# US 9,519,822 B2 · Determination method, determination device, determination system, and computer program

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- Number: US 9,519,822 B2
- Document ID: US9519822B2
- Status: Granted
- Kind: B2
- Application number: 14/383,601
- Filed: 2013-03-05
- Published: 2015-04-09
- Granted: 2016-12-13
- Earliest claimed: 2012-03-08
- Assignees: Toray Industries, Inc.
- Applicants: Toray Industries, Inc.
- Inventors: Kunihisa Nagino; Hiromichi Sasamoto; Shigeru Suzue
- Web page: https://patentgrep.com/patent/US9519822B2

## CPC

- G01N 33/5308: {for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites}
- G06T 7/0012: {Biomedical image inspection}
- G06V 20/695: {Preprocessing, e.g. image segmentation}
- G01N 21/6458: {Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16)}
- G06T 2207/10056: Microscopic image
- G06T 2207/10064: Fluorescence image
- G06T 2207/30072: Microarray; Biochip, DNA array; Well plate
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- G06T: IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
- G06V: IMAGE OR VIDEO RECOGNITION OR UNDERSTANDING

## Abstract

A determination method for determining reliability of a selective binding amount of a substance to be examined obtained as detection intensity of a label when a labeled substance to be examined binds to a selective binding substance fixed as a spot on a carrier includes: determining a position of the spot in image data obtained by imaging the detection intensity in the carrier and extracting a pixel group corresponding to the spot; calculating a ratio or a difference between a median value of the detection intensity of the pixel group extracted at the determining and a median value of the detection intensity of the pixel group excluding a certain top proportion of and/or a certain bottom proportion of pixels; and determining quality of the reliability based on the ratio or the difference calculated at the calculating and a certain reference value.

## Claims

### Claim 1

A method of determining reliability of a selective binding amount of a substance to be examined obtained as detection intensity of a label when a labeled substance to be examined binds to a selective binding substance fixed as a spot on a carrier, comprising:
a pixel group extracting step of determining a position of the spot in image data obtained by imaging the detection intensity in the carrier and extracting a pixel group corresponding to the spot;
a median calculating step of calculating a ratio of or a difference between 1) a median value of the detection intensity of the pixel group extracted at the pixel group extracting step and 2) a median value of the detection intensity of the pixel group excluding a certain top proportion of and/or a certain bottom proportion of pixels; and
a reliability determining step of determining quality of the reliability based on the ratio or the difference calculated at the median calculating step and a certain reference value.

### Claim 2 (depends on 1)

The method according to claim 1, wherein
a value of a ratio obtained from expression (1) and/or expression (2) is calculated at the median calculating step, the expressions (1) and (2) being as follows:
|X−Xt|/X (1)
|X−Xb|/X (2)
where X is a median value of the detection intensity of the pixel group, Xt is a median value of the detection intensity of the pixel group excluding the certain top proportion of pixels, and Xb is a median value of the detection intensity of the pixel group excluding the certain bottom proportion of pixels,
the reliability determining step determines that the reliability is defective when the value of the ratio obtained from at least one of expressions (1) and (2) is equal to or larger than the reference value.

### Claim 3 (depends on 1)

The method according to claim 1, wherein
the reference value is a value obtained from expression (3) as follows:
S=C+Z/X (3)
where S is the reference value, C is a constant, Z is an offset value corresponding to a sensitivity setting of a device that detects the detection intensity of the label, and X is the median value of the detection intensity of the pixel group.

### Claim 4 (depends on 3)

The method according to claim 3, wherein
the device that detects the detection intensity of the label is a photomultiplier, and
the offset value is a value obtained from expression (4) as follows:
Z=X^(A)*B (4)
where Z is the offset value, X is a gain voltage of the photomultiplier, and A and B are constants.

### Claim 5 (depends on 1)

The method according to claim 1, wherein
the carrier is a microarray,
the label is a fluorescent label,
the detection intensity is a fluorescence amount, and
the reliability determining step determines quality of the spot as the quality of the reliability.

