Summary
Now that automated image-acquisition instruments (high-throughput microscopes) are commercially available and becoming more widespread, hundreds of thousands of cellular images are routinely generated in a matter of days. Each cellular image generated in a high-throughput screening experiment contains a tremendous amount of information; in fact, the name high-content screening (HCS) refers to the high information content inherently present in cell images (J Biomol Screen 2:249–259, 1997). Historically, most of this information is ignored and the visual information present in images for a particular sample is often reduced to a single numerical output per well, usually by calculating the mean per-cell measurement for a particular feature. Here, we provide a detailed protocol for the use of open-source cell image analysis software, CellProfiler, to measure hundreds of features of each individual cell, including the size and shape of each compartment or organelle, and the intensity and texture of each type of staining in each subcompartment. We use as an example publicly available images from a cytoplasm-to-nucleus translocation assay.
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Acknowledgments
The author is grateful to David M. Sabatini and his laboratory members at the Whitehead Institute for Biomedical Research, and Polina Golland and Thouis R. Jones at the Massachusetts Institute of Technology for their support and intellectual contributions to the methods described in this work. Images from Ilya Ravkin and testing by Adam Papallo are also much appreciated. The methods described in this work were also developed with the support of academic grants from the Society for Biomolecular Sciences and L’Oreal for Women in Science, and a Novartis fellowship from the Life Sciences Research Foundation.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Carpenter, A.E. (2009). Extracting Rich Information from Images. In: Clemons, P., Tolliday, N., Wagner, B. (eds) Cell-Based Assays for High-Throughput Screening. Methods in Molecular Biology, vol 486. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-545-3_14
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DOI: https://doi.org/10.1007/978-1-60327-545-3_14
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