Image analysis stain controls

A related approach was used by Ranefall et al. (1998) to quantify images of immunohistochemically stained cell nuclear Ki-67 antigen and cyclin A protein in bladder carcinoma tissue. They combined automatic, computerized image analysis with appropriate controls and reference material. This approach is superior to the automatic method without... 

The growing availability of image analysis systems has encouraged manufacturers to now offer a range of double or triple stains, often as RTU reagents for specific applications. In our experience, many of these reagents performed superbly however, it is important to note that each part of the double or triple stain must itself be separately validated, and proper controls must be included. 

Quantification by Two-Dimensional Gel Electrophoresis 2-DE is still considered the gold standard for quantitative proteomics [57,67]. This procedure relies on differential expression of proteins in control and test samples. Proteins from the two samples are separated by the 2-DE protocol, followed by staining with a suitable dye (e.g., CCB or SYPRO Ruby). Next, images of the stained spots are acquired, and spot densities are measured and compared by image analysis software to provide differential expression of proteins. Gel-to-gel variations can affect the precision of quantification by this procedme. 

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