Imaging systems and methods useful for patterned structures
Disclosed herein, inter alia, are methods and systems of image analysis useful for identifying and/or quantifying features in patterns.
1 . A method of imaging a solid support comprising a plurality of features, the method comprising:
obtaining an image of the solid support using a detection apparatus comprising at least one camera, wherein the solid support comprises a first feature comprising a first fluorescent emission and a second feature comprising a second fluorescent emission;
providing the image or image-related data to a computer, wherein the computer has parameter data that describes a position of the features; and
executing the following on the computer:
performing a chromatic correction;
partitioning the image or the image-related data into a plurality of registration subimages;
detecting features for each registration subimage;
assigning an index address for each feature; and
quantifying a signal level of each feature, wherein quantifying the signal level of each feature comprises building a model of the subimage incorporating known feature locations and corresponding unknown intensities and fitting the model to the image of the solid support, and wherein the model of the subimage comprises a matrix that is pre-computed and stored in computer memory or a non-transitory computer-readable medium.
2 . The method of claim 1 , wherein the solid support comprises has a repeating pattern of features in a two-dimensional plane.
3 . The method of claim 2 , wherein the object solid support comprises polynucleotide clusters.
4 . The method of claim 1 , wherein the parameter data relates to
a grid orientation angle,
an apparent pitch in pixel units, and/or
a phase of a feature grid at a fixed pixel location of the image.
5 . The method of claim 1 , wherein the matrix is reused for a different subimage.
6 . The method of claim 1 , wherein each index address is a unique address.
7 . The method of claim 1 , wherein each unique address is an integer vector of length 2.
8 . The method of claim 1 , wherein the detection apparatus comprises a first camera configured to obtain an image of two different color emission wavelength ranges; and a second camera configured to obtain an image of two different emission wavelength ranges; wherein each emission wavelength range is spectrally distinct.
9 . The method of claim 8 , wherein each emission wavelength range coincides with a different label used to distinguish one nucleotide base type from another nucleotide base type.
10 . The method of claim 8 , wherein the first camera collects an image of the first fluorescent emission and the second fluorescent emission and the second camera collects an image of a third fluorescent emission and a fourth fluorescent emission.
11 . The method of claim 1 , wherein the chromatic correction comprises an affine transform.
12 . The method of claim 11 , wherein the affine transform is calculated from a maximum intensity projection of an image.
13 . A non-transitory computer-readable medium containing instructions to configure a processor to perform operations comprising:
obtaining an image of a solid support using a detection apparatus comprising at least one camera, wherein the solid support comprises a first feature comprising a first fluorescent emission and a second feature comprising a second fluorescent emission;
providing the image or image-related data to a computer, wherein the computer has parameter data that describes a position of the features; and
executing the following on the computer:
performing a chromatic correction;
partitioning the image or the image-related data into a plurality of registration subimages;
detecting features for each registration subimage;
assigning an index address for each feature; and
quantifying a signal level of each feature, wherein quantifying the signal level of each feature comprises building a model of the subimage incorporating known feature locations and corresponding unknown intensities and fitting the model to the image of the solid support, and wherein the model of the subimage comprises a matrix that is pre-computed and stored in computer memory or a non-transitory computer-readable medium.
14 . A system comprising:
a processor; and
a memory, wherein the processor and the memory are configured to perform operations comprising:
obtaining an image of the solid support using a detection apparatus comprising at least one camera, wherein the
solid support comprises a first feature comprising a first fluorescent emission and a second feature comprising a second fluorescent emission;
providing the image or image-related data to a computer, wherein the computer has parameter data that describes a position of the features; and
executing the following on the computer:
performing a chromatic correction;
partitioning the image or the image-related data into a plurality of registration subimages;
detecting features for each registration subimage;
assigning an index address for each feature; and
quantifying a signal level of each feature, wherein quantifying the signal level of each feature comprises building a model of the subimage incorporating known feature locations and corresponding unknown intensities and fitting the model to the image of the solid support, and wherein the model of the subimage comprises a matrix that is pre-computed and stored in computer memory or a non-transitory computer-readable medium.