Software that measures the subcellular distribution of proteins, termed Quantitative Subcellular Compartmentalization Analysis, or Q-SCAn.
- Provides a quantitative measurement of target molecules in subcellular compartments.
- Method is rapid and automated and improves subcellular measurements.
- Other methods have not been extensively developed and existing methods are generally laborious and/or subjective.
In eukaryotes, there are many significant examples of proteins that are localized in distinct subcellular compartments such as the pathway responsible for repairing base damage in the nucleus and mitochondria. Tools to measure the subcellular distribution of a specific protein have not been extensively developed, and existing methods are generally laborious, subjective, or both. To address this issue, Emory researchers developed a rapid and automated software method, termed Quantitative Subcellular Compartmentalization Analysis (Q-SCAn), to measure these distributions quantitatively that works within the capabilities and functions of an external open-source program called CellProfiler. Specifically, the method allows for individual subcellular compartments to be defined using fluorescence micrographs with a specific marker, then the intensity of a target, fluorescently-tagged protein can be quantified within each compartment. Q-SCAn has been validated via examination of subcellular localization of various proteins and localization mutants. These findings demonstrate that Q-SCAn is a functional and flexible method for measuring subcellular protein distribution.
Method has been developed for and implemented in the open-source software package, CellProfiler, maintained by the Broad Institute.