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Methods and systems for digitally counting features on arrays

  • US 9,582,877 B2
  • Filed: 10/07/2014
  • Issued: 02/28/2017
  • Est. Priority Date: 10/07/2013
  • Status: Active Grant
First Claim
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1. An array reader system comprisinga computer system comprising a digital processor configured to execute computer-executable instructions to at least:

  • (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 absolute 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.

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