A method for improved clustering and classification of microscopy images using quantitative co-localization coefficients.
BACKGROUND
The localization of proteins to specific subcellular structures in eukaryotic cells provides important information with respect to their function. Fluorescence microscopy approaches to determine localization distribution have proved to be an essential tool in the characterization of unknown proteins, and are now particularly pertinent as a result of the wide availability of fluorescently-tagged constructs and antibodies. However, there are currently very few image analysis options able to effectively discriminate proteins with apparently similar distributions in cells, despite this information being important for protein characterization.


RESULTS
We have developed a novel method for combining two existing image analysis approaches, which results in highly efficient and accurate discrimination of proteins with seemingly similar distributions. We have combined image texture-based analysis with quantitative co-localization coefficients, a method that has traditionally only been used to study the spatial overlap between two populations of molecules. Here we describe and present a novel application for quantitative co-localization, as applied to the study of Rab family small GTP binding proteins localizing to the endomembrane system of cultured cells.


CONCLUSIONS
We show how quantitative co-localization can be used alongside texture feature analysis, resulting in improved clustering of microscopy images. The use of co-localization as an additional clustering parameter is non-biased and highly applicable to high-throughput image data sets.
School of Biology and Environmental Science & Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Dublin 4, Belfield, Ireland.
Vasanth R
VR
Singan
Kenan
K
Handzic
Kathleen M
KM
Curran
Jeremy C
JC
Simpson
D016000:Cluster Analysis; D006367:HeLa Cells; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000145:classification* / Q000379:methods*; D008853:Microscopy / Q000379:methods*; D021381:Protein Transport; D013347:Subcellular Fractions / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000378:metabolism
D020691:rab GTP-Binding Proteins