Multicolor Fluorescence Analysis Device

Patent metadata

Field	Value
Publication number	US-2018017492-A1
Application number	US-201515545422-A
Country	US
Kind code	A1
Filing date	Feb 2, 2015
Priority date	Feb 2, 2015
Publication date	Jan 18, 2018
Grant date	—

Abstract

A multicolor fluorescence analysis device 1 according to the present invention is provided with: an irradiation unit 41 for irradiating excitation light having a plurality of excitation wavelength bands onto a sample s; a first fiberoptic plate 424 for guiding light including fluorescence emitted from the sample s as a result of the irradiation of the excitation light and emitting the same from a first emission part 425 b ; a second fiberoptic plate 431 for receiving light emitted from the first emission part 425 b at a second incidence part 434 a , guiding the same, and emitting the same from a second emission part 434 b ; a single multilayer dielectric interference film filter 432 that is provided on an end surface of the second emission part 434 b , 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; and a two-dimensional detection unit 433 that is disposed so as to be adhered to the multilayer dielectric interference film filter 432 and detect light having passed through the multilayer dielectric interference film filter 432.

First claim

1 . A multicolor fluorescence analysis device detecting fluorescent light which is emitted from a plurality of types of fluorescent pigments having different fluorescence wavelengths contained in a sample by being irradiated with excitation light, the device comprising:
an irradiation unit which irradiates the sample with excitation light having a plurality of excitation wavelength bands different from each other; 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 by being irradiated with the excitation light from 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 from 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 excitation wavelength band; and a two-dimensional detection unit which is disposed to be closely attached to the dielectric multilayer film interference filter, and detects light transmitted through the dielectric multilayer film interference filter.

2 . The multicolor fluorescence analysis device according to claim 1 , wherein the dielectric multilayer film interference filter is a first dielectric multilayer film interference filter, and the irradiation unit includes a light source and a second dielectric multilayer film interference filter which selectively transmits excitation light having a predetermined excitation wavelength band from light emitted from the light source.

3 . The multicolor fluorescence analysis device according to claim 2 , wherein a rising wavelength (λ 1i ) satisfying a relationship in which a transmittance (α 1 ) of light in a transmission wavelength band of the first dielectric multilayer film interference filter is less than a predetermined value (α), and a falling wavelength (λ 2j ) satisfying a relationship in which a transmittance (α 2 ) of light in a transmission wavelength band of the second dielectric multilayer film interference filter is less than the predetermined value (α) satisfy a relationship represented by Expression (1) described below,
[Expression 1]
λ 2j ≦λ 1i √{square root over ( 1 −(sin θ max /n eff ) 2 )}  (1)

in Expression (1), i and k are respectively integers greater than or equal to 1, 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.

4 . The multicolor fluorescence analysis device according to claim 3 , wherein the predetermined value (α) is 0.1%.

5 . The multicolor fluorescence analysis device according to claim 3 , 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.

6 . The multicolor fluorescence analysis device according to claim 3 , wherein the maximum incident angle (θ max ) of the light incident on the first dielectric multilayer film interference filter is 30°.

7 . 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.

8 . The multicolor fluorescence analysis device according to claim 3 , wherein the excitation light in each excitation wavelength band is sequentially emitted.

9 . The multicolor fluorescence analysis device according to claim 3 , wherein excitation light having at least two excitation wavelength bands are simultaneously emitted.

10 . The multicolor fluorescence analysis device according to claim 1 , wherein the sample is a DNA segment containing a fluorescent pigment, and the multicolor fluorescence analysis device is used for analyzing a base sequence of the DNA segment.

11 . The multicolor fluorescence analysis device according to claim 10 , wherein the fluorescent pigment is Alexa405, FAM, Texas Red, and Cy5.5.

12 . The multicolor fluorescence analysis device according to claim 11 ,
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.

13 . The multicolor fluorescence analysis device according to claim 11 , 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.

14 . The multicolor fluorescence analysis device according to claim 12 , 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.

15 . The multicolor fluorescence analysis device according to claim 10 , wherein an end surface of a core portion of the optical fiber is subjected to an aminosilane treatment, and a surface other than the end surface of the core portion of the optical fiber is subjected to an HMDS treatment, on a surface of the first fiber optic plate on a side which is in contact with the sample.

Assignees

• Hitachi High Tech Corp

Inventors

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

Classifications

• G01N21/64 Primary

  Fluorescence; Phosphorescence · CPC title

• G01N2021/6471

  Special filters, filter wheel · CPC title

• G01N2021/6484

  Optical fibres · CPC title

• G01N21/645 Primary

  Specially adapted constructive features of fluorimeters · CPC title

• G01N2021/6419

  Excitation at two or more wavelengths · CPC title