JP2003090963A - Microscope sample and its manufacturing method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a microscope sample which makes it efficient and accurate to evaluate the performance of a microscope using a differential interference contract method and a phase difference contrast method and also measure the size of the microstructure of a transparent living body sample as an object to be observed, and its manufacturing method. SOLUTION: The microscope sample 10 has a resin-made phase body 11 having an unevenness pattern 16 of predetermined size in a predetermined shape and support members 12 and 13 which support the phase body.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a microscope sample suitable for transmission observation of a microscope using a differential interference contrast method or a phase difference contrast method, and a manufacturing method thereof.

[0002]

2. Description of the Related Art Conventionally, in order to observe transparent biological specimens (living tissues, cells, etc.) without staining, the Differential Interference Contrast method and Phase Contrast method have been used. The old microscope is used. With such a microscope, slight unevenness (difference in thickness) and difference in refractive index of a transparent biological specimen, that is, the phase difference of light transmitted through the transparent biological specimen is converted into light-dark contrast and visualized. be able to. Then, based on the obtained contrast of light and dark, transmission observation of the fine structure of the transparent biological specimen is performed.

Further, the above-mentioned microscope is required to evaluate its performance (detection limit and resolution of phase difference) in advance and to quantitatively measure the size of the fine structure confirmed during transmission observation. It At this time, a sample having a test pattern of a known size (a skeleton extracted from diatom, a fiber extracted from muscle, etc.) is used. The size of the test pattern is measured using a well-known electron microscope or the like. The size of the test pattern is on the submicron level (1 μm or less).

A sample having a test pattern (hereinafter referred to as "test sample") is transmitted and observed using the above-mentioned microscope, as in the transmission observation of a transparent biological sample as an observation target. As a result, the phase difference of the light transmitted through the test sample is converted into the contrast of light and dark, and the test pattern is visualized.

Therefore, the performance of the microscope using the differential interference contrast method or the phase difference contrast method (detection limit and resolution of the phase difference) is observed, for example, at what contrast a test pattern of known size is observed. It can be evaluated by Further, the size of the fine structure of the transparent biological specimen to be observed can be measured by comparing the size with the test pattern.

Further, a glass plate made of quartz or the like is partially removed by ion beam processing or dry etching processing,
It was studied to form a desired number of test patterns each having a fine uneven structure of a desired size (submicron level). Hereinafter, the test pattern having the concavo-convex structure will be referred to as a “concavo-convex pattern”.

[0007]

However, since the above-mentioned conventional test pattern is a skeleton or a fiber taken out from a living organism, it has a desired size suitable for the performance evaluation of the microscope and the size measurement of the fine structure. It is difficult to prepare a test pattern or a plurality of test patterns having a uniform size. Therefore, it has been impossible to accurately evaluate the performance of the microscope and measure the size of the fine structure.

In the ion beam processing, the ion beam can be focused down to the submicron level, but in reality, it was not possible to form a fine uneven pattern on the glass plate at the submicron level. For example, if you try to form recesses at a constant pitch like lines and spaces,
The walls between adjacent recesses disappear. There is also a problem that the bottom surface of the concave portion becomes rough and the edge of the concave portion rises.

On the other hand, in the dry etching process,
It is possible to form a concavo-convex pattern of a submicron level on a glass plate with a rectangular cross section. However, it was not possible to accurately control the depth of the concave portions (height of the convex portions) of the uneven pattern. The depth of the concave portion (height of the convex portion) is a factor that determines the phase difference of the light transmitted through the test sample, and the fact that it cannot be controlled accurately is unsuitable as a method for processing the test sample.

As described above, in ion beam processing or dry etching processing on a glass plate, a concavo-convex pattern of a desired size optimum for performance evaluation of a microscope or size measurement of a fine structure is formed, or a uniform size is formed. It was difficult to form a plurality of uneven patterns. The size of the concavo-convex pattern means the depth of the concave portion (height of the convex portion) and the width of the concave portion and the convex portion.

