Computer method for ELISA spot assay analysis

Chih Yang Lin*, Yu-Tai Ching, Betty A. Wu-Hsieh

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


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 L*u*v* 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 L*u*v* 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.

Original languageEnglish
Article number083604
JournalOptical Engineering
Issue number8
StatePublished - 1 Aug 2005


  • CIE
  • Color analysis
  • Matched filter
  • Segmentation
  • Spot

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