JPH11202215A - Method and device for sticking cover glass for microscope sample - Google Patents

Abstract

PROBLEM TO BE SOLVED: To prevent a sample from being damaged and a sealing agent from being protruded from the downside of cover glass and to stick this cover glass to slide glass by covering the sample while more surely preventing the formation of air bubbles between the cover glass and this slide glass. SOLUTION: While proximately opposing one terminal part on one side to each other of cover glass 7 and slide glass 3 through a fine gap, in the state of upward spreading the gap between each one side of both the glass 7 and 3, a sealing agent 2 is held at the section of the said fine gap. By turning an arm 14 and an oscillating stand 12, the angle formed between each one side of both the glasses 7 and 3 is gradually reduced, all the parts of each one side of both the glass 7 and 3 are proximately opposed to each other through the fine gap and both the glass 7 and 3 are turned into a horizontal state. Thus, the sealing agent 2 is gradually dispersed from one end of a section between both the glass 7 and 3 toward the other end of the section by a capillary phenomenon.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a method of attaching a sample to be prepared to a microscope specimen to a thin slice, attaching the cover glass to the stained slide glass with an encapsulant, and attaching the cover glass to the stained slide glass. Related to the device.

[0002]

2. Description of the Related Art In order to prepare a sample to be prepared on a microscope sample, for example, a tissue piece cut from an affected part of a patient, the sample is sliced, attached to a slide glass, degreased and stained. Thereafter, a cover glass for covering the sample is attached to the slide glass with an encapsulating agent. When attaching the cover glass to the slide glass, make sure that no air bubbles that inhibit microscopic observation are formed between the cover glass and the slide glass, and that no scratched sample enters under the cover glass. It is important to do.

As a method for preventing bubbles from being formed between the cover glass and the slide glass, for example, Japanese Patent Publication No.
There is a method as described in JP-A-0367. FIG.
1 is a front view schematically illustrating a method described in this publication. To prepare a microscope specimen, first, a slide glass 3 on which a sliced sample 1 is attached and a mounting medium 2 is dropped is placed on a table 4. Then, the suction pad 6 provided on the holder 5 so as to be able to move up and down is lowered toward the cover glass 7 placed horizontally at a different position from the illustration, and one end in the longitudinal direction of the cover glass 7 (the left end in FIG. 8). ) Is adsorbed. At this time, the other end in the longitudinal direction of the cover glass 7 (the right end in FIG. 8) is pressed downward by a pressing body 9 provided with a downwardly elastic force by a compression spring 8,
The cover glass 7 is held in a state of being inclined in the length direction. The downward projecting amount of the pressing body 9 is regulated by a nut 10 screwed to a portion of the upper end of the pressing body 9 projecting above the holding body 5. Next, the holder 5 is moved above the slide glass 3 with the cover glass 7 held in an inclined state, and then the holder 5 is lowered toward the slide glass 3. Based on this lowering, first, the other end in the length direction of the cover glass 7 pressed by the pressing body 9 abuts on the upper surface of the end of the slide glass 3, and then, the portion near the one end in the length direction of the cover glass 7 sequentially becomes the slide glass. Approach the top of 3. Along with this, the mounting medium 2
The slide glass 3 is pushed and spread to one end side. After the cover glass 7 is overlaid on the slide glass 3 and pressed by the suction pad 6 in this manner, the suction by the suction pad 6 is released, and the holder 5 is lifted up. Away from the top of As a result, the sample 1 on the slide glass 3 was placed on the upper surface of the table 4.
Is covered with a cover glass 7 to leave a microscope specimen. The air interposed between the slide glass 3 and the cover glass 7 is filled with the encapsulant 2 that spreads from the other end to one end of the cover glass 7 as the cover glass 7 is pressed.
While being pushed toward one end from between the two glasses 3 and 7.

[0004]

As described above, when the cover glass 7 is tilted in the longitudinal direction and the cover glass 7 is pressed against the upper surface of the slide glass 3 from the other end to one end, the cover glass 7 If a part of the sample is hung on the other edge of the sample, the sample may be damaged. Further, when the cover glass 7 is pressed against the upper surface of the slide glass 3 while being curved as described above, the encapsulant 2 may protrude from under the cover glass 7. The mounting medium 2 protruding from under the cover glass 7 adheres to the upper surface of the cover glass 7, which makes it difficult to handle (move, store, etc.) the microscope specimen, or hinders microscopic observation. Absent.

