DY
Lee
Joo-Hyun
JH
Worman
Howard J
HJ
Dauer
William
W
Grailhe
Regis
R
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
D018832:Molecular Chaperones