JP4990653B2 - Confocal optical scanner detection device, optical scanner and nipou disk used therefor - Google Patents

Description

  The present invention relates to a confocal optical scanner detection device used in a confocal microscope or the like, an optical scanner, and an improved Nipow disk used therein.

  Conventionally, a confocal light scanning method is applied as a function measurement method of a living body by optical imaging. As one of the confocal light scanning methods, there is a nipou disc method using a nipou disc.

A conventional nipou disc has a large number of pinholes (the nipou disc is also called a pinhole disc). Here, FIG. 8 is a plan view showing an example of a conventional nipou disc 100. This Nipou disc 100 has 12 (12) pinhole rows PL in which pinholes P having a uniform diameter (50 microns) are arranged spirally from the center toward the periphery. Further, the Niipou disc 100 has 20,000 pinholes P, and the aperture ratio (ratio of pinholes per predetermined area of the disc) when viewed as a whole of the disc is about 3%. As for a microscope using this type of nipou disc, a confocal microscope shown in Patent Document 1 is known.
JP 2006-201207 A JP 2004-93721 A

  In the above configuration, only about 3% of the light irradiated to the Niipou disc can be used, and there is a problem that the efficiency is poor. As an improvement of this point, an optical scanner that uses a microlens disk together with a Niipou disk is known. In this configuration, since the irradiated light is received by the microlens disk and condensed on the Nipo disk, there is an advantage that about 40% of the irradiated light can be used. These microlens disk and nipou disk are integrally rotated at a constant speed by a motor.

  In the optical scanner using the Niipou disc 100, when the Nipo disc 100 rotates, a large number of light spots are scanned to form a confocal image, and about 1000 images per second can be obtained at the highest speed. As for this type of optical scanner, for example, a confocal optical scanner shown in Patent Document 2 is known.

  However, in the conventional optical scanner using the Niipou disk 100, the pinholes formed in the Niipou disk 100 have the same size and the same number, and the aperture ratio of the entire disk is constant and cannot be changed.

  Further, in order to increase the resolution of the optical scanner, it is better to reduce the pinhole size as much as possible to reduce the aperture ratio. However, in this case, the obtained image tends to be dark. In order to brighten the obtained image, it is necessary to increase the size of the pinhole as much as possible, but in this case, the resolution of the optical scanner cannot be increased.

  As described above, the conventional optical scanner must take into account the contradictory parameters of resolution and image brightness, and it is desirable that either of them can be used with priority depending on the measurement target. This was not possible because of the constant. For example, an optical scanner using the Niipou disc 100 has spatial resolution, but has low fluorescence acquisition efficiency, and obtains an image with a disc aperture ratio close to 100%, such as an incident image (non-confocal image). It was difficult to do. Moreover, the intermediate image between the confocal image and the epi-illumination image, which has gained brightness even at the expense of some spatial resolution, could not be obtained.

  Therefore, the present invention has been made to solve the above-described problem, and a confocal optical scanner detection device, an optical scanner, and an aperture ratio that can be changed and an appropriate image according to a measurement target can be acquired. An object of the present invention is to provide a nipou disk used for the same.

In order to solve the above-mentioned problems, the present invention takes an optical scanner that scans a sample by rotating a nippo disk, a light source that irradiates the sample through the nippo disk, and takes an image of the sample by inputting light emitted from the optical scanner. The NIPOW disc has a plurality of pinhole rows in which a plurality of pinholes are arranged from the center toward the periphery, and one pinhole row or two or more pinhole rows The size of the pinholes constituting each pinhole row is different for each pinhole row, and the number of pinhole rows with the same size pinhole row is the ascending order of the pinhole size. It features a confocal optical scanner detection device that has been reduced in number.

  In this confocal optical scanner detection device, the size of the pinhole constituting each pinhole row is different for each pinhole row or every two or more pinhole rows in the Nipkow disk. The image can be changed, and an image corresponding to a plurality of optical characteristics can be taken.

  Further, the confocal optical scanner detection device may be configured such that the pinhole size is different for each of two or three pinhole rows and is changed in ascending or descending order along the circulation direction. .

