Image data analysis in qPCR: A method for smart analysis of DNA amplification

Abstract

In this paper, a method for the direct quantitative analysis of amplified DNA via q-Polymerase Chain Reaction (qPCR) in miniaturised silicon-based chip system is presented. The designed tool presented here allows the automatic extraction of meaningful information from input fluorescent images by means of digital image processing algorithm. In particular, a smart mathematical model, optimizing the integration of all the analysis steps of the fluorescence data from on chip multiple real time PCR, is described. Such a tool is able to load the digital input images, select and smartly detect the region of interest for fluorescence, elaborate the data input, calculate the average fluorescence values and finally, plot the fitted curve as output, giving also, for each well, both the Cycle Threshold (CT) and Slope parameters. Keywords: Image analysis, qPCR, LabonChip, Edge detection, Hough transform