JP3525158B2 - Observation sample support method - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an observation sample supporting method for supporting a sample, which is an object of observation of a cut surface, so as to withstand cutting.

[0002]

2. Description of the Related Art As one of sample observation methods, while sequentially cutting a sample from one end side, each cut surface is imaged by a CCD camera, and this image data is processed into data suitable for observation, A method of displaying a cut surface of a sample and a stereoscopic image obtained by synthesizing these on a monitor is known.

The sample is composed of a sample body and a resin, an embedding agent such as paraffin, which embeds the sample body, and the sample body is a living organism appropriately selected from the medical field, pharmaceutical field, food field, agricultural field, biological field and the like. , Animals, plants, foods, etc. are used.

Further, a device for sequentially cutting the sample uses a feeding mechanism for feeding a required amount of the sample and a cutting mechanism for cutting the end portion of the sample after the feeding.

As shown in FIG. 9, the feed mechanism includes a plate P that supports the lower surface of the sample S, a pressing rod R that presses the plate P upward, and a drive source that moves the pressing rod R upward. , And a holding cylinder H through which the upper part of the sample S is inserted. The sample S receives pressure from the pressing rod R, rises in the holding cylinder H, is pushed out from the upper end of the holding cylinder H, and projects upward. On the other hand, the cutting mechanism includes a cutting blade Cb capable of cutting the sample end portion protruding upward from the upper end of the holding cylinder H during rotation, and a drive source for rotating the cutting blade Cb in a predetermined direction.

That is, the sample S is pushed upward from the upper end of the holding cylinder H by a necessary amount and protrudes, the protruding portion is cut by the cutting blade Cb, and the cut surfaces are sequentially observed.

[0007]

DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention As shown in FIG.
Since the rake angle θ1 and the clearance angle θ2 are set in the cutting blade Cb in addition to the cutting edge angle, the force for trying to lift the sample S according to the rake angle θ1 is applied to the sample S at the time of cutting. Fy works.

In the conventional case, since the lower surface of the sample S is only supported by the plate P, the above force F at the time of cutting is used.
When y acts, the sample S floats up and slips out upward from the holding cylinder H, which causes a problem that desired cutting cannot be performed satisfactorily.

In order to solve this, it is conceivable to add a sample tightening function to the holding cylinder H to counteract the force Fy, but it is extremely difficult to fix and support the sample S only by the tightening, and it is often the case. There is a problem that the tightening force becomes too strong and the sample S is crushed and deformed, making it unusable as an observation target.

The present invention has been made in view of the above circumstances, and an object of the present invention is to provide an observation sample supporting method capable of satisfactorily cutting a sample.

[0011]

In order to achieve the above-mentioned object, the invention of claim 1 uses a reverse taper as a movable stage.
, Which has a concave portion provided on its inner side surface, inserts the end portion of the sample into the concave portion of the movable stage in a heated state and presses it to melt the contact portion, and then solidify the molten portion. The sample is closely fixed to the movable stage, and the other end of the sample is inserted into a holding cylinder having an inner hole aligned with the cross-sectional shape of the sample.

The invention of claim 2 includes a movable stage and
And having a recess with a reverse taper on the inner surface
Using concave of the movable stage the ends of the sample in a cooled state
And press it to melt the contact part, then solidify the melted part to tightly fix the sample to the movable stage, and hold the other end of the sample with an inner hole that matches the cross-sectional shape of the sample. The feature is that it is inserted into the cylinder.

Further, the invention of claim 3 is the movable stage.
As a reverse taper has a recess provided on the inner surface
Using, the end portion of the sample in the cooled state is inserted into the concave portion of the movable stage in the heated state and pressed to melt the contact portion, and the molten portion is solidified to firmly fix the sample to the movable stage, It is characterized in that the other end of the sample is inserted into a holding cylinder having an inner hole aligned with the cross-sectional shape of the sample.

Further, according to the invention of claim 6, the end portion of the sample is clamped by a clamp member provided on the movable stage to be closely fixed to the movable stage, and the other end portion of the sample is aligned with the cross-sectional shape of the sample. It is characterized in that it is inserted into a holding cylinder having an inner hole.

