JP2009282103A - Confocal scanner microscope - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a compact confocal scanner microscope by eliminating the need of a dichroic mirror of a light source part , and eliminating an influence of a pointing stability, while facilitating incidence of a laser beam into an optical fiber. <P>SOLUTION: This confocal scanner microscope includes: the light source part emitting exciting light for exciting a sample; a scanner part scanning the sample using the exciting light; and a light detecting part including an imaging camera capturing an image of the sample on receiving fluorescence emitted from the sample. The light source part includes a plurality of light sources oscillating laser beams different in wavelength and configured to input the light beams from the plurality of light sources to the scanner part through a wavelength selective optical fiber coupler. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a confocal scanner microscope, and more particularly to an improvement of a light source unit that makes light from a light source incident on an optical fiber.

  A confocal scanner microscope to which the present invention is applied is for observing a sample by scanning a condensing point on the sample by a laser beam and imaging a return fluorescence from the sample to obtain an image. It is used for observing physiological reactions and morphology of living cells in the fields of technology, etc., or for observing LSI surfaces in the semiconductor market.

First, the confocal scanner microscope will be briefly described.
FIG. 3 is a functional block diagram showing a configuration example of a conventional confocal scanner microscope. The light source unit 10 includes a plurality of multi-wavelength light sources 11a, 11b, and 11c, switching means 14 that controls passage and blocking of light from the light sources, and filters 12a, 12b, and 12c having different light transmission bands.

  The light that has passed through the switching means 14 passes through the filters 12a, 12b, and 12c, enters the optical fiber bundle 15, exits from the other end of the optical fiber 15, and enters the mirror 16. The light reflected by the mirror 16 enters a scanner unit 20 including a collimator lens 17, a microlens array disk 2, a pinhole disk 3, an objective lens 5, and the like.

  The light detection unit 30 includes dichroic mirrors 22a, 22b, and 22c, barrier filters 25a, 25b, and 25c, lenses 24a, 24b, and 24c, and imaging cameras 8a, 8b, and 8c. Is received.

  The barrier filter 25 is composed of a plurality of filters having different transmission wavelength characteristics. The output light that has passed through the barrier filter 25 is collected by the lens 24 and enters the imaging camera 8. The imaging camera 8 amplifies the captured image, converts it into an electrical signal, and outputs it.

The image collecting device 30 converts the electrical signal output from the imaging camera 8 into image data and stores it.
The image display device 40 is usually a computer, reads an image stored in the image collection device 30, performs appropriate processing, and displays it on a display screen.

The operation in such a configuration will be described next.
In the light source unit 10, light emitted from the three multi-wavelength light sources 11a, 11b, and 11c is incident on one end of the optical fiber 15 through the switching means 14 and the filters 12a, 12b, and 12c, respectively.

  The light emitted from the optical fiber 15 is reflected by the mirror 16 and enters the collimator lens 17. The light that has been collimated by the collimator lens 17 enters the microlens array disk 2. If the output light of the optical fiber 15 can be directly incident on the lens 17, the mirror 16 is not necessary.

  The fluorescence emitted from the sample 6 usually includes a plurality of wavelengths. For example, the first dichroic mirror 22a transmits light of wavelength λ1, and reflects light of other wavelengths. Next, the second dichroic mirror 22b receives light reflected by the first dichroic mirror 22a, reflects light of wavelength λ2, and transmits light of other wavelengths. The third dichroic mirror 22c receives the light transmitted through the second dichroic mirror 22b and reflects the light having the wavelength λ3.

  In this way, the light is branched by the three dichroic mirrors 22a, 22b, and 22c, and the three wavelengths of fluorescence are individually output. The output from the dichroic mirror 22 is input to the barrier filters 25a, 25b, and 25c. By narrowing the transmission wavelength region with this barrier filter 25, it is possible to remove the wavelength other than the target wavelength and make only the target wavelength incident on the imaging camera 8, and to improve the SN ratio of the image.

