Laser scanning microscope

• US 6,094,300 A
• Filed: 11/18/1997
• Issued: 07/25/2000
• Est. Priority Date: 11/21/1996
• Status: Expired due to Term

• Current Assignee: Olympus Optical Corporation Limited (Olympus Corporation)
• Original Assignee: Olympus Optical Corporation Limited (Olympus Corporation)
• Inventors: Kishi, Yosuke, Kashima, Shingo
• Primary Examiner(s): Spyrou, Cassandra
• Assistant Examiner(s): WINSTEDT, JENNIFER E
• Application Number:
  US08/972,591
• Time in Patent Office: 980 Days
• Field of Search: 359/368, 359/385, 359/389, 359/201, 359/204, 356/317, 356/318, 356/417, 356/73
• US Class Current: 359/385
• CPC Class Codes: G02B 21/002 Scanning microscopes scanning near field optical microscopes G01Q60/18
• CPC Class Codes: G02B 21/16 adapted for ultraviolet illumination ; Fluorescence microscopes G02B21/0076 takes precedence

Abstract

A laser scanning microscope having a first scanning optical system that causes coherent light projected from a first laser light source to scan a specimen, and a second scanning optical system that projects coherent light output from a second laser light source at a given position on the specimen. Fluorescence from the specimen is measured by a photoelectric conversion element.

Claims

1. A laser scanning microscope comprising:

at least two laser light sources that output coherent light;

at least two scanning optical systems corresponding respectively to said at least two laser light sources, each of said at least two scanning optical systems separately causing rays of the coherent light outputted from the at least two laser light sources to scan a specimen; and

a sense optical system that senses at least one of;

(i) transmitted light produced by said coherent light passing through said specimen, (ii) reflected light produced by said coherent light being reflected by said specimen, and (iii) fluorescence from said specimen, when at least one of the scanning optical systems is causing said coherent light to scan said specimen.

2. A laser scanning microscope according to claim 1, further comprising:

an optical element that causes an optical axis of each of the rays of said coherent light caused to scan the specimen to coincide with each other; and

an objective that is placed on said coinciding optical axis and that gathers the rays of said coherent light at a same position on said specimen.

3. A laser scanning microscope according to claim 1, further comprising:

control means for providing scanning control with respect to each of said at least two scanning optical systems.

4. A laser scanning microscope according to claim 1, further comprising:

image processing means for forming images from each ray of said transmitted light, each ray of said reflected light sensed by said sense optical system for each of said scanning optical systems, and from each ray of said fluorescence, and for superimposing the images on at least a same image, when said at least two scanning optical systems cause said coherent light to scan said specimen.

5. A laser scanning microscope according to any one of claims 1, 2, 3, or 4, further comprising:

an optical adjustment mechanism that shifts along the optical axis of said coherent light a light convergence position on said specimen of at least one of the rays of said coherent light projected on said specimen by the scanning of said at least two scanning optical systems.

6. A laser scanning microscope according to claim 5, wherein said optical adjustment mechanism includes:

a fixed lens for gathering said coherent light; and

a movable lens that is placed so as to move freely along the optical axis of said coherent light, and that receives said coherent light gathered by said fixed lens and emits the coherent light at a given emission angle.

7. A laser scanning microscope according to claim 5, further comprising:

a position check optical system that projects position check light on said specimen, wherein a position on said specimen at which light is projected coincides with a light convergence position on said specimen of said coherent light projected on said specimen by the scanning of at least one of said scanning optical systems.

8. A laser scanning microscope according to any one of claims 1, 2, 3 or 4, further comprising:

a position check optical system that projects position check light on said specimen, wherein a position on said specimen at which light is projected coincides with a light convergence position on said specimen of said coherent light projected on said specimen by the scanning of at least one of said scanning optical systems.

9. A laser scanning microscope according to claim 1, wherein:

said at least two laser light sources comprise a first laser light source and a second laser light source, and

said at least two scanning optical systems comprise a first scanning optical system and a second scanning optical system corresponding respectively to said first laser light source and said second laser light source.

10. A laser scanning microscope according to claim 9 wherein:

said first scanning optical system causes said coherent light outputted from the first laser light source to scan a first side of a specimen;

said second scanning optical system causes said coherent light outputted from the second laser light source to scan a given region on a second side of said specimen opposite to said first side; and

said sense optical system senses the fluorescence from said specimen when said first scanning optical system scans the first side of said specimen, and senses at least one of the transmitted light and the fluorescence from said specimen when said second scanning optical system scans the given region on the second side of said specimen.

