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Babu Raman , Agnes Cheung, Mark R. Marten : Quantitative comparison and evaluation of two commercially available, two dimensional electrophoresis image analysis software packages, Z3 and Melanie,
Electrophoresis, 23 (2002) 2194-2202

University of Maryland, Baltimore County (UMBC)
Department of Chemical and Biochemical Engineering
1000 Hilltop Circle
Baltimore, MD 21250
 

• Image specifications:
• .tiff 
• 8-bit grayscale
  
• Gel (a): ~ 0.5 MB
• Gel (b): ~ 1.0 MB
  

• Downloads:
• Gel (a)
• Gel (b)

• Gel (a) with manually counted spots marked
• Gel (b) with manually counted spots marked
  

• Manual spot counting guidelines:

• Any small distinctly darkened and circular area was counted as a spot.
• Odd shaped features were carefully examined for spots.
• Tiny specks were not counted. 
• Large spots were carefully checked for clusters.
• In spot clusters, protrusions in the boundary were counted as spots.
• In streaked regions, again, protrusions were counted a spots.
• In faded regions of the gel, if the spots were round, they were assumed to be spots and were counted.

• "Center Pull" and "Height decrease" distortions were done using Adobe PhotoDeluxe Business Edition (PBE) version 1.0, Adobe Systems Inc., USA.
• % center pull distortion was calculated using Quantity One (Q1) version 4.2.2, Bio-Rad laboratories, California, USA.
• Downloads: 
• Center Pull distorted images
  
• Gel (a): 1.7 %
• Gel (a): 3.9 %
• Gel (a): 5.8 %
• Gel (a): 7.4 %

• Gel (b): 1.6 %
• Gel (b): 2.9 %
• Gel (b): 4.4 %
• Gel (b): 6.6 %     

• Distorted images: Height decreased in SDS PAGE dimension
  
• Gel (a): 1.0 %
• Gel (a): 2.0 %
• Gel (a): 2.5 %
• Gel (a): 4.0 %
• Gel (a): 5.0 %

• Gel (b): 1.0 %
• Gel (b): 2.0 %
• Gel (b): 2.5 %
• Gel (b): 4.0 %
• Gel (b): 5.0 %     

• The images were generated using MATLAB version 6.01 programming language, The MathWorks Inc., Massachusetts, USA
• Image specifications
• .tiff 
• 300 dpi
• 8-bit grayscale
• The gaussian volume of the spot in the center was varied across gels. 
• The peak height of the gaussian was the same for all the spots, corresponding to the highest gray value of 255.
  
• The volume ratio of the center spot in gels (b), (c), (d), (e), (f), (g), (h), (i), (j), (k), as compared to the center spot in gel (a) was 2, 4, 6, 10, 14, 18, 22, 26, 30, 40 respectively.
• Downloads:
• MATLAB program
• Image (a)
• Image (b)
  
• Image (c)
  
• Image (d)
  
• Image (e)
  
• Image (f)
  
• Image (g)
  
• Image (h)
  
• Image (i)
  
• Image (j)
  
• Image (k)