JP2016085173A - Thin section sample preparation device and thin section sample preparation method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a thin section sample preparation device and method which can uniformly apply an appropriate amount of an adhesive liquid on a slide glass and which suppress failure in sticking a thin section on the slide glass and failure in extending the stuck thin section.SOLUTION: A thin section sample preparation device includes: cutting means 31 for cutting a sample block 10 in which a sample 11 is embedded in an embedding agent 12 so as to prepare a thin section 16; temporal holding means 33 for temporally holding the thin section; supply means for supplying an adhesive liquid 19 on a slide glass 17; transfer means 54 for relatively transferring the slide glass and an adhesive liquid supply port 53 of the supply means; position setting means for setting a position of supplying the adhesive liquid on the slide glass; and displacement means for displacing the thin section from the temporal holding means onto the slide glass to which the adhesive liquid has been supplied.SELECTED DRAWING: Figure 1

Description

  The present invention relates to an apparatus and a method for producing a thin slice sample used for physicochemical sample analysis, microscopic observation of a biological sample and the like.

  2. Description of the Related Art Conventionally, a microtome is widely known as an apparatus for producing a thin slice sample used for physicochemical sample analysis or microscopic observation of a biological sample. A microtome is a device that produces a thin slice sample by slicing a sample block (embedded) in an embedding agent such as paraffin into a specimen block (embedded) such as a biological sample of an animal. . For example, Patent Documents 1 to 5 describe a thin-section sample preparation device that performs a series of operations such as slicing a sample block, sticking to a slide glass, stretching, and drying.

JP2012-229993A JP2012-229994A JP 2012-229995 A JP2012-229996A JP 2012-229997 A

  The thin slice cut by the cutter is attached to the slide glass using an adhesive liquid such as water. At this time, an appropriate amount of the adhesive liquid needs to be uniformly applied on the slide glass. If the adhesive liquid is too much or the coating is not uniform, the thin slice may slide on the glass slide and shift its position. If the position of the thin section is shifted, it is inconvenient when the sample is analyzed and observed later. On the other hand, when there is too little adhesive liquid, a thin section may not be stuck well on a slide glass, or a sample may not be extended uniformly in an extension process. Since many animal biological samples are difficult to collect again, it is necessary to avoid wasting samples as much as possible due to poor pasting or poor stretching.

  In applying the adhesive liquid on the slide glass, if the adhesive liquid is supplied from one point on the slide glass by a nozzle or the like, it is difficult to spread the adhesive liquid uniformly on the slide glass. In order to improve the uniformity of the adhesive liquid application, for example, the adhesive liquid can be simultaneously supplied from a plurality of fixed parallel nozzles. However, even in this case, the supply amount of the adhesive liquid varies among the plurality of nozzles, and there is a limit to improving the uniformity of the adhesive liquid application, and there is still room for improvement.

  The present invention has been made in consideration of the above, and it is possible to uniformly apply an appropriate amount of an adhesive liquid on a slide glass. It is an object of the present invention to provide an apparatus and method for preparing a sliced piece sample which is unlikely to cause poor extension.

  The thin-section sample preparation device of the present invention includes a cutting means for cutting a sample block in which a sample is embedded in an embedding agent to prepare a thin section, and a temporary holding means for temporarily holding the thin section. A supply means for supplying an adhesive liquid onto the slide glass, a moving means for relatively moving the slide glass and an adhesive liquid supply port of the supply means, and a supply position of the adhesive liquid supplied on the slide glass. Position setting means for setting, and transposing means for transferring the thin slice onto the slide glass supplied with the adhesive liquid from the temporary holding means.

  Preferably, the supply unit is operable in synchronization with the moving unit.

  More preferably, the moving means is capable of relatively intermittently moving the slide glass and the adhesive liquid supply port, and the supply means is configured such that the slide glass is stationary with respect to the adhesive liquid supply port. The adhesive liquid can be supplied in a state.

  Alternatively, more preferably, the moving means is capable of relatively continuously moving the slide glass and the adhesive liquid supply port, and the supply means is configured such that the slide glass moves relative to the adhesive liquid supply port. The adhesive liquid can be supplied in a relatively moving state.

  Preferably, the supply means has a positive displacement pump.

