Quantitative analysis of subcellular distributions with an open-source, object-based tool

The subcellular localization of objects, such as organelles, proteins, or
other molecules, instructs cellular form and function. Understanding the
underlying spatial relationships between objects through colocalization
analysis of microscopy images is a fundamental approach used to inform
biological mechanisms. We generated an automated and customizable
computational tool, the SubcellularDistribution pipeline, to facilitate
object-based image analysis from 3D fluorescence microcopy images. To test
the utility of the SubcellularDistribution pipeline, we examined the
subcellular distribution of mRNA relative to centrosomes within Drosophila
embryos. Centrosomes are microtubule-organizing centers, and RNA
enrichments at centrosomes are of emerging importance. Our open-source and
freely available software detected RNA distributions comparably to
commercially available image analysis software. The
SubcellularDistribution pipeline is designed to guide the user through the
complete process of preparing image analysis data for publication, from
image segmentation and data processing to visualization.