When the pressing force of the cover glass 7 against the slide glass 3 is reduced to prevent the mounting medium 2 from protruding from under the cover glass 7, the mounting medium 2 is placed on the upper surface of the slide glass 3. There is a possibility that air bubbles are not formed evenly over the entire surface of the portion covered by the cover glass 7 and between the two glasses 3 and 7. In view of the above-described circumstances, the present invention prevents damage to the sample and protrusion of the encapsulant 2 from under the cover glass 7 and prevents the formation of air bubbles between the glasses 3 and 7. It is intended to provide a method of attaching a cover glass 7 to a slide glass 3 while preventing the slide glass 3 more reliably.

[0006]

According to the method for attaching a cover glass to a microscope specimen of the present invention, one end of one surface of a slide glass on which a sample is placed and one end of one surface of a cover glass are attached to each other via a minute gap. When the slide glass and the cover glass are inclined or erected while the space between one side of the slide glass and the cover glass is extended upward or to the side, the micro glass The mounting medium is held in the gap. Then, in this state, the angle between one surface of the slide glass and one surface of the cover glass is gradually reduced, and the thickness of the one surface of the slide glass and one surface of the cover glass is substantially reduced over the entire surface. The encapsulant held in the minute gap between one end of one side of the slide glass and the cover glass by bringing the slide glass and the cover glass into a horizontal state while making the slide glass and the cover glass horizontal is placed through a uniform minute gap. Then, the light is diffused from one end between the slide glass and the cover glass to the other end between the portions.

[0007]

According to the cover glass attaching method of the present invention, when the cover glass is attached to the slide glass, these two glasses do not come into contact with each other. Therefore, it is possible to prevent the sample from being damaged by the edge of the cover glass. Furthermore, in order to diffuse the encapsulant held in the minute gap between one end of one side of these two glasses from one end of the part between these two glasses to the other end of the interpart, the encapsulant is covered with one side of the slide glass and the cover It can be uniformly spread over the entire surface of the minute gap with one side of the glass. Therefore, it is possible to effectively prevent bubbles from being formed between the two glasses, and to prevent the encapsulant from protruding from under the cover glass. As a result, observation of the microscope specimen is not hindered, and handling of the microscope specimen is not troublesome.

[0008]

1 to 6 schematically show a cover glass attaching apparatus according to the present invention. The cover glass sticking device is
An oscillating table 12 serving as a slide glass supporting member, an arm 14 having a suction pad 15 attached to a tip as a cover glass supporting member, and oscillating driving means for oscillating the oscillating table 12 and the arm 14 respectively. First and second motors 1
3 and 16, an encapsulating agent supply device 17, and a controller (not shown). Among these, the swing table 12 is supported on a part of the fixed base 11 so as to be freely displaceable about a first shaft 19 in a horizontal direction. The slide glass 3 can be placed on the upper surface of the swing table 12 in a state where the length direction of the slide glass 3 and the horizontal direction coincide with each other. Further, the swing table 12 can be freely swung about the first shaft 19 by the first motor 13. In the example shown, the output shaft of the first motor 13 is the first shaft 19, and the swing table 1
2 is made to match the rotation speed of the first motor 13, but if necessary, a speed reduction mechanism composed of a combination of gears or the like is incorporated so that the swing speed of the Restrict to value.

A base end portion of the arm 14 (a lower end portion in FIGS. 1 to 3) is provided on one side portion (the right side portion in FIG. 2) of the swing table 12 near the first shaft 19. 5 to 6) are swingably supported about a second shaft 20 provided on the swing table 12. This arm 14 is connected to the second motor 16
Thereby, the rocking displacement about the second shaft 20 is enabled. In the illustrated example, the output shaft of the second motor 16 is the second shaft 20 and the swing speed of the arm 14 is made to match the rotation speed of the second motor 16. In response to the above, a swinging speed of the arm 14 is regulated to a desired value by incorporating a speed reduction mechanism formed by combining gears and the like. A suction pad 15 made of an elastic material such as rubber or vinyl is attached to the tip of the arm 14, and the inside of the suction pad 15 can be freely communicated with a suction port of a vacuum pump 23 by a flexible conduit 22. An electromagnetic switching valve is provided in the middle of the flexible conduit 22 so that the inside of the suction pad 15 is
It can freely communicate with one of the intake port and the atmosphere.
With this configuration, a part of the cover glass 7 can be freely sucked on the suction surface of the suction pad 6. or,
The encapsulant supply device 17 is capable of dropping an appropriate amount of encapsulant downward in a timely manner by a discharge nozzle 24, and is capable of moving the discharge nozzle 24 in the horizontal direction. or,
Starting and stopping the first and second motors 13 and 16 and changing the rotation direction, and supplying and discharging the vacuum pump 23;
The control of the operation of the encapsulating agent supply device 17 is performed by the controller (not shown). The operation of attaching the cover glass 7 to the upper surface of the slide glass 3 by the cover glass attaching apparatus configured as described above is performed as follows.

