Automatic analysis of dividing cells in live cell movies to detect mitotic delays and correlate phenotypes in time.
Live-cell imaging allows detailed dynamic cellular phenotyping for cell biology and, in combination with small molecule or drug libraries, for high-content screening. Fully automated analysis of live cell movies has been hampered by the lack of computational approaches that allow tracking and recognition of individual cell fates over time in a precise manner. Here, we present a fully automated approach to analyze time-lapse movies of dividing cells. Our method dynamically categorizes cells into seven phases of the cell cycle and five aberrant morphological phenotypes over time. It reliably tracks cells and their progeny and can thus measure the length of mitotic phases and detect cause and effect if mitosis goes awry. We applied our computational scheme to annotate mitotic phenotypes induced by RNAi gene knockdown of CKAP5 (also known as ch-TOG) or by treatment with the drug nocodazole. Our approach can be readily applied to comparable assays aiming at uncovering the dynamic cause of cell division phenotypes.
University of Heidelberg, IPMB, BIOQUANT, and DKFZ Heidelberg, Department of Bioinformatics and Functional Genomics, Biomedical Computer Vision Group, D-69120 Heidelberg, Germany.
Nathalie
N
Harder
Felipe
F
Mora-Bermúdez
William J
WJ
Godinez
Annelie
A
Wünsche
Roland
R
Eils
Jan
J
Ellenberg
Karl
K
Rohr
D000465:Algorithms; D002453:Cell Cycle / Q000502:physiology; D002455:Cell Division / Q000502:physiology*; D019070:Cell Lineage; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006657:Histones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007700:Kinetics; D008677:Metaphase / Q000502:physiology; D018613:Microscopy, Confocal / Q000379:methods*; D008869:Microtubule-Associated Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D008938:Mitosis / Q000502:physiology*; D015739:Nocodazole / Q000494:pharmacology; D034622:RNA Interference; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D013997:Time Factors; D050257:Tubulin Modulators / Q000494:pharmacology
C454180:CKAP5 protein, human; D006657:Histones; D008869:Microtubule-Associated Proteins; D011993:Recombinant Fusion Proteins; D050257:Tubulin Modulators; D049452:Green Fluorescent Proteins; D015739:Nocodazole