title: A novel approach for high-quality microarray processing using third-dye array visualization technology.
abstract: Historically, microarray image processing has been technically challenging in obtaining quality gene expression data. After hybridization of Cy3- and Cy5-labeled samples, images are collected and processed to obtain gene expression ratio measurements for each of the elements on the array. The hybridization process often brings in contaminating noise, which can make correct identification of the signal difficult. In addition, spot intensity levels are highly variable due to the expression differences of different genes, and weak spots are often difficult to detect. These conditions are further complicated by inherent irregularities in spot position, shape, and size commonly found on high-density microarrays, making image processing an often labor-intensive task that is difficult to reliably automate. We previously reported a novel third-dye array visualization (TDAV) technology that allows prehybridization visualization and quality control of printed arrays. Here, we present a new microarray image processing approach utilizing TDAV. By incorporating the third-dye image, we show that overall quality of the microarray data is significantly improved, and automation of processing is feasible and reliable. Furthermore, we demonstrate use of the third-dye image to better quality control microarray image analysis. Both the principle and implementation of the approach are presented in detail, with experimental results.
authors.affiliation: Max McGee National Research Center for Juvenile Diabetes, Department of Pediatrics, Medical College of Wisconsin, Milwaukee, WI 53226, USA. xujing@mcw.edu
authors.forename: Xujing
authors.initials: X
authors.lastname: Wang
authors.forename: Nan
authors.initials: N
authors.lastname: Jiang
authors.forename: Xin
authors.initials: X
authors.lastname: Feng
authors.forename: Yizhou
authors.initials: Y
authors.lastname: Xie
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Tonellato
authors.forename: Soumitra
authors.initials: S
authors.lastname: Ghosh
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Hessner
mesh_terms: D000465:Algorithms; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D005456:Fluorescent Dyes; D020869:Gene Expression Profiling / Q000379:methods; D007089:Image Enhancement / Q000379:methods*; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D036103:Nanotechnology / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010363:Pattern Recognition, Automated; D015203:Reproducibility of Results; D012371:Robotics / Q000379:methods; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D004247:DNA