JPH0743278A - Method and apparatus for dyeing microscopic sample - Google Patents

Abstract

PURPOSE:To eliminate the deterioration of processing liquid while reducing consumption thereof. CONSTITUTION:A plurality of slide glasses 16 applied with sample are supported vertically through a microgap 17. In this regard, the upper slide glass 16 is projected longer, at one end thereof, than the lower one in the longitudinal direction. Processing liquid is dispensed by a dispensation nozzle to one longitudinal end of the uppermost slide glass 16. The processing liquid spreads over the upper face of the slide glass 16 and then intrudes by capillary phenomenon into the microgap 17 between the slide glasses 16 and spreads over the upper face of the lower slide glass 16. The slide glasses 16 are then supported while matching each other through a constant gap larger than the microgap 17. The slide glass 16 is rotated about the vertical axis under that state to remove excess processing liquid. The series of steps are repeated in predetermined order.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION The method for staining a microscope specimen and the apparatus therefor according to the present invention provide a piece of tissue or a cell attached to a slide glass for microscopic observation in a hospital or various laboratories. In this specification, these are collectively referred to as a sample)
It is used for dyeing.

[0002]

2. Description of the Related Art In hospitals and the like, it is widely practiced to observe a sample excised from an affected area with a microscope to diagnose a disease. In order to facilitate the diagnosis of diseases by such microscopic observation, the sample attached to the slide glass is stained, and various automatic staining devices that automatically perform this staining work are also provided. Has been done.

FIG. 15 shows an automatic dyeing device disclosed in Japanese Patent Laid-Open No. 61-235734. First, the automatic dyeing device shown in FIG. 15 will be briefly described.

On the upper surface of the substrate 1 which also serves as the bottom plate of the case,
A large number of containers 2, each of which stores a processing liquid and has an open top,
2 are arranged neatly. Above the substrate 1 on which a large number of containers 2 and 2 are arranged, two guide rails 3 and 3 parallel to each other are provided.
Is provided, and the support beam 4 hung on both guide rails 3 and 3 extends in the longitudinal direction of both guide rails 3 and 3 (arrow a in the drawing).
Direction). Further, the support beam 4 is provided with a moving tool 5 that moves in a direction perpendicular to the length direction of the guide rails 3, 3. An amount of movement of the support beam 4 along the guide rails 3 and 3 and an amount of movement of the moving tool 5 along the support beam 4 are separately provided controllers (not shown).
The support beam 4 and the moving tool 5 move in the horizontal direction based on a command from the controller. The moving tool 5 is provided with a hanging arm 6 that is moved up and down by a lifting mechanism attached to the moving tool 5. Then, a basket 7 containing a slide glass on which a sample is attached, as shown in FIG. 16, is suspended by a locking tool provided on the suspension arm 6 and driven by a solenoid.

When performing a staining process on a sample attached to a slide glass by the conventional automatic staining apparatus for a microscope specimen constructed as described above, the basket 7 containing the slide glass on which the sample is attached is suspended from the arm. The cage 7 is hung on the locking tool 6 and the moving tool 5 is horizontally moved in accordance with a predetermined order in accordance with a command from the controller, whereby the basket 7 is sequentially moved onto the predetermined container 2. When the basket 7 moves to a predetermined container 2, the hanging arm 6 is lowered and the slide glass housed in the basket 7 is immersed in the treatment liquid in the container 2. At this time, a shaking operation for vibrating the hanging arm 6 vertically is performed to make the treatment liquid and the sample fit together. When the immersion for a predetermined time is completed, the hanging arm 6 is lifted to pull out the basket 7 from the container 2, move the basket 7 toward the next container 2, and perform this operation a predetermined number of times. The sample attached to the inner slide glass is dyed by a desired dyeing method.

The automatic dyeing apparatus other than the automatic dyeing apparatus shown in FIG. 15 described above has the same basic structure and operation as the automatic dyeing apparatus shown in FIG. A container in which the liquid to be used is stored and whose upper part is opened is arranged on the upper surface of the base, and the slide glass on which the sample is adhered is dipped in the liquid in the container in a predetermined order.

[0007]

However, the conventional automatic dyeing apparatus described above has the following problems to be solved. That is, when performing a dyeing operation, a basket 7 containing a slide glass to which a sample to be dyed is attached.
Are sequentially immersed in the containers 2 and 2 in which the treatment liquids are stored. At this time, it is inevitable that one treatment liquid attached to the basket 7 is mixed with other treatment liquids during the dipping operation. For example, 150
When the cage 7 containing 20 slide glasses is dipped in ml of the treatment liquid, and then the cage 7 is dipped in the next container 2, the treatment liquid in the container 2 contains There is also reported an example in which about 3 ml of the treatment liquid of 1.

As described above, the treatment liquids are gradually deteriorated by mixing the treatment liquids of different types, but when the treatment liquids deteriorate, the desired dyeing cannot be performed. Various measures have been proposed in order to prevent such deterioration of the processing liquid, but the present situation is that there is no deciding factor.

Further, when the processing liquid deteriorates, this processing liquid is discarded and a new processing liquid is poured into the container 2. Since the treatment liquid is expensive, the treatment liquid deteriorates quickly and it is uneconomical to frequently replace the treatment liquid. Moreover, in the above-mentioned conventional apparatus, in order to improve the efficiency of the dyeing process, a plurality of containers 2 and 2 storing the same processing liquid (for example, 5 containers storing xylene and 8 containers storing alcohol) are used. Since the basket 7 was prepared and immersed in each of the containers 2 and 2, it was unavoidable that the consumption of the treatment liquid increased. Therefore, it has been desired to establish a technique capable of dyeing as much slide glass as possible with the smallest possible treatment liquid.

