Microarray analysis represents a powerful way to test scientific hypotheses on the functionality of cells. The measurements consider the whole genome, and the large number of generated data requires sophisticated analysis. To date, no gold-standard for the analysis of microarray images has been established. Due to the lack of a standard approach there is a strong need to identify new processing algorithms.

Microarray image processing leads to the characterization of gene expression levels simultaneously, for all cellular transcripts (mRNAs) in a single experiment. The calculation of expression levels for each microarray spot/gene is a crucial step to extract valuable information. By measuring the mRNA levels for the whole genome, the microarray experiments are capable to study functionality, pathological phenotype, and response of cells to a pharmaceutical treatment. The processing of the extensive number of non-homogeneous data contained in microarray images is still a challenge. We propose a density based spatial clustering procedure driven by a level-set approach for microarray spot segmentation together with a complete set of quality measures used to evaluate the proposed method compared with existing approaches for gene expression levels estimation. The set of quality measures used for evaluation include: regression ratios, intensity ratios, mean absolute error, coefficient of variation and fold change factor. We applied the proposed image processing pipeline to a set of microarray images and compared our results with the ones delivered by Genepix, using the aforementioned quality measures. The advantage of our proposed method is highlighted by a selection of up-regulated genes that had been identified exclusively by our approach. These genes prove to add valuable information regarding the biological mechanism activated as a response of Arabidopsis T to pathogen infection.