A Time-Series Method for Automated Measurement of Changes in Mitotic and Interphase Duration from Time-Lapse Movies

Background Automated time-lapse microscopy can visualize proliferation of large numbers of individual cells, enabling accurate measurement of the frequency of cell division and the duration of interphase and mitosis. However, extraction of quantitative information by manual inspection of time-lapse movies is too time-consuming to be useful for analysis of large experiments. Methodology/Principal Findings Here we present an automated time-series approach that can measure changes in the duration of mitosis and interphase in individual cells expressing fluorescent histone 2B. The approach requires analysis of only 2 features, nuclear area and average intensity. Compared to supervised learning approaches, this method reduces processing time and does not require generation of training data sets. We demonstrate that this method is as sensitive as manual analysis in identifying small changes in interphase or mitotic duration induced by drug or siRNA treatment. Conclusions/Significance This approach should facilitate automated analysis of high-throughput time-lapse data sets to identify small molecules or gene products that influence timing of cell division.

[1]  K. Greulich,et al.  Wavelength dependence of laser-induced DNA damage in lymphocytes observed by single-cell gel electrophoresis. , 1995, Journal of photochemistry and photobiology. B, Biology.

[2]  H. Erfle,et al.  High-throughput RNAi screening by time-lapse imaging of live human cells , 2006, Nature Methods.

[3]  T. Kanda,et al.  Histone–GFP fusion protein enables sensitive analysis of chromosome dynamics in living mammalian cells , 1998, Current Biology.

[4]  R. Gupta Species-specific differences in toxicity of antimitotic agents toward cultured mammalian cells. , 1985, Journal of the National Cancer Institute.

[5]  Qinghua Shi,et al.  Chromosome nondisjunction yields tetraploid rather than aneuploid cells in human cell lines , 2005, Nature.

[6]  Joshua T. Jones,et al.  Probing the precision of the mitotic clock with a live-cell fluorescent biosensor , 2004, Nature Biotechnology.

[7]  B. Godley,et al.  Blue Light Induces Mitochondrial DNA Damage and Free Radical Production in Epithelial Cells* , 2005, Journal of Biological Chemistry.

[8]  R Eils,et al.  Automated classification of mitotic phenotypes of human cells using fluorescent proteins. , 2008, Methods in cell biology.

[9]  Meng Wang,et al.  Novel cell segmentation and online SVM for cell cycle phase identification in automated microscopy , 2008, Bioinform..

[10]  Fiorenza Ianzini,et al.  The Large‐Scale Digital Cell Analysis System: an open system for nonperturbing live cell imaging , 2007, Journal of microscopy.

[11]  Xiaobo Zhou,et al.  Automatic Segmentation of High-Throughput RNAi Fluorescent Cellular Images , 2008, IEEE Transactions on Information Technology in Biomedicine.

[12]  C. Hager,et al.  Mechanisms of mitotic cell death induced by chemotherapy-mediated G2 checkpoint abrogation. , 2007, Cancer research.

[13]  Sachihiro Matsunaga,et al.  Development of a multistage classifier for a monitoring system of cell activity based on imaging of chromosomal dynamics , 2007, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[14]  Yi Wen Kong,et al.  The mechanism of micro-RNA-mediated translation repression is determined by the promoter of the target gene , 2008, Proceedings of the National Academy of Sciences.

[15]  T. Mitchison,et al.  Cell type variation in responses to antimitotic drugs that target microtubules and kinesin-5. , 2008, Cancer research.

[16]  Xiaobo Zhou,et al.  Automated segmentation, classification, and tracking of cancer cell nuclei in time-lapse microscopy , 2006, IEEE Transactions on Biomedical Engineering.

[17]  Meng Wang,et al.  Context based mixture model for cell phase identification in automated fluorescence microscopy , 2007, BMC Bioinformatics.

[18]  Bernd Fischer,et al.  CellCognition: time-resolved phenotype annotation in high-throughput live cell imaging , 2010, Nature Methods.

[19]  Stephen S. Taylor,et al.  Cancer cells display profound intra- and interline variation following prolonged exposure to antimitotic drugs. , 2008, Cancer cell.

[20]  Xiaobo Zhou,et al.  Multiple Nuclei Tracking Using Integer Programming for Quantitative Cancer Cell Cycle Analysis , 2010, IEEE Transactions on Medical Imaging.

[21]  Kannappan Palaniappan,et al.  Cell Segmentation Using Coupled Level Sets and Graph-Vertex Coloring , 2006, MICCAI.

[22]  Viji M. Draviam,et al.  Timing and checkpoints in the regulation of mitotic progression. , 2004, Developmental cell.

[23]  William J. Godinez,et al.  Automatic analysis of dividing cells in live cell movies to detect mitotic delays and correlate phenotypes in time. , 2009, Genome research.

[24]  Timothy A. Skimina,et al.  Activation of flavin-containing oxidases underlies light-induced production of H2O2 in mammalian cells. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[25]  Ram Dixit,et al.  Cell damage and reactive oxygen species production induced by fluorescence microscopy: effect on mitosis and guidelines for non-invasive fluorescence microscopy. , 2003, The Plant journal : for cell and molecular biology.

[26]  R. Kiss,et al.  Videomicroscopic extraction of specific information on cell proliferation and migration in vitro. , 2008, Experimental cell research.