Expression profiling using DNA microarrays is an important new method for
analyzing cellular physiology. In "spotted" microarrays, fluorescently
labeled cDNA from experimental and control cells is hybridized to arrayed
target DNA and the arrays imaged at two or more wavelengths. Statistical
analysis is performed on microarray images and show that non-additive
background, high intensity fluctuations across spots, and fabrication
artifacts interfere with the accurate determination of intensity
information. The probability density distributions generated by
pixel-by-pixel analysis of images can be used to measure the precision
with which spot intensities are determined. Simple weighting schemes
based on these probability distributions are effective in improving
significantly the quality of microarray data as it accumulates in a
multi-experiment database. Error estimates from image-based metrics
should be one component in an explicitly probabilistic scheme for the
analysis of DNA microarray data.