Evaluation of focusing performance in an assay analysis system

Patent metadata

Field	Value
Publication number	US-9719855-B2
Application number	US-201414267693-A
Country	US
Kind code	B2
Filing date	May 1, 2014
Priority date	Jul 16, 2010
Publication date	Aug 1, 2017
Grant date	Aug 1, 2017

Abstract

Methods, storage mediums and systems (MS&S) are provided which successively image an imaging region of an assay analysis system (AAS) as particles are loaded into the imaging region, generate a frequency spectrum of each image via a discrete Fourier transform, integrate a same coordinate portion of each frequency spectrum and terminate the loading of particles upon computing an integral which meets preset criterion. In addition, MS&S are provided which send a signal indicative of whether enough particles are in an imaging region for further processes by an AAS based on the magnitude of integral calculated from an image's frequency spectrum. MM&S are also provided such that the steps of generating a frequency spectrum of each image and integrating a portion of each frequency spectrum are replaced by generating a convolved spatial image with a filter kernel and integrating a same coordinate portion of each convolved spatial image.

First claim

What is claimed is:

1. A method, comprising:
performing an automated routine to adjust a focal position of a photosensitive detection subsystem of an assay analysis system;
subsequent to performing the automated routine, imaging particles arranged within an imaging chamber of the assay analysis system to create an image;
generating a frequency spectrum of the image via a discrete Fourier transform; and
comparing a width of a primary lobe of the frequency spectrum at a selected brightness value to a benchmark to evaluate a performance of the automated routine to adjust the focal position of the photosensitive detection subsystem.

2. The method of claim 1 , wherein the benchmark is specific to the assay analysis system.

3. The method of claim 1 , wherein the benchmark is based on a historic record of primary lobe widths at the selected brightness value of frequency spectra generated from images previously obtained by the assay analysis system.

4. The method of claim 1 , further comprising repeating, at a selected frequency based on a number of times the automated routine is performed for the assay analysis system, the steps of imaging, generating a frequency spectrum, and comparing a width of a primary lobe of the generated frequency spectrum to the benchmark.

5. The method of claim 1 , wherein the frequency spectrum is generated via a fast Fourier transform.

6. The method of claim 1 , wherein the frequency spectrum is generated via a discrete cosine transform.

7. The method of claim 1 , wherein the image is based on a fluorescent material associated with the particles.

8. An assay analysis system, comprising:
an imaging chamber configured to accept a plurality of particles to be imaged; and
a photosensitive detection system configured to receive light emitted by the plurality of particles;
wherein the assay analysis system is configured to:
perform an adjustment of a focal position of the photosensitive detection system;
create an image of the plurality of particles in the imaging chamber;
generate a frequency spectrum of the image via a discrete Fourier transform; and
performing a comparison of a width of a primary lobe of the frequency spectrum to a benchmark to evaluate the adjustment of the focal position of the photosensitive detection system.

9. The assay analysis system of claim 8 , wherein the frequency spectrum of the image is a two-dimensional frequency spectrum.

10. The assay analysis system of claim 8 , wherein the discrete Fourier transform is selected from the group consisting of fast Fourier transform and discrete cosine transform.

11. The assay analysis system of claim 8 , further comprising an illumination subsystem.

12. The assay analysis system of claim 11 , wherein the image is based on fluorescence emitted by the plurality of particles in response to illumination by the illumination subsystem.

13. The assay analysis system of claim 8 , wherein the adjustment is based on an automated adjustment routine.

14. The assay analysis system of claim 8 , wherein the comparison is based on a selected brightness value.

15. An assay analysis system, comprising:
an imaging chamber configured to accept a plurality of particles to be imaged; and
a photosensitive detection system configured to receive light emitted by the plurality of particles;
wherein the assay analysis system is configured to:
create an image of the plurality of particles in the imaging chamber;
generate a frequency spectrum of the image; and
evaluate a focal position of the photosensitive detection system by comparing a width of a primary lobe of the frequency spectrum to a benchmark.

16. The assay analysis system of claim 15 , wherein the assay analysis system is further configured to perform an automated focusing routine to adjust the focal position of the photosensitive detection system.

17. The assay analysis system of claim 15 , further comprising a storage medium having a database stored thereon, wherein the database includes a historic record of primary lobe widths, and wherein the benchmark is based on the database.

18. The assay analysis system of claim 15 , wherein the frequency spectrum is based on a fast Fourier transform of the image.

19. The assay analysis system of claim 15 , wherein the frequency spectrum is based on a discrete cosine transform of the image.

Assignees

• Luminex Corp

Inventors

• Arab Nicolas

Classifications

• G01N21/6458

  Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16) · CPC title

• G01J3/42

  Absorption spectrometry; Double beam spectrometry; Flicker spectrometry; Reflection spectrometry (beam switching arrangements G01J3/08) · CPC title

• G01N15/06

  Investigating concentration of particle suspensions (by weighing G01N5/00; investigating sedimentation of particle suspensions G01N15/04; investigating individual particles G01N15/10) · CPC title

• G01J3/2823 Primary

  Imaging spectrometer · CPC title

• G01N15/1463

  Physics · mapped topic