  Preferably, the supply means is configured such that the adhesive liquid supply port is a single nozzle.

  The thin-section sample preparation method of the present invention includes a step of cutting a sample block in which a sample is embedded in an embedding agent to prepare a thin section, a step of holding the thin section in a temporary holding means, and a slide A moving process for relatively moving the glass and the adhesive liquid supply port, and the adhesive liquid supply port on the slide glass based on a preset position on the slide glass in synchronization with the moving process. And a step of transferring the thin section from the temporary holding means onto the slide glass.

  Preferably, in the step of supplying the adhesive liquid, when the adhesive liquid supply port is on the slide glass, the slide glass and the adhesive liquid supply port are relatively intermittently moved, and the slide glass is supplied. Supplies the adhesive liquid in a stationary state with respect to the adhesive liquid supply port.

  Alternatively, preferably, in the step of supplying the adhesive liquid, when the adhesive liquid supply port is on the slide glass, the slide glass and the adhesive liquid supply port are moved relatively continuously, You may supply the said adhesive liquid in the state which the slide glass is moving relatively with respect to this adhesive liquid supply port.

  Another thin-section sample preparation method of the present invention includes a step of cutting a sample block in which a sample is embedded in an embedding agent, a step of holding the sliced sample in a temporary holding unit, A moving step of relatively moving the slide glass and the adhesive liquid supply port, and the adhesive liquid supply port on the slide glass based on a preset position on the slide glass in synchronization with the moving step And a step of transferring the sliced sample from the temporary holding means onto the slide glass.

  In any one of the above thin-section sample preparation methods, the step of supplying the adhesive liquid is preferably performed using a positive displacement pump. Piezo pumps are also called piezoelectric pumps.

  In any one of the above-described thin-section sample preparation methods, preferably, the step of supplying the adhesive liquid is performed using a single nozzle.

  According to the thin-section sample preparation apparatus or method of the present invention, it is possible to uniformly apply an appropriate amount of the adhesive liquid on the slide glass. Inferior section extension is unlikely to occur.

It is a figure which shows schematic structure of the thin slice sample preparation apparatus of one Embodiment of this invention. 2A is a cross-sectional view of the sample block before rough cutting as viewed from the front direction, FIG. 2B is a plan view of the sample block after rough cutting, and FIG. 2C is a cross-sectional view of the sample block after rough cutting as viewed from the front direction. is there. It is a figure which shows the adhesive liquid supply part of the thin slice sample preparation apparatus of one Embodiment of this invention. It is a figure for demonstrating the adhesive liquid application | coating method in the thin slice sample preparation method of one Embodiment of this invention. It is a figure for demonstrating the other adhesive liquid application | coating method in the thin slice sample preparation method of one Embodiment of this invention. It is a figure which shows the example of the parameter setting screen in the thin section sample preparation method of one Embodiment of this invention.

  An outline of the configuration and operation of a thin-section sample preparation apparatus and method according to an embodiment of the present invention will be described with reference to FIGS.

  In FIG. 1, a thin-section sample preparation apparatus 100 cuts out a thin section 16 from a sample block 10 with a cutter 31 as a cutting means, affixes this to a glass slide 17, and automatically and continuously slides with a thin section. This is an apparatus for producing the glass 18. In the present specification, the slide glass with a thin section may be simply referred to as a “thin section sample”. The thin-section sample preparation device 100 includes a sample block transport unit 20, a sample block storage unit 21, a height detection unit 22, an imaging unit 23, a cutting unit 30, an adhesive liquid application unit 50, a thin section application unit 60, an extension unit 65, A slide glass conveyance unit 70 and a control unit 80 for controlling the entire apparatus are provided.

  The sample block 10 is obtained by embedding a sample 11 in an embedding agent 12 such as paraffin. Examples of the sample 11 that is the subject include biological samples such as human and animal tissues. The sample block 10 generally has a surface size of 24 mm × 24 mm, 24 mm × 30 mm, 24 mm × 37 mm. Further, the height of the sample block 10 is generally about 5 mm.