The object of the present invention is to accurately evaluate the performance of a microscope using the differential interference contrast method or the phase contrast method, and to accurately measure the size of the fine structure of a transparent biological specimen to be observed. It is intended to provide a microscope specimen and a manufacturing method thereof.

[0012]

A microscope sample according to claim 1 is provided with a resin phase object having a concavo-convex pattern of a predetermined shape and size, and a support member for supporting the phase object. Is.

The microscope specimen according to claim 2 is the same as that according to claim 1.
In the microscope sample described in the paragraph 1, the phase object is made of two or more thin plate-shaped resins having different refractive indexes, and the concavo-convex pattern is formed on a boundary surface of the two or more thin plate-shaped resins. . The microscope sample according to claim 3 is the microscope sample according to claim 2, wherein the supporting member includes a cover glass and a slide glass, and the phase object is between the cover glass and the slide glass. It is sandwiched and supported in the thickness direction.

According to a fourth aspect of the present invention, there is provided a method of manufacturing a microscope specimen, which comprises a first step of preparing an original plate having an uneven pattern of a predetermined shape and size formed on a surface thereof, and a first step for the uneven pattern of the original plate. The second step of applying the resin material of No. 1 and molding the thin plate resin in which the concavo-convex pattern is transferred to the surface, and the third step of peeling the thin plate resin from the original plate
A fourth step of applying a second resin material having a refractive index different from that of the first resin material to the step and the concave-convex pattern transferred to the surface of the thin plate resin peeled in the third step And a fifth step of adhering the cover glass to the second resin material while pressing and spreading the second resin material into a thin plate shape using a cover glass.

A method for manufacturing a microscope sample according to a fifth aspect is the method for manufacturing a microscope sample according to the fourth aspect, wherein in the second step, the first resin material is applied to the concavo-convex pattern of the original plate. And a step of spreading the first resin material into a thin plate shape using a slide glass and adhering the slide glass to the first resin material, and the third step includes: The thin plate resin is peeled off with the slide glass adhered.

[0016]

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described in detail below with reference to the drawings. (First Embodiment) A first embodiment of the present invention corresponds to claims 1 to 4. The microscope sample 10 of the first embodiment is
As shown in FIG. 1, it is composed of a transparent phase object 11 made of resin, a slide glass 12 supporting the phase object 11 and a cover glass 13 (support member).

Further, the phase object 11 is composed of two thin-film resins 14 and 15 having different refractive indexes, and an uneven pattern 16 is formed on the boundary surface between them. The thickness of the phase object 11 is about several μm. The concavo-convex pattern 16 has a rectangular cross section, and has a plurality of concave portions (or convex portions) of uniform size (concave depth, concave width, convex width) formed at equal intervals. The recess depth (height of the protrusion) A of the concavo-convex pattern 16 is about 50 nm, the width B of the recess and the width C of the protrusion are 0.2.
μm.

Of the two thin-film resins 14 and 15 constituting the phase object 11, the thin-film resin 14 on the slide glass 12 side is made of a UV curable resin material (first resin material) in the form of a thin plate. It is molded and has a refractive index of 1.47.
˜1.48 (25 ° C.). The thin film resin 15 on the cover glass 13 side is formed by molding an epoxy-based or silicon-based adhesive material (second resin material) into a thin plate shape, and has a refractive index close to that of the thin film resin 14. In the first embodiment, the difference in refractive index between the two thin film resins 14 and 15 is
It is about 0.1.

The phase object 11 having the above structure is sandwiched between the slide glass 12 and the cover glass 13 and supported in the thickness direction. The thickness of the slide glass 12 is
It is 1.0 to 1.2 mm. The thickness of the cover glass 13 is
It is 0.17 mm. The microscope sample 10 configured as described above is used to evaluate the performance (detection limit) of a microscope using the differential interference contrast method or the phase contrast method, and to observe a transparent biological sample (living tissue or cell etc.). ) Used to measure the size of the microstructure.

