# US 10,481,086 B2 · Mass spectrometer and spectrometry, which use near infrared fluorescence

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- Number: US 10,481,086 B2
- Document ID: US10481086B2
- Status: Granted
- Kind: B2
- Application number: 16/088,432
- Filed: 2016-10-28
- Published: 2019-04-18
- Granted: 2019-11-19
- Earliest claimed: 2016-03-31
- Assignees: Asta Co., Ltd.
- Applicants: Asta Co., Ltd.
- Inventors: Jooyeon Oh; Yang Sun Kim
- Web page: https://patentgrep.com/patent/US10481086B2

## CPC

- G01N 21/359: using near infrared light
- G01N 21/64: Fluorescence; Phosphorescence
- G01N 27/62: by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
- H01J 49/0004: {Imaging particle spectrometry}
- H01J 49/164: {Laser desorption/ionisation, e.g. matrix-assisted laser desorption/ionisation [MALDI] (sample holders H01J49/0418)}
- H01J 49/26: Mass spectrometers or separator tubes
- G01N 21/6456: {Spatial resolved fluorescence measurements; Imaging}
- 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)
- H01J: ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS (spark-gaps H01T; electric arc lamps H05B31/00; particle accelerators H05H)

## Abstract

A mass spectrometer using near infrared (NIR) fluorescence can comprise: a plate formed such that a sample, which includes a matrix for ionization and a fluorescent material having an excitation wavelength of an NIR wavelength band, is loaded thereon; a fluorescence detection unit formed so as to acquire a fluorescence image by using the fluorescent material from the sample on the plate; a light emission unit formed so as to emit a laser beam for ionization at the sample on the plate; and an ion detection unit formed so as to detect, by using the laser beam, ions generated from the sample. By using the NIR fluorescence, fluorescence interference due to a matrix used for MALDI imaging can be excluded even after coating the matrix, and since fluorescence interference due to auto-fluorescence inside tissue is low, fluorescence can be measured even for a thick sample.

## Claims

### Claim 1

A mass spectrometer using near infrared (NIR) fluorescence for combination of NIR fluorescence imaging and matrix assistant laser desorption/ionization (MALDI) imaging, comprising:
a plate on which a sample is loaded, wherein the sample includes a fluorescent material having an excitation wavelength of an NIR wavelength band and a matrix for ionization, the sample comprising human tissue or animal tissue and a thickness of at least 10 μm;
a fluorescence detection unit comprising a light source that irradiates the sample on the plate with excitation light to excite the fluorescent material included in the sample, the fluorescence detection unit acquiring a fluorescence image from a first surface of the plate responsive to the irradiation of the sample on the plate with the excitation light;
a light emission unit configured to emit a laser beam for ionization at the sample on the plate, the laser beam emitted to a second surface of the plate that is opposite to the first surface of the plate;
a moveable stage on which the plate is mounted, the moveable stage configured to move with respect to the fluorescence detection unit and the laser beam; and
an ion detection unit configured to acquire a mass image of the sample by detecting ions generated from the sample due to the laser beam being emitted to the sample while moving the plate with respect to the laser beam using the moving stage,
wherein the fluorescence detection unit and the ion detection unit are on a same axis but located opposite to each other with the plate between the fluorescence detection unit and the ion detection unit.

### Claim 2 (depends on 1)

The mass spectrometer using NIR fluorescence according to claim 1, wherein the first surface of the plate includes an optically transparent region, and
the fluorescence detection unit is further configured to acquire the fluorescence image through the optically transparent region.

### Claim 3 (depends on 1)

The mass spectrometer using NIR fluorescence according to claim 1, further comprising:
a vacuum chamber to receive the plate, the fluorescence detection unit and the ion detection unit.

### Claim 4 (depends on 1)

The mass spectrometer using NIR fluorescence according to claim 1, wherein the ion detection unit is further configured to calculate a mass-to-charge ratio of the ion using a time taken for the ion generated from the sample by the laser beam to reach the ion detection unit.

### Claim 5

Mass spectrometry using near infrared (NIR) fluorescence for combination of NIR fluorescence imaging and matrix assistant laser desorption/ionization (MALDI) imaging, comprising:
placing a sample, which includes a fluorescent material having an excitation wavelength of an NIR wavelength band and a matrix for ionization, on a plate, the sample comprising human tissue or animal tissue and a thickness of at least 10 μm;
acquiring, by a fluorescence detection unit that comprises a light source that irradiates the sample on the plate with excitation light to excite the fluorescent material included in the sample, a fluorescence image from a first surface of the plate responsive to the irradiation of the sample on the plate with the excitation light;
emitting, by a light emission unit, a laser beam for ionization at the sample on the plate, the laser beam emitted to a second surface of the plate that is opposite to the first surface of the plate; and
acquiring, by an ion detection unit, a mass image of the sample by detecting ions generated from the sample due to the laser beam being emitted to the sample while moving the plate with respect to the laser beam using the moving stage,
wherein the fluorescence detection unit and the ion detection unit are on a same axis but located opposite to each other with the plate between the fluorescence detection unit and the ion detection unit.

### Claim 6 (depends on 5)

The mass spectrometry using NIR fluorescence according to claim 5, wherein the mass spectrometry further comprises, after the acquiring the fluorescence image, determining a target region to perform mass spectrometry based on the fluorescence image, and
the emitting the laser beam comprises emitting the laser beam to the determined target region.

### Claim 7 (depends on 5)

The mass spectrometry using NIR fluorescence according to claim 5, wherein the acquiring the fluorescence image from the first surface of the plate comprises acquiring the fluorescence image through an optically transparent region provided in the first surface of the plate.

### Claim 8 (depends on 5)

The mass spectrometer using NIR fluorescence according to claim 5, wherein the mass spectrometry comprises, before the acquiring the fluorescence image, placing the plate, the fluorescence detection unit, and the ion detection unit in a vacuum chamber.

### Claim 9 (depends on 5)

The mass spectrometry using NIR fluorescence according to claim 5, wherein the mass spectrometry further comprises calculating a mass-to-charge ratio of the ion using a time taken for the ion generated from the sample by the laser beam to reach the ion detection unit.