First, the suction surface of the suction pad 15 attached to the tip of the arm 14 and the upper surface of the swing table 12 on which the slide glass 3 is placed are opposed to each other with a certain distance. Then, in this state, one side surface of the cover glass 7 (the right side surface in FIGS.
6 (upper side surface) are adsorbed in a state where the length direction of the cover glass 7 is aligned with the horizontal direction. In order to cause a part of the cover glass 7 to be sucked by the suction pad 6 in this manner, one of the suction pad 6 and the cover glass 7 is directed toward the other of the suction pad 6 and the cover glass 7. It is done by moving. For example,
The cover glass 7 is inserted between the suction surface of the suction pad 6 and the upper surface of the swing table 12, and the suction surface of the suction pad 6 is brought close to one side surface of the cover glass 7 by the swing of the arm 14. . It is to be noted that the swing table 12 is swung so that the upper surface of the swing table 12 is inclined.
2 is carried out within a range where the slide glass 3 does not slide down. Further, a material having a large frictional resistance is used on the upper surface of the rocking table 12 or a locking portion is provided, so that the slide glass 3 is hard to slide down from the upper surface of the rocking table 12.

When the cover glass 7 is sucked to the suction pad 6, the arm 14 and the swing table 12 are moved to the position shown in FIG.
And one end of the upper surface of the slide glass 3 (the lower end in FIGS. 1 to 4 and the right end in FIGS. 5 to 6) and one end of the other side which is a side surface which is not sucked by the suction pad 15 of the cover glass 7 (FIG. 1 to 4 and the right end in FIGS. 5 to 6). Further, the glasses 3 and 7 are held in a state where the space 21 between the opposing surfaces of the glasses 3 and 7 is spread upward. In this way, the arm 1
While holding the swing table 4 and the swing table 12, the inclination angle α (FIG. 1) of the swing table is set to about 75 °. In this state, a minute gap of 1 mm or less is interposed between one end of the upper surface of the slide glass 3 and the other end of the side surface of the cover glass 7 which are opposed to each other.

As shown in FIG. 3, a suitable amount of the discharge nozzle 24 of the encapsulant supply device 17 is provided from above the space 21 existing between the opposing surfaces of the glasses 3 and 7. The encapsulating agent 2 is dropped toward a portion near the other end of the upper surface of the slide glass 3. The encapsulant 2 flows down in the width direction of the slide glass 3 with the upper surface of the slide glass 3 directed from the other end to the one end, and is held in the minute gap. That is, since the encapsulant 2 has a viscous tone (has a high viscosity), it is held in the minute gap. At this time, FIG.
As shown by the arrow in FIG. 3, the discharge nozzle 24 of the encapsulating agent supply device 17 drops the encapsulating agent 2 while
It is moved in the horizontal direction (in the length direction of both glasses 3 and 7 in the horizontal direction of FIG. 4) along the space 21 existing between the 7. Therefore, on the upper surface of the slide glass 3, the encapsulant 2
Can be applied almost uniformly in the horizontal direction.