Next, a method of manufacturing the above-described microscope sample 10 will be described with reference to FIGS. First, a quartz or ceramic substrate 21 is prepared, and a chromium film 22 is vapor-deposited on the substrate 21 (a). Chrome film 2
The vapor deposition of 2 is performed in the vacuum vapor deposition device until the film thickness D reaches a desired value (for example, 50 nm). The accuracy of the film thickness D (thickness unevenness) is 5% or less of the film thickness D.

Then, the resist film 2 is formed on the chromium film 22.
3A is formed (a), and a desired pattern is exposed on the resist film 23 using an electron beam exposure apparatus. As a result, a resist pattern including a plurality of grooves 23a is formed on the resist film 23 (b). The width E of each groove 23a and the interval F between the grooves 23a are 0.2 μm. Next, the resist pattern of the resist film 23 (the plurality of grooves 23
The chromium film 22 is etched by a chemical treatment through a). As a result, the chromium film 22 also has a plurality of grooves 22.
A chrome pattern of a is formed (c). The width of the groove 22a of the chrome pattern and the interval between the grooves 22a are
It is the same as the width E of the resist pattern and the distance F (0.2 μm).

Then, the resist film 23 is removed.
(d). As a result, it is possible to obtain the original plate 20 in which the chrome pattern (the plurality of grooves 22a) of the chrome film 22 is formed on the substrate 21. The original plate 20 has an uneven pattern 20a formed of the surface of the substrate 21 and the surface of the chrome pattern. 2A to 2D correspond to the first step of preparing the original plate 20.

Here, the concavo-convex pattern 20a of the original plate 20.
Has a rectangular cross section. The depth of the concave portion of the concave-convex pattern 20a (height of the convex portion) is about 50 nm, which is the same as the film thickness D of the chromium film 22. The width of the concave portions and the width of the convex portions of the concave-convex pattern 20a are determined by the exposure pattern for the resist film 23 and are 0.2 μm. Next, an ultraviolet curable resin material 24 is applied to the uneven pattern 20a of the prepared original plate 20 (e), and the viscous resin material 24 is pressed into a thin plate shape using a glass pressing plate 25. spread
(f). At this time, it is preferable to pressurize the pressing plate 25 so that the resin material 24 spreads over the entire concavo-convex pattern 20a while maintaining parallel to the surface of the substrate 21. Also, be careful not to mix bubbles in the resin material 24.

Then, the thin plate-shaped resin material 24 sandwiched between the pressing plate 25 and the original plate 20 is irradiated with ultraviolet rays from a mercury lamp for about 5 to 60 seconds. as a result,
The resin material 24 is cured, and the uneven pattern 20 of the original plate 20 is obtained.
It becomes the thin plate resin 14 in which a is transferred to the surface. Figure 2
(e) and (f) are the second step of molding the thin film resin 14 using the original plate 20.

Next, when the thin film resin 14 is obtained by curing the resin material 24, the pressing plate 25 is separated from the thin film resin 14 and the thin film resin 14 is separated from the original plate 20 (g). At the time of this peeling, care must be taken not to damage the thin film resin 14. FIG. 2G corresponds to the third step.

Here, the ultraviolet curable resin material 24, which is the material of the thin film resin 14, is acetal glycol diacrylate, urethane acrylate, and 1-hydroxycyclohexylcyclohexyl phenyl ketone (trade name: Irgacure 184; Ciba).・ Mixed with Geigy Co., Ltd. In consideration of heat and light absorption characteristics, releasability, light resistance, durability, and hardness, the number of colors (APHA)
Is 30 to 50, and the refractive index at 25 ° C. is about 1.4 to 1.8.