  In FIG. 1, a plurality of sample blocks 10 can be stored in the sample block storage unit 21. One sample block 10 selected from the plurality of sample blocks 10 stored in the sample block storage unit 21 passes through the position A (height detection unit 22) and the position B (imaging unit 23) by the sample block transport unit 20. It is conveyed to position C (cutting unit 30). The sample block transport unit 20 is configured to be able to reciprocate the sample block 10 between positions A to C after taking out the sample block 10 and transporting it to the position A. Further, the sample block transport unit 20 is configured to be able to adjust the inclination and height position of the sample block 10. After the inclination (inclination) of the sample block 10 with respect to the horizontal plane is first adjusted, the conveyance unit 20 controlled by the control unit 80 adjusts the height of the sample block 10 appropriately according to the cutting amount.

  At position A, a height detector 22 for detecting the inclination and height position of the sample block 10 is arranged. As the height detection part 22, what detects the surface of the sample block 10 with the at least 3 contact-type sensor which is described in patent document 4 and is not on the same straight line can be used, for example. Based on the sensor information, the control unit 80 controls the transport unit 20 to adjust the inclination and height position of the sample block 10.

  An imaging unit 23 is disposed at the position B. As the imaging unit 23, for example, as described in Patent Document 2, a unit having a light source and a CCD camera can be used. By processing the image data obtained by the imaging unit 23, the control unit 80 determines whether the area of the portion of the sample 11 exposed on the surface of the sample block 10 is sufficient to cut out the thin slice. . Hereinafter, exposing the sample 11 to the surface of the sample block 10 by cutting is referred to as “surface exposure”, and determining whether the area of the exposed sample 11 is sufficient for cutting out a thin slice is “surface determination”. That's it. As a reference for the surface determination, for example, when the exposed area of the sample 11 on the cutting surface of the sample block 10 is larger than a preset area ratio, it can be determined that the surface is sufficient.

  The cutting part 30 is arranged at the position C. The cutting unit 30 has a cutter 31 that can slice a surface layer portion of the sample block 10. The cutter 31 is held such that the cutting edge extends in a direction perpendicular to the paper surface in FIG. In this embodiment, the cutter 31 is fixed, and the surface layer portion of the sample block 10 is sliced by moving the sample block 10 in the right direction in FIG. The cutter 31 may be configured to be movable in a direction perpendicular to the paper surface. In that case, the surface of the sample block 10 is sliced by moving the sample block 10 in the right direction in FIG.

  Here, the cutting method of the sample block 10 is summarized as follows. In FIG. 2, a sample block 10 is obtained by embedding a sample (subject) 11 in an embedding agent 12 such as paraffin. FIG. 2A is a cross-sectional view of the sample block before rough cutting as viewed from the front, and the sample 11 is embedded in the embedding agent 12 so as not to be exposed to the outside (or slightly exposed). The sample block 10 stored in the storage unit 21 is normally in this state. In the sample block 10, the surface layer portion is roughly cut by the cutting unit 30 until the exposed area on the surface of the sample 11 is equal to or larger than a preset area (dotted line 14 in FIG. 2A). The roughened sample block is subjected to surface determination by the imaging unit 23 (FIG. 2B). When the surface is insufficient, rough cutting and surface determination are repeated. When the chamfering is sufficient, the cutting unit 30 performs discard cutting or main cutting for cutting out the thin section 16 (dotted line 15 in FIG. 2C). Note that the sample block 10 is transported on a sample block table 13 not shown in FIG. It should also be noted that the scale of FIG. 2 is not accurate and is stretched in the height direction for ease of explanation.

  The thin slice 16 cut out by the cutting unit 30 has a very thin thickness of about 3 to 10 μm. For this reason, usually, the main cutting is performed after performing a plurality of times of discarding with the same thickness and conditions as the main cutting. This is because the thickness of the thin section 16 is stabilized by performing the discarding. In particular, when different cutters 31 are used for rough cutting and main cutting, when different positions of the cutting edge of one cutter 31 are used, or when the thickness of rough cutting differs from the thickness of main cutting, a thin slice to be produced In order to improve the thickness accuracy, it is preferable to perform several times of cutting with the thickness of the main cutting at the position of the cutter and the blade edge used for the main cutting. In some cases, discarding may be omitted.

