LASER EXCITATION FLUORESCENT MICROSCOPE

• US 20100182683A1
• Filed: 03/26/2010
• Published: 07/22/2010
• Est. Priority Date: 10/31/2007
• Status: Active Grant

• Current Assignee: Nikon Corporation
• Original Assignee: Nikon Corporation
• Inventors: SATO, Masatoshi, OKUGAWA, Hisashi, AIKAWA, Naoshi
• Granted Patent: US 8,310,754 B2
• Time in Patent Office: Days
• US Class Current: 359/389
• CPC Class Codes: G02B 21/0076 arrangements using fluorescence or luminescence
• CPC Class Codes: G02B 5/0833 comprising inorganic materials only
• CPC Class Codes: G02B 5/285 comprising deposited thin solid films G02B5/281 - G02B5/289 take precedence; multilayered film filters for fibre optic multiplexing G02B6/29361

Abstract

The present application has a proposition to provide a highly efficient laser excitation fluorescent microscope. Accordingly, a laser excitation fluorescent microscope of the present application includes a laser light source part radiating at least two types of excitation lights having different wavelengths; a light collecting part collecting the two types of excitation lights on a sample; a high-functional dichroic mirror, disposed between the laser light source part and the light collecting part, reflecting the two types of excitation lights to make the excitation lights incident on the light collecting part, and transmitting two types of fluorescence generated at the sample; and a detecting part detecting light transmitted through the high-functional dichroic mirror, in which an incident angle θ of the excitation lights and the fluorescence to the high-functional dichroic mirror satisfies a formula of 0°<θ<45°.

Claims

1. A laser excitation fluorescent microscope, comprising:

a laser light source part radiating at least two types of excitation lights having different wavelengths;

a light collecting part collecting the two types of excitation lights radiated by the laser light source part on a sample;

a high-functional dichroic mirror, disposed between the laser light source part and the light collecting part, reflecting the two types of excitation lights radiated by the laser light source part to make the excitation lights incident on the light collecting part, and transmitting two types of fluorescence generated at the sample in accordance with the two types of excitation lights; and

a detecting part detecting light transmitted through the high-functional dichroic mirror, wherein

an incident angle θ

of the excitation lights and the fluorescence to the high-functional dichroic mirror satisfies a formula of 0°

<

θ

<

45°

.

2. The laser excitation fluorescent microscope according to claim 1, wherein

the incident angle θ

of the excitation lights and the fluorescence to the high-functional dichroic mirror satisfies a formula of 10°

<

θ

<

25°

.

3. The laser excitation fluorescent microscope according to claim 1, wherein

the incident angle θ

of the excitation lights and the fluorescence to the high-functional dichroic mirror satisfies a formula of 10°

<

θ

<

15°

.

4. The laser excitation fluorescent microscope according to claim 1, wherein

the incident angle θ

of the excitation lights and the fluorescence to the high-functional dichroic mirror is 12°

.

5. The laser excitation fluorescent microscope according to claim 1, wherein

a separating layer of the high-functional dichroic mirror is formed by a dielectric multilayer.

6. A laser excitation fluorescent microscope, comprising:

a laser light source part radiating at least two types of excitation lights having different wavelengths;

a light collecting part collecting the two types of excitation lights radiated by the laser light source part on a sample;

a high-functional dichroic mirror, disposed between the laser light source part and the light collecting part, reflecting the two types of excitation lights radiated by the laser light source part to make the excitation lights incident on the light collecting part, and transmitting two types of fluorescence generated at the sample in accordance with the two types of excitation lights, the high-functional dichroic mirror being formed by a dielectric multilayer; and

a detecting part detecting light transmitted through the high-functional dichroic mirror, wherein

a wavelength characteristic curve of a reflectivity and a transmittance of the high-functional dichroic mirror has;

a first reflecting band covering a wavelength band of one of the two types of excitation lights;

a first transmitting band covering a wavelength band of one of the fluorescence generated in accordance with said one of the two types of excitation lights;

a second reflecting band covering a wavelength band of other one of the two types of excitation lights; and

a second transmitting band covering a wavelength band of one of the fluorescence generated in accordance with said other one of the two types of excitation lights, wherein;

a reflectivity in each of the first reflecting band and the second reflecting band is 95% or more;

a transmittance in each of the first transmitting band and the second transmitting band is 95% or more;

a wavelength width T₁ of the first transmitting band and a wavelength width T₂ of the second transmitting band are each 25 nm or more; and

a rising width A₁ from the first reflecting band to the first transmitting band and a rising width A₂ from the second reflecting band to the second transmitting band are each 6 nm or less.

7. The laser excitation fluorescent microscope according to claim 6, wherein

the transmittance in each of the first transmitting band and the second transmitting band indicates a value of 98% or more for a range of 90% or more of the wavelength width.

8. The laser excitation fluorescent microscope according to claim 6, wherein

a gap B between the first transmitting band and the second transmitting band is 20 nm or less.

9. The laser excitation fluorescent microscope according to claim 6, wherein

an incident angle θ

of the excitation lights and the fluorescence to the high-functional dichroic mirror satisfies a formula of 0°

<

θ

<

45°

.

10. The laser excitation fluorescent microscope according to claim 6, wherein

the incident angle θ

of the excitation lights and the fluorescence to the high-functional dichroic mirror satisfies a formula of 10°

<

θ

<

25°

.

11. The laser excitation fluorescent microscope according to claim 6, wherein

the incident angle θ

of the excitation lights and the fluorescence to the high-functional dichroic mirror satisfies a formula of 10°

<

θ

<

15°

.

12. The laser excitation fluorescent microscope according to claim 6, wherein

the incident angle θ

of the excitation lights and the fluorescence to the high-functional dichroic mirror is 12°

.

13. The laser excitation fluorescent microscope according to claim 6, wherein

the detecting part detects a spectrum of light transmitted through the high-functional dichroic mirror.