In the first embodiment, in consideration of transferability in the step of FIG. 2 (f), a resin material having a specific gravity of about 0.8 to 1.3 at 25 ° C. and a viscosity of about 10 to 4800 CPS at 25 ° C. 24 was used. Further, considering the releasability in FIG. 2 (g), the number of colors is 40, and the refractive index is 1.47 to 1.48 at 25 ° C.
The resin material 24 of was used. Therefore, it is possible to obtain the thin film resin 14 in which the shape and the dimensions D, E, and F of the uneven pattern 20a of the original plate 20 are directly transferred to the surface. Therefore, the concavo-convex pattern 14a of the thin film resin 14 has a rectangular cross section, and the depth A of the concave portion (height of the convex portion) is about 50 n.
m, the width B of the concave portion, and the width C of the convex portion are 0.2 μm.

Further, in the first embodiment, a material that does not cause distortion inside the resin material 24 during curing in the step of FIG. 2 (f) is used. Next, the flat glass 14 side of the thin film resin 14 peeled from the original 20 is placed on the slide glass 1
Adhere to 2 (h). Further, the adhesive material 26 is applied to the concavo-convex pattern 14a of the thin film resin 14 (fourth step).

The cover glass 13 is used to spread the adhesive material 26 into a thin plate shape, and the cover glass 13
Are bonded (fifth step). At this time, the cover glass 13
It is preferable to pressurize the cover glass 13 so that the adhesive material 26 spreads over the entire concavo-convex pattern 14a while maintaining parallel to the surface of the cover glass 13. Also, be careful not to mix bubbles in the adhesive material 26.

The adhesive material 26 is a material that does not cause internal strain during curing, and is used for bonding lenses in an objective lens incorporated in a microscope using the differential interference contrast method or the phase difference contrast method, for example. It is a material. The adhesive material 26 is cured to form the thin plate resin 15, and the thin plate resin 15 causes the thin film resin 14
When the concavo-convex pattern 14a is sealed, the microscope sample 10 of the first embodiment shown in FIG. 1 is completed. The structure of the completed microscope sample 10 is as described above.

Using the microscope sample 10 of the first embodiment,
Evaluation of microscope performance (detection limit of phase difference, resolution) using differential interference contrast method and phase difference contrast method,
When measuring the size of the fine structure of a transparent biological specimen (living tissue, cells, etc.) to be observed, the microscope specimen 10 is placed on the stage of a microscope and is observed by transmission.

At the time of transmission observation, a phase difference corresponding to the concavo-convex pattern 16 of the phase object 11 is generated in the light transmitted through the microscope sample 10 in the thickness direction. Then, the phase difference of the light transmitted through the microscope sample 10 is converted into the contrast of light and dark, and the concavo-convex pattern 16 of the phase object 11 is visualized. In addition, there is no distortion inside the thin film resins 14 and 15 that form the phase object 11, and the polarization characteristics are uniform. Naturally, the polarization characteristics of the slide glass 12 and the cover glass 13 are also uniform.
Therefore, during transmission observation of the microscope sample 10, the microscope sample 1
The phase of the light passing through 0 is the concavo-convex pattern 1 of the phase object 11.
Structures other than 6 do not change. Therefore, only the uneven pattern 16 can be visualized.

Here, in the first embodiment, the phase object 11
The difference Δn in the refractive index between the thin film resins 14 and 15 constituting the
1, the depth of the concave portion of the concave-convex pattern 16 (height of the convex portion) A
Is 50 nm, the phase difference (= Δn × A) of light passing through the microscope sample 10 is 5 nm. Phase difference 5nm
Corresponds to the detection limit of the phase difference of the microscope. Also, the first
In the embodiment, the width B of the concave portion and the width C of the convex portion of the concave-convex pattern 16 of the phase object 11 are set to 0.2 μm. This corresponds to the resolution of the microscope (when the numerical aperture of the objective lens is 1.4).

That is, the concavo-convex pattern 1 of the microscope sample 10
Reference numeral 6 is a test pattern having a size optimal for performance evaluation of a microscope and size measurement of a fine structure. Therefore, the performance of the microscope (detection limit of phase difference, resolution) can be efficiently and accurately evaluated depending on how much contrast the concavo-convex pattern 16 of the microscope sample 10 is observed.