  The respective output lights that have passed through the barrier filter are condensed by the lenses 24a, 24b, and 24c, and enter the imaging cameras 8a, 8b, and 8c. Each of the three imaging cameras converts the captured image into an electrical signal and outputs it. The image acquisition device 30 converts this electrical signal into digital image data, and stores it in association with the fluorescence color branched by the dichroic mirror.

The image display device 40 reads out image data and filter information stored in the image collection device 30, reconstructs the original fluorescence color (color reconstruction), and displays the fluorescence image on the screen. It is of course possible to display a fluorescent image without color reconstruction.
In this way, it is possible to simultaneously measure fluorescent images of wavelengths branched by the dichroic mirror.

JP 2004-212434 A

  According to the above configuration, a confocal scanner microscope capable of observing fluorescence with a plurality of excitation lights without attaching or removing a light source and an optical fiber can be realized without changing the optical fiber inlet of the confocal scanner microscope. be able to.

  However, in such a conventional apparatus, there is a problem that the dichroic mirrors 13a and 13b and the mirror 18 are required in the light source section, and the apparatus becomes large. In addition, the distance between the light source and the fiber becomes long, and it is easily affected by the pointing stability of the light source (the stability of the position where the laser spot hits), so there is a problem that the secular change of the optical fiber transmittance is large. .

  Furthermore, there is a problem that skill is required to make the laser accurately enter the optical fiber. An object of the present invention is to provide a confocal scanner microscope capable of reducing the size of the apparatus by eliminating the need for a dichroic mirror of a light source unit, removing the influence of pointing stability, and allowing a laser to be easily incident on an optical fiber. To do.

In order to achieve this object, a confocal scanner microscope according to the present invention comprises:
A light source unit that emits excitation light that excites a sample, a scanner unit that scans the sample using the excitation light, and a light detection unit that includes an imaging camera that receives fluorescence emitted from the sample and captures an image of the sample In a confocal scanner microscope,
The light source unit is configured to input a plurality of light sources that oscillate lasers having different wavelengths and input light from the plurality of light sources to the scanner unit via a wavelength-selective optical fiber coupler. .

In the confocal scanner microscope according to claim 2,
A light source unit that emits excitation light that excites a sample, a scanner unit that scans the sample using the excitation light, and a light detection unit that includes an imaging camera that receives fluorescence emitted from the sample and captures an image of the sample In a confocal scanner microscope,
The light source unit includes a set of two light sources that oscillate lasers having different wavelengths, and a plurality of sets of the light sources are formed. An optical fiber that emits and propagates transmitted light from the plurality of optical fiber couplers is bundled into an optical fiber bundle, and the emitted light from the optical fiber bundle is input to the scanner unit. .

In claim 3, in the confocal scanner microscope according to claim 1 or 2,
Switching means and a filter are provided between each of the light source units and the wavelength selective optical fiber coupler.

As apparent from the above description, according to the first to third aspects of the present invention, each light from a plurality of light sources is configured to be input to the scanner unit via the wavelength selective optical fiber coupler. A dichroic mirror or mirror is not required in the light source section, and the apparatus can be downsized. In addition, the distance between the light source and the fiber can be shortened, and it becomes difficult to be influenced by pointing stability, and the secular change of the optical fiber transmittance can be reduced.
In addition, the laser can be easily incident on the optical fiber.

Hereinafter, the present invention will be described in detail with reference to the drawings. FIG. 1 is a functional block diagram showing an embodiment of a confocal scanner microscope according to the present invention.
In FIG. 1, the same parts as those in FIG. The difference from FIG. 3 is the light source unit 10a (in the embodiment, an example in which two types of laser beams are input to the scanner unit 20 is shown).

The light source unit 10a includes two multi-wavelength light sources 11a and 11b, switching means 14 for controlling passage and blocking of light from the light sources, and filters 12a and 12b having different light transmission bands (until heretofore, Same as example).
In the present invention, each of the light emitted through the switching means 14 and the filters 12a and 12b is received by the wavelength selective optical fiber coupler 50.

