JP2014006291A - Microscope, microscope system and image synthesis method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a microscope capable of imaging a desired range on a sample in a short time.SOLUTION: The microscope includes a plurality of microscope objective lenses and an imaging element. Each of the plurality of microscope objective lenses has a sample-side imaging range having a predetermined area; and the microscope objective lenses adjoining to each other are arranged to partially overlap the sample-side imaging ranges. Each of the plurality of microscope objective lenses preferably comprises a reduction optical system.

Description

  The present invention relates to a microscope, a microscope system, and an image composition method.

  2. Description of the Related Art Conventionally, there is a microscope provided with at least two microscope objective lenses as a microscope capable of simultaneously imaging a plurality of locations of a specimen (Patent Document 1). In the microscope disclosed in Patent Document 1, two microscope objective lenses are arranged at a predetermined interval.

JP 2008-76530 A

  In the microscope disclosed in Patent Document 1, the interval between the two microscope objective lenses is an integral multiple of the well pitch. In this case, in order to image the well between the two microscope objective lenses, the well plate and the microscope objective lens must be moved relative to each other. Therefore, the problem that imaging takes time arises.

  The present invention has been made in view of the above, and an object of the present invention is to provide a microscope, a microscope system, and an image composition method that can image a wide range on a specimen in a short time.

  In order to solve the above-described problems and achieve the object, a microscope according to the present invention is a microscope having a plurality of microscope objective lenses and an imaging device, and each of the plurality of microscope objective lenses is a predetermined one. The microscope-side objective lens having a wide specimen-side imaging range is arranged so that a part of the specimen-side imaging range overlaps.

  A microscope system according to the present invention includes a microscope and an image processing device, and the image processing device synthesizes images obtained from each of a plurality of microscope objective lenses.

  In addition, the image composition method according to the present invention sets a plurality of specimen-side imaging ranges having a predetermined width, uses an imaging step for simultaneously imaging a plurality of specimen-side imaging ranges, and a plurality of images obtained in the imaging step. An image generation step for generating a composite image, and in the imaging step, imaging is performed so that a part of the specimen-side imaging range overlaps, and the image generation step extracts duplicate images from a plurality of images, Then, a composite position of a plurality of images is determined, and a composite image is generated based on the composite position information.

  The present invention has an effect of providing a microscope, a microscope system, and an image composition method that can image a wide range on a specimen in a short time.

It is a schematic diagram which shows the structure of the microscope of this embodiment. It is a schematic diagram which shows the structure of another microscope of this embodiment. It is a figure which shows the structure of a microscope objective lens unit. It is the figure which looked at the sample side imaging range within the surface orthogonal to an optical axis. It is a figure which shows the structure of another microscope objective-lens unit. It is a figure which shows a light-shielding plate. It is a figure which shows arrangement | positioning of a microscope objective lens, (a) is a figure which shows the arrangement | sequence which becomes dense, (b) is the figure which has arrange | positioned the light source for illumination by the arrangement | sequence which becomes dense. It is a schematic diagram which shows the structure of the microscope system of this embodiment. It is a flowchart which shows the procedure of the process which synthesize | combines an image. It is a section lineblock diagram of the 1st microscope objective lens. FIG. 6 is an aberration diagram of the first microscope objective lens. It is a section lineblock diagram of the 2nd microscope objective lens. It is an aberration diagram of the second microscope objective lens. It is a lens section lineblock diagram of an imaging lens. It is an aberration diagram of the imaging lens. It is lens sectional drawing when a 2nd microscope objective lens and an imaging lens are combined.

  Hereinafter, embodiments of a microscope, a microscope system, and an image composition method according to the present invention will be described in detail with reference to the drawings. In addition, this invention is not limited by this embodiment.

  The microscope of the present embodiment is a microscope having a plurality of microscope objective lenses and an image sensor, and each of the plurality of microscope objective lenses has a specimen-side imaging range having a predetermined width, and is an adjacent microscope. The objective lens is arranged so that a part of the specimen-side imaging range is overlapped.

  FIG. 1 is a schematic diagram showing the configuration of the microscope of the present embodiment. As shown in FIG. 1, the microscope 1 includes a microscope body 2, a light source 3, a stage 4, an XY handle 5, a condenser lens 6, an aiming mechanism 7, and a microscope objective lens unit 10.

  A light source 3 and a condenser lens 6 are attached to the microscope body 2. Illumination light from the light source 3 enters the condenser lens 6. A stage 4 is arranged in the microscope body 2. A specimen S is placed on the stage 4. Illumination light from the light source 3 is applied to the sample S via the condenser lens 6.

  A microscope objective lens unit 10 is disposed at a position facing the condenser lens 6 with the stage 4 interposed therebetween. As will be described later, the microscope objective lens unit 10 has a plurality of microscope objective lenses. The microscope objective lens unit 10 is attached to the microscope main body 2. Therefore, the image of the specimen S is picked up by the microscope objective lens unit 10. Note that the microscope 1 may include a display device 8. In this way, the image of the sample S that has been picked up is displayed on the display device 8, so that the observer can see the image of the sample S on the display device 8.