According to each of the first to third aspects of the invention, the end portion of the sample is melted and solidified to be closely fixed to the movable stage, and the other end portion of the sample is inserted into the holding cylinder for observation. The sample of interest is supported to withstand cutting. Further, according to the invention of claim 6 , the sample to be observed is obtained by tightening the end of the sample with the clamp member so as to be closely fixed to the movable stage and inserting the other end of the sample into the holding cylinder. Supported to withstand cutting.

[0016]

DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG. 1 shows a sample observation device suitable for carrying out the present invention. This sample observing apparatus is composed of a sample observing machine 1 capable of observing a sample by image processing, and a sample cutting machine 11 for feeding and cutting the sample.

The sample observer 1 includes a color or monochrome camera 2 having a two-dimensional CCD 2a, a mirror case 3 having a mirror 3a such as a half or dichroic, an objective lens 4, and a ring illuminator 5. Light source 6a
A first lamp house 6 with a built-in, a second lamp house 7 with a built-in light source 7a, a frame 8 for supporting the device in the sample cutter 1 1 above <br/>, the imaging control unit not shown, the image It is composed of a processing unit and a monitor.

In this sample observing device 1, the light from the first lamp house 6 is applied to the sample cut surface via the mirror 3a and the objective lens 4, or the light from the second lamp house 7 is illuminated by the ring illuminator 5. By irradiating the sample cut surface via the, the reflected light from the sample cut surface is incident on the camera 2 via the objective lens 4 and the mirror 3a, and the image (image data) of the sample cut surface is captured in the memory. be able to.

Further, data processing such as area division, contour deformation processing, and synthesis is performed on the image data taken in for each cut surface, and a cut surface image of a sample and a solid obtained by combining these are obtained as necessary. The image (three-dimensional image) can be displayed on the monitor.

On the other hand, as shown in FIG. 2, the sample cutting machine 11 includes a frame 12 that can be placed on a floor surface, and a frame 1.
The sample holder 13 is arranged on one side of the upper surface of the sample 2, the sample supporter 14 is arranged below the sample holder 13, and the sample cutting section 15 is arranged at the center of the upper surface of the frame 12.

The sample holder 13 comprises a holding cylinder 13a having an inner hole aligned with the cross-sectional shape of the sample S, and a Peltier element 13b capable of cooling the holding cylinder 13a. The holding cylinder 13a is made of a metal having high thermal conductivity, and has a tapered surface at the lower end of the inner hole for facilitating the insertion of the sample S from below. Further, the holding tube 13a is divided into a plurality of pieces in the circumferential direction, and by driving at least a part of the divided pieces by utilizing spring pressure, air pressure, or the like, the sample can be tightened by the divided pieces.

For example, in the case of using the sample S in the shape of a quadrangular prism, as shown in FIG. 3A, the holding cylinder 13a is provided on the fixed divided piece 13a1 facing the two side faces of the sample S and the other two side faces. The movable divided pieces 13a2 are opposed to each other, and the movable divided pieces 13a2 are moved toward the center of the sample S so that the sample can be tightened. Alternatively, FIG. 3 (b)
As shown in, the holding cylinder 13a is composed of a fixed divided piece 13a3 facing two side surfaces of the sample S and two movable divided pieces 13a4, 13a5 facing the other side surfaces,
By moving each of the movable divided pieces 13a4 and 13a5 at right angles to the side surface of the sample, the sample can be clamped.

In the sample holder 13, the sample S inserted in the holding cylinder 13a can be tightened by the divided pieces, and the sample S in the holding cylinder 13a is cooled by the cooling action of the Peltier element 13b as necessary. can do. In the present embodiment, since the sample S is not fixedly supported only by the tightening by the divided pieces, the tightening force may be a weak force that does not crush the sample S.

