Apparatus, system, and method for image normalization using a gaussian residual of fit selection criteria

Patent metadata

Field	Value
Publication number	US-9984279-B2
Application number	US-201615004485-A
Country	US
Kind code	B2
Filing date	Jan 22, 2016
Priority date	Jun 15, 2012
Publication date	May 29, 2018
Grant date	May 29, 2018

Abstract

An apparatus and method for image normalization using a Gaussian residual of fit selection criteria. The method may include acquiring a two-dimensional image of a plurality of particles, where the plurality of particles comprises a plurality of calibration particles, and identifying a calibration particle by correlating a portion of the image corresponding to the calibration particle to a mathematical model (e.g. Gaussian fit). The measured intensity of the calibration particle may then be used to normalize the intensity of the image.

First claim

What is claimed is:

1. A method comprising:
a computing system acquiring a two-dimensional image of a plurality of particles in a first channel, wherein the plurality of particles have been exposed to an analyte, wherein the plurality of particles includes a plurality of calibration particles having internal distributions of a first fluorescent material and a plurality of non-calibration particles having surface distributions of a second fluorescent material;
the computing system identifying a calibration particle of the plurality of calibration particles by analyzing first pixel values of first pixels that correspond to the calibration particle in the first channel to correlate intensity values of the first pixels to a Gaussian function;
the computing system identifying a non-calibration particle of the plurality of non-calibration particles by analyzing second pixel values of second pixels that correspond to the non-calibration particle in the first channel to determine that intensity values of the second pixels do not correlate to a Gaussian function;
the computing system measuring an intensity of the calibration particle based on the first pixel values; and
the computing system saving a new version of the two-dimensional image, wherein the new version of the two-dimensional image includes third pixels with third pixel values that are normalized based at least in part on the intensity of the calibration particle.

2. The method of claim 1 , wherein the acquiring includes capturing the two-dimensional image via an imaging detector.

3. The method of claim 2 , wherein the imaging detector includes a charge coupled device (CCD) sensor.

4. The method of claim 1 , wherein the plurality of calibration particles are distributed into a plurality of regions of the two-dimensional image, and wherein the third pixel values for pixels in a particular region are normalized based on intensities of the plurality of calibration particles in the particular region.

5. The method of claim 1 , further comprising normalizing an intensity of a second two-dimensional image of the plurality of particles in a second channel based at least in part on the intensity of the calibration particle.

6. The method of claim 5 , wherein the second channel is different from the first channel, and wherein the second channel is a classification channel.

7. The method of claim 1 , wherein the new version of the two-dimensional image is usable to combine a plurality of images of the particles.

8. The method of claim 1 , wherein the first fluorescent material is the same as the second fluorescent material.

9. A non-transitory, computer-readable medium having instructions stored thereon that, in response to execution by a computing system, cause the computing system to carry out operations comprising:
acquiring a two-dimensional image of a plurality of particles in a first channel, wherein the plurality of particles have been exposed to an analyte, wherein the plurality of particles includes a plurality of calibration particles having internal distributions of a first fluorescent material and a plurality of non-calibration particles having surface distributions of a second fluorescent material;
identifying a calibration particle of the plurality of calibration particles by analyzing pixel values of pixels that correspond to the calibration particle in the first channel to correlate brightness values of the pixels to a Gaussian function, wherein at least one non-calibration particle has brightness values in the first channel that do not correlate to a Gaussian function;
measuring a brightness of the calibration particle based on the pixel values; and
saving a new version of the two-dimensional image, wherein the new version of the two-dimensional image includes new pixels with new pixel values that are normalized based at least in part on the brightness of the calibration particle.

10. The medium of claim 9 , wherein the plurality of calibration particles include more than one first fluorescent material.

11. The medium of claim 9 , wherein the first channel is a reporter channel, wherein the two-dimensional image is a reporter image, and wherein the new version of the two-dimensional image is usable to normalize pixel values in a classification image.

12. The medium of claim 9 , wherein measuring the brightness of the calibration particle includes detecting a peak of the calibration particle.

13. The medium of claim 9 , wherein measuring the brightness of the calibration particle includes integrating an area of the two-dimensional image around a center of the calibration particle.

14. The medium of claim 9 , wherein the operations further comprise: determining a background brightness level for the two-dimensional image.

15. The medium of claim 14 , wherein the operations further comprise: subtracting the background brightness level from the pixel values before identifying the calibration particle.

16. An apparatus, comprising:
at least one processor; and
an imaging device including an imaging region;
wherein the imaging device is configured to operate in conjunction with a processor of the at least one processors to:
accept, into the imaging region, a plurality of particles that includes a plurality of calibration particles having internal distributions of a first fluorescent material and a plurality of non-calibration particles having non-internal distributions of a second fluorescent material;
supply light to the imaging region, wherein the light corresponds to a first channel; and
capture an image of the plurality of particles in the first channel; and

wherein the at least one processor is configured to:
identify a calibration particle of the plurality of calibration particles by analyzing first pixel values of first pixels that correspond to the calibration particle to correlate intensity values of the first pixels in the first channel to a Gaussian function;
identify a non-calibration particle of the plurality of non-calibration particles by analyzing second pixel values of second pixels that correspond to the non-calibration particle to determine that intensity values of the second pixels in the first channel do not correlate to a Gaussian function;
measure an intensity of the calibration particle; and
normalize an intensity of the image based at least in part on the intensity of the calibration particle.

17. The apparatus of claim 16 , wherein the identifying the calibration particle includes approximating the Gaussian function with a quadratic function.

18. The apparatus of claim 16 , wherein the plurality of calibration particles are distributed into a plurality of regions of the image, and wherein the intensity of the image is normalized in a particular region based on intensities of the plurality of calibration particles in the particular region.

19. The apparatus of claim 18 , wherein the at least one processor is further configured to partition the image into the plurality of regions based on a grid.

20. The apparatus of claim 18 , wherein the plurality of regions is a continuous plurality of regions.

Assignees

• Luminex Corp

Inventors

• Fisher Matthew S
• Arab Nicolas

Classifications

• G01N15/1012 Primary

  Calibrating particle analysers; References therefor · CPC title

• G01N15/1463

  Physics · mapped topic

• G01N15/1434

  Optical arrangements · CPC title

• G06K9/00134

  Physics · mapped topic

• G06K9/0014 Primary

  Physics · mapped topic