  Note that the stage 4 is movable in the XY directions within a plane orthogonal to the optical axis of the condenser lens 6 (or the microscope objective lens unit 10). By operating the XY handle 5, the stage 4 can be moved in the XY direction. The microscope body 2 is provided with an aiming mechanism 7. By operating this aiming mechanism 7, the stage 4 can be moved in the vertical direction. Thereby, focusing with respect to the sample S can be performed. Further, a revolver 9 may be provided as in the microscope 1 'shown in FIG.

  FIG. 2 is a schematic diagram showing the configuration of another microscope of the present embodiment. In addition, the same number is attached | subjected about the same structure as FIG. 1, and description is abbreviate | omitted.

  As shown in FIG. 2, the microscope 1 ′ includes a microscope body 2, a light source 3, a light projecting tube 3 ′, a stage 4, an aiming mechanism 7, a revolver 9, and a microscope objective lens unit 10. .

  One end of a light projecting tube 3 ′ is attached to the upper part of the microscope body 2. The light projecting tube 3 ′ is provided with a light source 3. In the microscope body 2, a stage 4 is arranged at a position facing the other end of the light projecting tube 3 '. A specimen S is placed on the stage 4. The microscope body 2 is provided with an aiming mechanism 7. By operating this aiming mechanism 7, the stage 4 can be moved in the vertical direction. Thereby, focusing with respect to the sample S can be performed. Further, similarly to the microscope 1, the stage 4 may be moved in the XY directions.

  A revolver 9 is attached to the bottom of the light projecting tube 3 'on the other end side. A microscope objective lens unit 10 and other microscope objective lenses are attached to the revolver 9. By rotating the revolver 9, the microscope objective lens unit 10 and other microscope objective lenses can be positioned on the specimen S.

  By positioning the microscope objective lens unit 10 on the specimen S, an image of the specimen S is picked up by the microscope objective lens unit 10. Note that the microscope 1 ′ may include the display device 8 as with the microscope 1. In this way, the image of the sample S that has been picked up is displayed on the display device 8, so that the observer can see the image of the sample S on the display device 8.

  In the microscope 1 ′, the lens barrel 30 can be attached to the upper part on the other end side of the light projecting tube 3 ′. A binocular unit 31 can be attached to the lens barrel 30. By locating the microscope objective lens on the sample S with the binocular unit 31 attached, the observer can observe the optical image of the sample S with the naked eye.

  FIG. 3 is a diagram illustrating a configuration of the microscope objective lens unit. The microscope objective lens unit 10 includes a plurality of microscope objective lenses, that is, microscope objective lenses 11a, 11b, and 11c. Each of the microscope objective lenses 11a, 11b, and 11c is disposed to face the sample S. In FIG. 3, the microscope objective lenses 11a, 11b, and 11c are arranged in a line at equal intervals. In addition, the below-mentioned 1st microscope objective lens is used for each of the microscope objective lenses 11a, 11b, and 11c. In FIG. 3, the number of microscope objective lenses is 3, but it may be 2 or 4 or more.

  Each of the microscope objective lenses 11a, 11b, and 11c has a specimen-side imaging range La, Lb, and Lc (hereinafter simply referred to as imaging ranges La, Lb, and Lc) having a predetermined width. Here, the microscope objective lenses 11a and 11b are adjacent to each other. The microscope objective lenses 11a and 11b are arranged so that a part of the imaging range La and the imaging range Lb overlap each other. Similarly, the microscope objective lenses 11b and 11c are adjacent to each other, and are arranged so that a part of the imaging range Lb and the imaging range Lc overlap.

  An image sensor 12a is disposed at the image position of the microscope objective lens 11a. Similarly, the image sensor 12b is disposed at the image position of the microscope objective lens 11b, and the image sensor 12c is disposed at the image position of the microscope objective lens 11c. The image sensors 12a, 12b, and 12c are also arranged in a line at equal intervals.

  Here, as shown in FIG. 3, an overlapping region X1 exists in the imaging range La and the imaging range Lb. In addition, an overlapping region X2 exists in the imaging range Lb and the imaging range Lc. Therefore, the image of the overlapping region X1 is captured by the image sensors 12a and 12b, and the image of the overlapping region X2 is captured by the image sensors 12b and 12c.

  As described above, in the microscope of this embodiment, a part of the two imaging ranges overlaps, so that no gap (an area where no imaging is performed) occurs between one imaging range and the other imaging range. Therefore, a wide range on the specimen can be photographed at once. As a result, according to the microscope of the present embodiment, it does not take time to capture a wide range on the specimen S (image acquisition).

  As shown in FIG. 3, in the microscope of the present embodiment, the imaging range (La, Lb, Lc) is larger (wider) than the outer diameter of the microscope objective lens (11a, 11b, 11c). Therefore, as described above, overlapping regions (X1, X2) can be generated by bringing adjacent microscope objective lenses closer to each other. In addition, the area of the overlapping region can be changed by changing the interval between adjacent microscope objective lenses.

