JP2015099256A - Confocal microscope system - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a small-size confocal microscope system that is more compact than conventional systems and occupies no large installation space.SOLUTION: In a confocal microscope system comprising: a scanner unit that is configured of a micro-lens disk and a pinhole disk and scans laser beams; an optical projection subsystem that projects light on this scanner unit; an optical exciting subsystem that excites a sample by projecting laser beams scanned by the scanner unit onto the sample; and an optical imaging subsystem that separates light having again passed the scanner unit, out of fluorescent light emitted from the sample, and causes an image to be formed on a photographic device, the optical projection subsystem and optical exciting subsystem are arranged in parallel.

Description

  The present invention relates to a confocal microscope system, and more particularly to a confocal microscope system in which installation space is improved by downsizing.

  2A and 2B are configuration explanatory views showing an example of a conventional confocal microscope system described in Patent Document 1. FIG. 2A is a plan view and FIG. 2B is an enlarged front view. In FIG. 2, the confocal microscope system 1 is covered with a protective housing 2. A temperature control chamber 3 capable of adjusting the temperature is provided inside the protective housing 2, and the inside of the temperature control chamber 3 is adjusted to have a substantially constant temperature (for example, 30 ° C.). The protective housing 2 is arranged so that the right side surface in FIG. 2A is the front surface and the right side surface of the temperature control chamber 3 is along the front surface of the protective housing 2.

  An XY driving device 4 is disposed on the front side inside the temperature control chamber 3, and a moving table 5 that is driven in the XY directions is provided on the XY driving device 4. A sample cassette 7 having a glass bottom dish 6 in which a sample is stored and a light source 8 are mounted on the moving table 5. The sample cassette 7 moves on the XY driving device 4 in the XY direction as the moving table 5 moves.

  A condenser lens 19 and a light source 8 are disposed above the sample cassette 7 so as to face the sample cassette 7, and an objective lens 10 is disposed below the sample cassette 7 so as to face the sample cassette 7. The objective lens 10 is driven by the drive unit 11 in the Z direction indicated by the arrow.

  A scanner unit 12 is disposed on the left side of the XY driving device 4. The scanner unit 12 includes a microlens disk 13 and a pinhole disk 14 connected by a hub 15, and is rotated by a motor 16. A dichroic mirror 17 is disposed between the microlens disk 13 and the pinhole disk 14 constituting the scanner unit 12.

  A mirror 18 is disposed below the objective lens 10, and an imaging lens 19 is disposed between the mirror 18 and the scanner unit 12.

  A relay lens 20 and a mirror 21 are arranged along the optical axis reflected in the depth direction by a dichroic mirror 17 provided in the scanner unit 12.

  A filter wheel 23 rotated by a motor 22, a relay lens 24, a CCD 25 and a laser unit 26 as an imaging device are arranged along an optical axis reflected leftward by the mirror 21. The relay lens 24 is disposed in the temperature control chamber 3.

  The laser unit 26 on which the laser oscillator is mounted is disposed so as to face the scanner unit 12 in the depth direction viewed from the front surface of the temperature control chamber 3.

  The confocal microscope system configured as described above projects the laser unit 26, the scanner unit 12 that scans the laser beam output from the laser unit 26, and the laser beam scanned by the scanner unit 12 onto the sample cassette 7. A projection optical system for exciting the sample, an XY driving device 4 for XY driving the sample cassette 7 arranged opposite to the objective lens 10 on the table, and the scanner unit again among the fluorescence emitted from the excited sample A separation optical system that separates the light that has passed through 12, an imaging optical system that forms an image of the separated fluorescence again, and an imaging device disposed on the imaging surface of the imaging optical system, It is used for observation of physiological reactions and morphology of living cells in fields such as biotechnology, and LSI surface observation in the semiconductor market.

JP 2011-180411 A

  However, in such a conventional confocal microscope system, the laser light output from the laser unit 26 and the excitation light projected onto the sample are arranged in a straight line, and the fluorescence from the sample again passes through the scanner unit 12. Since an optical path for passing through is necessary, there is a problem that the depth of the entire apparatus becomes very large, and a considerably large space is required to accommodate the entire apparatus.

