BACKGROUND: The combination of time-lapse imaging of live cells with high-throughput perturbation assays is a powerful tool for genetics and cell biology. The Mitocheck project employed this technique to associate thousands of genes with transient biological phenotypes in cell division, cell death and migration. The original analysis of these data proceeded by assigning nuclear morphologies to cells at each time-point using automated image classification, followed by description of population frequencies and temporal distribution of cellular states through event-order maps. One of the choices made by that analysis was not to rely on temporal tracking of the individual cells, due to the relatively low image sampling frequency, and to focus on effects that could be discerned from population-level behaviour. RESULTS: Here, we present a variation of this approach that employs explicit modelling by dynamic differential equations of the cellular state populations. Model fitting to the time course data allowed reliable estimation of the penetrance and time of appearance of four types of disruption of the cell cycle: quiescence, mitotic arrest, polynucleation and cell death. Model parameters yielded estimates of the duration of the interphase and mitosis phases. We identified 2190 siRNAs that induced a disruption of the cell cycle at reproducible times, or increased the durations of the interphase or mitosis phases. CONCLUSIONS: We quantified the dynamic effects of the siRNAs and compiled them as a resource that can be used to characterize the role of their target genes in cell death, mitosis and cell cycle regulation. The described population-based modelling method might be applicable to other large-scale cell-based assays with temporal readout when only population-level measures are available.

We investigate different approaches for efficient feature space reduction and compare different methods for cell classification. The application context is the development of automatic methods for analysing fluorescence microscopy images with the goal to identify those genes that are involved in the mitosis of human cells (cell division). We distinguish four cell classes comprising interphase cells, mitotic cells, apoptotic cells, and cells with clustered nuclei. Feature space reduction was performed using the Principal Component Analysis and Independent Component Analysis methods. Six classification methods were examined including unsupervised clustering algorithms such as K-means, Hard Competitive Learning, and Neural Gas as well as Hierarchical Clustering, Support Vector Machines, and Random Forests classifiers. Detailed results on the cell image classification accuracy and computational efficiency achieved using different feature sets and different classification methods are reported.