  FIG. 4 is a view of the specimen-side imaging range in a plane orthogonal to the optical axis. In FIG. 4, the circle is a real field of the microscope objective lens, that is, a range on the specimen S side where the microscope objective lens can form an image. Further, the square is the specimen-side imaging range, that is, the range of the imaging surface of the imaging device projected by the microscope objective lens. Here, the broken lines indicate the real field of view and the specimen-side imaging range in the microscope objective lens 11a. The solid line indicates the real field of view and the sample-side imaging range in the microscope objective lens 11b. A two-dot chain line indicates the real field of view of the microscope objective lens 11c and the sample-side imaging range.

  As shown in FIG. 4, each of the microscope objective lenses 11a, 11b, and 11c is not only arranged so that the real fields of adjacent microscope objective lenses overlap, but also the specimen side imaging range (imaging of the projected imaging device). Are arranged so as to overlap.

  FIG. 5 is a schematic diagram showing a configuration of another microscope objective lens unit. The microscope objective lens unit 13 includes a plurality of microscope objective lenses 14a, 14b, 14c and a plurality of imaging lenses 15a, 15b, 15c. Each of the microscope objective lenses 14a, 14b, and 14c is disposed to face the sample S. Each of the imaging lenses 15a, 15b, and 15c is disposed to face each of the imaging elements 16a, 16b, and 16c.

  A parallel light beam is emitted from the microscope objective lens 14a. This parallel light beam enters the imaging lens 15a and is condensed at a predetermined position. In this way, the space between the microscope objective lens 14a and the imaging lens 15a is afocal. Between the microscope objective lens 14b and the imaging lens 15b, and between the microscope objective lens 14c and the imaging lens 15c are also afocal.

  Since an image of the sample S is formed at the condensing position by the imaging lens 15a, the image sensor 16a is disposed at this condensing position. Similarly, the image sensor 16b is disposed at the condensing position by the imaging lens 15b, and the image sensor 16c is disposed at the condensing position by the imaging lens 15c.

  Further, each of the microscope objective lenses 14a, 14b, and 14c is arranged in a line at equal intervals. Similarly, the imaging lenses 15a, 15b, and 15c are also arranged in a line at equal intervals. Note that a second microscope objective described later is used for each of the microscope objectives 14a, 14b, and 14c. In addition, an imaging lens described later is used for each of the imaging lenses 15a, 15b, and 15c.

  Although not shown, each of the microscope objective lenses 14a, 14b, and 14c also has an imaging range (specimen side imaging range). The microscope objective lenses 14a and 14b and the microscope objective lenses 14b and 14c are arranged so that a part of the imaging range overlaps. The technical significance (action and effect) of the microscope objective lens unit 13 is the same as that of the microscope objective lens unit 10, and a detailed description thereof will be omitted.

  Further, in the microscope objective lens units 10 and 13, a light shielding plate may be disposed between adjacent imaging elements. If it demonstrates using the microscope objective lens unit 10, as shown in FIG. 6, the light-shielding plate 18 will be provided between the microscope objective lens 11a and the microscope objective lens 11b, and between the microscope objective lens 11b and the microscope objective lens 11c. ing. One end of the light shielding plate 18 is in the vicinity of the objective lens, and the other end of the light shielding plate 18 is in the vicinity of the imaging elements 12a, 12b, and 12c.

  The light from the microscope objective lens 11a is imaged (incident on) the adjacent imaging element 12b by the light shielding plate 18 disposed between the microscope objective lens 11a and the microscope objective lens 11b, and the microscope objective lens It can be surely limited that the light from 11b is imaged on the adjacent image sensor 12a. By doing in this way, generation | occurrence | production of the crosstalk which the image imaged by the adjacent microscope objective lens mixes can be reduced. In the microscope objective lens unit 13, a light shielding plate 18 may be used.

  In the microscope objective lens units 10 and 13, three image sensors are used, but one image sensor may be used.

  In the microscope of this embodiment, each of the plurality of microscope objective lenses is preferably a reduction optical system.

  As shown in FIG. 3, the width of the imaging range La is wider than the width of the imaging surface of the imaging element 12a. Similarly, the width of the imaging range Lb is wider than the width of the imaging surface of the imaging device 12b, and the width of the imaging range Lc is wider than the width of the imaging surface of the imaging device 12c. As described above, in the microscope according to the present embodiment, the imaging range with a large area is reduced and projected onto the imaging surface with a small area by the microscope objective lens. This indicates that each of the microscope objective lenses 11a, 11b, and 11c is a reduction optical system.

  As described above, by making each of the microscope objective lenses 11a, 11b, and 11c a reduction optical system, the microscope objective lens 11a is provided on the imaging element side, although an overlapping region is generated in the imaging range on the specimen S side. , 11b and 11c can be spatially separated. Therefore, each of the spatially separated images can be picked up by each of the image pickup devices 12a, 12b, and 12c.