Further, the concavo-convex pattern 1 of the microscope sample 10
The size of the fine structure of the transparent biological specimen (living tissue, cells, etc.) to be observed can be efficiently and accurately quantitatively measured by comparing the size with that of No. 6. The size of the fine structure can be measured, for example, by (1) capturing an image of the concavo-convex pattern 16 obtained during transmission observation of the microscope sample 10 with a television camera, and importing it into an image processing device with a built-in dimension measurement function. Concavo-convex pattern 1 according to the procedure of the device
6) Design data (dimensions A, B, C) are input, (3) Microscopic specimens are exchanged for transparent biological specimens to be observed, images are captured, and (4) Concavo-convex pattern 16 is used as a "measurer". It is done by doing.

In the microscope sample 10 of the first embodiment described above, the depth (height of the convex portion) A of the concave portion of the concave-convex pattern 16 is set to 5
0 nm, the width B of the concave portion and the width C of the convex portion are 0.2 μm,
The invention is not limited to this size. The dimensions (A, B, C) of the concavo-convex pattern 16 of the microscope sample 10 are the dimensions (D, E, F) of the concavo-convex pattern 20a of the original plate 20 (FIG. 2 (d)) used when manufacturing the microscope sample 10. Therefore, by changing the dimensions (D, E, F) of the concavo-convex pattern 20a, the concavo-convex pattern 16 having a desired dimension (A, B, C) can be obtained.

Here, the dimensions (D,
E, F), the depth D of the concave portion (height of the convex portion) is nothing but the film thickness D of the chromium film 22, and is accurately within the range of 10 nm to 100 nm depending on the time for depositing the chromium film 22. It is controllable.

Further, the dimensions (D, E,
In F), the width E of the concave portion and the width F of the convex portion are
However, when the electron beam exposure apparatus is used as in the first embodiment, the minimum size can be about 0.1 μm. Therefore,
In preparing the original plate 20, by changing the deposition time of the chromium film 22 and the exposure pattern for the resist film 23, and changing the dimensions (D, E, F) of the concavo-convex pattern 20a,
The depth A of the concave portion of the concave-convex pattern 16 (height of the convex portion) is set to 10 n.
The width B of the concave portion and the width C of the convex portion can be set to an arbitrary value of 0.1 μm or more with an arbitrary value of m to 100 nm.

That is, it is possible to freely obtain a test pattern having a desired size (A, B, C) which is optimum for performance evaluation of a microscope and size measurement of a fine structure. For example, the detection limit of the phase difference of the microscope (the depth (height) A when Δn = 0.1 is 5
It is also possible to obtain a test pattern suitable for a condition of a resolution of the microscope (0 nm) or less and a resolution of the microscope (0.2 μm when the numerical aperture of the objective lens is 1.4).

With the recent development of television cameras and image processing technology, it is possible to capture a dark image, which has been difficult to detect by visual observation or photography so far, to enhance brightness and contrast, and to detect the phase difference of the microscope below the detection limit. As described above, the depth A of the concave portion (height of the convex portion) is 50 nm or less, and the width B of the concave portion is 0 and the width C of the convex portion is 0. It is very effective that a test pattern of 0.2 μm or less can be obtained.

(Second Embodiment) Next, a second embodiment of the present invention will be described. The second embodiment of the present invention corresponds to claims 1 to 5. In the second embodiment, another example of the method for manufacturing the above-described microscope sample 10 will be described. The manufacturing method of the second embodiment is the same as the manufacturing method of the first embodiment (FIGS. 2 (a) to 2 (h)), and FIGS.
3A to 3C are adopted instead of the process of FIG.

That is, after the ultraviolet curable resin material 24 is applied to the uneven pattern 20a of the prepared original plate 20 (FIG. 2 (e)), the resin material 24 is pressed into a thin plate shape using the slide glass 12. While unfolding, the slide glass 12 is adhered (FIG. 3 (a)). At this time, the slide glass 12 is pressed so that the resin material 24 spreads over the entire concavo-convex pattern 20a while maintaining parallel to the surface of the substrate 21.

