Dynamic analysis of the silver staining gel image of 2-DE for protein spots segmentation

Abstract

SUMMARYA novel image analysis method combined with recently developed powerful computing hardware is proposed, in which the time course for the image data of silver staining gel taken during the developmental process was used for the analysis of protein spots in two-dimensional polyacrylamide gel electrophoresis. The alignment procedure of the time course gel images using the landmark spots of plastic fragments was a critical step in the precise analysis of protein spots. The merged, or adjacent, protein spots that inhibit the quantification and identification of the proteins were successfully segmented from regions that were automatically determined from the previous time gel image, which effectively removed operator subjectivity. Furthermore, the effect of termination time of the developing process on the gel image analysis was not a concern, because the present analysis used the whole-time course of the gel image.

Proteomics Analysis of SsNsd1-Mediated Compound Appressoria Formation in Sclerotinia sclerotiorum

“…The silver staining has a low dynamic range, which has been criticized for the quantitative analyses of spots. However, the silver staining has very high sensitivity, which allows for a detection of very low protein amounts [ 20 ]; the improved and advanced image processing method could become feasible in better quantification of protein spots [ 57 ]. Besides, to obtain more data of the exact protein abundance, only the differential proteins with ≥3-fold changes were accepted for further study in this research.…”

Phase analysis in single-chain variable fragment production by recombinant Pichia pastoris based on proteomics combined with multivariate statistics

“…The recognition of culture phases in a culture process would be important if the determination of different culture operations in each phase had a critical advantage for the improvement of bioproduction (3,4). We have developed a novel image analysis method combined with recently developed powerful computing hardware, in which the time series of the image data of silver staining gel taken during the developmental process was used for analysis (5). This image analysis method can check the spots of the same protein in each image at different times of development, and was modified in the present study in order to compare corresponding protein spots from two different gel images.…”