  Moreover, when cutting the sample block 10, the humidity of a sample block is also important, and it is preferable to humidify before cutting. From the viewpoint of processing time (cycle time), etc., humidification is performed by providing a humidifier (not shown) between position B and position C in FIG. 1, and the sample block is transported to position C for scraping or main cutting. It is preferable to perform at the timing.

  In FIG. 1, a carrier tape 33 that temporarily holds a thin section 16 obtained by slicing the surface layer portion of the sample block 10 with a cutter 31 is supplied to a position C. The carrier tape 33 is fed out from the supply reel 32, guided by the guide rollers 34 and 35, and supplied above the sample block 10. The carrier tape 33 holding the thin section 16 is guided by the guide rollers 36 to 39 and taken up on the take-up reel 40.

  The thin slice 16 produced by the main cutting operation is attached to the carrier tape 33. At this time, it is preferable that the surface of the carrier tape is subjected to treatments such as humidification, cooling, and charging so that the thin section 16 is more reliably attached to the carrier tape 33.

  On the other hand, the slide glass 17 is conveyed from the slide glass storage unit (not shown) to the adhesive liquid application unit 50. In the application unit 50, the adhesive liquid 19 such as water is sent from the tank 51 by the pump 52, supplied from the nozzle 53, and applied onto the slide glass 17. The slide glass moves relative to the nozzle by the moving means 54. Details of the application unit 50 will be described later.

  The slide glass 17 to which the adhesive 19 is applied moves to the pasting part 60. The thin section 16 held on the carrier tape 33 is pasted on the slide glass 17 by the thin section pasting section 60 disposed between the guide rollers 38 and 39. The thin-section pasting portion 60 includes a pair of guide rollers 61 and 61 disposed on the upstream side of the traveling path of the carrier tape 33, and a pair of guide rollers 62 and 62 disposed on the downstream side of the traveling path of the carrier tape 33. It has. The thin section pasting portion 60 is bent downward with the carrier tape 33 sandwiched between the pair of guide rollers 61 and 61 and the pair of guide rollers 62 and 62, and the thin section 16 held by the carrier tape 33 is bent. Then, it is brought into contact with the slide glass 17 coated with the adhesive liquid 19. As a result, the thin slice is transferred from the carrier tape onto the slide glass, that is, transferred. At this time, the thin slice is transferred from the carrier tape onto the slide glass by the surface tension of the adhesive liquid. Therefore, if the amount of the adhesive liquid is insufficient or the coating is not uniform, the carrier tape cannot be successfully transferred to a predetermined position on the slide glass, and sticking failure tends to occur.

  The slide glass 18 with a thin section is conveyed to the extension unit 65 by the slide glass conveyance unit 70. The extension unit 65 includes a heating plate (not shown), extends the heel of the thin section 16, evaporates moisture on the slide glass 17, and tightly fixes the thin section 16 to the slide glass 17. Here, since the thin slice extends by being heated through the adhesive liquid 19, if the adhesive liquid does not exist uniformly between the slide glass and the thin slice, the extension is insufficient or non-uniform. Prone to poor stretch.

  The operation of each component of the thin-section sample preparation device 100 of this embodiment is controlled by the control unit 80. The control unit 80 controls the operation of each component based on information input to an input unit (not shown). The input unit is configured to be able to input, for example, the thickness of the thin section, the number of manufactured slide glasses with thin sections, and the like.

  The device 100 includes a housing and a cooler (not shown), and the inside of the device surrounded by the housing is cooled by the cooler and air-conditioned so as to have a constant temperature. This is to prevent unevenness in the thickness of the thin section because the sample block is easily expanded or contracted by a subtle change in temperature. The temperature setting of the air conditioning is appropriately set according to the type of the sample 11 and the embedding agent 12 used as the sample block 10. The set temperature is typically in the range of 10 ° C to 25 ° C. If the temperature of the sample block 10 is too high, there arises a problem that the quality of the thin slice sample is deteriorated due to the soft embedding agent. In addition, if the temperature difference between the temperature inside the apparatus and the sample block 10 is large, the sample block 10 contracts and expands due to a temperature change, which causes a problem that the variation in thickness between thin slices produced continuously increases. Furthermore, if the temperature of the sample block 10 is too low, a problem such as breaking of a thin section occurs.