  The present invention solves such a problem, and an object thereof is to realize a compact confocal microscope system that is compact and does not require a large installation place.

In order to achieve such a problem, the invention according to claim 1 of the present invention is:
A scanner unit composed of a microlens disk and a pinhole disk, which scans the laser beam, a projection optical system that projects the laser beam on the scanner unit, and a laser beam scanned by the scanner unit is projected onto the sample. In a confocal microscope system including an excitation optical system that excites a sample and an imaging optical system that separates light that has passed through the scanner portion again from fluorescence emitted from the sample and forms an image on an imaging device.
In the confocal microscope system, the projection optical system, the excitation optical system, and the imaging optical system are arranged in parallel.

The invention of claim 2 is the confocal microscope system according to claim 1,
Bending mirrors are respectively disposed on the incident side and the exit side of the scanner unit, and a dichroic mirror is disposed between a microlens disk and a pinhole disk constituting the scanner unit.

The invention of claim 3 is the confocal microscope system according to claim 1,
A mirror for adjusting the output optical axis of the light projecting optical system with respect to the scanner unit is disposed on the front surface of the scanner unit.

The invention of claim 4 is the confocal microscope system according to claim 1,
A mirror for adjusting the fluorescence optical axis with respect to the photographing apparatus is disposed on the front surface of the photographing apparatus.

  As a result, a compact and compact confocal microscope system that does not require a large installation place can be realized.

It is a configuration explanatory view showing an embodiment of the present invention. FIG. 10 is a configuration explanatory view showing an example of a conventional confocal microscope system described in Patent Document 1.

  Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings. FIG. 1 is a structural explanatory view showing an embodiment of the present invention. FIG. 1 (a) is a plan view, FIG. 1 (b) is a front view of a main part, and parts common to FIG. ing. In FIG. 1, the confocal microscope system 1 is covered with a protective housing 2. In addition, the lower side of FIG.

  An XY driving device 27 is provided in the depth direction at the center inside the protective housing 2, and a moving table 28 that is driven in the XY directions is provided on the XY driving device 27. A sample can be stored in the moving table 28, and the sample moves on the XY driving device 27 in the XY direction according to the movement of the moving table 28.

  An objective lens 29 is arranged so as to face the sample. The objective lens 29 is driven in the arrow Z direction by the drive unit 30 as in the conventional case.

  A scanner unit 35 is disposed in front of the left side of the XY drive device 27 and is connected to a microlens disk 31 and a pinhole disk 32 by a hub 33 and rotated by a motor 34, as in the prior art.

  A mirror 36 is disposed below the objective lens 29 as in the prior art, and an imaging lens 37 is disposed between the mirror 36 and the scanner unit 35. A dichroic mirror 38 is disposed between the microlens disk 31 and the pinhole disk 32 constituting the scanner unit 35 as in the conventional case.

  A relay lens 39, a filter wheel 41 rotated by a motor 40, a relay lens 41, and a mirror 43 are arranged on the optical axis reflected rightward by a dichroic mirror 38 provided in the scanner unit 35. Has been.

  A camera 44 having an image sensor such as a CCD or CMOS is disposed on the back side of the right hand of the XY drive device 27, and the reflected light of the mirror 43 is incident on the camera 44.

  A laser unit 45 is disposed on the front side of the filter wheel 41. The output light of the laser unit 45 is incident on and transmitted through a dichroic mirror 38 provided in the scanner unit 35 via the mirror 46, and is further reflected by the mirror 47 and incident on the imaging lens 37. These mirrors 43, 46 and 47 function as mirrors for bending the optical axis.

  Here, the mirror 47, the imaging lens 37, and the objective lens 29 constitute a projection optical system, the dichroic mirror 38 functions as a separation optical system, and the relay lenses 39, 42, the filter wheel 41, and the mirror 43 are imaging optics. The laser unit 45 and the mirror 46 constitute an excitation optical system.

  The light projecting optical system, the excitation optical system, and the imaging optical system are arranged so that their optical axes are parallel to each other.

