Scanning microarrays: current methods and future directions.

The microarray platform is a powerful tool for conducting large-scale, high-throughput gene expression experiments. However, careful attention to detail throughout the five major steps in the microarray process--design, printing, hybridization, scanning, and analysis--must be used to ensure that reliable and accurate conclusions are obtained from data. The act of scanning the array has received the least attention of all parts of the microarray process, despite it being a critical quality-limiting component. This chapter specifically addresses the effects of scan parameters and limitations of the scanning technology divided into two categories: instrumentation effects (those that arise from the scanning instrumentation itself) and user-controller parameters (those that an operator chooses) for the most common microarray platform--the two-color cDNA microarray printed on a glass substrate. Significant research efforts have gone into developing microarray analysis techniques, but the field is ripe for research to characterize the variability and errors introduced by the scanning process itself, the scanner instrumentation, and the user. Implications of these errors for large-scale, multiple slide and multiple laboratory experiments are discussed. Wise choices for scanning parameters and consideration of instrument specifics will ultimately increase data reliability and reduce the need for complex preprocessing mechanisms prior to the extraction of expression information. In addition, emerging technologies such as surface plasmon imaging, resonance light scattering, and hyperspectral imaging are presented briefly as promising, complementary techniques to traditional scanning methods.