  Note that when each of the microscope objective lenses 11a, 11b, and 11c is a magnifying optical system, the microscope objective lens even when the overlapping region is zero (the imaging ranges La, Lb, and Lc are adjacent to each other). A part of the image formed by 11a and a part of the image formed by the microscope objective lens 11b overlap on the image plane. Further, a part of the image formed by the microscope objective lens 11b and a part of the image formed by the microscope objective lens 11c overlap on the image plane.

  Thus, when the microscope objective lens is a magnifying optical system, the images formed by each of the microscope objective lenses 11a, 11b, and 11c cannot be spatially separated even if the overlapping region is zero. . Therefore, when the microscope objective lens is a magnifying optical system, it is not possible to provide an overlapping region.

  Further, when each of the microscope objective lenses 11a, 11b, and 11c is an equal-magnification optical system, when the overlapping region is zero, no overlapping occurs between adjacent images. However, if an interval between adjacent objective lenses is narrowed so as to provide an overlapping region, overlapping occurs between adjacent images. Therefore, when the microscope objective lens is an equal magnification optical system, it is not possible to provide an overlapping region.

  Note that each of the microscope objective lenses 14a, 14b, and 14c is also a reduction optical system. Therefore, the same effect as the microscope objective lenses 11a, 11b, and 11c is produced.

  Moreover, in the microscope of this embodiment, it is preferable that each of the plurality of microscope objective lenses is a non-telecentric optical system on the specimen side.

  As shown in FIG. 3, the off-axis chief rays CL1 and CL2 are not parallel to the normal of the surface of the sample S. This is because each of the microscope objective lenses 11a, 11b, and 11c is a non-telecentric optical system on the specimen side.

  Thus, by making each of the microscope objective lenses 11a, 11b, and 11c an optical system that is non-telecentric on the specimen side, the imaging range can be made wider than the outer shape (diameter) of the microscope objective lens. Therefore, an overlapping area can be generated in adjacent imaging ranges.

  Each of the microscope objective lenses 14a, 14b, and 14c is a non-telecentric optical system on the specimen side. Therefore, the same effect as the microscope objective lenses 11a, 11b, and 11c is produced.

  In the microscope according to the present embodiment, it is preferable that each of the plurality of microscope objective lenses has a negative refractive power at the most specimen side.

  By doing in this way, in each of the microscope objective lenses 11a, 11b, and 11c, the off-axis principal rays CL1 and CL2 toward the sample S can be expanded in a diverging direction (directed away from the optical axis). Thereby, the imaging range can be made wider than the outer diameter (diameter) of the microscope objective lens. Therefore, an overlapping area can be generated in adjacent imaging ranges.

  In each of the microscope objective lenses 14a, 14b, and 14c, the lens closest to the specimen S has a negative refractive power. Therefore, the same effect as the microscope objective lenses 11a, 11b, and 11c is produced.

In the microscope of this embodiment, it is preferable that each of the plurality of microscope objective lenses satisfies the following conditional expression (1).
Dmm <A <3Dmm (1)
here,
A is the diameter of the sample-side imaging range,
D is the outer diameter of the lens on the most specimen side,
It is.

  In the microscope objective lens unit 10 shown in FIG. 3, lens data will be described later, but A = 6 mm and D = 3 mm. In the microscope objective lens unit shown in FIG. 5, A = 11.3 mm and D = 7.5 mm. Therefore, both satisfy the conditional expression (1).

  Each of the microscope objective lenses 11a, 11b, 11c, 14a, 14b, and 14c satisfies the conditional expression (1), thereby generating an appropriate overlapping region and obtaining an image with less peripheral light attenuation. it can. When the sample-side imaging range is rectangular, A is the side length (long side or short side).

  If the lower limit value of conditional expression (1) is not reached, an overlapping area cannot be generated in adjacent imaging ranges. If the upper limit value of conditional expression (1) is exceeded, the diameter of the off-axis light beam becomes too small (thinner) compared to the diameter of the on-axis light beam, so that the peripheral attenuation becomes large. In addition, since the angle of the light beam passing through the lens increases, the possibility of occurrence of flare increases.

  In the microscope according to the present embodiment, each of the plurality of microscope objective lenses is preferably a microscope objective lens having the same magnification.

  By making each of the microscope objective lenses 11a, 11b, and 11c a microscope objective lens having the same magnification, images with the same magnification can be obtained, so that observation becomes easy. In addition, when image processing (image synthesis) described later is performed, image processing becomes easy.

  Note that each of the microscope objective lenses 14a, 14b, and 14c is preferably a microscope objective lens having the same magnification.

  Moreover, in the microscope of this embodiment, it is preferable that each of the plurality of microscope objective lenses is arranged so that the interval between adjacent microscope objective lenses is minimized.

  By doing in this way, adjacent imaging ranges can be efficiently overlapped. 7A and 7B are diagrams showing the arrangement of microscope objective lenses, where FIG. 7A is a diagram showing a dense arrangement, and FIG. 7B is a diagram in which illumination light sources are arranged in a dense arrangement. When the holding frame for holding each of the microscope objective lenses is a cylinder, as shown in FIG. 7A, each microscope objective lens is attached so that six microscope objective lenses are in contact with one microscope objective lens. It is good to arrange.