  When such a Niipou disc is used, it is possible to take an image in which the aperture ratio is sequentially changed.

  In addition, the confocal optical scanner detection device may be provided with a microlens disk that condenses the light from the light source in a pinhole facing the nipo disk.

  By providing the microlens disk in this way, a brighter image can be taken.

The present invention is an optical scanner that scans a sample by rotating a nippo disk, and the nipou disk has a plurality of pinhole rows in which a plurality of pinholes are arranged from the center toward the periphery. The size of the pinholes constituting each pinhole row is different for each row or two or more pinhole rows, and the number of pinhole rows with the same size pinholes is the pinhole disc It features an optical scanner that decreases in ascending order of size .

  In the case of this optical scanner, since the size of the pinhole constituting each pinhole row is different for each pinhole row or two or more pinhole rows in the Nipkow disk, the aperture ratio is changed. Can do.

  The optical scanner may have different pinhole sizes for each of two or three pinhole rows, and may be changed in ascending or descending order along the circumferential direction.

  When such a Niipou disc is used, it becomes possible to take an image in which the aperture ratio is changed sequentially.

  Furthermore, it is preferable that the nipou disk further has an opening portion opened so that the opening ratio is 100% between adjacent pinhole rows.

  This is suitable for photographing an image (non-confocal image such as an epi-illumination image) in which the aperture ratio is 100%.

  Further, the optical scanner can be provided with a microlens disk that opposes the nipou disk and collects light from the light source in a pinhole.

  By providing the microlens disk in this way, a brighter image can be taken.

  In addition, the Nipkow disk can be configured such that the number of pinhole rows having the same size pinholes decreases in ascending order of pinhole size.

  Further, it is preferable that the pinhole disk has each pinhole arranged in a spiral shape.

  By doing this, it is possible to prevent unevenness in light and darkness during optical scanning on the inner and outer circumferences of the Nipkow disc.

The present invention is a nipou disk used in an optical scanner, which has a plurality of pinhole rows in which a plurality of pinholes are arranged from the center toward the periphery, and one pinhole row or two or more pinholes. The size of the pinholes constituting the pinhole row is different for each row, and the number of pinhole rows having the same size pinholes is reduced in ascending order of the pinhole size . Provide Nipkow disc.

  As described above in detail, according to the present invention, the confocal optical scanner detection device, the optical scanner, and the nipou disc used therein can change the aperture ratio and acquire an appropriate image according to the measurement target. Can be obtained.

  Embodiments of the present invention will be described below. In addition, the same code | symbol is used for the same element and the overlapping description is abbreviate | omitted.

(Configuration of confocal optical scanner detection device and optical scanner)
FIG. 1 is a diagram showing a schematic configuration of a confocal optical scanner detection device 30 according to an embodiment of the present invention. The confocal optical scanner detection device 30 includes the optical scanner 1, light sources 20 and 21, a relay lens 22, and an objective lens 23, and further includes an imaging device 24 and a controller 25.

  The optical scanner 1 includes a microlens disk 2, an improved nippo disk 3, a motor 4, a fixing member 5 and a beam splitter 6. In this optical scanner 1, a microlens disk 2 and an improved nippo disk 3 are integrated by a fixing member 5, and the integrated microlens disk 2 and the improved nippo disk 3 receive a driving force of a motor 4 to rotate. 7 is rotated at a constant speed.

  The microlens disk 2 has a microlens 2a. The microlens 2a collects the irradiation light L1 from the light source 20, and pinholes (minute openings) P1 and P2 described later provided in the improved Nipo disk 3 are provided. , P3 or the opening 8 is irradiated.

  The improved Nipkow disk 3 has a plurality of pinhole rows PL1, PL2, and PL3 described later in which a plurality of pinholes P1, P2, and P3 are arranged from the center toward the periphery. The improved nippo disk 3 is an improved version of the conventional nippo disk, and its configuration will be described in detail later.

