System, method, and product for imaging probe arrays with small feature size
First Claim
Patent Images
1. A method for resolving features on a probe array, comprising:
- acquiring images of a region of a probe array with an image acquisition device comprising an objective lens and a sensing element;
shifting the images by less than one half a size of a pixel of the sensing element while maintaining a fixed spatial relationship between the probe array and the objective lens to produce micro-shifted images;
reconstructing pixels of the micro-shifted images by unboxing the micro-shifted images, wherein unboxing comprises deconvolving the micro-shifted images using the Richardson-Lucy algorithm, wherein the Richardson-Lucy algorithm is controlled by at least three independent parameters, and wherein the at least three independent parameters comprise over-relaxation or acceleration, momentum, and total variation control; and
deriving intensity values for one or more features from the reconstructed pixels of the micro-shifted images, thereby resolving features on the probe array.
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Abstract
An embodiment of a method for resolving features on a probe array is described that, comprises acquiring a plurality of micro-shifted images of a region of a probe array; reconstructing an image of the probe array using the micro-shifted images; and deriving intensity values for one or more probe features disposed on the probe array from the reconstructed image.
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Citations
11 Claims
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1. A method for resolving features on a probe array, comprising:
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acquiring images of a region of a probe array with an image acquisition device comprising an objective lens and a sensing element; shifting the images by less than one half a size of a pixel of the sensing element while maintaining a fixed spatial relationship between the probe array and the objective lens to produce micro-shifted images; reconstructing pixels of the micro-shifted images by unboxing the micro-shifted images, wherein unboxing comprises deconvolving the micro-shifted images using the Richardson-Lucy algorithm, wherein the Richardson-Lucy algorithm is controlled by at least three independent parameters, and wherein the at least three independent parameters comprise over-relaxation or acceleration, momentum, and total variation control; and deriving intensity values for one or more features from the reconstructed pixels of the micro-shifted images, thereby resolving features on the probe array. - View Dependent Claims (2, 3, 4, 5, 6, 8, 9)
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7. A method of deriving signal intensity values in a probe array experiment, which comprises:
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hybridizing a labeled sample to a probe array; acquiring images of the labeled sample on the probe array with an image acquisition device comprising an objective lens and a sensing element; shifting the images by less than one half a size of a pixel while maintaining a fixed spatial relationship between the probe array and the objective lens to create micro-shifted images; tiling a single pixel from a common location within each of the micro-shifted images to provide a reconstructed image; unboxing the reconstructed image by applying a set of linear equations to determine subpixel values, thereby providing a final image, wherein unboxing comprises using the Richardson-Lucy algorithm, wherein the Richardson-Lucy algorithm is controlled by at least three independent parameters, wherein the at least three independent parameters comprise over-relaxation or acceleration, momentum, and total variation control; and deriving intensity values for one of more probe features disposed on the probe array from the final image. - View Dependent Claims (10, 11)
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Specification