The dyeing method and apparatus of the present invention have been invented in view of the above-mentioned circumstances, and the treatment liquids of different types do not mix with each other, and therefore the treatment liquids do not deteriorate, and It enables dyeing work with a small amount of processing liquid.

[0011]

Among the method and apparatus for staining a microscope specimen according to the present invention, the staining method according to claim 1 is a plurality of slide glasses having a sample to be stained attached to the upper surface thereof. In the vertical direction with a minute gap between them, and the upper slide glass is supported with its one end in the length direction protruding more than the one end in the length direction of the slide glass immediately below. , By selecting and dispensing a predetermined processing liquid to one end in the length direction of the uppermost slide glass, this processing liquid flows down from the upper slide glass toward the lower slide glass, and the processing liquid is sequentially processed. Process of permeating the sample into the sample, and after this permeating process, the plurality of slide glasses are vertically aligned and aligned with each other, and are supported with a constant gap larger than the minute gap. Then By rotating the plurality of glass slides around a vertical axis in the state, and a removal step for removing excess treatment liquid. Then, each of these steps is sequentially repeated a predetermined number of times to stain the sample.

Further, the dyeing device according to a second aspect of the present invention comprises a frame-shaped storage container which is composed of upper and lower horizontal sides and left and right vertical sides, and axially supports end portions of the mutually butting sides. In order to support both ends in the length direction of the slide glass on which the sample to be dyed is adhered, the inner surfaces of the left and right vertical sides facing each other are formed at the same pitch in the up and down direction, respectively. The same number of first and second support pieces,
One of the upper and lower horizontal sides can be pivoted to one side in the width direction at the same inclination as the vertical side in the direction to open the widthwise one side opening of the storage container by the pivoting means. The first supporting member is pivotally supported on the other side, and the engaging portion formed on either one of the upper and lower end portions and the upper or lower other side portion and the receiving portion formed on the other side are freely engageable with each other. The side plate, the other side of the upper and lower horizontal sides in the width direction, the second side plate that pivotally supports the upper and lower ends at the same inclination as the vertical side, and the outer surface of the central portion of either one of the upper and lower sides of the storage container. A provided joint member, and a detachable and attachable member to the joint member. When attached, the storage container supports the storage container from one of the upper and lower ends, and rotates the storage container and the storage container. In this case, a supporting means for supporting the outer surface of the central portion of either the upper or lower end of the storage container And deforming means for deforming the posture of the container, it is arranged in the longitudinal direction end portion upper container,
It is equipped with a dispensing nozzle that can dispense the treatment liquid toward one end of the uppermost slide glass in the length direction. And
The storage container has the first posture in which the vertical side is parallel to the vertical axis and the vertical side is inclined by an angle θ with respect to the vertical axis by the deforming means, so that one end of the upper horizontal side in the longitudinal direction is It is freely deformable to any one of the second postures in which the lower lateral side projects from the one end in the length direction. In addition, the first and second support pieces are arranged so that the support pieces on one end side in the length direction of the storage container are at an angle substantially equal to the angle θ so that they are horizontal when the storage container is in the second posture. The support piece is formed upward, and the support piece on the other end side in the length direction of the storage container is formed downward at an angle substantially equal to the angle θ. Furthermore, the dispensing nozzle is
Based on a command from the controller, it can be connected to one of a plurality of processing liquid containers via a liquid feed pump and a rotary valve.

[0013]

The operation when the sample is dyed using the apparatus of the present invention constructed as described above is as follows: First, the first side plate is rotated to move the width of the storage container. The slide glass is allowed to pass through the opening on one end side in the direction.
In this state, a plurality of slide glasses with the sample to be dyed attached to the upper surface are placed in a storage container so that the samples are on the upper surface, and both ends in the length direction of the slide glass are opposed to each other. Place it so that it hangs over the second support piece. When a desired number of slide glasses are stored in the storage container, the first side plate is rotated in the opposite direction, and the other end of the first side plate in the up-down direction and the lateral side facing the other end. The engaging portion and the receiving portion, which are respectively provided in the above, are engaged with each other. Next, the storage container is supported by the rotation support member in the casing. In this state, the storage container remains in the first position. From this state, the displacement means is operated to deform the storage container from the first posture to the second posture.

Then, based on a command from the controller, a rotary nozzle and a liquid feed pump are used to insert a plurality of processing liquids into one of the processing liquid containers, and a dispensing nozzle connected to the processing liquid container into the storage container. The treatment liquid is dispensed to one end of the stored slide glass in the longitudinal direction of the uppermost slide glass. The treatment liquid dispensed to one end of the uppermost slide glass in the length direction first spreads on the upper surface of the slide glass to permeate the sample. Then, the treatment liquid flows down toward the slide glass immediately below, but at this time, there is a minute gap between the lower surface of the slide glass at the uppermost position and the upper surface of the slide glass immediately below, and the flowed-down processing is performed. The liquid enters the minute gap due to a capillary phenomenon and reaches the upper surface of the slide glass immediately below. In this way, the treatment liquid sequentially reaches the lower slide glass (permeation process).

When the processing liquid reaches the upper surface of the lowermost slide glass, the deforming means is operated in reverse to return the storage container from the second position to the first position.

Next, either the upper or lower side of the storage container is supported by the support means, and the rotation support member is rotationally driven. Along with this, the storage container rotates, but at this time, the excess processing liquid that is not used for dyeing the sample on the upper surface of the slide glass is scattered into the casing by the centrifugal force of rotation.
As a result, the excess processing liquid not used for dyeing is removed from the upper surface of the slide glass (removal process).

Further, the controller dyes the sample by repeating each of the steps a predetermined number of times according to a program stored in advance.