  The beam splitter 6 is installed between the microlens disk 2 and the improved Nipow disk 3. The beam splitter 6 causes an irradiation light L1 (described later) that has passed through the microlens disk 2 to enter the improved Nipkow disk 3. Further, the beam splitter 6 reflects return light L <b> 3 (described later) or transmitted illumination light L <b> 2 (described later) from the sample 31 so as to enter the relay lens 22.

  The light source 20 outputs laser light (or white light) as irradiation light L 1 and makes the irradiation light L 1 incident on the microlens disk 2 of the optical scanner 1. The light source 21 outputs laser light (or white light) as transmitted illumination light L2 and irradiates the sample 31.

  The relay lens 22 causes the imaging device 24 to form an image of the return light L3 or the transmitted illumination light L2 reflected by the beam splitter 6. The objective lens 23 inputs the irradiation light L <b> 1 output from the optical scanner 1 and forms a minute spot on the sample 31.

  The imaging device 24 enters the return light L3 or the transmitted illumination light L2 output from the beam splitter 6 and takes an image of the sample 31.

  The controller 25 has at least a CPU, a ROM, and a RAM, and the CPU operates according to a control program stored in the ROM. Then, the controller 25 synchronizes the shooting timing of the image pickup device 24, the shutter (not shown) inside the image pickup device 24, and the position of the pinholes P1, P2, P3 or the opening 8 of the improved improved nippo disk 3 or the like. To control the shooting timing.

(Configuration of improved Niipou disc 3)
The improved Niipou disc 3 is a substrate in which a surface of a circular transparent substrate made of glass or the like is coated with a light-shielding film, etc., and has a size partitioned at a predetermined ratio along the circumferential direction as shown in FIG. And is divided into four zones Z1, Z2, Z3, and Z4 having different aperture ratios. In the zones Z1, Z2, and Z3, pinhole rows PL1, PL2, and PL3 are formed, respectively, and in the zone Z4, openings 8 are formed (pinholes P1, P2 in the pinhole rows PL1, PL2, and PL3). , P3 and the opening 8 are etched so that the light-shielding film is removed and light is transmitted).

  Since this improved Nipow disc 3 has three (three) pinhole rows PL1, PL2 and PL3 in zones Z1, Z2 and Z3, respectively, it has nine pinhole rows as a whole. In the pinhole row PL1, a plurality of pinholes P1 are spirally arranged from the center toward the periphery. Similarly, the pinhole row PL2 has a plurality of pinholes P2, and the pinhole row PL3 has a plurality of pinholes P3 arranged in a spiral from the center toward the periphery.

  Further, the pinhole rows PL1, PL2, and PL3 have different sizes of the pinholes P1, P2, and P3 constituting the pinhole rows PL1, PL2, and PL3, and the pinhole rows PL1, PL2, and PL3 are along the circumferential direction f of the disk. The pinholes P1, P2, and P3 are arranged in descending order.

  That is, the diameters of the pinholes P1, P2, and P3 constituting the pinhole rows PL1, PL2, and PL3 are reduced in order (P1> P2> P3). Therefore, the aperture ratios (α1, α2, α3) for the zones Z1, Z2, and Z3 are also α1> α2> α3, and the aperture ratio for each zone changes in three stages.

  The opening 8 is disposed in a portion between the pinhole row PL1 and the pinhole row PL3, and is formed by removing the coating on that portion. The opening 8 has an aperture ratio of 100% because the light shielding film in the zone Z4 is completely removed.

  An example of the microlens disk 2 is shown in FIGS. On the microlens disc 2, microlenses 2a are laid out at positions corresponding to the pinholes P1, P2, and P3. Each microlens 2a forms a lens array 2L similar to the pinholes P1, P2, and P3. 6 are arranged in a portion corresponding to the opening 8 so that the microlenses 2a are closest to each other, and the microlens disc 32 shown in FIG. The micro lens 2a is not disposed.

