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Microarray features that received a "−100" flag by the scanning software GenePix or that had a median fluorescence intensity <300 were removed from the analysis [44].
However, the scan images included beam-hardening artifacts that could not be fully removed with corrections provided by the scanning software.
Data processing included the removal of spots that were flagged by the scanning software [ 28] or did not surpass a minimum threshold of 200, and log2 transformation of the background-subtracted foreground intensities.
Briefly, fluorescence data processing encompassed the filtering of spots marked as unreliable by the scanning software or weak (when compared to control elements) and loess normalization before fitting a two-stage, mixed-model analysis.
For these analyses, spot values from the raw data were flagged if any of the following were observed: > 30% of the pixels were saturated in either channel, spot diameter < 70 um in either channel, or spot was not found by the scanning software.
Binary idat files are a convenient starting point, as they are routinely output by the scanning software, provide a compact representation of the data and have a consistent format (unlike output from Illumina's BeadStudio/GenomeStudio software, which is exported at the user's discretion, meaning the raw signals needed for the analysis are not always available).
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Best, Kasper D. Hansen The article describes illuminaio, an R package to process the raw data files produced by the Illumina scanning software.
Here, W represents the range of possible values for observations, i.e. all positive real-valued intensities generated by the array scanning software (in our case [0, 2 − 1]).
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The intensities of the bands were estimated by densitometric scanning software from Alpha Innotech Corporation (San Leandro, CA, USA).
The waveforms driving the piezo disk are generated by MATLAB scanning software.
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