Method of and system for enhanced dynamic range assay analysis

1. A method, comprising:

(a) obtaining multiple images of pixels representing light intensity output from a chemiluminescent assay, wherein the light intensity output indicates a presence and an amount of a substance within wells of a test plate, and wherein the multiple images of pixels are generated by using a detector to detect the light intensity of the pixels of the wells using:

a standard exposure time; and

a longer-than-standard exposure time; and

(b) generating a composite image using the multiple images of pixels, including:

determining an exposure compensation ratio based on the standard exposure time and the longer-than-standard exposure time;

determining a presence of out-of-range pixels in the image that used the longer-than-standard exposure time, wherein the out-of-range pixels have light intensities that exceed a threshold based on a maximum capacity of the detector;

placing out-of-range pixels in the composite image at a same relative position as the out-of-range pixels occurred in the image that used the longer-than-standard exposure time, wherein the light intensities of the out-of-range pixels are based on the light intensities from the image that used the standard exposure time and the exposure compensation ratio;

determining a presence of in-range pixels in the image that used a longer-than-standard exposure time, wherein the in-range pixels have light intensities that are below the threshold; and

placing in-range pixels in the composite image at a same relative position as said in-range pixels occurred in the image that used the longer-than-standard exposure time, wherein the light intensities of said in-range pixels are based on the light intensities from the image that used the longer-than-standard exposure time adjusted for light intensity decay due to a passage of time between the image that used the standard exposure time and the image that used the longer-than-standard exposure time.

2. The method of claim 1 , further comprising:

using the composite image to determine the presence and the amount of the substance within the wells.

3. The method of claim 1 or claim 2 , wherein the longer-than-standard exposure time is about 2 to about 10 times the standard exposure time.

4. The method of claim 3 , further comprising:

applying an image calibration procedure to the multiple images of pixels generated in step (a), wherein the image calibration procedure includes subtracting dark noise and applying flat field correction.

5. The method of claim 3 , further comprising:

obtaining one or more of the composite images;

combining the pixels in one or more of the composite images with pixels at a same relative position in other composite images to generate a template image of the wells, wherein the composite pixels at the same relative positions represent anticipated positions of printed features in a test plate; and

using the template image to find a location of a feature within the test plate, wherein the location indicates presence and amount of a substance in the test plate.

6. A method, comprising:

obtaining multiple pixel images representing light intensity output from one or more chemiluminescent assays, wherein wells of a testing substrate include features printed in the wells;

combining the multiple pixel images with pixels at a same relative position in other images to generate a template image of the wells, wherein pixels of the template image at the same relative positions represent anticipated positions of printed features in a test plate; and

using the template image to find a location of a feature within the test plate, wherein the location indicates a presence and an amount of a substance in the test plate.

7. A system, comprising:

an image detector;

a timer for tracking exposure time of the image detector; and

computer readable medium, including instructions that when executed cause a computer system to generate a composite image using the multiple images of pixels by:

(a) obtaining multiple images of pixels representing light intensity output from a chemiluminescent assay, wherein the light intensity output indicates a presence and an amount of a substance within wells of a test plate, and wherein the multiple images of pixels are generated by using a detector to detect the light intensity of the pixels of the wells using:

a standard exposure time; and

a longer-than-standard exposure time; and

(b) generating a composite image using the multiple images of pixels, including:

determining an exposure compensation ratio based on the standard exposure time and the longer-than-standard exposure time;

determining a presence of out-of-range pixels in the image that used the longer-than-standard exposure time, wherein the out-of-range pixels have light intensities that exceed a threshold based on a maximum capacity of the detector;

placing out-of-range pixels in the composite image at a same relative position as the out-of-range pixels occurred in the image that used the longer-than-standard exposure time, wherein the light intensities of the out-of-range pixels are based on the light intensities from the image that used the standard exposure time and the exposure compensation ratio;

determining a presence of in-range pixels in the image that used a longer-than-standard exposure time, wherein the in-range pixels have light intensities that are below the threshold; and

placing in-range pixels in the composite image at a same relative position as said in-range pixels occurred in the image that used the longer-than-standard exposure time, wherein the light intensities of said in-range pixels are based on the light intensities from the image that used the longer-than-standard exposure time adjusted for light intensity decay due to a passage of time between the image that used the standard exposure time and the image that used the longer-than-standard exposure time.

8. The system of claim 7 , wherein the composite image is used to determine the presence and amount of the substance within the wells.

9. The system of claim 7 or claim 8 , wherein the longer-than-standard exposure time is about 2 to about 10 times the standard exposure time.

10. The system of claim 9 , wherein the multiple images of pixels are processed using a calibration procedure, including subtracting dark noise and applying flat field correction.