(Description of operation of confocal optical scanner detection device and optical scanner)
The confocal optical scanner detection device 30 operates as follows. First, the irradiation light L <b> 1 from the light source 20 enters the optical scanner 1. The irradiation light L1 incident on the optical scanner 1 is condensed by the microlens 2a of the microlens disk 2, passes through the beam splitter 6, and is formed on the improved nipou disk 3 by the pinholes P1, P2, P3. Alternatively, the light is condensed at the opening 8. The irradiation light L1 passing through the pinholes P1, P2, P3 or the opening 8 is irradiated onto the sample 31 by the objective lens 23.

  On the other hand, the return light L3 from the sample 31 passes through the objective lens 23 and the improved nippo disk 3 again, and is reflected by the beam splitter 6 (bent in a direction perpendicular to the optical axis direction of the irradiation light L1), and a relay lens. The light enters the imaging device 24 via 22. In this way, an image of the sample 31 is formed on the imaging device 24. Here, the controller 25 synchronizes the shooting timing of the imaging device 24 and the shutter (not shown) inside the imaging device 24 so that the images from the pinholes P1, P2, P3 or the opening 8 are different from each other. take.

  In this confocal optical scanner detection device 30, the microlens disk 2 and the improved nippo disk 3 of the optical scanner 1 are rotated at a constant speed by the motor 4, and the pinholes P 1, P 2, A light spot from P3 or the opening 8 scans the sample 31.

  Further, the pinholes P1, P2, and P3 on the improved nippo disk 3 and the light spot on the sample 31 are in a confocal relationship, and both the irradiation light L1 from the light source 20 and the return light L3 from the sample 31 are both. Since it passes through the pinholes P1, P2, and P3, resolution due to the confocal effect corresponding to each pinhole diameter can be obtained.

  Further, when the transmitted illumination light L2 from the light source 21 is transmitted through the sample 31 and then enters the optical scanner 1, the transmitted illumination light L2 is transmitted through the objective lens 23, and then the pinholes P1 and P2 on the improved Niipou disc 3 are used. , P3 or the opening 8 is transmitted. Further, the transmitted illumination light L <b> 2 is reflected by the beam splitter 6 and enters the imaging device 24 via the relay lens 22. Also in this case, an image of the sample 31 is formed on the imaging device 24.

  In the above case, the improved nippo disk 3 has three types of pinholes P1, P2 and P3 having different sizes (diameters), and the aperture ratio can be changed in three stages. Further, since the improved Nipo disk 3 has the opening 8, the opening ratio can be changed in four stages as a whole. Therefore, the confocal optical scanner detection device 30 can change the resolution and the brightness of the image in four patterns, and can shoot four types of images having different optical characteristics with one unit.

In this case, if the improved nippo disk 3 is formed with X-holes of pinholes and is configured so that one image can be acquired with n lines, the optical characteristics of the image that can be acquired per one rotation of the disk are expressed in the Y stage. (N, X, Y are integers)
It is only necessary to form a pinhole row having the number of lines obtained by X / (nY) in each zone.

  This improved nipou disk 3 has nine pinhole rows PL1, PL2 and PL3 formed therein, but instead of the opening 8 in the zone Z4, a pinhole row having a pinhole larger than the pinhole P3 is arranged. If it is provided (not shown), it has 12 pinhole rows as a whole. Further, it is assumed that one image can be acquired by the three items.

  In this case, if the confocal light scanner detection device 30 is configured such that one image can be obtained with three pinholes using such an improved nippo disk, four images can be obtained in one rotation.

  Since the standard disk rotation speed of the optical scanner 1 is 80 rotations per second, 320 images can be taken every second when using the improved nippo disk having 12 pinhole rows. The image forming speed for each aperture ratio (zone) is 80 sheets per second.

  Furthermore, assuming that the size of the pinhole is changed in eight stages in the entire improved nippo disk 3, 40 images per second can be taken for each aperture ratio. Even in this case, the number of images that can be taken is larger than 30 images per second, which is the standard speed in real-time imaging. Therefore, the shooting speed (number of shots per unit time) of the confocal optical scanner detection device 30 is comparable to the standard speed.

  In addition, the confocal optical scanner detection device 30 can set the aperture ratio to 100% when photographing using the aperture 8, so that not only the confocal image but also the non-confocal image such as the epi-illumination image. It is also suitable for taking images.