  The wavelength selective optical fiber coupler 50 has a port A and a port B on the input side, the wavelength λ1 from the multiwavelength light source 11a is incident on the port A, and the wavelength λ2 from the multiwavelength light source 11b is input to the port B. Is incident. The lights having the wavelengths λ1 and λ2 are branched and coupled by the coupler, and the combined light (λ1 + λ2) having a transmittance of about 90% is emitted from the port C.

The combined light (λ1 + λ2) is incident on the lens 17 through the optical fiber introduction port 60 and the mirror 16. Note that about 10% of light exits from port D, but the light from this port is terminated.
Since the operation after entering the lens 17 is the same as that of the conventional example, the description thereof is omitted here.

FIG. 2 shows another embodiment, and shows an example in which eight lasers (11a to 11h) are used as a multi-wavelength light source. The configuration of the scanner unit 20 and the light detection unit 30 is the same as that of the conventional example of FIG.
In this example, the lasers 11a and 11b are set as one set, and four sets of light sources are provided, each set including 11c and 11d, 11e and 11f, and 11g and 11h.

Also in this example, the light emitted from each set of light sources enters the ports A and B of the wavelength-selective optical fiber couplers 50 (a to d) through the switching means 14 (a to h) and the filters 12 (a to h). . Then, the combined lights (λ1 + λ2), (λ3 + λ4), (λ5 + λ6), and (λ7 + λ8) emitted from each port C are bundled and emitted to the mirror 16 (see FIG. 1) side as an optical fiber bundle 15a.
According to such a configuration, it is possible to simultaneously irradiate the sample with light of various types of wavelengths.

The foregoing description of the present invention has only shown certain preferred embodiments for purposes of illustration and illustration. Accordingly, it will be apparent to those skilled in the art that the present invention can be modified and modified in many ways without departing from the essence thereof.
The scope of the present invention defined by the description in the appended claims is intended to include modifications and variations within the scope.

It is a functional block diagram which shows one Example of the confocal scanner microscope which concerns on this invention. It is a functional block diagram which shows another Example. It is a functional block diagram which shows an example of the conventional confocal scanner microscope.

Explanation of symbols

2 Microlens array disk 3 Pinhole disk 4 Dichroic mirror 5 Objective lens 6 Sample 8 Imaging camera 10 Light source unit 11 Multi-wavelength light source 12 Filter 13, 22 Dichroic mirror 14 Switching means 15 Optical fiber 16 Mirror 17 Lens 20 Scanner unit 24 Lens 25 Barrier filter 30 Image acquisition device 40 Image display device 50 Wavelength selective optical fiber coupler

Claims (3)

A light source unit that emits excitation light that excites a sample, a scanner unit that scans the sample using the excitation light, and a light detection unit that includes an imaging camera that receives fluorescence emitted from the sample and captures an image of the sample In a confocal scanner microscope,
The light source unit is configured to input a plurality of light sources that oscillate lasers having different wavelengths and input light from the plurality of light sources to the scanner unit via a wavelength-selective optical fiber coupler. Confocal scanner microscope. A light source unit that emits excitation light that excites a sample, a scanner unit that scans the sample using the excitation light, and a light detection unit that includes an imaging camera that receives fluorescence emitted from the sample and captures an image of the sample In a confocal scanner microscope,
The light source unit includes a pair of two light sources that oscillate lasers having different wavelengths. The light source unit includes a plurality of sets of the light sources, and emits each light from the plurality of sets of light sources through a wavelength selective optical fiber coupler. The optical fiber that propagates the transmitted light from the wavelength selective optical fiber coupler is bundled into an optical fiber bundle, and the emitted light from the optical fiber bundle is input to the scanner unit. Confocal scanner microscope.   3. The confocal scanner microscope according to claim 1, further comprising a switching unit and a filter provided between each of the light source units and the wavelength selective optical fiber coupler.

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