Flat field correction of two-dimensional biochemical assay images
First Claim
1. A method for detecting optically detectable biochemical assay results in a two-dimensional biochemical assay medium, said method comprising the following steps:
- (a) irradiating said assay medium with light from a light source in a two-dimensional optical imaging system to generate a two-dimensional image of said assay medium;
(b) irradiating a reference plate with light from said light source in said two-dimensional optical imaging system independently of said assay medium to generate a two-dimensional image of said reference plate, said reference plate being a plate that responds to incident light uniformly along the length and width of said reference it plate; and
(c) comparing said image of said assay medium to said image of said reference plate and correcting said image of said assay medium for nonuniformities in said optical imaging system indicated by said image of said reference plate;
steps (a) and (b) being performed in any order and (c) performed after both (a) and (b) are completed.
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Abstract
The optical imaging of two-dimensional solute zone arrays in electrophoresis gels is corrected for nonuniformities in the optical system such as those arising from the light source or from light dispersion underneath the gel. The correction is achieved by the use of a reference plate that responds to a light source uniformly along its length and width by being either uniformly light absorptive or uniformly light transmissive, or by emitting light upon excitation. Thus, any nonuniformities or deviations in the image of the reference plate arise only from nonuniformities or deviations within the optical system. Analogous corrections are made in other two-dimensional assay images, such as microarrays and microtiter plates.
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Citations
12 Claims
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1. A method for detecting optically detectable biochemical assay results in a two-dimensional biochemical assay medium, said method comprising the following steps:
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(a) irradiating said assay medium with light from a light source in a two-dimensional optical imaging system to generate a two-dimensional image of said assay medium;
(b) irradiating a reference plate with light from said light source in said two-dimensional optical imaging system independently of said assay medium to generate a two-dimensional image of said reference plate, said reference plate being a plate that responds to incident light uniformly along the length and width of said reference it plate; and
(c) comparing said image of said assay medium to said image of said reference plate and correcting said image of said assay medium for nonuniformities in said optical imaging system indicated by said image of said reference plate;
steps (a) and (b) being performed in any order and (c) performed after both (a) and (b) are completed. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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Specification