  In the improved Niipou disc 3, the size of the pinholes P1, P2, and P3 is changed in descending order (so as to decrease) along the circumferential direction f. Images that are sequentially changed (in this case, images in which the aperture ratio is sequentially changed to large, medium, and small) can be taken.

  When the imaging device 24 continues photographing while rotating the improved Niipou disc 3 with such a confocal light scanner detection device 30, images having different optical characteristics are sequentially obtained for each of the zones Z1, Z2, Z3, and Z4. It is done.

  Therefore, if images shot using the zones Z1, Z2, Z3, and Z4 are A, B, C, and D, respectively, and if one shot is taken in each of the zones Z1, Z2, and Z3, the shot image is A , B, C, D, A, B, C, D..., There is a possibility that four types of images having different optical characteristics may be mixed.

  Therefore, the controller 25 controls the shooting timing of the imaging device 24 to match the position of the zone used for shooting among the zones Z1, Z2, Z3, and Z4 (so that the images in each zone are different from each other). By synchronizing the shooting timing of the image pickup device 24 and the shutter inside the image device 24 and the rotation of the improved Niipou disc 3, an image shot with a required aperture ratio can be obtained, and the optical characteristics can be obtained. Mixing of different images can be avoided.

  For example, by synchronizing the shooting timing of the imaging device 24 and the position of the zone Z1, the shot images can be A, A, A, A, A,.

  In this case, the controller 25 detects the position of the zone Z1 in the improved nippo disk 3 with a predetermined sensor (the sensor performs an operation as a position detection means), and the controller 25 uses the imaging device 24 based on the detected position of the zone Z1. The operation as a synchronization control means for controlling the shooting timing of the image may be executed.

  As described above, the confocal optical scanner detection device 30 and the optical scanner 1 are provided with pinholes having different sizes in the improved Niipou disc 3, so that the aperture ratio can be changed stepwise. . Therefore, the confocal optical scanner detection device 30 can capture optical tomographic images that can be acquired by the confocal method with a plurality of types having different optical characteristics. In addition, since transmission and emission images can be taken at almost the same timing, the optical conditions can be flexibly changed according to the measurement target.

  Further, the confocal optical scanner detection device 30 acquires a confocal image using a desired pinhole by synchronizing the shooting timing of the imaging device 24 and the position of the shutter and the zone inside the imaging device 24. You can also.

  Further, since the pinholes P1, P2, and P3 are arranged in a spiral shape, unevenness in light scanning at the inner and outer circumferences of the improved nippo disk 3 does not occur by appropriately controlling the pinhole pitch. ing.

(Modification)
The above-described confocal optical scanner detection device 30 may use the improved nipou disc 13 shown in FIG. 3 instead of the improved nippo disc 3. The improved nippo disc 13 is different from the improved nippo disc 3 in the following points and the other points are the same. In other words, the improved Niipou disc 13 has one pinhole row PL1, PL2, PL3 in each of the zones Z1, Z2, Z3.

  Therefore, the improved Niipou disc 13 has different pinhole sizes for each pinhole row. Even with the improved Niipou disc 13, the resolution and the brightness of the image can be changed in stages. When this improved Niipou disc 13 is used, a confocal image can be taken with a desired aperture ratio by synchronizing the shooting timing of the image pickup device 24 and the position of the pinhole row.

  Further, the confocal light scanner detection device 30 may use an improved nipou disk 14 shown in FIG. 4 instead of the improved nippo disk 3. The improved nippo disk 14 is different from the improved nippo disk 3 in the following points and the other points are the same. In other words, the improved Niipou disc 14 has two, three, and four pinhole rows PL1, PL2, and PL3 for each of the zones Z1, Z2, and Z3, and has the size of the pinholes P1, P2, and P3. The number of pinhole rows PL1, PL2, and PL3 increases in descending order.

  Note that the effect of the position of the lens 2a in the microlens disc 2 is the same as in FIG. 7 in addition to FIG.

