Method of calibrating and correcting color-bleed factors for color separation in DNA analysis

Patent metadata

Field	Value
Publication number	US-9611199-B2
Application number	US-201514867366-A
Country	US
Kind code	B2
Filing date	Sep 28, 2015
Priority date	Oct 20, 2010
Publication date	Apr 4, 2017
Grant date	Apr 4, 2017

Abstract

A method includes calibrating color bleed factors of optical detector channels of a sample processing apparatus through processing a color bleed calibration substance which includes a plurality of different size fragments replicated from different groups of DNA loci, wherein fragments in a same group are labeled with a same fluorescent dye, and fragments in different groups are labeled with different fluorescent dyes having different emission spectra, wherein the different size fragments are processed during different acquisition times.

First claim

What is claimed is:

1. A method, comprising:
generating a first signal indicative of a reference gain of optical detectors of a sample processing apparatus based on a first emission from a gain-monitoring material of the sample processing apparatus in response to illuminating the gain-monitoring material;
generating a second signal indicative of a subsequent gain of the optical detectors based on a second emission from a gain-monitoring material in response to illuminating the gain-monitoring material; and
scaling at least one of color bleed factors of the sample processing apparatus or data acquired by the sample processing apparatus based on a signal indicative of a difference between the reference gain and the subsequent gain.

2. The method of claim 1 , wherein the second signal is determined at least one of before, during or after an acquisition time in which one or more DNA samples are processed by the sample processing.

3. The method of claim 1 , wherein the gain-monitoring material is part of the sample processing apparatus.

4. The method of claim 1 , wherein the gain-monitoring material is part of a sample carrier received by the sample processing apparatus, wherein the sample carrier includes one or more channels for carrying samples to be processed by the sample processing apparatus.

5. The method of claim 1 , wherein the gain-monitoring material includes one or more fluorescent materials that have a known emission spectrum within a predetermined emission spectrum of interest.

6. The method of claim 1 , wherein the gain-monitoring material is illuminated by a source of the sample processing apparatus used to illuminates separated DNA fragments.

7. The method of claim 1 , wherein the gain-monitoring material is illuminated by a source of the sample processing apparatus that is different from a sample source used to illuminates separated DNA fragments.

8. An apparatus, comprising:
memory storing color bleed factors;
optical detectors; and
a processor configured to:
generate a first signal indicative of a reference gain of the optical detectors based on a first emission from a gain-monitoring material in response to illuminating the gain-monitoring material;
generate a second signal indicative of a subsequent gain of the optical detectors based on a second emission from a gain-monitoring material in response to illuminating the gain-monitoring material; and
scale at least one of the color bleed factors or acquired data based on a signal indicative of a difference between the reference gain and the subsequent gain.

9. The apparatus of claim 8 , wherein the second signal is determined at least one of before, during or after an acquisition time in which one or more DNA samples are processed by the sample processing.

10. The apparatus of claim 8 , wherein the gain-monitoring material is part of the apparatus.

11. The apparatus of claim 8 , further comprising:
a sample carrier with one or more channels configured to carry samples, wherein the gain-monitoring material is part of the sample carrier.

12. The apparatus of claim 11 , further comprising:
one or more fluorescent materials that have a known emission spectrum within a predetermined emission spectrum of interest, wherein the gain-monitoring material includes the one or more fluorescent materials.

13. The apparatus of claim 8 , further comprising:
an illumination source configured to illuminate separated DNA fragments, wherein the illumination source illuminates the gain-monitoring material.

14. The apparatus of claim 8 , further comprising:
a first illumination source configured to illuminate separated DNA fragments; and
a second illumination source configured to illuminate the gain-monitoring material, wherein the first and second illumination sources are different sources.

Assignees

• Analogic Corp

Inventors

• Lai Ching Ming

Classifications

• C12Q1/686

  Polymerase chain reaction [PCR] · CPC title

• C07C17/206

  the other compound being HX · CPC title

• C07C17/25 Primary

  by splitting-off hydrogen halides from halogenated hydrocarbons · CPC title

• G01N21/6486

  Measuring fluorescence of biological material, e.g. DNA, RNA, cells (G01N21/6428 takes precedence) · CPC title

• C07C17/087

  to unsaturated halogenated hydrocarbons · CPC title