Current approaches often require specialized imaging or high magnification confocal images acquired in multiple planes, limiting the duration or throughput of experiments . Furthermore, current analysis methods depend on supervised learning, such as support-vector machine -based image classification , which is computationally intensive, requires extensive training, and may not be robust when applied across LY294002 PI3K inhibitor different cell lines or under changing experimental conditions. Our goal in this study was to develop a fully automated method that could measure changes in interphase and mitotic duration using simple wide-field fluorescence imaging. Because most cultured cells have a cell cycle duration of more than 18 hours, we utilized a single-plane, wide-field fluorescence imaging approach that enables long-term imaging of cells. Based on these imaging parameters, we developed a time-series approach to determine cell cycle phase duration, which does not require a WZ4002 EGFR/HER2 inhibitor training data set, and is computationally rapid. The software is integrated into a complete analysis platform that is publicly available. We show that this approach can accurately determine small changes in mitosis or interphase duration induced by a variety of different perturbations. DCELLIQ is a computer package that we have developed for automated analysis of timelapse movies of cells expressing the fluorescent nuclear marker H2B-GFP . This program automatically segments the images and identifies nuclei by local adaptive thresholding and seeded watershed segmentation with fragment merging . This process yields a binary image that represents the location of each nucleus, designating the region for subsequent feature extraction. Nuclei are then tracked from frame to frame by finding best matches for each nucleus based upon area, grey value histogram, XY displacement, speed, direction, shape similarity and Delaunay triangulation . A trace is then defined as a single nucleus tracked over time, with each trace including only one daughter cell when divisions occur .