  In FIG. 7A, a solid circle indicates a microscope objective lens (holding frame), and a broken circle indicates a real field of view of the microscope objective lens. Further, although the imaging range (sample-side imaging range) is not shown, the imaging ranges are overlapped.

  Further, as shown in FIG. 7B, an illumination light source 17 may be disposed between adjacent microscope objective lenses. In FIG. 7B, six light sources 17 are arranged around one microscope objective lens, but the number of light sources is not limited to this. Also in the microscope objective lens units 10 and 13 (FIGS. 3 and 5), the light source 17 can be disposed between adjacent microscope objective lenses.

  As the microscope objective lens, a microscope objective lens having a small F number is preferably used. In the above description, imaging using the bright field method has been described. However, imaging using a phase difference method or a differential interference method may be performed.

  The microscope system of the present embodiment includes the above-described microscope and an image processing device, and the image processing device synthesizes images obtained from each of a plurality of microscope objective lenses.

  FIG. 8 is a schematic diagram showing the configuration of the microscope system of the present embodiment. As shown in FIG. 8, the microscope system 100 includes a microscope 1 ′ and an image processing device 20. Since the microscope 1 'has already been described, the description thereof is omitted. Note that the microscope 1 may be used instead of the microscope 1 '.

  The image processing apparatus 20 combines images obtained from each of the plurality of microscope objective lenses. The image processing apparatus 20 includes an image processing unit 21, a control unit 22, a display unit 23, and a storage unit 24. In addition, the image processing unit 21 includes a duplicate image extraction unit 211 and an observation image synthesis unit 213. In addition, the duplicate image extraction unit 211 includes a synthesis position determination unit 212.

  The image processing apparatus 20 is realized by a general-purpose computer such as a workstation or a personal computer. As described above, the microscope 1 'captures a plurality of images. Image data (digital data) of the captured image is sent to the image processing apparatus 20 via the control unit 22. The image processing apparatus 20 performs image processing on a plurality of image data, combines them into one observation image, and displays the resultant on the display unit 23. The control unit 22 gives an operation timing instruction, data transfer, and the like to the image processing unit 21, the display unit 23, and the storage unit 24, and comprehensively controls the operation of each unit. Further, the control unit 22 performs data transfer with the microscope 1.

  The display unit 23 is realized by an LCD or LED display device. The display unit 23 displays each image obtained by imaging, a composite image, a control screen, and the like based on the display signal input from the control unit 22.

  The storage unit 24 is realized by an information storage medium such as various IC memories, a hard disk, and a CD-ROM, and a reading device thereof. The storage unit 24 stores a program related to the operation of the image processing apparatus 20, a program for realizing various functions of the image processing apparatus 20, data related to the execution of these programs, and the like.

  The image processing unit 21 acquires a plurality of image data from the microscope 1 via the control unit 22 and generates one composite image from the plurality of image data. For example, when the microscope objective lens unit 10 captures an image of the specimen S, the image Ia acquired via the microscope objective lens 11a, the image Ib acquired via the microscope objective lens 11b, and the microscope objective lens 11c are acquired. Obtained image Ic. Each image data of the images Ia, Ib, and Ic is input to the image processing unit 21.

  The image composition method of the present embodiment uses a plurality of specimen-side imaging ranges having a predetermined area, an imaging step for simultaneously imaging the plurality of specimen-side imaging ranges, and a plurality of images obtained in the imaging step. An image generation step for generating a composite image, and in the imaging step, imaging is performed so that a part of the specimen-side imaging range overlaps, and the image generation step extracts duplicate images from a plurality of images, Then, a composite position of a plurality of images is determined, and a composite image is generated based on the composite position information.

  FIG. 9 is a flowchart illustrating a processing procedure for synthesizing an image. First, the number n of input images is set (step S1). For example, in the configuration of FIG. 3, since the image is acquired by three microscope objective lens units, 3 is set as the number n of input images. Subsequently, an initial value 1 of the processing count N is set (step S2).

  Subsequently, in the image processing unit 21, the overlapping image extraction unit 211 extracts an overlapping region from the image data of the image Ia and the image data of the image Ib (step S3). Then, the composite position determination unit 212 compares the images of the overlapping areas while changing the relative positions of the images Ia and Ib, and determines the relative positions of the images Ia and Ib when the overlapping areas match (step S4). .

  When the process of step S4 ends, it is determined whether or not the number of processes has reached a specified number (step 5). If the number of processes has not reached the specified number, the process returns to step 3 and the same process is performed on the images Ib and Ic. Thereby, the relative positions of the images Ia, Ib, and Ic are determined.

  When the number of processing times reaches the specified number, the observation image synthesis unit 213 combines the images Ia, Ib, and Ic based on the information of the synthesis position determination unit 212 to synthesize three images into one image (step S6). In this way, a composite image of the sample S is generated.