### Claim 6

A determination device comprising at least a controller configured to determine reliability of a selective binding amount of a substance to be examined obtained as detection intensity of a label when a labeled substance to be examined binds to a selective binding substance fixed as a spot on a carrier, the control unit controller comprising a processor configured to:
determine a position of the spot in image data obtained by imaging the detection intensity in the carrier and extract a pixel group corresponding to the spot;
calculate a ratio of or a difference between 1) a median value of the detection intensity of the pixel group and 2) a median value of the detection intensity of the pixel group excluding a certain top proportion of and/or a certain bottom proportion of pixels; and
determine quality of the reliability based on the ratio or the difference and a certain reference value.

### Claim 7

A determination system configured by connecting a detection device that reads detection intensity of a label, which is obtained when a labeled substance to be examined binds to a selective binding substance fixed as a spot on a carrier, to a controller configured to determine reliability of the selective binding amount of the substance to be examined obtained as the detection intensity, wherein
the controller comprises a processor configured to:
acquire, as imaged image data, the detection intensity in the carrier read via the detection device;
determine a position of the spot in the acquired image data and extract a pixel group corresponding to the spot;
calculate a ratio of or a difference between 1) a median value of the detection intensity of the pixel group and 2) a median value of the detection intensity of the pixel group excluding a certain top proportion of and/or a certain bottom proportion of pixels; and
determine quality of the reliability based on the ratio or the difference and a certain reference value.

### Claim 8

A non-transitory computer readable recording medium on which an executable program is recorded, the program instructing a processor to execute determining reliability of a selective binding amount of a substance to be examined obtained as detection intensity of a label when a labeled substance to be examined binds to a selective binding substance fixed as a spot on a carrier, wherein the determining includes:
a pixel group extracting step of determining a position of the spot in image data obtained by imaging the detection intensity in the carrier and extracting a pixel group corresponding to the spot;
a median calculating step of calculating a ratio of or a difference between 1) a median value of the detection intensity of the pixel group extracted at the pixel group extracting step and 2) a median value of the detection intensity of the pixel group excluding a certain top proportion of and/or a certain bottom proportion of pixels; and
a reliability determining step of determining quality of the reliability based on the ratio or the difference calculated at the median calculating step and a certain reference value.

### Claim 9 (depends on 2)

The determination method according to claim 2, wherein
the reference value is a value obtained from expression (3) as follows:
S=C+Z/X (3)
where S is the reference value, C is a constant, Z is an offset value corresponding to a sensitivity setting of a device for detecting the detection intensity of the label, and X is the median value of the detection intensity of the pixel group.

### Claim 10 (depends on 9)

The determination method according to claim 9, wherein
the device that detects the detection intensity of the label is a photomultiplier, and
the offset value is a value obtained from expression (4) as follows:
Z=X^(A)*B (4)
where Z is the offset value, X is a gain voltage of the photomultiplier, and A and B are constants.

### Claim 11 (depends on 2)

The determination method according to claim 2, wherein
the carrier is a microarray,
the label is a fluorescent label,
the detection intensity is a fluorescence amount, and
the reliability determining step determines quality of the spot as the quality of the reliability.

### Claim 12 (depends on 3)

The determination method according to claim 3, wherein
the carrier is a microarray,
the label is a fluorescent label,
the detection intensity is a fluorescence amount, and
the reliability determining step determines quality of the spot as the quality of the reliability.

### Claim 13 (depends on 4)

The determination method according to claim 4, wherein
the carrier is a microarray,
the label is a fluorescent label,
the detection intensity is a fluorescence amount, and
the reliability determining step determines quality of the spot as the quality of the reliability.

### Claim 14 (depends on 9)

The determination method according to claim 9, wherein
the carrier is a microarray,
the label is a fluorescent label,
the detection intensity is a fluorescence amount, and
the reliability determining step determines quality of the spot as the quality of the reliability.

### Claim 15 (depends on 10)

The determination method according to claim 10, wherein
the carrier is a microarray,
the label is a fluorescent label,
the detection intensity is a fluorescence amount, and
the reliability determining step determines quality of the spot as the quality of the reliability.