Adaptive contrast enhancement of two-dimensional electrophoretic protein gel images facilitates visualization, orientation and alignment
2‐DE is a powerful technique to discriminate post‐translationally modified protein isoforms. However, all steps of 2‐DE preparation and gel‐staining may introduce unwanted artefacts, including inconsistent variation of background intensity over the entire 2‐DE gel image. Background intensity variati...
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Published in | Electrophoresis Vol. 27; no. 20; pp. 4086 - 4095 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
Published |
Weinheim
WILEY-VCH Verlag
01.10.2006
WILEY‐VCH Verlag |
Subjects | |
Online Access | Get full text |
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Summary: | 2‐DE is a powerful technique to discriminate post‐translationally modified protein isoforms. However, all steps of 2‐DE preparation and gel‐staining may introduce unwanted artefacts, including inconsistent variation of background intensity over the entire 2‐DE gel image. Background intensity variations limit the accuracy of gel orientation, overlay alignment and spot detection methods. We present a compact and efficient denoising algorithm that adaptively enhances the image contrast and then, through thresholding and median filtering, removes the gray‐scale range covering the background. Applicability of the algorithm is demonstrated on immunoblots, isotope‐labeled gels, and protein‐stained gels. Validation is performed in contexts of (i) automatic gel orientation based on Hough transformation, (ii) overlay alignment based on cross correlation and (iii) spot detection. In gel stains with low background variability, e.g. Sypro Ruby, denoising will lower the spot detection sensitivity. In gel regions with high background levels denoising enhances spot detection. We propose that the denoising algorithm prepares images with high background for further automatic analysis, without requiring manual input on a gel‐to‐gel basis. |
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Bibliography: | ArticleID:ELPS200500925 ark:/67375/WNG-4M9M4DMM-X istex:E5B29D65CE22A8B85053A8A2B55767C390B02CC8 Additional corresponding author ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0173-0835 1522-2683 |
DOI: | 10.1002/elps.200500925 |