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Determining the distribution of probes between different subcellular locations through automated unmixing of subcellular patterns

热4 dechang 添加于 2010-4-6 22:41 | 1260 次阅读 | 2 个评论

作者
Peng T, Bonamy GMC, Glory-Afshar E, Rines DR, Chanda SK, Murphy RF
摘要
Many proteins or other biological macromolecules are localized to more than one subcellular structure. The fraction of a protein in different cellular compartments is often measured by colocalization with organelle-specific fluorescent markers, requiring availability of fluorescent probes for each compartment and acquisition of images for each in conjunction with the macromolecule of interest. Alternatively, tailored algorithms allow finding particular regions in images and quantifying the amount of fluorescence they contain. Unfortunately, this approach requires extensive hand-tuning of algorithms and is often cell type-dependent. Here we describe a machine-learning approach for estimating the amount of fluorescent signal in different subcellular compartments without hand tuning, requiring only the acquisition of separate training images of markers for each compartment. In testing on images of cells stained with mixtures of probes for different organelles, we achieved a 93% correlation between estimated and expected amounts of probes in each compartment. We also demonstrated that the method can be used to quantify drug-dependent protein translocations. The method enables automated and unbiased determination of the distributions of protein across cellular compartments, and will significantly improve imaging-based high-throughput assays and facilitate proteome-scale localization efforts.
详细资料
- 文献种类:期刊
- 期刊名称: Proceedings of the National Academy of Sciences of the United States of America
- 期刊缩写: Proc Natl Acad Sci U S A
- 期卷页: 2010年第107卷第7期 2944-2949页
- 地址: Center for Bioimage Informatics and Department of Biomedical Engineering, Carnegie Mellon University, Pittsburgh, PA 15213, USA
- ISBN: 0027-8424
- 备注:PMID:20133616
标签

unmixing of subcellular patterns distribution of probes
相关链接 DOI URL
附件
unmixing

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