Computer method for ELISA spot assay analysis

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

doi:10.1117/1.2009767

Computer method for ELISA spot assay analysis

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

^*Corresponding author for this work

Department of Computer Science

Research output: Contribution to journal › Article › peer-review

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 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 language	English
Article number	083604
Journal	Optical Engineering
Volume	44
Issue number	8
DOIs	https://doi.org/10.1117/1.2009767
State	Published - 1 Aug 2005

Keywords

CIE
Color analysis
ELISA
Matched filter
Segmentation
Spot

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10.1117/1.2009767

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title = "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 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.",

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N2 - 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.

AB - 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.

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