[0018]

1 to 11 show an embodiment of a dyeing apparatus according to the present invention. The storage container 8 constituting the present invention is constructed as shown in FIG. The storage container 8 includes horizontal plates 9 and 10 which are upper and lower horizontal sides, and vertical plates 11 and 1 which are vertical left and right sides.
2 and 2 are combined in a rectangular frame shape, and the portions of the vertical and horizontal plates 9 to 12 that are in contact with each other are pivotally supported. In this embodiment, the protruding pieces 13 and 13 are provided at the four corners of both ends of the upper and lower horizontal plates 9 and 10.
Is formed, and each of the plates 9 to 12 is formed through the protruding pieces 13 and 13.
Is pivotally supported. Therefore, the storage container 8 is the vertical plate 1
By applying an external force from the outer surface side of 1 and 12, FIG.
The first posture shown in Fig. 9 can be freely transformed into the second posture shown in Fig. 9.

The first posture is a state in which the vertical plates 11 and 12 are parallel to the vertical axis a. On the other hand, the second posture is the vertical plate 11,
12 is inclined by an angle θ (Fig. 1) with respect to the vertical axis a,
The lengthwise one end portion (the right end portion in FIG. 1) of the upper horizontal plate 9 is in a state of projecting from the lengthwise one end portion (the right end portion in FIG. 1) of the lower horizontal plate 10.

Of the vertical plates 11 and 12 which constitute the storage container 8,
First and second support pieces 14 and 15 are formed on the inner side surfaces facing each other in the vertical direction at the same pitch.
The slide glass 1 is attached to the first and second support pieces 14 and 15.
It is provided to support both ends of the lengths 6 and 16 (left and right direction in FIGS. 5 and 7) respectively. Therefore, the pair of first and second support pieces 14 and 15 are formed at the same height position. Along with this, when the storage container 8 is deformed from the first posture to the second posture, the first support pieces 14, 14 formed on the inner side surface of the vertical plate 11 on the displacement direction side (the left side in FIG. 1), Above angle θ
Only tilt upwards. Also, the other second support piece 1
Nos. 5 and 15 are inclined downward by the angle θ. As a result, when the storage container 8 is in the second posture, the first and second support pieces 14 and 15 are in a horizontal state. In this state, vertically adjacent support pieces 14, 14 (and 1
5 and 15), the upper slide glass 15
Between the lower surface of the and the upper surface of the lower slide glass 16,
Preferably, a minute gap 17, 1 of about 0.1 mm to 0.5 mm
7 is formed in consideration of the thickness of the slide glass 16. However, since the thickness of the sample attached to the upper surface of the slide glass 16 is about several μm, it is not necessary to consider the thickness of this sample.

One end side in the width direction of the storage container 8 (lower side in FIG. 2)
Two first side plates 19 and 19 are provided in the. The first side plates 19, 19 are provided with an inclination equal to the inclination of the vertical plates 11, 12. Then, the upper end portions of the first side plates 19 are pivotally supported on the lateral plate 9.

Further, through holes 20 are formed in the lower end portions of the first side plates 19 as shown in FIG. 8 (A).
At the same time, on the side surface of the lower horizontal plate 10, at a position facing the through hole 20, there is provided a locking projection 21 whose tip is a spherical portion having a diameter slightly larger than the inner diameter of the through hole 20. There is.
A notch 22 serving as an opening / closing means is formed on the front surface or the back surface of the upper end portion of the first side plate 19, 19.
9 and 19 are rotatable in the direction of arrow b in FIG. 8A so that the one side opening (right side in FIG. 8) of the storage container 8 can be opened and closed. By swinging the first side plates 19 and 19 away from the storage container 8, the slide glasses 16 and 16 can be freely inserted and removed from the one side opening, and the slide glasses 16 and 16 can be rotated in the opposite direction to the through hole. By engaging 20 with the locking projection 21, the one side opening can be closed. The through hole 20 serves as a receiving portion, and the locking projection 21 serves as a locking portion.

Two first side plates 1 are provided on the other end side (upper side in FIG. 2) of the storage container 8 in the width direction (vertical direction in FIG. 2).
8 and 18 are provided. The second side plates 18, 18 are provided at the same inclination as the vertical plates 11, 12. Second side plate 1
The upper and lower ends of 8 and 18 are pivotally supported by a pair of horizontal plates 9 and 10.

A joint member 23 is provided at the central position of the upper lateral plate 9 constituting the storage container 8 and can be connected to the lower end of a rotary shaft 26 described later. The joint member 23 has a conventionally known structure. In addition, the small concave portion 2 is provided at the center position of the outer surface of the lower horizontal plate 10 that constitutes the storage container 8.
4 is forming. The small recessed portion 24 is provided for abutting a tip end portion of a support shaft 25 described later.

Cylindrical case 3 in which the storage container 8 is disposed
Above the center of 3, there is provided a rotary shaft 26 whose distal end enters the case 33. The rotating shaft 26 is extendable and contractible in the vertical direction by an air cylinder 32, and the base end portion thereof is rotatably supported by fixed portions via bearings 27a and 27b. Further, a pulley 28 is provided at an intermediate portion, and is freely rotatable by a belt 31 which is hung between a pulley 30 provided on an output shaft of a motor 29 which is a driving device. Then, as described above, the tip of the rotary shaft 26 can be connected to the joint member 23 of the storage container 8.

Below the center of the case 33, there is provided a support shaft 25 whose front end has entered the case 33. This support shaft 25, which is a supporting means, is also the same as the rotary shaft 26.
It is displaceable in the up-down direction, and when it rises toward the storage container 8, its tip comes into contact with the inner surface of the small recess 24 of the storage container 8 and prevents the storage container 8 from shaking when the storage container 8 is rotated. .

