Mathematical Algorithms for Automatic Search, Recognition, and Detection of Spots in ELISPOT Assay

Accuracy of spot detection and quantification plays a critical role in the analysis of ELISPOT data. Differences in staining intensities of spots and their morphological variations make it difficult developing a reliable software application. We have developed an image recognition method allowing for the automatic detection of round objects (spots) on ELISPOT images independently of the registration conditions. The emphasis is done on objects of elliptical shape which is typical for a wide range of spots that can be analyzed by both monochrome and a dual-color version of our software. The method of subdivision of objects into groups is also described which is based on color attributes of spots.