Further, in order to bond the slide glass 12 in the step of FIG. 3 (a), one surface of the slide glass 12 is previously treated with a primer. Then, with the surface subjected to the primer treatment facing the resin material 24 side, as shown in FIG.
The step (a) is executed. Then, next, a thin plate-shaped resin material 24 sandwiched between the slide glass 12 and the original plate 20.
On the other hand, ultraviolet rays are irradiated for about 5 to 60 seconds. As a result, the resin material 24 is hardened, and the uneven pattern 20a of the original plate 20 becomes the thin plate resin 14 transferred to the surface. Figure 2
(e) and FIG. 3 (a) correspond to the second step of claim 5.

Next, when the thin film resin 14 is obtained by curing the resin material 24, the thin film resin 14 is peeled off from the original plate 20 with the slide glass 12 adhered to the thin film resin 14 ( Fig. 3 (b)). By this peeling, it is possible to obtain the thin film resin 14 in which the shape and the dimensions D, E, and F of the uneven pattern 20a of the original plate 20 are directly transferred to the surface. FIG. 3B corresponds to the third step of claim 5.

Next, the adhesive material 26 is applied to the concavo-convex pattern 14a of the thin film resin 14 peeled from the original plate 20, and the adhesive material 26 is spread into a thin plate shape by using the cover glass 13, and the cover glass 13 is used. Glue (Fig. 3
(c)). At this time, the cover glass 13 is pressed so that the adhesive material 26 spreads over the entire concavo-convex pattern 14a while maintaining parallel to the surface of the cover glass 13.

When the adhesive material 26 is cured to become the thin plate resin 15, and the uneven pattern 14a of the thin film resin 14 is sealed by the thin plate resin 15, the microscope sample 10 shown in FIG. 1 is completed. As described above, according to the manufacturing method of the second embodiment, after the thin film resin 14 and the slide glass 12 are integrated beforehand by adhesion, the thin glass resin 14 is peeled off from the original plate 20 together with the slide glass 12. It is possible to prevent the thin film resin 14 from being damaged at the time of peeling.

In the second embodiment described above, the surface of the slide glass 12 is subjected to the primer treatment, but if a glass plate made of a material to which the thin film resin 14 is adhered is used instead of the slide glass 12, the primer is used. Processing can be omitted. As the glass plate used at that time, one having the same thickness as the slide glass 12 is preferable. Also,
In the above-described first and second embodiments, the microscope sample 10
The concavo-convex pattern 16 (FIG. 1) is composed of a plurality of concave portions (or convex portions) of uniform size and arranged at equal intervals, that is, the width B of the concave portion and the width C of the convex portion are only one kind. Although a configuration has been described, the present invention is not limited to this configuration. The present invention can be applied to a configuration in which two or more types of concave portions (or convex portions) having different widths are arranged at arbitrary intervals.

Two or more types of concave portions (or convex portions) having different widths
As an example of combining, the width is 0.1 μm, 0.15
μm, 0.2 μm, 0.25 μm, 0.3 μm, 0.33 μ
m, 0.4 μm, 0.5 μm, 0.6 μm, 1.0 μm,
A configuration in which 11 types of concave portions (or convex portions) of 2.0 μm are combined is conceivable. As an example of the concavo-convex pattern in which the concave portions (or convex portions) are arranged at arbitrary intervals, as shown in FIG. 4, a concave-convex pattern 31 in which one concave portion (or convex portion) having a specified width is independently formed, 2 at the same interval as the width of the specified dimension
The concave / convex pattern 32 that forms the concave portions (or convex portions) of the book, and the concave portions (or convex portions) having the width of the designated dimension are arranged at intervals of 0.05 mm.
The concavo-convex pattern 33 and the like formed by repeating this process are conceivable.

