Microarray technology has made the possibility in analyzing expression profiling, monitoring tens of thousands of genes simultaneously. Microarray image analysis is complicated, time consuming task and requires manual intervention. Gridding of Microarray image is performed before spot segmentation. The spatial non-uniformity of spots results in non-uniform grid alignment, which leads to inconsistency of extracted feature from the spot. This paper presents a novel automatic approach to locate the spots without the formation of grids. An approach using Genetic algorithm is proposed, in which searching for the spots in sub image is done and the relevant information of the spot is extracted. The process is repeated for the entire image. This algorithm is novel as it is independent of the spot size, colour, and location. The code is developed using MATLAB and test is carried on oligonucleotide array. From the results it is observed that even low intensity spots can be determined accurately.