  Further, it is possible to determine whether or not the sample block 10 is facing and to check whether or not the surface temperature of the sample block is within a predetermined range. For example, a non-contact temperature sensor or the like may be provided between position B to position C, the surface temperature of the sample block may be measured, and scraping or main cutting may be performed only when the temperature is within a predetermined temperature range. . When the surface temperature of the sample block is outside the predetermined range, for example, the sample block transport unit 20 can be stopped to wait for the surface temperature to enter the predetermined range by air conditioning. The reference temperature range can be set in advance according to the type of embedding agent such as paraffin.

  Next, the structure and operation | movement detail of the adhesive liquid application part 50 are demonstrated based on FIGS.

  In FIG. 3, the adhesive liquid application unit 50 of this embodiment includes a tank 51, a pump 52, and a nozzle 53. A tank, a pump, and a nozzle comprise adhesive liquid supply means with other peripheral members, such as piping. The adhesive liquid 19 is supplied onto the slide glass 17 from a tank, which is an adhesive liquid supply port, via a pump. The positions of the slide glass and the nozzle can be relatively moved by the moving means 54. The adhesive liquid supply means and the movement means operate in synchronization.

  As the adhesive liquid 19, water is usually used. In addition, a weakly acidic liquid such as a 0.1% aqueous acetic acid solution may be used depending on the type of sample. However, the adhesive liquid used in the thin-section sample preparation apparatus or method of the present embodiment is not particularly limited, and an appropriate one can be selected according to the type of sample.

  As the tank 51, various containers that are not affected by the adhesive liquid 19 and do not contaminate the adhesive liquid can be used.

  The pump 52 sends the adhesive liquid 19 from the tank 51 to the nozzle 53. The amount of the adhesive liquid supplied on the slide glass 17 is typically several tens of μL. The amount of the adhesive liquid supplied is adjusted by the control unit (80 in FIG. 1) controlling a pump, an on-off valve (not shown), and the like. For example, the type of the pump is not particularly limited, and the adhesive liquid supply amount can be adjusted by controlling only the opening and closing of the valve. Preferably, a positive displacement pump is used as the pump. This is because the supply amount of the adhesive liquid can be easily controlled by turning on and off the operation of the pump by the control unit. Examples of positive displacement pumps include piston pumps, plunger pumps, diaphragm pumps, gear pumps, and the like. Among them, the diaphragm pump is preferable because it has a simple structure and can adjust the supply amount of the adhesive liquid with a small amount even with high accuracy. Furthermore, a piezo pump in which the diaphragm (partition) of the diaphragm pump is vibrated by a piezoelectric element is particularly preferable. Piezo pumps can be reduced in size, and diaphragms with a small area can be vibrated at tens of Hz, which is a higher frequency than other diaphragm pumps. This is because it can be adjusted with high accuracy. In addition, the piezo pump has an advantage that it is easy to calibrate the amount of adhesive liquid supplied, and this is also suitable for accurately supplying an appropriate amount of a small amount of adhesive liquid.

  It is also preferable to use a syringe pump as a special example of the positive displacement pump. A syringe pump is a device capable of continuously or intermittently feeding liquid filled in a syringe (syringe) by controlling a piston with a stepping motor or the like and accurately feeding the liquid. The syringe pump is also suitable for the pump of this embodiment because it can accurately adjust the supply amount of a very small amount of liquid.

  Of the members constituting the pump 52 and the pipe, the member in contact with the adhesive liquid preferably has resistance to the adhesive liquid. For example, when using an acetic acid aqueous solution, it is preferable to have acetic acid resistance. In the case of using a diaphragm pump, it is preferable that members such as a pump chamber wall, a diaphragm, and a check valve at the entrance and exit of the chamber have resistance to the adhesive liquid. This is because, when these members are eroded, the adhesive liquid supply amount changes with time. As such a member, it is possible to use polyether ether ketone (PEEK); polyphenylene sulfide (PPS); polytetrafluoroethylene (PTFE), fluororesin such as perfluoroelastomer.

  The shape of the nozzle 53 is not particularly limited. Preferably, in order to supply an accurate amount of the adhesive liquid, the liquid has a good shape. For example, liquid breakage can be improved by using a small-diameter nozzle or by making the tip of the nozzle tapered. Moreover, as a material of the nozzle, it is preferable to use a synthetic resin resistant to various chemicals.

