# US 10,871,447 B2 · Bleaching of dyes in luminescent detection

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- Number: US 10,871,447 B2
- Document ID: US10871447B2
- Status: Granted
- Kind: B2
- Application number: 15/736,457
- Filed: 2016-06-30
- Published: 2018-06-21
- Granted: 2020-12-22
- Earliest claimed: 2015-06-30
- Assignees: Imec Vzw
- Applicants: Imec Vzw
- Inventors: Peter Peumans; Liesbet Lagae; Willem Van Roy; Tim Stakenborg; Pol Van Dorpe
- Web page: https://patentgrep.com/patent/US10871447B2

## CPC

- G01N 21/6408: {with measurement of decay time, time resolved fluorescence}
- G01N 21/6428: {Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53)}
- G01N 21/648: {using evanescent coupling or surface plasmon coupling for the excitation of fluorescence}
- G01N 21/274: {Calibration, base line adjustment, drift correction}
- G01N 21/645: {Specially adapted constructive features of fluorimeters}
- G01N 2021/6413: {Distinction short and delayed fluorescence or phosphorescence}
- G01N 2021/6439: {with indicators, stains, dyes, tags, labels, marks}
- 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)

## Abstract

Sensor devices for quantifying luminescent targets are described herein. An example device comprises a light source for exciting the targets, thus generating luminescence signals and a detector for detecting these signals, resulting in a detected signal which comprises a desired signal originating from the targets and a background signal. It moreover comprises a bleaching device for bleaching of at least part of the sources generating the background signal and a processor configured to trigger the bleaching device to start bleaching, and to trigger the light source for exciting the remaining luminescent targets which are not bleached, and to trigger the detector for detecting the luminescence signal of the remaining luminescent targets, so as to generate a measurement signal representative for the quantification of the luminescent targets.

## Claims

### Claim 1

A sensor device for determining presence of luminescent targets, wherein the sensor device comprises:
a light source for exciting the luminescent targets, thus generating luminescence signals;
a detector for detecting luminescence signals of the luminescent targets, resulting in a detected signal which comprises a desired signal originating from the luminescent targets and a background signal;
a bleaching device for bleaching at least part of sources generating the background signal; and
a processor having stored thereon program instructions that, upon execution by the processor, cause performance of a set of acts comprising:
triggering the bleaching device to start bleaching and stop bleaching;
triggering the light source, after bleaching, for exciting remaining luminescent targets, wherein the remaining luminescent targets are those luminescent targets that are not bleached; and
triggering the detector for detecting the luminescence signal of the remaining luminescent targets, so as to generate a measurement signal representative for quantification of the luminescent targets.

### Claim 2 (depends on 1)

The sensor device according to claim 1, wherein the bleaching device is adapted for selective bleaching, and wherein the sources generating the background signal are bleached more than the luminescent targets.

### Claim 3 (depends on 1)

The sensor device according to claim 1, wherein the processor is configured to obtain a luminescent intensity using the detector during bleaching and to stop the bleaching device from bleaching when the obtained luminescent intensity has dropped below an adjustable threshold.

### Claim 4 (depends on 1)

The sensor device according to claim 1, wherein the bleaching device is a light source adapted for generating a high intensity pulse causing bleaching of at least part of the sources generating the background signal.

### Claim 5 (depends on 1)

The sensor device according to claim 1, wherein the luminescent targets are present in a fluid and wherein the bleaching device is adapted for applying a chemical plug which causes bleaching of at least part of the sources generating the background signal.

### Claim 6 (depends on 1)

The sensor device according to claim 1, the sensor device further comprising
a surface or a three dimensional volume for binding the luminescent targets; and
an evanescent field generating structure,
wherein the light source is coupled to the evanescent field generating structure, and
wherein, the evanescent field generating structure is adapted for generating an evanescent field at the surface or in the three dimensional volume.

### Claim 7 (depends on 2)

A diagnostic device comprising:
a sensor device according to claim 2 for sensing an analyte and generating a sensing signal; and
an output unit for providing an output of the sensor device on which a diagnosis can be based.

### Claim 8 (depends on 7)

The diagnostic device according to claim 7, wherein the output unit is adapted for outputting a signal indicative of presence/absence of the analyte or a concentration of the analyte.

### Claim 9 (depends on 1)

A method for quantifying luminescent targets using the sensor device of claim 1, the method comprising:
providing, at a first time, a sample comprising the luminescent targets or injecting a blocking material;
bleaching at least part of sources generating a background signal;
providing a sample comprising the luminescent targets if this was not done when injecting the first time; and
measuring a luminescence signal exciting the luminescent targets which are not bleached, thus generating a luminescence signal, detecting the luminescence signal of the remaining luminescent targets which are not bleached,
wherein the measured luminescence signal comprises a desired signal emanating from the luminescent targets and a background signal, so as to generate a measurement signal representative for the quantification of the luminescent targets.

### Claim 10 (depends on 9)

The method according to claim 9, further comprising determining a moment in time when bleaching should start, and starting bleaching at that moment.

### Claim 11 (depends on 10)

The method according to claim 10, further comprising a measure and compare step after injecting the sample or blocking material,
the measure and compare step comprising
exciting the sample or the blocking material, thus generating a luminescence signal, detecting the luminescence signal coming from the sample or blocking material;
analyzing the measured luminescence signal so as to obtain a moment to start bleaching; and
repeating the detecting and analyzing until the moment to start bleaching is determined.

### Claim 12 (depends on 9)

The method according to claim 9, comprising determining a duration of the bleaching step, and stopping bleaching after the determined duration has expired.

### Claim 13 (depends on 9)

The method according to claim 9, the method further comprising a feedback loop during the bleaching step, the feedback loop comprising:
detecting a luminescence signal during bleaching;
analyzing the detected signal and comparing it with a luminescence signal obtained before bleaching started; and
stopping the bleaching when the detected signal drops below a predetermined fraction of the luminescence signal obtained before bleaching started, or continuing bleaching and repeating the detection and analyzing steps if the detected signal has not dropped below the predetermined fraction of the luminescence signal obtained before bleaching started.