Next, the swing table 12 is swung in the counterclockwise direction in FIGS.
As shown in (1), the swing table 12 is set in a horizontal state. While the swing table 12 swings in this manner, the arm 14 is not swung with respect to the swing table 12. Therefore, the above both glasses 3,
The angle between the 7 does not change. After the swing table 12 is in a horizontal state, the arm 14 is swinged in the counterclockwise direction indicated by the arrow in FIG. 5 by the second motor 16. Then, the angle between the two glasses 3 and 7 is gradually reduced, and the cover glass 7 is kept horizontal as shown in FIG. , And close to each other via a minute gap. Then, the thickness of the minute gap is made substantially uniform over the entire surface. In this way, by making the minute gaps between the two glasses 7 and 3 uniform, the encapsulant 2 held in the minute gap portion between the ends of the opposing surfaces of the glasses 7 and 3 can be used. From the one end of the portion between the glasses 7 and 3 toward the other end of the portion between the two glasses, the light is diffused by the capillary phenomenon over the width direction of the glasses 7 and 3. Therefore, the encapsulant 2 can be evenly spread over the entire surface of the slide glass 3 covered by the cover glass 7. In this manner, the encapsulant 2 is uniformly spread between the two glasses 3 and 7, and the formation of air bubbles between the two glasses 3 and 7 can be effectively prevented. Also, during the enclosing operation, the cover glass 7 is not pressed against the upper surface of the slide glass 3. Therefore, the amount of the encapsulant 2 interposed between the two glasses 7 and 3 depends on the size of the minute gap when the two glasses 7 and 3 are opposed to each other, the size of the cover glass 7, and the like. If done properly,
The encapsulant can be spread over the entire surface of the minute gap, and the encapsulant 2 can be prevented from protruding from under the cover glass 7. As a result, observation of the microscope specimen is not hindered, and handling of the microscope specimen is not troublesome. Further, the cover glass 7 for the slide glass 3
Since the two glasses 3 and 7 do not come into direct contact with each other during the attaching operation, the sample can be prevented from being damaged by the edge of the cover glass 7.

In the illustrated example, after the slide glass 3 is set in a horizontal state, the angle between the slide glass 3 and the cover glass 7 is gradually reduced by swinging the arm 14. The arm 14 and the rocking table 12 may be simultaneously rocked to bring the slide glass 3 to a horizontal state, and at the same time, the angle between the two glasses 3 and 7 may be gradually reduced. Further, the arm 14 is swung to cover the cover glass 7.
After all the other side faces are made to approach a part of the upper surface of the slide glass 3 in an inclined state with a minute gap therebetween, the rocking table 12 is rocked to bring the both glasses 7 and 3 into a horizontal state. You may do it.

Further, the encapsulant supply device 17 is not limited to the above-described type in which the encapsulant 2 is dropped downward.
The mounting medium 2 may be ejected in a horizontal direction. For example, as shown in FIG. 7, one end of the upper surface of the slide glass 3 in the horizontal state (the right end in FIG. 7) and one end of the lower surface of the cover glass 7 in the inclined state (the right end in FIG. 7) are minute. In a state where the two glasses 7 and 3 are opposed to each other with a gap therebetween, the space between the opposing surfaces of the two glasses 7 and 3 is widened from one (left side in FIG. 7) to the other (right side in FIG. 7). For example, the encapsulant 2 is ejected by the ejection nozzle 25 toward the second direction.

In the illustrated example, the encapsulating agent supply device 1 is used.
7, the encapsulant 2 is not dropped directly onto the minute gap between the one ends of the mutually facing surfaces of the glasses 3 and 7,
The encapsulant 2 is caused to flow down along the upper surface of the slide glass 3 inclined by an angle α (70 °), and then held in the minute gap. The reason for this is that the kinetic energy of the falling prevents the encapsulant 2 from flowing down without stopping at the minute gap, and the slide glass 3
This is to improve the conformity between the upper surface of the glass and the encapsulant 2 and to more reliably prevent the formation of air bubbles between the glasses 3 and 7.

Further, in the case of this embodiment, the arm 14 as the cover glass support member is swung with respect to the swing table 12 as the slide glass support member, so that the other side of the cover glass 7 is moved to the slide glass 3. I tried to get closer to the top,
Conversely, the upper surface of the slide glass may be brought closer to the other side surface of the cover glass by swinging the slide glass support member relative to the cover glass support member.

For example, first, one end of the upper surface of a slide glass in a horizontal state and one end of one surface of an inclined or upright cover glass are closely opposed to each other with a small gap therebetween. In part, the encapsulant is evenly applied horizontally. Then, the slide glass support member is swung with respect to the cover glass support member in a direction in which the angle between the slide glass and the cover glass decreases. As a result, the encapsulant flows down the upper surface of the slide glass toward the minute gap portion, and is held in the minute gap portion. In this state, by further swinging the slide glass support member, the angle between the slide glass and the cover glass is further reduced, and these two glasses are made to have a very uniform thickness over the entire surface. It is made to face and close via a gap. Thus, the encapsulant held in the minute gap is diffused from one end between the two glasses to the other end between the two glasses, and the encapsulant is uniformly spread over the entire surface of the minute gap between the two glasses. Can be.

