The Impact of Photobleaching on Microarray Analysis

摘要: DNA-Microarrays have become a potent technology for high-throughput analysis of genetic regulation. However, the wide dynamic range signal intensities fluorophore-based microarrays exceeds single array scan by far, thus limiting key benefit microarray technology: parallelization. The implementation multi-scan techniques represents promising approach to overcome these limitations. These are, in turn, limited fluorophores’ susceptibility photobleaching when exposed scanner’s laser light. In this paper characteristics cyanine-3 and cyanine-5 as part solid state DNA are studied. effects initial fluorophore intensity well scanner dependent variables such photomultiplier tube’s voltage on bleaching imaging investigated. resulting data is used develop model capable simulating expected degree reduction caused each individually, allowing removal photobleaching-induced, systematic bias procedures. Single-scan applications also they rely pre-scans determine optimal settings. findings constitute step towards standardization experiments may help increase lab-to-lab comparability experiment results.