As shown in FIGS. 4 (a) and 4 (b), the sample support portion 14 has a movable stage 14a having a recess 14a1 on the upper surface, which is aligned with the cross sectional shape of the sample S, and the movable stage 1a.
Peltier element 14b capable of heating and cooling 4a, and Peltier element 14b and movable stage 1 via heat insulating material 14c.
A nut cylinder 14d for supporting the nut 4a, a guide cylinder 14e for guiding the nut cylinder 14d in the vertical direction, a ball screw 14f screwed into the nut cylinder 14d, and a motor 14g capable of rotating the ball screw 14f are provided. Movable stage 1
Reference numeral 4 is made of a metal having a high thermal conductivity, and a reverse taper is provided on the inner surface of the recess 14a1 to enhance the sample fixing force.

The sample support portion 14 is a movable stage 14
The sample S is supported upright on a and the motor 14
Rotation of the ball screw 14f by g causes the nut cylinder 14
The movable stage 14a can be moved up and down together with d. Further, the sample S contacting the movable stage 14a can be heated by the heat generating action of the Peltier element 14b or cooled by the cooling action as required. By the way, switching between the heat generating action and the cooling action that can occur in the Peltier element 14b can be easily performed by changing the polarity of the applied voltage.

The sample cutting section 15 is a cutting blade 15 shown in FIG.
a rotating plate 15b having a detachably attached a and the rotating plate 15b
A shaft 15c connected to the shaft 15c, and a motor 15e with a speed reducer capable of rotating the shaft 15c via a belt 15d. As shown in FIG. 5, the cutting blade 15a includes a main body 15a1, a cutter blade 15a2, and a blade retainer 15a3 for fixing the cutter blade 15a2 to the main body 15a1, and has a predetermined cutting edge angle, rake angle and clearance angle (see FIG. 10 ). See).

The sample cutting unit 15 rotates the rotary plate 15b (one rotation) via the shaft 15c by the motor 15e, and thereby the portion of the sample S protruding upward from the upper end of the holding cylinder 13a is cut by the cutting blade. It is possible to cut parallel to the end face by 15a.

The sample S is formed in the shape of a quadrangular prism in the illustrated example from the sample main body and the embedding agent such as resin or paraffin that encapsulates the sample main body. As the sample body, an organism, an animal, a plant, a food or the like appropriately selected from the medical field, the pharmaceutical field, the food field, the agricultural field, the biological field, etc. is used.

Here, some methods of fixing the sample S to the movable stage 14a will be described. First, the first fixing method will be described with reference to FIG. First, Peltier element 1
6b, the movable stage 14a is heated to a predetermined temperature (a temperature at which the embedding agent of the sample S can be melted).
As shown in (a), the recess 14 of the movable stage 14a
The lower end of the sample S is inserted into and supported by a1 and the lower end surface of the sample S is pressed against the movable stage 14a to melt the contact portion. In parallel with this, the holding cylinder 13a is cooled by the Peltier element 13b, and as shown in FIG.
The movable stage 14a is raised to hold the sample S in the holding cylinder 13a.
Insert inside. As a result, the sample S is cooled and the above-mentioned molten component is cooled and solidified, and the molten component plays the role of an adhesive, so that the sample S is closely fixed to the movable stage 14a.

In the first fixing method, the sample holder 13
The sample S is cooled by the Peltier element 13b of FIG. 6 to solidify the molten component. Instead of this, when the sample S is inserted into the holding cylinder 13a or thereafter, the Peltier element 14b of the sample support portion 14 is used. The molten component may be cooled and solidified.

Next, the second fixing method will be described with reference to FIG. First, as shown in FIG. 7A, the holding cylinder 13a
The sample S is inserted therein and held by tightening with the divided pieces, and in the same state, the sample S in the holding cylinder 13a is cooled in advance by the Peltier element 13b, and after the sample is cooled, the cooling by the Peltier element 13b is stopped. Then, as shown in FIG. 3B, the movable stage 14a is raised and the lower end of the sample S is inserted into the recess 14a1 of the movable stage 14a. As a result, the lower end surface of the sample S, which has been cooled in advance, is pressed against the movable stage 14a at room temperature and the contact portion is melted, and the melted portion is gradually solidified by the cold heat of the sample S, and the melted portion becomes the adhesive. The sample S plays a role and is closely fixed to the movable stage 14a.

