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

• DOI: 10.1073/pnas.0912090107
• Publication Date: 2010-02-02T03:51:30Z
• Authors: Tao Peng; Ghislain M. C. Bo...; Estelle Glory-Afshar; Daniel R. Rines; Sumit K. Chanda; Robert F. Murphy
• Keywords: Colocalization; Cellular compartment; Compartment (ship); Organelle; Proteome; Protein Sorting Signals

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 probes for each compartment and acquisition images conjunction the macromolecule interest. Alternatively, tailored algorithms allow finding particular regions quantifying amount fluorescence they contain. Unfortunately, this approach requires extensive hand-tuning cell type-dependent. Here we describe machine-learning estimating signal without hand tuning, only separate training markers compartment. In testing on cells stained mixtures organelles, achieved 93% correlation between estimated expected amounts We also demonstrated that method can be used quantify drug-dependent translocations. enables automated unbiased determination distributions across compartments, will significantly improve imaging-based high-throughput assays facilitate proteome-scale localization efforts.