In this embodiment, the first and second motors 13 and 16 are used as the oscillating drive means. However, in the present invention, other than the above, a means utilizing gravity-induced natural fall or air. And the like utilizing the spraying of the above can be adopted.

[0020]

According to the method and the apparatus for attaching a cover glass of the present invention constructed and functioning as described above, damage to the sample, protrusion of the encapsulant from under the cover glass, and the cover glass and the slide glass can be prevented. The cover glass can be stuck to the slide glass while preventing the formation of bubbles between the slide glasses. Furthermore, in the method in which the cover glass is pressed while being curved on the upper surface of the slide glass as in the above-described conventional example, the speed at which the cover glass is pressed against the upper surface of the slide glass is high. In some cases, bubbles may be formed between the two glasses. On the other hand, according to the present invention, there is no such disadvantage. In addition, since the cover glass can be attached not in the length direction but in the width direction whose dimension is shorter than the length direction, the time required for attaching the cover glass can be reduced.

[Brief description of the drawings]

FIG. 1 is a front view schematically showing a cover glass sticking apparatus of the present invention.

FIG. 2 is a view seen from the right side of FIG. 1;

FIG. 3 shows a state in which an encapsulant is dropped by a dropping device.
The figure similar to FIG.

FIG. 4 is a right side view of FIG. 3, showing a state in which a discharge nozzle of a dropping device is moved, with a part thereof being omitted;

FIG. 5 is a view similar to FIG. 3, but showing a state in which a rocking table is swung to set a slide glass in a horizontal state.

FIG. 6 is a view similar to FIG. 5, showing a state in which an arm is swung so that an upper surface of a slide glass and one surface of a cover glass are closely opposed to each other.

FIG. 7 is a schematic front view showing another example of an encapsulating agent supply device.

FIG. 8 is a schematic front view showing an example of a conventional cover glass sticking apparatus.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Sample 2 Encapsulant 3 Slide glass 4 stand 5 Holder 6 Suction pad 7 Cover glass 8 Compression spring 9 Pressing body 10 Nut 11 Base 12 Swing table 13 First motor 14 Arm 15 Suction pad 16 Second motor 17 Encapsulant supply device 19 First shaft 20 Second shaft 21 Space 22 Flexible conduit 23 Vacuum pump 24 Discharge nozzle 25 Spray nozzle

Claims (2)

[Claims] An end of one side of a slide glass on which a sample is placed and an end of one side of a cover glass are closely opposed to each other via a minute gap, and the one side of the slide glass and the one side of the cover glass are opposed to each other. The space between them is spread upward or to the side, the slide glass and the cover glass are inclined or erected, and the encapsulant is held in the minute gap portion, and one side of the slide glass and the By gradually reducing the angle formed by one side of the cover glass and one side of the slide glass and one side of the cover glass to closely approach each other via a minute gap having a substantially uniform thickness over the entire surface, The encapsulant held in the minute gap between one end portions of the slide glass and the cover glass is applied from one end of the portion between the slide glass and the cover glass. A method of attaching a cover glass to a microscope specimen, which is diffused toward the other end of the part. 2. A slide glass support member on one side capable of supporting a slide glass, a cover glass support member on one side opposite to the slide glass capable of freely supporting a cover glass, and a slide glass support member and a cover glass support. And a swing drive means for swinging and displacing at least one of the members about an axis provided in a horizontal direction, wherein the swing drive means includes one end on one surface of the slide glass and the cover. One end of one side of the glass is closely opposed to each other with a minute gap therebetween, and the space between one side of the slide glass and the cover glass is spread upward or to the side, and these slide glasses are The slide glass support member and the cover glass are tilted or erected, and the encapsulant is held in the minute gap. At least one of the cover glass support member is swung about an axis provided in the horizontal direction, gradually reducing the angle between one surface of the slide glass and one surface of the cover glass, A cover glass adhering device having a function of causing one surface of the slide glass and one surface of the cover glass to closely face each other via a minute gap having a substantially uniform thickness over the entire surface.