Scanning optical microscope and method of acquiring image

• US 6,433,929 B1
• Filed: 06/08/2001
• Issued: 08/13/2002
• Est. Priority Date: 06/12/2000
• Status: Expired due to Term

• Current Assignee: Olympus Optical Company Ltd. (Olympus Corporation)
• Original Assignee: Olympus Optical Company Ltd. (Olympus Corporation)
• Inventors: Sasaki, Hiroshi
• Primary Examiner(s): Spyrou, Cassandra
• Assistant Examiner(s): Robinson, Mark A.
• Application Number:
  US09/877,767
• Publication Number:
  US 20020036824A1
• Time in Patent Office: 431 Days
• Field of Search: 359/212, 359/234, 359/223, 359/368, 359/385, 359/388, 250/461.1, 250/461.2, 356/320
• US Class Current: 359/388
• 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 scanning optical microscope comprises a light source configured to selectively output to a sample dyed with two or more types of fluorescent dyes an excitation light having an excitation wavelength according to the each fluorescent dye, scanner configured to scan the excitation light outputted from the light source, an objective lens configured to condense the excitation light scanned by the scanner on the sample, a detector configured to detect a fluorescence of the fluorescent light dye according to the excitation light by the excitation light condensed by the objective lens, one confocal pinhole whose pinhole diameter arranged in front of the detector is adjustable, and controller configured to adjust the pinhole diameter of the confocal pinhole to a diameter suitable for the fluorescence emitted from the sample by using the excitation light in synchronization with switching of the excitation lights from the light source when acquiring one image by detecting each fluorescence according to each excitation light in the time division manner through the confocal pinhole by switching the excitation lights with which the sample is irradiated in synchronization with scanning by the scanner.

Claims

1. A scanning optical microscope comprising:

a light source configured to selectively output to a sample dyed with two or more types of fluorescent dyes an excitation light having an excitation wavelength according to each of said fluorescent dyes;

scanner configured to scan said excitation light outputted from said light source;

an objective lens configured to condense said excitation light scanned by said scanner on said sample;

a detector configured to detect a fluorescence from said fluorescent dye according to said excitation light by using said excitation light condensed by said objective lens;

one confocal pinhole whose pinhole diameter arranged in front of said detector is adjustable; and

controller configured to adjust said pinhole diameter of said confocal pinhole to a diameter suitable for said fluorescence emitted from said sample by said excitation light in synchronization with switching of said excitation light from said light source, when acquiring one image by detecting each fluorescence according to said each excitation light in a time division manner through said confocal pinhole by switching said excitation light with which said sample is irradiated in synchronization with scanning by scanner.

2. The scanning optical microscope according to claim 1, wherein

said confocal pinhole is a minute device group having a plurality of minute devices and

said plurality of said minute devices are controlled by a minute device group controller.

3. The scanning optical microscope according to claim 1, wherein said confocal pinhole includes a minute device group configured by arranging a plurality of minute deflecting mirrors in a two-dimensional matrix form, further comprising minute device group controller configured to control an angle of each minute deflecting mirror within a diffraction diameter so as to reflect a light spot in an arrangement direction of said detector, and to control an angle of said each minute deflecting mirror outside said diffraction diameter to an angle different from said angle of said each minute deflecting mirror within said diffraction diameter.

4. The scanning optical microscope according to claim 1, wherein said controller switches said excitation light in synchronization with scanning by said light scanner in accordance with each one line with respect to said sample.

5. The scanning optical microscope according to claim 4, wherein

said confocal pinhole is a minute device group having a plurality of minute devices, and

said plurality of said minute devices are controlled by a minute device group controller.

6. The scanning optical microscope according to claim 4, wherein said confocal pinhole includes a minute device group configured by arranging a plurality of minute deflecting mirrors in a two-dimensional matrix form, further comprising a minute device controller configured to control an angle of said each minute deflecting mirror within a diffraction diameter so as to reflect a light spot in an arrangement direction of said detector, and to control an angle of said each minute deflecting mirror outside said diffraction diameter to an angle different from said each angle of said each minute deflecting mirror in said diffraction diameter.

7. The scanning optical microscope according to claim 1, wherein said controller switches said excitation lights in synchronization with scanning by said scanner in accordance with each one frame.

8. The scanning optical microscope according to claim 7, wherein said confocal pinhole is a minute device group having a plurality of minute devices, and

said plurality of said minute devices are controlled by a device controller.

9. The scanning optical microscope according to claim 7, wherein said confocal pinhole includes a minute device group configured by arranging a plurality of minute deflecting mirrors in a two-dimensional matrix form, further comprising minute device group controller configured to control an angle of said each minute deflecting mirror within a diffraction diameter so as to reflect a light spot in an arrangement direction of said detector, and to control an angle of said each minute deflecting mirror out side said diffraction diameter to an angle different from said angle of said each minute deflecting mirror in said diffraction diameter.

10. The scanning optical microscope according to claim 1, wherein said controller switches said excitation lights in units of pixel during scanning with respect to said sample by said scanner.

11. The scanning optical microscope according to claim 10, wherein

said confocal pinhole is a minute device group having a plurality of minute devices, and

said plurality of said minute devices are controlled by a minute device controller.

12. The scanning optical microscope according to claim 10, wherein said confocal pinhole includes a minute device group configured by arranging a plurality of minute deflecting mirrors in a two-dimensional matrix form, further comprising minute device group controller to control an angle of said each minute deflecting mirror within a diffraction diameter so as to reflect a light spot in an arrangement direction of said detector, and to control an angle of said each minute deflecting mirror outside said diffraction diameter to an angle different from said angle of said each minute deflecting mirror within said diffraction diameter.

13. The scanning optical microscope according to claim 1, wherein said microscope includes a single detector.

14. The scanning optical microscope according to claim 13, further comprising a barrier filter which is fixed in front of said detector, blocks two or more types of excitation lights for exciting said two or more types of fluorescences, and transmits therethrough said two or more types of fluorescences emitted by said sample by said excitation lights.

15. The scanning optical microscope according to claim 13, further comprising:

a first barrier filter for blocking a first excitation light of said excitation lights and transmitting therethrough said fluorescence emitted from said sample by said first excitation light;

a second barrier filter for blocking a second excitation light of said excitation lights and transmitting therethrough said fluorescence emitted from said sample by said second fluorescence; and

means for switching said first barrier filter and said second barrier filter between said confocal pinhole and said detector in synchronization with change of said excitation lights.

16. The scanning optical microscope according to claim 1, wherein said microscope includes a plurality of detectors.