In the second fixing method, the melted portion is solidified by the cold heat of the sample S that has been cooled in advance, but after the stage contact portion of the sample S is melted, the Peltier element 14b of the sample support portion 14 is melted. The molten component may be cooled and solidified by, of course, the molten component can be solidified even when the sample S is cooled by the Peltier element 13b of the sample holder 13.

Next, the third fixing method will be described with reference to FIG. First, as shown in FIG. 7A, the holding cylinder 13a
The sample S is inserted therein and held by tightening with the divided pieces, and in the same state, the sample S in the holding cylinder 13a is cooled in advance by the Peltier element 13b, and after the sample is cooled, the cooling by the Peltier element 13b is stopped. Subsequently, the Peltier element 14b heats the movable stage 14a to a predetermined temperature (a temperature at which the embedding agent of the sample S is mainly meltable), and the movable stage 14a is raised as shown in FIG. The lower end of the sample S is inserted into the recess 14a1 of the movable stage 14a. As a result, the lower end surface of the sample S that has been cooled in advance is pressed against the movable stage 14a in the heated state, and the contact portion is melted. When the heating by the Peltier element 14b is stopped before or after the stage contact portion of the sample S is melted, the melted component is gradually solidified by the cold heat of the sample S, and the melted component plays the role of an adhesive to move the sample S to the movable stage. It is tightly fixed to 14a.

In the third fixing method, the molten component is solidified by the cold heat of the pre-cooled sample S. However, after the stage contact portion of the sample S is melted, the Peltier element 14b of the sample support portion 14 is melted. The molten component may be cooled and solidified by, of course, the molten component can be solidified even when the sample S is cooled by the Peltier element 13b of the sample holder 13.

By the way, in the above-mentioned second fixing method and third fixing method, since the end of the sample S can be closely attached and fixed to the movable stage 14a while the sample S is held in the holding portion 13a, the sample S itself can be fixed. Even if there is a slight bend, the sample S can be fixed to the movable stage 14a without bending, and an unreasonable force against the bend is applied to the sample S even when the movable stage 14a moves up and down. Can be prevented.

After the sample S is fixed on the movable stage 14a by the above-mentioned fixing method, the movable stage 14a is lifted by a predetermined amount while the tightening by the divided pieces is released, and the sample S is moved by a predetermined amount from the upper end of the holding cylinder 13a. The sample S is projected, and then the sample S is clamped and held by the divided pieces of the holding cylinder 13a. As described above, in the present embodiment, the sample S is not fixedly supported only by the tightening by the divided pieces, so that the tightening force is sufficient so that the sample S is not crushed.

Then, the rotating plate 15b is rotated once to cut the protruding portion of the sample S by the cutting blade 15a thereof in parallel with the end face (see FIG. 10).

After the cutting, the camera 2 of the sample observing machine 1 takes an image while illuminating the cut surface of the sample S. The cutting of the sample S and the imaging of the cut surface are sequentially repeated.
The images of the cut surfaces are repeated between the cuts while rotating at a constant speed, whereby the images (image data) of the cut surfaces are captured in the memory.

The image data for each cut taken in the memory is subjected to data processing such as area division, contour deformation processing, and synthesis, and the cut surface image of the sample S and this are synthesized in response to a request from the outside. The three-dimensional image (three-dimensional image) obtained by this is displayed on the monitor, and based on this, various observations such as pathological examination in the medical field, toxicity examination in the pharmaceutical field, food inspection in the food field, crop inspection in the agricultural field, etc. Plant structure analysis and biological sample inspection in the biological field will be conducted.