  In addition, the microscope system of the present embodiment has a moving mechanism that moves the plurality of microscope objective lenses and the sample relatively in the optical axis direction, and combines them while moving the plurality of microscope objective lenses and the sample relatively in the optical axis direction. It is preferable to acquire a plurality of images and combine a plurality of composite images.

  When the imaging range is expanded, it may be difficult to obtain an image that is in focus throughout the imaging range with a single imaging. For example, the sample surface of the sample S may be inclined (not orthogonal) with respect to the optical axis of the microscope objective lens unit 11. In such a case, the image Ib (the microscope objective lens 11b) is an in-focus image, but the images Ia and Ic (the microscope objective lenses 11a and 11c) are images that are not in focus.

  Therefore, an image is acquired while changing the interval between the microscope objective lens unit 11 and the sample S. Specifically, the aiming mechanism 7 is operated, and the stage is moved to a position where the focused image Ia can be acquired. Then, the stage is gradually brought closer to or away from the microscope objective lens from this position. At this time, the operation of the aiming mechanism 7 is performed in a step-like manner (the stage is moved and stopped alternately). In each step, the images Ia, Ib, and Ic are acquired and the images are combined (the images Ia, Ib, and Ic are combined into one image).

  Subsequently, an image area with the highest contrast is extracted from each of the composite images. Then, the extracted images are synthesized. By doing in this way, even if the in-focus position of the specimen S is shifted in the optical axis direction due to the inclination of the specimen S, it is possible to obtain a focused image in the entire imaging range.

  Such processing may be performed in the image processing unit 21. A stepping motor or a piezoelectric element can be used for the operation of the aiming mechanism 7.

  Next, numerical examples of the first microscope objective lens, the second microscope objective lens, and the imaging lens will be described. FIG. 10 shows a lens cross-sectional view of the first microscope objective lens, FIG. 12 shows a cross-sectional view of the second microscope objective lens, and FIG. 14 shows a lens cross-sectional view of the imaging lens. FIG. 16 is a lens cross-sectional view when the second microscope objective lens and the imaging lens are combined.

  In the following lens cross-sectional views, L1, L2, L3, L4, L5, L11, L12, L13, L14, and L15 are lenses. Moreover, OBJ is an object surface and corresponds to a sample surface. Further, IMG is an image plane and corresponds to the imaging plane of the imaging element. Each of GC1 and GC2 indicates a cover glass.

  As shown in FIG. 10, the first microscope objective lens includes, in order from the object side, a biconcave negative lens L1, a positive meniscus lens L2 having a convex surface facing the object side, and a positive meniscus lens having a convex surface facing the object side. L3, a negative meniscus lens L4 having a convex surface facing the image side, and a biconvex positive lens L5.

  The aspheric surface has 10 surfaces including both surfaces of the biconcave negative lens L1, both surfaces of the positive meniscus lens L2, both surfaces of the positive meniscus lens L3, both surfaces of the negative meniscus lens L4, and both surfaces of the biconvex positive lens L5. It is used.

  FIG. 11 shows aberration diagrams of the first microscope objective lens described above. In the aberration diagrams, each of (a), (b), (c), and (d) includes spherical aberration (SA), astigmatism (AS), distortion (DT), off-axis lateral aberration (coma aberration, Chromatic aberration of magnification, DY). In each figure, NA represents the numerical aperture on the object side, and FIY represents the maximum image height. These reference numerals are the same in all aberration diagrams hereinafter.

  As shown in FIG. 12, the second microscope objective lens includes, in order from the object side, a negative meniscus lens L1 having a convex surface facing the image side, a positive meniscus lens L2 having a convex surface facing the image side, and a convex surface facing the object side. And a negative meniscus lens L4 having a convex surface facing the object side. Here, the negative meniscus lens L1 and the positive meniscus lens L2 are cemented.

  The aspherical surface is used for one surface of the object side surface of the negative meniscus lens L1.

FIG. 13 shows aberration diagrams of the second microscope objective lens.
As shown in FIG. 14, the imaging lens has a biconvex positive lens L11, a negative meniscus lens L12 having a convex surface facing the object side, a biconvex positive lens L13, and a convex surface facing the image side, in order from the image side. And a negative meniscus lens L14 and a biconcave negative lens L15.

  The aspherical surface has 10 surfaces including both surfaces of the biconvex positive lens L11, both surfaces of the negative meniscus lens L12, both surfaces of the biconvex positive lens L13, both surfaces of the negative meniscus lens L14, and both surfaces of the biconcave negative lens L15. It is used for.

  FIG. 15 shows aberration diagrams of the imaging lens.

  Below, the numerical data of the above-mentioned microscope objective lens and imaging lens are shown. r is the radius of curvature of each lens surface, d is the distance between the lens surfaces, nd is the refractive index of the d-line of each lens, and νd is the Abbe number of each lens. * Represents an aspherical surface. The focal length is the focal length of the entire system, and NA is the numerical aperture on the object side.