11. A laser scanning microscope according to claim 9, wherein:

said first scanning optical system causes said coherent light outputted from the first laser light source to scan the specimen;

said second scanning optical system causes said coherent light outputted from the second laser light source to scan a given region of said specimen;

said sense optical system senses the fluorescence from said specimen when said first scanning optical system scans said specimen, and senses at least one of the transmitted light and the reflected light from said specimen when said second scanning optical system scans the given region of said specimen; and

said laser microscope further comprises;

(i) a dichroic mirror that causes an optical axis of said coherent light from said first scanning optical system to coincide with an optical axis of said coherent light from said second scanning optical system; and

(ii) an objective that is placed on said coinciding optical axis and that gathers the rays of said coherent light on a same cross section of said specimen.

12. A laser scanning microscope according to claim 9, wherein:

said specimen is dyed with fluorescence pigment,

said first laser light source outputs coherent light including a wavelength that excites said fluorescence pigment to generate fluorescence, and

said scanning optical system projects said coherent light from said second laser light source on a given region of said specimen to cause said specimen to react.

13. A laser scanning microscope according to claim 9, wherein:

said first laser light source outputs one of visible continuous laser light and IR pulse laser light, and

said second laser light source outputs one of ultraviolet pulse laser light and IR pulse laser light.

14. A laser scanning microscope according to claim 13, wherein said IR pulse laser light is projected on a fluorescence pigment with which said specimen has been dyed, to induce two-photon absorption.

15. A laser scanning microscope according to claim 9, wherein:

said specimen is a workpiece,

said first laser light source outputs coherent light for observing said workpiece, and

said second laser light source outputs high-power coherent light for machining said workpiece.

16. A laser scanning microscope according to claim 9, wherein said first and second scanning optical systems each have a scanning mirror that scans in a direction perpendicular to each other and that deflects said coherent light in a direction perpendicular to each other.

17. A laser scanning microscope according to claim 9, further comprising:

control means for providing scanning control of said first and second scanning optical systems,

image processing means for arranging images obtained when said first and second scanning optical systems each scan said specimen, and

a monitor for displaying a result of the image processing from the image processing means.

18. A laser scanning microscope according to claim 9, further comprising an optical adjustment mechanism that shifts along an optical axis of said coherent light a light convergence position on said specimen of said coherent light projected on said specimen by the scanning of said second scanning optical system.

19. A laser scanning microscope according to claim 18, wherein said optical adjustment mechanism includes:

a fixed lens for gathering said coherent light; and

a movable lens that is placed so as to move freely along the optical axis of said coherent light, and that receives said coherent light gathered by said fixed lens and emits the coherent light at a given emission angle.

20. A laser scanning microscope according to claim 9, further comprising:

a light source for outputting position check light; and

mirror that directs the position check light onto an optical axis of said coherent light in said second scanning optical system, and projects said position check light at a same position at which said coherent light is projected on said specimen by the scanning said second scanning optical system.

21. A laser scanning microscope according to claim 20, wherein said position check light source outputs laser light having a wavelength that has no effect on said specimen.

22. A laser scanning microscope comprising:

a laser light source for outputting sub-picosecond monochromatic coherent light pulses with a wavelength in a near infrared range;

an objective system for gathering the monochromatic coherent light pulses from the laser light source on a specimen;

at least one bundle-of-rays conversion optical system for collimating a bundle of rays of monochromatic coherent light pulses outputted from said laser light source to a desired size;

optical-path branching means for at least one of;

(i) branching an optical path of the monochromatic coherent light pulses from the bundle-of-rays conversion optical system into two optical paths, and (ii) switching the optical path;

a first scanning optical system that is placed on a first branch optical path via the optical-path branching means and that gathers said monochromatic coherent light pulses on an image surface in said objective system and causes said monochromatic coherent light pulses to scan the image surface;

a sense optical system for sensing fluorescence from said specimen that is excited by a light convergence spot of the gathered monochromatic coherent light;

a second scanning optical system that is placed on a second branch optical path via the optical-path branching means and that gathers said monochromatic coherent light pulses on the image surface in said objective system and causes said monochromatic light pulses to scan the image surface; and

optical-path coupling means for introducing said light convergence spot into said objective system,

wherein said objective system is shared by said first and second optical paths.