In the state where the tip of the rotary shaft 26 is connected to the joint member 23 of the storage container 8 and the tip of the support shaft 25 is brought into contact with the small recess 24 of the storage container 8, the rotary shaft 26 is shown. The support shaft 25 is collinear with the central axis of the storage container 8. At this time, the storage container 8 is in the first posture.

On the side of the case 33, a pressing rod 34 as a deforming means is provided so as to be freely displaceable by, for example, a rack and pinion. That is, the vertical plate 12 that constitutes the storage container 8
The pressing rod 34 is provided so as to face the lower end of the storage container 8, and the storage container 8 is pressed by displacing the pressing rod 34 toward the storage container 8, and the storage container 8 is moved from the first position shown by the solid line in FIG. It is deformed to the second posture shown by the chain line in the figure.

In the present embodiment, as shown in FIG. 3, the displacement of the pressing rod 34 and the support shaft 25 is interlocked.
That is, a first sensor such as a proximity switch for detecting the lowering of the support shaft 25 is provided around or near the support shaft 24.
When the first sensor detects the lowering of the support shaft 25, the pinion is rotationally driven to move the pressing rod 34.
To displace. Further, a second sensor for detecting that the pressing rod 34 is displaced to a position where the storage container 8 is deformed from the second posture to the first posture is provided at the portion where the pressing rod 34 is installed. The support shaft 25 is raised according to the detection values of the second sensor. However, when the pressing rod 34 is displaced to the above-mentioned position, the pressing rod 34 is briefly (for example, 3
After resting, the displacement is restarted in the same direction. This is to ensure that the tip of the support shaft 25 is brought into contact with the small recess 24.

A dispensing nozzle 35 is provided above the inside of the case 33 and on the side of the storage container 8 supported between the support shaft 25 and the rotary shaft 26 opposite to the pressing rod 34. The dispensing nozzle 35 is connected to one end of a first connecting pipe 37 provided with a roller pump 36 on the way. The other end of the first connecting pipe 37 is connected to the discharge port of a rotary valve 38 which is conventionally known. Further, one end of a second connecting pipe 39 is connected to the suction port of the rotary valve 38. The other end of the second connecting pipe 39 has branch pipes 41, 41 having one end connected to the processing liquid containers 40, 40 storing various processing liquids.
Connected to.

The operation of the roller pump 36 and the rotary valve 38 is controlled by a controller (not shown). That is, this controller causes the rotary valve 38 to be switched, the roller pump 36 to be driven, to be stopped, and the like according to the processing procedure stored in advance in this controller.

One end of the bottom of the case 33 has a waste liquid container 42.
The other end of the discharge pipe 43 connected to is connected. The waste liquid container 42 and the discharge pipe 43 are provided for discharging the processing liquid accumulated on the bottom surface of the case 33 in the process of dyeing the sample. In the present embodiment, the bottom surface of the case 33 is inclined toward the connecting portion of the discharge pipe 43 so as to be lowered so as to expedite the discharge of the processing liquid. A seal member 44 is provided in a through hole portion formed in the center of the bottom surface of the case 33 to allow the support shaft 25 to pass therethrough, and the liquid tightness of this portion is maintained.

The operation of staining a sample by using the microscope specimen staining apparatus of the present invention configured as described above is as follows.

First, the through holes 20 of the first side plates 19, 19.
And the engaging projection 21 on the side surface of the lower horizontal plate 10 are disengaged, the first side plates 19 and 19 are rotated, and the slide glasses 16 and 16 pass through one side opening of the storage container 8. Set it in a free state. In this state, slide glass 16, 16 having the sample to be dyed adhered to the upper surface is inserted from this one side opening,
The first and second support pieces 14 and 15 are supported so as to be bridged between them. After supporting a desired number of slide glasses 16 and 16, a lid (not shown) of the case 33 is opened, and the storage container 8 is put into the case 33 through the opening. At this time, the rotary shaft 26 is in a retracted state and the support shaft 25 is in a lowered state. And the storage container 8
The joint member 23 is opposed to the tip of the rotary shaft 26, and further the rotary shaft 26 is directed toward the joint member 23 and lowered (extended) to connect the rotary shaft 26 and the storage container 8. In this state, the storage container 8 is supported by the rotary shaft 26 with the vertical plates 11 and 12 in a first posture parallel to the vertical axis.

Next, the driving means for expanding and contracting the pressing rod 34 is energized to displace the pressing rod 34 and press the lower end of the vertical plate 12 of the storage container 8. This allows
The storage container 8 has the vertical plates 11 and 12 from the first posture shown in FIG.
Becomes a second posture inclined by an angle θ with respect to the vertical axis a. Since the pressing rod 34 is gradually displaced by the rack and pinion, the storage container 8 is gradually deformed from the first posture to the second posture, and the storage container 8 does not swing due to this deformation. The same applies to the reverse deformation.

As the storage container 8 is moved to the second position, the first and second support pieces 14 and 15 become horizontal, and the first and second support pieces 14 and 14 and the second support pieces 15 and 15 are placed. The slide glasses 16 and 16 are supported in the up-and-down direction between and. At this time, as shown in FIG. 9, the slide glasses 16 and 16 supported in the storage container 8 are directly above the lower slide glass 16 at one end in the length direction (the left end in FIGS. 1 and 9). The slide glass 16 is in a state of protruding from one end in the length direction. Further, a minute gap 17 exists between the vertically adjacent slide glasses 16, 16.