As described above, the sample S is fixedly supported by closely fixing the end of the sample S to the movable stage 14a and tightening the other end of the sample by the divided piece of the holding cylinder 13a. Therefore, even when the force to lift the sample S is exerted when the edge of the sample S is cut by the cutting blade 15a, it is possible to reliably prevent the sample S from rising and maintain a stable posture. The sample S can be satisfactorily cut as it is, and by extension, the sample observation based on the cut surface imaging after the cutting and the image processing can be appropriately performed.

Further, since the sample S is brought into close contact with and fixed to the movable stage 14a by melting and solidifying the stage contact portion of the sample S by using the melted portion as an adhesive, the sample is fixed to the movable stage 14a. Because of this, there is no need for special glue or equipment.

Moreover, the recess 14a of the movable stage 14a
Since the solidified portion of the sample S is engaged with the reverse taper provided on the inner side surface of the sample 1, the sample S can be firmly fixed to the movable stage 14a.

Further, since the sample S is closely fixed to the movable stage 14a, it is possible to move the sample S downward as well as upward by the movable stage 14a, and the cut surface of the sample S can be moved more than the cutting position. or if you need to retract, useful when issuing <br/> unplug downwardly sample S from the holding cylinder 13a, the handling of the sample S is extremely easy.

That is, when it takes time to image the sample cut surface due to the amount of light, for example, when a fluorescence image is taken by a camera having a built-in CCD, the rotary plate 15b is rotated at a constant speed. In such a case, the movable stage 14a is not used because there is not enough time between cuttings.
If it the sample S by the downward movement retracted from the cutting position the cutting surface of the sample S by, buy time by be idle the cutting blade 15a becomes possible, scratches on the cut surface of the sample S in this case It can also prevent sticking.

The sample holder 13 used in the above embodiment
The Peltier element 13b may be replaced with other conventionally known cooling means, for example, a refrigerator using a refrigerant circuit. Further, the Peltier element 14b of the sample support portion 14 may be replaced with other heating / cooling means known in the related art, for example, a heat pump for both heating and cooling. If only heating is performed, an electric heater or the like may be used instead of the Peltier element 14b. It may be used instead.

In the above embodiment, the end of the sample S is melted and solidified so as to be closely fixed to the movable stage 14a. However, as shown in FIG. A cylindrical clamp member 14i may be provided on the upper surface, and the end portion of the sample S may be clamped by the clamp member 14i so as to be closely fixed to the movable stage 14h. The clamp member 14i includes a fixing piece 14i1 facing two side surfaces of the sample S, as shown in FIG.
And a movable piece 14i2 facing the other two side surfaces,
The sample can be tightened by moving the movable division piece 14i2 toward the center of the sample S by using spring pressure, air pressure, or the like. Of course, the number of movable pieces may be two or more, and if the inner surface of the clamp member is provided with a biting portion such as a protrusion, the sample S can be more firmly fixed to the movable stage 14h. .

[0047]

As described in detail above, according to the present invention,
Even if the force that lifts the end of the sample is exerted when it is cut, the sample can be reliably prevented from rising from the movable stage, and the sample is cut well while maintaining a stable posture. be able to. Also, since the sample is tightly fixed to the movable stage, and when it is necessary to retract from the cutting position the cutting surface of the sample, useful when out and disconnect downwardly sample from the holding cylinder, the sample handling It will be extremely easy.

[Brief description of drawings]

FIG. 1 is an overall view of a sample observation device according to the present invention

[Fig. 2] Detailed view of the sample cutting machine

FIG. 3 is a top view of a divided piece forming a holding cylinder.

FIG. 4 is a vertical sectional view and a top view of a movable stage.

FIG. 5 is a perspective view of a cutting blade and a sample.

FIG. 6 is a diagram showing a method of fixing a sample to a movable stage.

FIG. 7 is a diagram showing a method of fixing a sample to a movable stage.

FIG. 8 is a diagram showing a method of fixing a sample to a movable stage.

FIG. 9 is a diagram showing a conventional sample supporting method and sample cutting method.

FIG. 10 is a diagram showing a force acting on a sample during cutting.

