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A novel approach to spot detection for two‐dimensional gel electrophoresis images using pixel value collection
Author(s) -
Cutler Paul,
Heald Geoffrey,
White Ian R.,
Ruan Jason
Publication year - 2003
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200390054
Subject(s) - computer science , bottleneck , software , artificial intelligence , pixel , image processing , digitization , segmentation , image segmentation , sensitivity (control systems) , computer vision , pattern recognition (psychology) , image (mathematics) , data mining , engineering , electronic engineering , embedded system , programming language
Separation of complex mixtures of proteins by two‐dimensional gel electrophoresis (2‐DE) is a fundamental component of current proteomic technology. Quantitative analysis of the images generated by digitization of such gels is critical for the identification of alterations in protein expression within a given biological system. Despite the availability of several commercially available software packages designed for this purpose, image analysis is extremely resource intensive, subjective and remains a major bottleneck. In addition to reducing throughput, the requirement for manual intervention results in the introduction of operator subjectivity, which can limit the statistical significance of the numerical data generated. A key requirement of image analysis is the accurate definition of protein spot boundaries using a suitable method of image segmentation. We describe a method of spot detection applicable to 2‐DE image files using a segmentation method involving pixel value collection via serial analysis of the image through its range of density levels. This algorithm is reproducible, sensitive, accurate and primarily designed to be automatic, removing operator subjectivity. Furthermore, it is believed that this method may offer the potential for improved spot detection over currently available software.