As described above, the slide glasses 16, 16
If the one end in the length direction of the lower slide glass 16 is projected from the one end in the length direction of the slide glass 16 immediately above, the rotary valve 38 and the roller pump 36 are subsequently instructed by the controller. Of the plurality of treatment liquid containers 40, 40 storing various treatment liquids, a predetermined treatment liquid container 40 and the dispensing nozzle 35 are connected to each other. And
From the dispensing nozzle 35 by the operation of the roller pump 36,
The treatment liquid is supplied to the upper surface of one end of the slide glass 16 at the uppermost position.

The treatment liquid spreads on the upper surface of the slide glass 16 and permeates the sample attached to the upper surface of the slide glass 16, and the excess treatment liquid flows down toward the slide glass 16 below. And slide glass 1
The treatment liquid enters into the minute gaps 17 existing between 6 and 16 by the capillary phenomenon and permeates the sample attached to the upper surface of the slide glass 16 immediately below. Thereafter, the treatment liquid flows down in the same manner, and the sample finally attached to the upper surface of the slide glass 16 at the lowest position is permeated by the treatment liquid.

The amount of treatment liquid used in this case is
The required amount is obtained in advance by an experiment, and only the required amount is dispensed from the dispensing nozzle 35. The amount of the processing liquid supplied to the upper surface of one slide glass 16 depends on the size of the minute gap 17, but when the size of the minute gap 17 is 0.3 mm, it is about 0.5 ml. According to the calculation by the inventor of the present invention, in the conventional apparatus, about 1.5 ml is required for each sample, so that the consumption of the processing liquid is 1/3.
Becomes After dispensing the required amount of the processing liquid, the controller immediately stops the operation of the roller pump 36 to prevent the processing liquid from being wasted. The amount of the processing liquid dispensed from the dispensing nozzle 35 is not determined in advance by an experiment, but a photoelectric sensor is provided below the slide glass 16 at the lowermost position so that the processing liquid can be discharged from the slide glass 16. It is also possible to stop the operation of the roller pump 36 when it is detected that it is flowing down. Alternatively, if the roller pump 36 is operated for a certain period of time based on the liquid delivery performance of the roller pump 36, it is assumed that the required amount of processing liquid has been supplied and has reached the slide glass 15 at the lowest position. 36
May be stopped.

As described above, the predetermined treatment liquid is dispensed from the dispensing nozzle 35, and the treatment liquid is allowed to flow down from the uppermost slide glass 16 to the lowermost slide glass 16 in order, and all slide glasses The sample adhered to the upper surface of 16 and 16 is permeated (permeation step). Depending on the type of the sample, the procedure of the dyeing process, etc., there is a case where it is necessary to keep the sample in the treatment liquid for a while. In such a case, the controller, after dispensing the processing liquid, by stopping the processing for a desired time, during this desired time,
Permeate the sample into the process liquid. Further, at this time, it is also possible to displace the pressing rod 34, which is the deforming means, in small steps to improve the familiarity between the sample and the processing liquid.

When the infiltration process is completed, the pressing rod 34 is displaced to the right in FIG. 1 to deform the storage container 8 from the second posture to the first posture. At this time, as described above, the pressing rod 34 is gradually displaced by the rack and pinion, and the pressing rod 34 is temporarily stopped at the position where the storage container 8 is in the first posture, so that the storage container 8 is prevented from swinging. Thus, the work of bringing the tip of the support shaft 25 into contact with the small recess 24 after the storage container 8 is deformed from the second posture to the first posture is facilitated.

When the pressing rod 34 is displaced to a position where the storage container 8 is in the first posture, and the displacement is stopped at this position, the support shaft 25 is raised accordingly, and the small recess of the storage container 8 is raised. 24 and the tip of the support shaft 25 contact. After this, the pressing rod 34 restarts displacement in the same direction again, and the storage container 8
Wait at a position that does not interfere with the rotation of. Since the storage container 8 is in the first posture, the slide glasses 16 and 16 supported in the storage container 8 extend in the vertical direction at regular intervals sufficiently larger than the minute gap 17 and on the four sides of each other. Are in a consistent state.

Then, by driving the motor 29,
The rotating shaft 26 is rotated via the pulleys 30, 28 and the belt 31. The diameter of the pulley 28 is made larger than the diameter of the pulley 30, and the rotation speed of the rotating shaft 26 is increased to prevent the slide glasses 16, 16 in the storage container 8 from rattling with the rotation. The rotation speed is, for example, 10
It is about 0 to 200 rpm. In the present embodiment, an example in which the motor 29, the pulleys 28 and 30, and the belt 31 are used as the driving means for rotationally driving the rotary shaft 26 is shown, but the dyeing device of the present invention is not limited to this. There is no limitation, and various conventionally known means such as connecting the output shaft of the motor to the rotary shaft 26 via a known speed reducer can be adopted.

When the motor 29 is stopped, the storage container 8 is provided with notches 58, 58 at one end in the lengthwise direction of the upper horizontal plate 9 for allowing the processing liquid to pass therethrough. In the state of being located on the side opposite to the pressing rod (left side in FIGS. 1 and 2), stop as shown in FIGS. Therefore, a control mechanism for controlling the stop position of the storage container 8 is additionally provided, for example, a motor 29 for rotating the rotary shaft 26 is a stepping motor. A conventionally known mechanism can be adopted as this control mechanism.

When the rotary shaft 26 rotates, the storage container 8 connected and supported by the joint member 23 at the lower end of the rotary shaft 26 rotates about the rotary shaft 26. At this time, since the tip of the support shaft 25 is in contact with the small recess 24 in the center of the lower surface of the storage container 8, the storage container 8 does not shake as the storage container 8 rotates. Due to the centrifugal force caused by this rotation, the extra treatment liquid that has not been used for dyeing and is present on the upper surfaces of the slide glasses 16 and 16 is scattered. Therefore, by rotating the storage container 8, the excess processing liquid is removed from the upper surfaces of the slide glasses 16 and 16 (removal process). Excess processing liquid scattered from the upper surfaces of the slide glasses 16 and 16 flows on the bottom surface of the case 33 and is stored in the waste liquid container 42 via the discharge pipe 43.