[Explanation of symbols]

1 ... Sample observation machine, 11 ... Sample cutting machine, S ... Sample, 13 ...
Sample holder, 13a ... Holding cylinder, 13a1 to 13a5 ... Divided piece, 13b ... Peltier element, 14 ... Sample support, 14
a ... movable stage, 14b ... Peltier element, 14h ... movable stage, 14i ... clamp member, 15 ... sample cutting part, 15a ... cutting blade.

─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Kenichi Kudo 5-15-16 Sugamo, Toshima-ku, Tokyo (72) Inventor Hideo Yokota 2-15-7 Suga, Tama-ku, Kawasaki-shi, Kanagawa Capital Inada Tsutsumi 206 (72) Inventor Mitsunori Kokubo 2068-3 Ooka, Numazu-shi, Shizuoka Toshiba Machine Co., Ltd.Numazu office (72) Inventor Hiroshi Nanto 2068-3 Ooka, Numazu-shi, Shizuoka Toshiba Machine Co., Ltd.Numazu office (72) Inventor Shinsei Koichi 2068-3 Ooka, Numazu City, Shizuoka Prefecture Toshiba Machine Techno Co., Ltd. (56) Reference JP-A-8-35921 (JP, A) JP-A-6-265452 (JP, A) JP-A-3-249539 ( JP, A) JP-A-2-299643 (JP, A) Actual opening Sho 57-63375 (JP, U) Actual opening Sho 57-19973 (JP, U) Actual opening Sho 55-65543 (JP, U) Actual opening Flat 5-87555 (JP, U) Kosho 11-12727 (JP, Y1) Kenichi Kobayashi, Toshiro Higuchi, Isamu Aoki, Kenichi Kudo, Development of three-dimensional internal structure microscope, Japan Society for Precision Engineering, Japan, Precision Engineering Society, January 5, 1995, 1st Volume 61, Issue 1, p. 100-106 (58) Fields surveyed (Int.Cl. ⁷ , DB name) G01N 1/00-1/44 JISST file (JOIS)

Claims (6)

(57) [Claims] 1. In an observation sample supporting method for supporting a sample, which is sequentially cut from one end side and whose cut surface is observed, in a predetermined posture, a concave taper provided as an movable stage on the inner side surface is provided with a reverse taper.
Insert the end of the sample into the recess of the heated movable stage using
After pressing and melting the contact part, the melted part is solidified to firmly fix the sample to the movable stage, and the other end of the sample is placed in a holding cylinder having an inner hole aligned with the cross-sectional shape of the sample. An observation sample supporting method, characterized in that the observation sample is inserted. 2. An observing sample supporting method for supporting a sample, which is sequentially cut from one end side and whose cut surface is observed, in a predetermined posture, wherein a concave taper provided as an movable stage is provided with an inverse taper on the inner side surface.
Insert the end of the sample in the cooled state into the recess of the movable stage using
After pressing and melting the contact part, the melted part is solidified to firmly fix the sample to the movable stage, and the other end of the sample is placed in a holding cylinder having an inner hole aligned with the cross-sectional shape of the sample. An observation sample supporting method, characterized in that the observation sample is inserted. 3. An observation sample supporting method for supporting a sample, which is sequentially cut from one end side and whose cut surface is observed, in a predetermined posture, wherein a concave portion provided with an inverse taper on the inner side surface as a movable stage.
Using the one that has a section , insert the end of the sample in the cooled state into the concave part of the movable stage in the heated state and press it to melt the contact part, then solidify the melted portion and fix the sample closely to the movable stage At the same time, the other end of the sample is inserted into a holding cylinder having an inner hole aligned with the cross-sectional shape of the sample. 4. The method for supporting an observation sample according to claim 1, wherein the solidification of the molten component is caused by cooling. 5. The observation according to claim 1, wherein the sample comprises a sample body and an embedding agent which embeds the body, and the embedding agent is mainly melted. Sample support method. 6. The holding cylinder is composed of a plurality of divided pieces capable of tightening the sample, and the sample inserted into the holding cylinder is tightened by the divided pieces when cutting the sample. Item 6. A method for supporting an observation sample according to any one of items 1 to 5.