Acquiring Fluorescence Time-lapse Movies of Budding Yeast and Analyzing Single-cell Dynamics using GRAFTS
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Zopf-2013-Acquiring fluorescen.pdf
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786.66 KB
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Author(s)
Zopf, Christopher J.
Maheshri, Narendra
Date Issued
July 2013
Journal
Journal of Visualized Experiments
Publisher
MyJoVE Corporation
Citation
Zopf, Christopher J., and Narendra Maheshri. “Acquiring Fluorescence Time-Lapse Movies of Budding Yeast and Analyzing Single-Cell Dynamics Using GRAFTS.” JoVE no. 77 (2013).
Version
Final published version
Abstract
Fluorescence time-lapse microscopy has become a powerful tool in the study of many biological processes at the single-cell level. In particular, movies depicting the temporal dependence of gene expression provide insight into the dynamics of its regulation; however, there are many technical challenges to obtaining and analyzing fluorescence movies of single cells. We describe here a simple protocol using a commercially available microfluidic culture device to generate such data, and a MATLAB-based, graphical user interface (GUI) -based software package to quantify the fluorescence images. The software segments and tracks cells, enables the user to visually curate errors in the data, and automatically assigns lineage and division times. The GUI further analyzes the time series to produce whole cell traces as well as their first and second time derivatives. While the software was designed for S. cerevisiae, its modularity and versatility should allow it to serve as a platform for studying other cell types with few modifications.
MIT Department
Massachusetts Institute of Technology. Department of Chemical Engineering
Terms of Use
Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.3791/50456