The aspherical shape is expressed by the following equation, where x is an optical axis with the light traveling direction being positive, and y is a direction orthogonal to the optical axis.
^{x = (y 2 / R)} / [1+ {1- (k + 1) (y / R) 2} 1/2]
+ Ay ⁴ + by ⁶ + cy ⁸ + dy ¹⁰ + ey ¹² + fy ¹⁴ + gy ¹⁶
Where R is a paraxial radius of curvature, k is a conic coefficient, a, b, c, d, e, f, g are 4th, 6th, 8th, 10th, 12th, 14th, 16th, respectively. Aspheric coefficient. In the aspheric coefficient, “e−n” (n is an integer) indicates “10 ⁻ⁿ ”.

1st microscope objective lens unit mm

Surface data Surface number rd nd νd
Object ∞ 0.00
1 ∞ 0.17 1.5163 64.1
2 ∞ 9.28
3 * -25.64 0.41 1.5307 55.7
4 * 1.61 0.55
5 * 1.08 0.69 1.5307 55.7
6 * 1.16 0.31
7 * 1.27 0.55 1.5307 55.7
8 * 2.82 0.63
9 * -2.01 0.33 1.6349 23.9
10 * -7.77 0.04
11 * 8.27 0.70 1.5307 55.7
12 * -1.74 7.73
13 ∞ 0.30 1.5688 56.4
Image plane ∞

Aspheric data 3rd surface radius of curvature -25.64
k = 3.4042e + 002
a = -3.6894e-002, b = 8.9862e-003, c = 5.1275e-003,
d = -2.2514e-003
Fourth surface radius of curvature 1.61
k = -1.6171e + 000
a = -9.9451e-002, b = -3.8461e-003, c = 3.0399e-002,
d = -1.1887e-002
5th surface radius of curvature 1.08
k = -1.2590e + 000
a = -1.7655e-002, b = -2.5790e-003
6th curvature radius 1.16
k = -8.6895e-001
a = -1.0585e-001, b = 1.7929e-002, c = 1.6927e-003
7th surface radius of curvature 1.27
k = -1.5753e + 000
a = -4.1155e-002, b = 1.4678e-002
8th surface radius of curvature 2.82
k = 1.3514e + 000
a = 1.7655e-002, b = -3.3527e-003
9th surface radius of curvature -2.01
k = 1.5046e + 000
a = 5.3763e-002, b = -2.9705e-002, c = -6.9808e-004
10th surface radius of curvature -7.77
k = 3.5929e + 000
a = 3.4244e-002, b = -3.2268e-002, c = -2.1329e-003
Eleventh surface radius of curvature 8.27
k = -5.0000e + 000
a = -1.3838e-002, b = -5.7848e-004, c = 4.1265e-004
12th surface radius of curvature -1.74
k = -1.1539e + 000
a = -2.5780e-002, b = -2.2360e-003, c = 3.7672e-003

Second microscope objective unit mm

Surface data Surface number rd nd νd
Object ∞ 0.00
1 ∞ 0.17 1.5163 64.1
2 ∞ 6.42
3 * -4.61 0.65 1.8503 32.3
4 -150.74 1.63 1.7292 54.7
5 -4.75 0.06
6 6.07 0.90 1.5952 67.7
7 115.28 0.06
8 2.51 0.81 1.5952 67.7
9 2.09

Aspheric data 3rd surface radius of curvature -4.61
k = 9.1007e-001
a = -6.4093e-004, b = 5.5827e-005, c = 1.2432e-005

Imaging lens unit mm

Surface data Surface number rd nd νd
Object ∞ ∞
1 * 1.75 0.56 1.5346 56.2
2 * -8.78 0.05
3 * 5.32 0.35 1.6142 25.6
4 * 1.59 0.35
5 * 16.62 0.46 1.5346 56.2
6 * -7.74 0.45
7 * -2.04 0.47 1.5346 56.2
8 * -0.99 0.47
9 * -2.92 0.38 1.5346 56.2
10 * 2.43 0.93
11 ∞ 0.30 1.5688 56.4
Image plane ∞