  The optical axis of the laser light output from the laser unit 45 can be adjusted by changing the inclination of the mirror 46, and the optical axis of the fluorescence incident on the camera 44 is changed to the Z axis by changing the inclination of the mirror 43. XY direction optical axis adjustment can be performed.

  The θ direction of the camera 44 can be adjusted by rotating the camera 44.

  The excitation light beam emitted from the laser unit 45 is collected by the micro lens disk 31 of the scanner unit 35, passes through the dichroic mirror 38, passes through the pinhole disk 32, is reflected by the mirror 47, and is reflected by the imaging lens 37. It becomes parallel light.

  The parallel light reflected by the mirror 36 is converged by the objective lens 29 and irradiated onto the sample placed on the moving table 28. The sample generates a fluorescence signal when irradiated with parallel light.

  The fluorescence signal generated from the sample is condensed by the objective lens 29, converted into parallel light, reflected by the mirror 36, converged by the imaging lens 37, and forms a fluorescent image on the imaging surface. Since the pinhole disk 32 constituting the scanner unit 35 is arranged so that the pinhole surface coincides with the image formation surface, the fluorescent image becomes a confocal image.

  The confocal image is incident on the dichroic mirror 38 and reflected, is incident on the mirror 43 via the relay lenses 39 and 42, is reflected, and is then formed on the camera 44. Here, the dichroic mirror 38 transmits a predetermined fluorescence wavelength. Specifically, the transmission characteristics are switched by a band-pass filter mounted on the filter wheel 41, and only the fluorescence wavelength band corresponding to the wavelength of the excitation light beam is transmitted.

  The confocal microscope system configured in this way is arranged so that the three optical axes of the projection optical system, the imaging optical system, and the excitation optical system are parallel with the separation optical system as the center, The width of the protective housing 2 can be narrowed, and the installation space efficiency can be improved.

  Since the optical axis adjusting mirror 46 of the excitation optical system is disposed on the front surface of the apparatus, the laser light incident on the scanner unit 35 can be easily adjusted.

  Further, by arranging the optical axis adjusting mirror 43 of the imaging optical system on the right surface, the width of the protective housing 2 can be reduced and the excitation light entering the camera 44 can be easily adjusted.

  Furthermore, in adjusting the optical axis of the excitation light incident on the camera 44, the XY direction can be separated by the mirror 43, and the θ direction can be separated by the camera 44, so that the optical axis can be adjusted easily. .

  As described above, according to the present invention, it is possible to simplify the optical adjustment and realize a confocal microscope system that can reduce the influence of the disturbance after the adjustment.

DESCRIPTION OF SYMBOLS 1 Confocal microscope system 2 Protective housing 27 XY drive device 28 Moving table 29 Objective lens 30 Drive unit 31 Micro lens disk 32 Pinhole disk 33 Hub 34, 40 Motor 35 Scanner part 36, 43, 46, 47 Mirror 37 Imaging Lens 38 Dichroic mirror 39, 42 Relay lens 41 Filter wheel 44 Camera 45 Laser unit 48 Imaging lens

Claims (4)

A scanner unit composed of a microlens disk and a pinhole disk, which scans the laser beam, a projection optical system that projects the laser beam on the scanner unit, and a laser beam scanned by the scanner unit is projected onto the sample. In a confocal microscope system including an excitation optical system that excites a sample and an imaging optical system that separates light that has passed through the scanner portion again from fluorescence emitted from the sample and forms an image on an imaging device.
A confocal microscope system, wherein the projection optical system, the excitation optical system, and the imaging optical system are arranged in parallel.   The dichroic mirror is disposed between a microlens disk and a pinhole disk that constitute the scanner unit, and bending mirrors are disposed on the incident side and the exit side of the scanner unit, respectively. Confocal microscope system.   The confocal microscope system according to claim 1, wherein a mirror for adjusting an emission optical axis of the light projecting optical system with respect to the scanner unit is disposed on a front surface of the scanner unit.   The confocal microscope system according to claim 1, wherein a mirror for adjusting the fluorescence optical axis with respect to the imaging apparatus is disposed on a front surface of the imaging apparatus.