Further, in the above-described first and second embodiments, the ultraviolet curable resin material 24 is used as the material of the thin film resin 14, but a silicon-based transparent resin material may be used. Among the silicon-based resin materials, materials that have good transferability and releasability and do not generate internal strain during curing are preferable. Further, in the above-described first and second embodiments, the thin film resin 15 is provided between the thin film resin 14 and the cover glass 13. However, instead of the thin film resin 15, a liquid such as water or oil is sealed. Alternatively, air may be interposed. By changing the refractive index between the thin film resin 14 and the cover glass 13, the phase difference of the light transmitted through the microscope sample can be controlled. By the way, thin film resin 1
As the refractive index between 4 and the cover glass 13 is closer to the refractive index of the thin film resin 14, the phase difference of the transmitted light can be made smaller.

Further, in the above-described first and second embodiments, when preparing the original plate 20, the exposure pattern for the resist film 23 was formed by electron beam exposure. However, instead of electron beam exposure, reduction projection exposure or it is performed. Exposure methods other than the above can also be used. 0.1 in any way
If an exposure pattern of less than μm can be formed, a fine original plate 20 corresponding to the exposure pattern can be obtained, and by transferring the pattern to the resin material 24, it is possible to manufacture a transparent microscope sample on which a finer uneven pattern is formed. .

Further, in the above-mentioned first and second embodiments, the example of the original plate 20 in which the chromium film 22 is formed on the substrate 21 is explained, but instead of the chromium film 22, nickel, aluminum, silver, copper or the like is used. You may use the metal film of.

[0052]

As described above, according to the present invention,
It is possible to evaluate the performance of the microscope using the differential interference contrast method and the phase contrast method, and to improve the accuracy of the size measurement of the fine structure of the transparent biological specimen to be observed.

[Brief description of drawings]

FIG. 1 is a cross-sectional view showing a configuration of a microscope sample 10 according to a first embodiment.

FIG. 2 is a cross-sectional view illustrating a method for manufacturing a microscope sample 10.

FIG. 3 is a cross-sectional view illustrating another method for manufacturing the microscope sample 10.

FIG. 4 is a top view showing another example of the uneven pattern.

[Explanation of symbols]

10 microscope specimens 11 phase objects 12 slide glass 13 cover glass 14,15 Thin film resin 16, 31, 32, 33 Concavo-convex pattern 21 board 22 Chrome film 23 Resist film 24 Resin material 25 push plate 26 Adhesive material

Claims (5)

[Claims] 1. A microscope specimen, comprising: a resin phase object having a concavo-convex pattern of a predetermined shape and size; and a support member for supporting the phase object. 2. The microscope specimen according to claim 1, wherein the phase object is made of two or more thin plate-shaped resins having different refractive indexes, and the uneven pattern is formed on a boundary surface of the two or more thin plate-shaped resins. A microscopic specimen characterized by being formed. 3. The microscope specimen according to claim 2, wherein the support member includes a cover glass and a slide glass, and the phase object is sandwiched between the cover glass and the slide glass to have a thickness. A microscope specimen characterized by being supported in a direction. 4. A first step of preparing an original plate on a surface of which an uneven pattern having a predetermined shape and dimensions is formed, and a step of applying a first resin material to the uneven pattern of the original plate to form the uneven pattern. A second step of molding the thin plate resin transferred to the surface, a third step of peeling the thin plate resin from the original plate, and a transfer to the surface of the thin plate resin peeled in the third step. A fourth step of applying a second resin material having a refractive index different from that of the first resin material to the concavo-convex pattern, and spreading the second resin material into a thin plate shape using a cover glass. And a fifth step of adhering the cover glass to the second resin material, the method for manufacturing a microscope specimen. 5. The method for manufacturing a microscope specimen according to claim 4, wherein the second step includes a step of applying the first resin material to the concave-convex pattern of the original plate, and a slide glass. A step of spreading the first resin material into a thin plate shape and adhering the slide glass to the first resin material, and the third step is the step of adhering the slide glass to the thin plate. A method for manufacturing a microscope specimen, which comprises peeling a resinous material.