Computer method for ELISA spot assay analysis

Abstract

The ELISA (enzyme-linked immunosorbent assay) spot assay is a method widely used by immunologists to enumerate cytokine-producing cells within a specific cell population. The ELISA results are presented in an image containing numerous colored spots. We present a method to identify the spots in the image and report on important statistics regarding them. The proposed method employs color analysis in the CIE Luv color space and matched filter technique. The system is trained to obtain a standard color for the spots and calculate the color differences between the spots and background in the Luv space. Matched filters are then used to remove noise and enhance the spots in the color difference map. Intensity thresholding is applied to obtain a binary image in which the pixels in the spots have a grayscale of 1 while the grayscale of the other pixels is 0. A software system is implemented, based on this method, to help immunologists analyze the results obtained from the. ELISA. (c) 2005 Society of Photo-Optical Instrumentation Engineers.