Image based correction for unwanted light signals in a specific region of interest
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[0044] A real time PCR run for HIV was performed on an ABI Prism 7500 instrument (Applied Biosystems, Foster City, Calif.). This instrument utilizes a 96-well plate format with wells arranged in a 12×8 array. The run was configured so that there were 84 wells containing positive samples with a concentration of 1×106 copies / mL and 12 wells not containing positive samples, i.e., negative wells. The negative wells were distributed on the plate to maximize the potential cross-talk from the wells containing positive samples. FIG. 4 illustrates the layout of the plate.
[0045] The ABI Prism 7500 instrument uses a CCD camera and measures fluorescence in five wavelength bands. FIG. 5 shows one image from the end of the PCR run. FIG. 6 shows the same image with the reaction regions of interest and the regions of correction superimposed. In this example, a diagonal array of diamond-shaped regions of correction, each of which contained of 25 pixels, were used. The first reading in the PCR run w...
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