With respect to one treatment liquid, the above-mentioned penetration, removal,
After each step is performed, the controller operates the rotary valve 38 to communicate the processing solution container 40 storing the next processing solution with the dispensing nozzle 35, and again performs each step in sequence. Thereafter, various treatment liquids are sequentially permeated into the sample in the same manner to stain the sample.

In the above-described embodiment, as means for rotating the first side plates 19 and 19, as shown in FIG. 8A, the upper end portions of the first side plates 19 and 19 are arranged. The example in which the notch 22 is formed in the above has been described, but FIG.
A hinge 45 may be used as shown in (C). This Figure 8
In each of the examples shown in FIG. 8B or FIG. 8C, one half of the hinge 45 is pivotally supported by a pivot 46 and the other half to which the first side plate 19 is fixed is shown. 8 (B)
(C) is rotatable in the direction of arrow c. The lower end of the first side plate 19 and the lower lateral plate 10 facing the lower end.
It is the same as in the case of the example shown in FIG. 8A that either the locking portion or the receiving portion is provided on the side surface of each.

Further, as shown in FIGS. 10 to 11, an outward flange-like flange portion 47 is provided on the outer peripheral surface of the end of the joint member 23 provided on one end surface of the storage container 8 and the other end side surface of the storage container 8 is provided. A groove portion 48 having a T-shaped cross section into which the first half portion of the joint member 23 can be inserted is formed, and the groove portion 48 on the other surface side of the one storage container 8 is inserted into the joint member on the one surface side of the other storage container 8. It is also possible to connect the plurality of storage containers 8 in the vertical direction by inserting 23, further inserting the retaining member 49, and further fixing it by means such as screwing.
In this way, if the plurality of storage containers 8 are configured to be connectable,
By increasing the number of the dispensing nozzles 35, a larger number of samples can be dyed at one time, which improves the processing efficiency. The dispensing nozzle 35 faces the notches 58, 58 at the upper ends of the upper and lower storage containers 8, 8, respectively, and dispenses the treatment liquid from the notches 58, 58.

Next, FIG. 12 shows a second embodiment of the dyeing apparatus according to the present invention. In this embodiment, the upper and lower lateral sides are composed of a pair of strip-shaped plate members 50. A pair of plate members 50, which constitute the upper and lower lateral sides,
50 is axially supported by the vertical plates 11 and 12 so as to be parallel to each other. Further, flat plate portions 51 and 51 are provided in the middle portion of the pair of plate members 50 and 50, and the flat plate portions 51 and 51 are provided with the joint member 23, the small recess 24 or the groove portion 48, respectively.
Other configurations and operations are similar to those in the case of the first embodiment described above.

Next, FIG. 13 shows a third embodiment of the dyeing apparatus of the present invention. In the present embodiment, one end portion 5 of the vertical plates 11 and 12 having substantially U-shaped cross-sections that form the left and right vertical sides.
The dimensions of 2, 52 are relatively long. With this configuration, the one side opening of the storage container 8b is made shorter than the length dimension of the slide glasses 16 and 16, and the slide glasses 16 and 16 stored in the storage container 8b come out of this one side opening. Are prevented. That is, this one end 52,
52 functions as a second side plate. On the other hand, the vertical plates 11, 1
The other end portions 53 of the second portion 53 are formed to have a relatively short size. The dimension between the other end portions 53, 53 is made larger than the length dimension of the slide glasses 16, 16, and the slide glasses 16, 16 can be freely taken in and out from the other side opening.

At the upper and lower end portions of the other end portions 53, 53,
The left and right ends of the plate members 50, 50, which are the lateral sides, are axially supported. In the present embodiment, the first side plate is provided between the other end parts 53, 53 and the plate members 50, 50. 19a, 19a
Is provided. That is, the lower end portions of the first side plates 19a, 19a are connected to the lower end portions of the other end portions 53, 53 and the lower plate member 50.
It is rotatably supported between the left and right ends of the. Therefore, the first side plates 19a, 19a are rotatable about the axis support portion in the directions away from each other as shown by the arrow d in FIG.

The upper end portions of the other end portions 53, 53 and the left and right end portions of the upper plate member 50 are located between the surfaces of the upper end portion and the left and right end portions which face each other, and the first side plate. 1
The shaft is supported with a gap slightly larger than the thickness of 9a and 19a. The first side plates 19a, 19
An arcuate notch 54 centered on the shaft supporting portion is provided at the upper end of a so that the openings face each other. Therefore, this first side plate 19a, 19a
Is rotated to a state of standing upright in the vertical direction as shown in FIG. 13, the upper end portions of the first side plates 19a, 19a are
The other end 53, 5 while the pivot is entering the notch 54
3 and the left and right ends of the plate member 50. A small protrusion 55 is provided at an intermediate portion of the notch 54, and when the pivot gets over the small protrusion 55 and reaches the inner end of the notch 54, depending on the rotation of the storage container 8, the first side plate is rotated. The 19a and 19a do not rotate in a horizontal lying state. First side plates 19a, 19a
The width dimension of the first side plates 19a, 19a is set to be approximately the same as the width dimension of the one end portions 52, 52, and in the state where the first side plates 19a, 19a are erected, the slide glasses 16, 16 are not allowed to pass,
Conversely, when lying down, slide glass 16, 1
Allow 6 passes. Other configurations and operations are the same as those in the first and second embodiments described above.