  Further, the confocal optical scanner detection device 30 may use an improved nipou disc 15 shown in FIG. 5 instead of the improved nippo disc 3. The improved nippo disk 15 is different from the improved nippo disk 3 in the following points and the other points are the same. That is, in the improved Niipou disc 15, the pinholes P1, P2, and P3 of the pinhole rows PL1, PL2, and PL3 are linearly arranged, and each zone Z1, Z2, Z3, and Z4 is divided into four equal parts along the circumferential direction. It is a quadrant of equal size.

  The above description is the description of the embodiment of the present invention, and does not limit the present invention, and various modifications can be easily implemented. In addition, an apparatus or a method configured by appropriately combining components, functions, features, or method steps in each embodiment is also included in the present invention.

  For example, in the improved Niipou disc of the above embodiment, four zones are provided, but the number of zones may be two so that the aperture ratio can be changed in two steps, large or small. You may make it 5 or more and can change an aperture ratio in five steps or more. The size of the zones may be equal or different.

It is a figure which shows schematic structure of the confocal optical scanner detection apparatus 30 which concerns on embodiment of this invention. It is a top view which shows an example of an improved nippo disk. It is a top view which shows an example of the improved type nipo disk which concerns on a modification. It is a top view which shows an example of the improved nippo disk which concerns on another modification. It is a top view which shows an example of the improved nippo disk which concerns on another modification. It is a top view which shows an example of a microlens disc. It is a top view which shows the other example of a microlens disc. It is a top view which shows an example of the conventional nipou disk.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 ... Optical scanner, 2 ... Micro lens disk, 3 ... Improved Niipou disk 8 ... Opening part, 24 ... Optical imaging device, 25 ... Controller, 20, 21 ... Light source PL1, PL2, PL3 ... Pinhole row P1, P2, P3 …Pinhole

Claims (9)

An optical scanner that scans a sample by rotating a nipou disc, a light source that irradiates the sample via the nipou disc, and an imaging device that captures an image of the sample by inputting light emitted from the optical scanner. A focus light scanner detection device comprising:
The Nipkow disc has a plurality of pinhole rows in which a plurality of pinholes are arranged from the center toward the periphery, and each pinhole row is arranged for one pinhole row or two or more pinhole rows. The size of the pinhole to be configured is different ,
2. The confocal optical scanner detection apparatus according to claim 1, wherein the number of the pinhole rows having the same size pinholes is reduced in ascending order of the pinhole size .   2. The confocal optical scanner according to claim 1, wherein the size of the pinhole is different for each of the two or three pinhole rows, and is changed in ascending or descending order along the circumferential direction. Detection device.   The confocal optical scanner detection device according to claim 1, wherein a microlens disk facing the nipou disk and condensing light from the light source is provided in the pinhole. An optical scanner that scans a sample by rotating a Nipkow disc,
The Nipkow disc has a plurality of pinhole rows in which a plurality of pinholes are arranged from the center toward the periphery, and each pinhole row is arranged for one pinhole row or two or more pinhole rows. The size of the pinhole to be configured is different ,
2. The optical scanner according to claim 1, wherein the number of the pinhole rows having the same size pinhole is reduced in ascending order of the pinhole size .   The optical scanner according to claim 4, wherein the size of the pinhole is different for each of the two or three pinhole rows and is changed in ascending or descending order along the circumferential direction.   6. The optical scanner according to claim 4, wherein the Nipkow disk further has an opening that is opened so that an opening ratio is 100% between the adjacent pinhole rows.   The optical scanner according to any one of claims 4 to 6, wherein a microlens disk that condenses the light from the light source is provided in the pinhole so as to face the nipou disk. 8. The optical scanner according to claim 4 , wherein the pinholes are arranged in a spiral shape on each of the pinholes. 9. A nipou disc used in an optical scanner,
A plurality of pinhole rows in which a plurality of pinholes are arranged from the center toward the periphery, and one pinhole row or two or more pinhole rows are included in the pinhole row. A nipou disc characterized in that the number of pinhole rows having different sizes and having the same size pinholes decreases in ascending order of the size of the pinholes .

Images