For example, say that for one image, the maximum pixel value is 35 000 and the second image is 45 000. How can we be sure we can compare these two images with reliability?
Pixel values of 35 000 and 45 000 will cause no problem in the comparison of the images. However, it is always good to use as much of the dynamic range as possible as this will increase the sensitivity of the system. Typhoon™ imager has a very broad linear range, therefore it is possible to accurately detect even low signals. Of course it would be preferable to aim for similar signals for both images. This will improve data quality and reduce reliance on DeCyder Differential Analysis Software to perform normalization. However, DeCyder Differential Analysis Software will normalize images even if they have very different pixel values, and the outcome should not be affected by virtue of the internal standard present in each gel. Ideally, we recommend that the maximum pixel value is optimized on the first two gels. Aim to achieve a maximum pixel value between 50 000 and 70 000 in each of these gels. This will give scope in subsequent gels to avoid saturating spots (values above 100 000) or
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