Next, FIG. 14 shows a fourth embodiment of the present invention. In this embodiment, the rotation support member is a turntable 56 which is rotatable in the case 33 about the vertical axis. Positioning members such as an engaging projection and an engaging recess are provided on the upper surface of the turntable 56 and the lower surface of the storage container 8. Further, a small recess is provided in the center of the upper surface of the storage container 8, and the support shaft 5 is provided in this small recess.
The lower end of 7 can enter freely. The support shaft 57 is movable up and down and displaceable in the left-right direction in FIG. 14 by a conventionally known drive device, and the support shaft 57 can be lowered while the storage container 8 is in the first posture. Then, the tip of the support shaft 57 enters the small recess. Further, by sending a command to the controller, the controller descends the support shaft 57 from the solid line state in FIG. 14 to the chain line state in FIG. This allows
The storage container 8 is transformed from the first posture to the second posture, and waits for the processing liquid to be dispensed from the dispensing nozzle 35.

The storage container 8 is deformed to the second position, and one end of the slide glass 16 in the lower direction (left end in FIG. 4) is projected so that the glass slides 16 and 16 are lower, and then the treatment liquid is dispensed. After performing the stroke, the support shaft 57 is displaced from the chain line state in FIG. 14 to the solid line state. This allows
The storage container 8 is deformed from the second posture to the first posture, and the slide glasses 16 and 16 are vertically supported while being aligned with each other. After this, a removal process for removing the excess processing liquid is performed. In this embodiment, this removal process is performed by rotating the turntable 56 that supports and fixes the storage container 8. Other configurations and operations are similar to those of the first embodiment described above.

In the first and second embodiments described above, the upper ends of the first side plates 19 and 19 are pivotally supported, and the lower ends thereof are engageable with the storage container 8. On the contrary, as shown in the third embodiment, the lower end portions of the first side plates 19 and 19 may be pivotally supported, and the upper end portions thereof may be engaged with the storage container 8. Also in the third embodiment, similarly to the first and second embodiments, the upper ends of the first side plates 19a, 19a are pivotally supported, and the lower ends thereof are engaged with the storage container 8. You may be free. In any of the embodiments, when the upper end portions of the first side plates 19 and 19a are pivotally supported, the first side plates 19 and 19a are stored on the workbench for storing the slide glasses 16 and 16. If a holding arm that holds the slide glasses 16 and 16 in a freely rotatable state through the opening on the one end side of the container 8 is provided, the slide glass storing operation becomes easy. As the holding arm, a structure in which the first side plates 19 and 19a are placed on an arm piece provided on a stud so as to be able to move up and down can be considered.

Further, in the first to third embodiments, the example in which the pressing rod 34 which is displaced by the rack and pinion is adopted as the deforming means has been described, but this is displaced by using the solenoid. May be. Further, instead of the pressing rod 34, the storage container 8 may be a pull rod that pulls in the displacement direction side. Further, when the deforming means and the supporting means are interlocked with each other, the supporting means or the deforming means may be moved up and down, or the pressure fluid may be moved due to displacement, or electrically using a relay circuit or the like. .

[0057]

Since the method for staining a microscope specimen and the apparatus therefor according to the present invention are constructed and operate as described above, different kinds of treatment liquids do not mix and deteriorate. Moreover, since the amount of the processing liquid used is small, the consumption amount of the expensive processing liquid is reduced. Therefore, according to the staining method and the apparatus therefor of the present invention, a larger amount of sample can be treated with a small amount of the treatment liquid which is not deteriorated. Further, since the number of containers storing various processing liquids can be minimized, the management of the containers becomes easy. Moreover, the installation space is smaller than that of the conventional device.

[Brief description of drawings]

FIG. 1 is a schematic view showing a first embodiment of a dyeing device according to the present invention.

FIG. 2 is a plan view of the same main part.

FIG. 3 is a view showing a support shaft and a pressing rod which are interlocked with each other.

FIG. 4 is a schematic perspective view of a storage container.

FIG. 5 is a front view of the storage container with the first side plate omitted.

FIG. 6 is a plan view of the storage container.

7 is a cross-sectional view taken along the line AA of FIG.

FIG. 8 is a side view showing a mounting state of the first side plate.

FIG. 9 is a schematic front view showing only the upper half of the second posture of the storage container.

FIG. 10 is a perspective view showing a connecting portion between storage containers.

FIG. 11 is a bottom view of the same.

FIG. 12 is a schematic perspective view of a storage container with a part omitted, showing a second embodiment of the present invention.

FIG. 13 is a schematic perspective view of a storage container showing a third embodiment of the present invention.

FIG. 14 is a schematic front view of a main part, showing a fourth embodiment of the present invention.

FIG. 15 is a perspective view showing a conventional dyeing device.

FIG. 16 is a perspective view of a dyeing basket used in a conventional dyeing device.

[Explanation of symbols]

 1 Substrate 2 Container 3 Guide rail 4 Support beam 5 Moving tool 6 Suspending arm 7 Basket 8, 8a, 8b Storage container 9, 10 Horizontal plate 11, 12 Vertical plate 13 Projection piece 14 First support piece 15 Second support Piece 16 Slide glass 17 Minute gap 18 Second side plate 19, 19a First side plate 20 Through hole 21 Locking protrusion 22 Notch 23 Joint member 24 Small recess 25 Support shaft 26 Rotating shaft 27a, 27b Bearing 28 Pulley 29 Motor 30 Pulley 31 Belt 32 Air cylinder 33 Case 34 Push rod 35 Dispensing nozzle 36 Roller pump 37 First connecting pipe 38 Rotary valve 39 Second connecting pipe 40 Processing liquid container 41 Branch pipe 42 Waste liquid container 43 Discharge pipe 44 Sealing material 45 Hinge 46 Pivot 47 Collar 48 Groove 49 Stop member 50 Plate 51 Flat plate 52 One end 53 The other end 5 Notch 55 notch small projection 56 turntable 57 support shaft 58 cut

