Multicolor fluorescence analysis device

Patent metadata

Field	Value
Publication number	US-10168281-B2
Application number	US-201515539228-A
Country	US
Kind code	B2
Filing date	Nov 11, 2015
Priority date	Jan 27, 2015
Publication date	Jan 1, 2019
Grant date	Jan 1, 2019

Abstract

A multicolor fluorescence analysis device 11 is for detecting fluorescence emitted, as a result of excitation light irradiation, from a plurality of types of fluorophores included in a sample s, and is provided with an irradiation optical unit 520 for irradiating light emitted from a light source 510 onto a sample s as excitation light, a fluorescence condensation unit 530 having a fluorescence filter 531 that transmits light emitted from the sample s and transmits light of transmission wavelength bands different from the excitation wavelength bands, and a two-dimensional detector 554 that has a plurality of types of transmission filters 556 for transmitting prescribed wavelengths of light and detects the intensity of the light of the prescribed wavelength for each transmission filter 556 , and the light emitted from at least two fluorophores from among the plurality of types of fluorophores is detected simultaneously and the fluorophore types are identified accordingly.

First claim

The invention claimed is:

1. A multicolor fluorescence analysis device for detecting fluorescence emitted from a plurality of types of fluorescent dyes having different fluorescence wavelengths contained in a sample by irradiation with excitation light, the multicolor fluorescence analysis device comprising:
a light source for excitation;
an irradiation optical unit having an excitation filter that transmits light in a plurality of different excitation wavelength bands and irradiating the sample with light emitted from the light source as the excitation light through the excitation filter;
a fluorescence condensation unit having a fluorescence filter that transmits at least a portion of the fluorescence emitted from the sample by the irradiation of the excitation light and transmits light in a plurality of transmission wavelength bands not including the excitation wavelength band; and
a two-dimensional detector having a plurality of types of transmission filters that transmit light having a predetermined wavelength out of the light transmitted by the fluorescence filter and a two-dimensional sensor configured to detect an intensity of the light transmitted by each of the transmission filters for each of the transmission filters, the plurality of types of transmission filters being provided to each detection element of the two-dimensional sensor so to correspond to each detection element,
wherein light emitted from at least two types of fluorescent dyes out of the plurality of types of fluorescent dyes is simultaneously detected by using the two-dimensional detector, and the types of the fluorescent dyes are identified from intensities of the detected light, and
the transmission filters are configured with four types having different transmission wavelength bands.

2. The multicolor fluorescence analysis device according to claim 1 , wherein the four types of transmission filters are arranged by setting a block where the respective types of transmission filters are aligned one by one as one unit and repeating the unit in a plane direction.

3. The multicolor fluorescence analysis device according to claim 1 , wherein the fluorescent dyes are of four types, and intensity ratios of fluorescence from the fluorescent dyes are different among the respective transmission filters.

4. The multicolor fluorescence analysis device according to claim 3 ,
wherein the four types of fluorescent dyes are a first dye, a second dye, a third dye, and a fourth dye, and the four types of transmission filters are a first filter, a second filter, a third filter, and a fourth filter,
each of the fluorescence from the first dye and the fluorescence from the second dye has the highest intensity ratio in the first filter,
in the second filter, the intensity ratio of the fluorescence from the first dye is larger than the intensity ratio of the fluorescence from the second dye,
each of the fluorescence from the third dye and the fluorescence from the fourth dye has the highest intensity ratio in the third filter, and
in the fourth filter, the intensity ratio of the fluorescence from the fourth dye is larger than the intensity ratio of the fluorescence from the third dye.

5. The multicolor fluorescence analysis device according to claim 1 , wherein the excitation light to be irradiated on the sample is configured with at least two excitation wavelength bands.

6. The multicolor fluorescence analysis device according to claim 1 , wherein the excitation light to be irradiated on the sample is configured with at least three excitation wavelength bands.

7. The multicolor fluorescence analysis device according to claim 1 , further comprising a data processing unit identifying two or more types of fluorescent dyes from the intensity of light detected by the two-dimensional detector.

8. A multicolor fluorescence analysis device for detecting fluorescence emitted from a plurality of types of fluorescent dyes having different fluorescence wavelengths contained in a sample by irradiation with excitation light, the multicolor fluorescence analysis device comprising:
a light source for excitation;
an irradiation optical unit having a plurality of excitation filters that transmit light in a plurality of different excitation wavelength bands and are switchable and irradiating the sample with light emitted from the light source as the excitation light through the excitation filter;
a fluorescence condensation unit having a fluorescence filter that transmits at least a portion of the fluorescence emitted from the sample by the irradiation of the excitation light and transmits light in a plurality of transmission wavelength bands not including the excitation wavelength band; and
a two-dimensional detector having a plurality of types of transmission filters that transmit light having a predetermined wavelength out of the light transmitted by the fluorescence filter and a two-dimensional sensor configured to detect an intensity of the light transmitted by each of the transmission filters for each of the transmission filters, the plurality of types of transmission filters being provided to each detection element of the two-dimensional sensor so to correspond to each detection element,
wherein light emitted from at least two types of fluorescent dyes out of the plurality of types of fluorescent dyes is simultaneously detected by using the two-dimensional detector, and the types of the fluorescent dyes are identified from intensities of the detected light, and
the transmission filters are configured with four types having different transmission wavelength bands.

9. The multicolor fluorescence analysis device according to claim 8 , wherein the four types of transmission filters are arranged by setting a block where the respective types of transmission filters are aligned one by one as one unit and repeating the unit in a plane direction.

10. The multicolor fluorescence analysis device according to claim 8 , wherein the fluorescent dyes are of four types, and intensity ratios of fluorescence from the fluorescent dyes are different among the respective transmission filters.

11. The multicolor fluorescence analysis device according to claim 10 , wherein the four types of fluorescent dyes are a first dye, a second dye, a third dye, and a fourth dye, and the four types of transmission filters are a first filter, a second filter, a third filter, and a fourth filter,
each of the fluorescence from the first dye and the fluorescence from the second dye has the highest transmittance in the first filter,
in the second filter, the intensity ratio of the fluorescence from the first dye is larger than the intensity ratio of the fluorescence from the second dye,
each of the fluorescence from the third dye and the fluorescence from the fourth dye has the largest intensity ratio in the third filter, and
in the fourth filter, the intensity ratio of fluorescence from the fourth dye is larger than the intensity ratio of fluorescence from the third dye.

12. A multicolor fluorescence analysis device for detecting fluorescence emitted from a plurality of types of fluorescent dyes having different fluorescence wavelengths contained in a sample by irradiation with excitation light, the multicolor fluorescence analysis device comprising:
a plurality of light sources for excitation;
an irradiation optical unit having an excitation filter that is provided for each of the light sources to transmit light in a plurality of different excitation wavelength bands and irradiating the sample with light emitted from the light source as the excitation light through the excitation filter;
a fluorescence condensation unit having a fluorescence filter that

Assignees

• Hitachi High Tech Corp

Inventors

• Shoji Tomohiro
• Shiokawa Junji
• Nishizawa Akihito
• Kato Hirokazu
• Ishizuka Junji

Classifications

• G01J3/4406 Primary

  Fluorescence spectrometry · CPC title

• G01N21/64 Primary

  Fluorescence; Phosphorescence · CPC title

• G01N21/645 Primary

  Specially adapted constructive features of fluorimeters · CPC title

• G01N2021/6421

  Measuring at two or more wavelengths · CPC title

• G01N2021/6441

  with two or more labels · CPC title