Multicolor fluorescence analysis device

Patent metadata

Field	Value
Publication number	US-10908083-B2
Application number	US-201515545422-A
Country	US
Kind code	B2
Filing date	Feb 2, 2015
Priority date	Feb 2, 2015
Publication date	Feb 2, 2021
Grant date	Feb 2, 2021

Abstract

A multicolor fluorescence analysis device is provided. The device includes a first fiberoptic plate for guiding light including fluorescence emitted from a sample as a result of irradiation of excitation light and emitting the same from a first emission part. The device includes a second fiberoptic plate for receiving light emitted from the first emission part at a second incidence part, guiding the same, and emitting the same from a second emission part. The device includes a single multilayer dielectric interference film filter that is provided on an end surface of the second emission part, transmits at least a portion of the fluorescence, and transmits light of a plurality of transmission wavelength bands that do not include the excitation wavelength bands. The device includes a two-dimensional detection unit that is disposed so as to be adhered to the multilayer dielectric interference film filter.

First claim

The invention claimed is:

1. A multicolor fluorescence analysis device detecting fluorescent light which is emitted from a plurality of types of fluorescent pigments contained in a sample and having different fluorescence wavelengths by being irradiated with excitation light, the device comprising:
an irradiation unit which sequentially irradiates the sample with a plurality of excitation lights, the excitation lights having different excitation wavelength bands;
a first fiber optic plate which is in contact with at least a part of the sample, receives light including the fluorescent light emitted from the sample via a first incident portion and guides the light, and allows the light to exit from a first exiting portion on a side opposite to the first incident portion;
a second fiber optic plate which receives the light exiting from the first exiting portion via a second incident portion and guides the light, and allows the light to exit from a second exiting portion on a side opposite to the second incident portion;
a single dielectric multilayer film interference filter which is disposed on an end surface of the second exiting portion, and transmits at least a part of the fluorescent light and transmits light having a plurality of transmission wavelength bands not including the plurality of excitation wavelength bands; and
a two-dimensional detection unit which is disposed to be attached to the dielectric multilayer film interference filter and disposed downstream of the first and second fiber optic plates, and detects light transmitted through the dielectric multilayer film interference filter;
wherein the dielectric multilayer film interference filter is a first dielectric multilayer film interference filter having a first transmittance (α 1 ) for a first transmission wavelength band that is less than a predetermined value (α), and the irradiation unit includes a light source and a second dielectric multilayer film interference filter, having a second transmittance (α 2 ) for a second transmission wavelength band that is less than the predetermined value (α), which selectively transmits the excitation lights;
wherein a first transmission wavelength at a lower end of the first transmission wavelength band and a second transmission wavelength at an upper end of the second transmission wavelength band satisfy a relationship λ 2 ≤λ 1 (1−(sin θ max /n eff ) 2 ) 1/2 , such that the first transmission wavelength band and the second transmission wavelength band do not overlap,
wherein λ 2 is the second transmission wavelength, λ 1 is the first transmission wavelength, θ max is a maximum incident angle of light incident on the first dielectric multilayer film interference filter, and n eff is an effective refractive index of the first dielectric multilayer film interference filter.

2. The multicolor fluorescence analysis device according to claim 1 , wherein the predetermined value (α) is 0.1%.

3. The multicolor fluorescence analysis device according to claim 1 , wherein the effective refractive index (n eff ) of the first dielectric multilayer film interference filter is greater than or equal to 1.5 and less than or equal to 2.2.

4. The multicolor fluorescence analysis device according to claim 1 , wherein the maximum incident angle (θ max ) of the light incident on the first dielectric multilayer film interference filter is 30°.

5. The multicolor fluorescence analysis device according to claim 1 , wherein an outer diameter of an optical fiber configuring the second fiber optic plate is less than an outer diameter of an optical fiber configuring the first fiber optic plate.

6. The multicolor fluorescence analysis device according to claim 1 , wherein the irradiation unit is configured to simultaneously emit at least two of the excitation lights.

7. The multicolor fluorescence analysis device according to claim 1 , wherein the plurality of types of fluorescent pigments includes Alexa405, FAM, Texas Red, and Cy5.5.

8. The multicolor fluorescence analysis device according to claim 1 ,
wherein the second dielectric multilayer film interference filter includes a first filter, a second filter, a third filter, and a fourth filter,
a transmittance of the first filter is greater than or equal to 85% in a wavelength band of 380 nm to 396 nm, and is less than 0.1% in a wavelength band other than the wavelength band of 380 nm to 396 nm,
a transmittance of the second filter is greater than or equal to 85% in a wavelength band of 474 nm to 497 nm, and is less than 0.1% in a wavelength band other than the wavelength band of 474 nm to 497 nm,
a transmittance of the third filter is greater than or equal to 85% in a wavelength band of 561 nm to 575 nm, and is less than 0.1% in a wavelength band other than the wavelength band of 561 nm to 575 nm,
a transmittance of the fourth filter is greater than or equal to 85% in a wavelength band of 641 nm to 657 nm, and is less than 0.1% in a wavelength band other than the wavelength band of 641 nm to 657 nm, and
the first filter, the second filter, the third filter, and the fourth filter are used by being switched.

9. The multicolor fluorescence analysis device according to claim 8 , wherein a transmittance of the first dielectric multilayer film interference filter is greater than or equal to 85% in wavelength bands of 436 nm to 462 nm, 536 nm to 548 nm, 620 nm to 632 nm, and 713 nm to 800 nm, and is less than 0.1% in a wavelength band other than the aforementioned wavelength bands.

10. The multicolor fluorescence analysis device according to claim 1 , wherein the second dielectric multilayer film interference filter is a single filter, a transmittance of the second dielectric multilayer film interference filter is greater than or equal to 85% in the wavelength bands of 380 nm to 396 nm, 474 nm to 497 nm, 561 nm to 575 nm, and 641 nm to 657 nm, and is less than 0.1% in a wavelength band other than the aforementioned wavelength bands.

11. The multicolor fluorescence analysis device according to claim 1 , wherein
an end surface of a core portion of an optical fiber in the first fiber optic plate is subjected to an aminosilane treatment, and
a surface of the first fiber optic plate on a side which is in contact with the sample, wherein the surface is other than the end surface of the core portion of the optical fiber, is subjected to an HMDS treatment.

Assignees

• Hitachi High Tech Corp

Inventors

• Kato Hirokazu
• Shoji Tomohiro
• Takahashi Satoshi
• Maekawa Akira

Classifications

• G01N21/6454 Primary

  using an integrated detector array · CPC title

• G01N2021/6471

  Special filters, filter wheel · CPC title

• G01N2021/6419

  Excitation at two or more wavelengths · CPC title

• G01N2021/6421

  Measuring at two or more wavelengths · CPC title

• G01N2021/6484

  Optical fibres · CPC title