Claims (9)

[Claims] 1. A plurality of slide glasses having a sample to be dyed adhered to the upper surface thereof are vertically extended with a minute gap therebetween, and the upper slide glass is closer to one end in the longitudinal direction. This treatment is performed by supporting the slide glass immediately below it in a state of protruding from the one end in the length direction and selecting and dispensing a predetermined treatment liquid to the one end in the length direction of the uppermost slide glass. The liquid is made to flow down from the upper slide glass to the lower slide glass, and the treatment liquid is sequentially permeated into the sample.After this permeation stroke, the slide glasses are vertically aligned and aligned with each other. In a state where the slide glass is supported by interposing a fixed interval larger than the minute gap, and by rotating the plurality of slide glasses about the vertical axis in this state,
A staining method for a microscope specimen, which comprises a removal step for removing an excessive treatment liquid, and sequentially repeats each of these steps a predetermined number of times to stain the sample. 2. A frame-shaped storage container comprising upper and lower horizontal sides and left and right vertical sides and pivotally supporting the ends of the respective sides abutting each other, and a slide glass to which a sample to be dyed is attached. In order to respectively support both ends in the length direction, on the inner side surfaces facing each other of the left and right vertical sides, which are formed at the same pitch in the vertical direction, respectively, the same number of firsts,
The second support piece and one of the upper and lower lateral sides in the width direction one end side, one of the upper and lower end portions at the same inclination as the vertical side,
A rotation unit pivotally supports the storage container in a direction to open the opening in the one end in the width direction of the storage container, and is formed on one of the other upper and lower ends and the upper and lower horizontal sides. A first side plate in which the stop portion and the receiving portion formed on the other side are freely engageable with each other, and the upper and lower end portions thereof are axially supported at the other widthwise other end sides of the upper and lower horizontal sides at the same inclination as the vertical side. The second side plate, a joint member provided on the outer surface of the central portion of either one of the upper and lower ends of the storage container, and the storage member is detachably attached to the joint member, and the storage container is attached to the upper or lower end The rotation support member for rotating the storage container and the supporting means for supporting the outer surface of the central part of the upper or lower end of the storage container when rotating the storage container, and the posture of the storage container. Deformation means for deforming, and disposed above one end in the length direction of the storage container, A dispensing nozzle capable of dispensing the treatment liquid is provided toward one end in the length direction of the slide glass at the upper position, and the storage container has a first posture in which the vertical side is parallel to the vertical axis by the deforming means. , The vertical side is inclined by an angle θ with respect to the vertical axis, so that one end in the lengthwise direction of the upper horizontal side projects beyond the one end in the lengthwise direction of the lower horizontal side. The first and second support pieces are each horizontal when the storage container is in the second posture. The support piece on the other end side in the length direction of the storage container is formed downwards by an angle substantially equal to the angle θ while forming the same upwards at an angle substantially equal to each other. Based on this, it is possible to connect to one of the multiple processing liquid containers via the liquid delivery pump and rotary valve. Comprising Te, dyeing device of the microscopic specimen. 3. The first side plate is made of synthetic resin, and the rotating means is a notch formed at the end of the side plate on the shaft support side.
The staining device for a microscope specimen according to claim 2. 4. A hinge in which the rotating means has one side thereof fixed to one of the upper and lower ends of the first side plate, and the other side is pivotally supported to one side of the upper and lower lateral sides in the width direction. Yes, claim 2
The staining apparatus for microscope specimens according to 1. 5. The first side plate is composed of two plate members,
One end of each of these plate members in the vertical direction is rotatably supported by both ends of one of the upper and lower lateral sides in the width direction on both ends in the length direction, and the plate member is pivotally supported by a pivot shaft. At the other end in the up-down direction, an arcuate cutout having a small projection formed in the middle thereof is provided, and the cutout is provided with the above-mentioned horizontal side of a pivot shaft that axially supports the other horizontal side and the vertical side. The staining device for a microscope specimen according to claim 2, wherein the device is made enterable by passing over a small protrusion in a portion between the vertical side. 6. The microscope specimen according to claim 2, wherein the support means is a support shaft whose tip is abuttable with a small recess provided on the outer surface of the central portion at the other end of either the upper or lower side. Dyeing equipment. 7. A first sensor for detecting that the support means releases the support of the storage container, and a second sensor for detecting that the deformation means has returned the storage container from the second posture to the first posture. And the function of displacing the deforming means so that the storage container takes the second posture by the detection signal of the first sensor, and the support means by the support signal by the detection signal of the second sensor. 7. The staining device for a microscope specimen according to claim 2, which has a function of displacing the support means so as to support the. 8. A central part of the joint member, which has an outward flange-shaped collar portion at its tip, has a substantially T-shaped cross-section, and is provided on either side of the upper and lower sides of the storage container without the joint member. A plurality of storage containers are provided by providing an engaging groove having a substantially T-shaped cross section with a narrow opening side and a wide inner side on the side surface, inserting the joint member into the engaging groove, and further inserting and fixing the retaining member. The dyeing device for a microscope specimen according to claim 2, wherein the dyes are vertically connectable. 9. The rotation support member is a turntable that can fix the lower surface of the storage container and is rotatable about a vertical axis, and the supporting means is a support shaft that supports the central portion of the upper surface of the storage container. The staining device for a microscope specimen according to claim 2, wherein the support shaft is provided as a deformable means by being provided so as to be movable in a three-dimensional direction.