METHODS AND SYSTEMS FOR DIGITALLY COUNTING FEATURES ON ARRAYS
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Accused Products
Abstract
Methods, systems and platforms for digital imaging of multiple regions of an array, and detection and counting of the labeled features thereon, are described.
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Citations
40 Claims
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1-20. -20. (canceled)
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21. A method for determining the number of a target molecule on an array, comprising:
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(a) acquiring image data of a region of an array, wherein the array comprises a plurality of stochastically labeled and non-labeled features, and wherein the features comprise oligonucleotide probes; (b) measuring a signal intensity and a local background intensity for one of the plurality of stochastically labeled and non-labeled features based on the acquired image data; (c) calculating a local background corrected signal intensity for the one of the plurality of stochastically labeled and non-labeled features using the signal intensity and the local background intensity measured in (b); (d) transforming the local background corrected signal intensity calculated in (c) for the one of the plurality of stochastically labeled and non-labeled features to determine the number of stochastically labeled and non-labeled features in the of the array; and (e) calculating the number of a target molecule based on the number of stochastically labeled and non-labeled features determined in (d) within the region of the array. - View Dependent Claims (22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32)
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33. A method for performing local background correction of array signal intensity data, comprising:
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(a) centering a predefined data analysis window on a feature within a digital image of an array; (b) calculating an intensity value statistic for signal and background pixels based on a predefined pattern of pixels within or around the feature; and (c) utilizing the intensity value statistic for signal and background pixels to calculate a background corrected signal intensity for the feature. - View Dependent Claims (34, 35, 36, 37)
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38. A method for determining a dynamic image intensity threshold for use in discriminating between labeled and non-labeled features on an array comprising a plurality of labeled and non-labeled features, comprising:
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(a) measuring image intensity data for each of the plurality of labeled and non-labeled features of the array; (b) performing a local background correction on the measured image intensity data for each of the plurality of labeled and non-labeled features on the array to obtain local background corrected image intensity data; and (c) performing one or more statistical analyses of the local background corrected image intensity data for the plurality of labeled and non-labeled features, thereby determining a dynamic image intensity threshold for discrimination between the labeled and non-labeled features. - View Dependent Claims (39, 40)
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Specification