Aspheric data 1st surface radius of curvature 1.75
k = -6.6472e-001
a = 1.7752e-002, b = -7.8166e-003, c = 2.7602e-003,
d = -4.2651e-003, e = 4.0909e-003, f = 1.8911e-003,
g = 9.9189e-003
Second surface radius of curvature -8.78
k = -7.4134e + 001
a = -6.2175e-003, b = 3.3713e-002, c = -1.0145e-002,
d = 3.3040e-003, e = 6.2061e-003, f = 1.7435e-002,
g = -5.4679e-003
Third surface radius of curvature 5.32
k = -9.9352e + 001
a = -3.0063e-002, b = 6.8076e-002, c = -3.4746e-002,
d = 2.9098e-002, e = 5.6036e-003, f = 4.5813e-003,
g = -2.2911e-002
Fourth surface radius of curvature 1.59
k = 5.9510e-001
a = -1.4632e-001, b = 1.2471e-001, c = -6.3320e-002,
d = 1.2893e-002, e = -1.3638e-003, f = -3.8744e-003,
g = -1.6260e-003
5th curvature radius 16.62
k = -4.2834e + 002
a = -1.5273e-002, b = -9.0265e-003, c = 3.0649e-002,
d = 5.8923e-003, e = -1.7942e-003, f = -7.7961e-004,
g = -5.3525e-004
6th surface radius of curvature -7.74
k = -1.4002e + 002
a = -5.2445e-002, b = 3.5404e-002, c = -3.8982e-002,
d = 1.4522e-002, e = 4.4660e-003, f = 7.4253e-004,
g = -4.2146e-004
7th surface radius of curvature -2.04
k = 1.1476e + 000
a = -4.4695e-002, b = 1.2870e-001, c = -7.1027e-002,
d = 1.2830e-002, e = 1.4986e-003, f = 5.4195e-005,
g = -3.6272e-004
8th curvature radius -0.99
k = -1.0822e + 000
a = 8.2661e-002, b = -1.6805e-002, c = 1.8282e-002,
d = -2.7597e-003, e = -3.0741e-004, f = -2.4855e-005,
g = -6.5291e-005
9th surface radius of curvature -2.92
k = -6.1166e + 000
a = 2.3788e-002, b = -2.9091e-002, c = 5.6364e-003,
d = 4.1246e-004, e = -1.3461e-004, f = -2.1375e-007,
g = 5.3744e-007
10th surface radius of curvature 2.43
k = -2.0265e + 001
a = -4.3537e-002, b = 7.4732e-003, c = -2.7053e-003,
d = 3.9955e-004, e = -2.6524e-005, f = -4.6809e-007,
g = -1.2765e-008

  As described above, the microscope according to the present invention is useful when imaging a desired range on a specimen in a short time.

DESCRIPTION OF SYMBOLS 1, 1 'Microscope 2 Microscope main body 3 Light source 3' Projection tube 4 Stage 5 XY handle 6 Condenser lens 7 Aiming mechanism 8 Display apparatus 9 Revolver 10 Microscope objective lens unit 11a, 11b, 11c Microscope objective lens 12a, 12b, 12c Element 13 Microscope objective lens unit 14a, 14b, 14c Microscope objective lens 15a, 15b, 15c Imaging lens 16a, 16b, 16c Imaging element 17 Light source 18 Light shielding plate 20 Image processing device 21 Image processing unit 22 Control unit 23 Display unit 24 Storage Part 30 Lens barrel 31 Binocular part 100 Microscope system 211 Duplicate image extraction part 212 Composition position determination part 213 Observation image composition part CG1, CG2 Cover glass Ia, Ib, Ic image IMG image plane L1-L5, L11-L15 Lens La, Lb , Lc imaging Circumference OBJ object surface S samples X1, X2 overlap region

Claims (11)

A microscope having a plurality of microscope objective lenses and an image sensor,
Each of the plurality of microscope objective lenses has a specimen-side imaging range of a predetermined area,
The adjacent microscope objective lens is disposed so that a part of the specimen-side imaging range overlaps.   The microscope according to claim 1, wherein each of the plurality of microscope objective lenses is a reduction optical system.   The microscope according to claim 1 or 2, wherein each of the plurality of microscope objective lenses is a non-telecentric optical system on the specimen side.   The microscope according to any one of claims 1 to 3, wherein in each of the plurality of microscope objective lenses, a lens closest to the specimen has a negative refractive power. The microscope according to claim 4, wherein the following conditional expression (1) is satisfied.
Dmm <A <3Dmm (1)
here,
A is the diameter of the specimen-side imaging range,
D is the outer diameter of the lens on the most specimen side,
It is.   The microscope according to claim 1, wherein each of the plurality of microscope objective lenses is a microscope objective lens having the same magnification.   Each of these microscope objective lenses is arranged so that the space | interval of adjacent microscope objective lenses may become the minimum, The microscope of any one of Claim 1 to 6 characterized by the above-mentioned.   A microscope according to any one of claims 1 to 7 and an image processing device, wherein the image processing device synthesizes images obtained from each of the plurality of microscope objective lenses. A microscope system. A moving mechanism for moving the plurality of microscope objective lenses and the specimen relative to each other in the optical axis direction;
9. The microscope system according to claim 8, wherein a plurality of synthesized images are acquired while relatively moving the plurality of microscope objective lenses and the specimen in the optical axis direction, and the plurality of synthesized images are synthesized.   The microscope system according to any one of claims 1 to 8, further comprising a light-shielding plate disposed between the plurality of microscope objective lenses. An imaging step of setting a plurality of specimen-side imaging ranges of a predetermined area and simultaneously imaging the plurality of specimen-side imaging ranges;
Using a plurality of images obtained in the imaging step, an image generation step of generating a composite image,
In the imaging step, imaging is performed so that a part of the specimen-side imaging range overlaps,
The image generation step includes extracting a duplicate image from the plurality of images, determining a combination position of the plurality of images from the overlap image, and generating the combination image based on the information of the combination position. A characteristic image composition method.

Images