Protein microarray technology has recently emerged as a powerful tool for biomedical research. Before automatic analysis the protein microarray images, protein spots in the images must be determined appropriately by spot segmentation algorithm. In this paper, an improved seeded region growing (ISRG) algorithm for protein microarray segmentation is presented, the seeds are obtained by finding the positions of the printed spots, and the protein spot regions are grown through these seeds. The experiment results show that the presented algorithm is accurate for adaptive shape segmentation and robust for proteinmicroarray images contaminated by noise.