A novel tracking and analysis system for time-lapse cell imaging of Saccharomyces cerevisiae

抄録

Recent studies have revealed that tracking single cells using time-lapse fluorescence microscopy is an optimal tool for spatiotemporal evaluation of proteins of interest. Using this approach with Saccharomyces cerevisiae as a model organism, we previously found that heterochromatin regions involved in epigenetic regulation differ between individual cells. Determining the regularity of this phenomenon requires measurement of spatiotemporal epigenetic-dependent changes in protein levels across more than one generation. In past studies, we conducted these analyses manually to obtain a dendrogram, but this required more than 15 h, even for a single set of microscopic cell images. Thus, in this study, we developed a software-based analysis system to analyze time-lapse cellular images of S. cerevisiae, which allowed automatic generation of a dendrogram from a given set of time-lapse cell images. This approach is divided into two phases: a cell extraction and tracking phase, and an analysis phase. The cell extraction and tracking phase generates a set of necessary information for each cell, such as geometrical properties and the daughter–mother relationships, using image processing-based analysis techniques. Then, based on this information, the analysis phase allows generation of the final dendrogram by analyzing the fluorescent characteristics of each cell. The system is equipped with manual error correction to correct for the inevitable errors that occur in these analyses. The time required to obtain the final dendrograms was drastically reduced from 15 h in manual analysis to 0.8 h using this novel system.