  The moving means 54 relatively moves the slide glass 17 and the nozzle 53 (adhesive liquid supply port). In order to move both relatively, the slide glass may be fixed and the nozzle may be moved. In this case, the nozzle may be moved on the slide glass in the longitudinal direction of the slide glass along the center line in the short width direction of the slide glass. Alternatively, the nozzle may be fixed and the slide glass may be moved. In this case, the slide glass may be moved under the nozzle in the longitudinal direction of the slide glass. Alternatively, both the slide glass and the nozzle may be moved. In short, it is sufficient that the positional relationship between the slide glass and the nozzle can be relatively changed, and the position on the slide glass to which the adhesive liquid is supplied can be changed. Further, it is only necessary that the adhesive liquid can be sequentially supplied to a plurality of positions.

  Further, the moving means 54 can operate in synchronism with the aforementioned adhesive liquid supply means. Thereby, the adhesive liquid 19 can be supplied while moving the position of the nozzle 53 on the slide glass 17.

  FIG. 4 shows a method of supplying the adhesive liquid 19 while relatively moving the slide glass 17 and the nozzle 53 continuously in one direction. FIG. 4 shows an example in which the nozzle is fixed and the slide glass is moved. In FIG. 4A, the slide glass 17 is moving below the nozzle 53 in the longitudinal direction of the slide glass in the right direction in the figure. In FIG. 4B, when the nozzle reaches a predetermined position on the slide glass, the adhesive liquid is supplied from the nozzle. In FIG. 4C, the slide glass continuously moves to the right in the figure, and the adhesive liquid continues to be supplied from the nozzle. In FIG. 4D, when the nozzle reaches a predetermined position on the slide glass, the supply of the adhesive liquid from the nozzle is stopped. In FIG. 4E, the slide glass continues to move further to the right in the figure, during which the adhesive liquid on the slide glass spreads uniformly.

  Moreover, when the moving means 54 has the nozzle 53 on the slide glass 17, it is preferable that the slide glass and the nozzle can be moved relatively and intermittently. That is, it is preferable that the position on the slide glass facing the nozzle can be relatively moved and stopped. For example, it is preferable that the position of the nozzle can be repeatedly moved and stopped on the slide glass by a predetermined amount. At this time, the adhesive liquid can be sequentially supplied to a plurality of positions on the slide glass by supplying the adhesive liquid in a state where the nozzle position is stationary with respect to the slide glass. As the moving means or the supplying means, a means capable of three-dimensional movement such as a three-dimensional robot is used, and the nozzle or the slide glass is held by the robot arm and moved relatively to a plurality of positions on the slide glass. You may enable it to supply an adhesive liquid sequentially.

  FIG. 5 shows a method for supplying the adhesive liquid 19 while relatively moving the slide glass 17 and the nozzle 53 intermittently in one direction. FIG. 5 shows an example in which the nozzle is fixed and the slide glass is moved. In FIG. 5A, the slide glass 17 has moved below the nozzle 53 in the longitudinal direction of the slide glass and to the right in the figure. In FIG. 5B, the slide glass stops at a predetermined position, and the adhesive liquid 19 is supplied from the nozzle while the nozzle is stationary with respect to the slide glass. In FIG. 5C, the slide glass moves again in the right direction in the figure. In FIG. 5D, the slide glass stops again at the next predetermined position, and the adhesive liquid 19 is supplied from the nozzle while the nozzle is stationary with respect to the slide glass. Thereafter, the movement and stop of the slide glass and the supply of the adhesive liquid are repeated. In FIG. 5E, the slide glass stops at the last predetermined position, and the adhesive liquid 19 is supplied from the nozzle while the nozzle is stationary with respect to the slide glass. In FIG. 5F, the slide glass continues to move further to the right in the figure, during which the adhesive liquid on the slide glass spreads uniformly.

  As shown in FIGS. 4 and 5, the moving means 54 and the adhesive liquid supply means are operated in synchronization to supply the adhesive liquid while relatively moving the slide glass 17 and the nozzle 53, thereby sliding the slide. The adhesive liquid can be more uniformly applied on the glass. In addition, since the adhesive liquid can be applied only to the necessary part on the slide glass, by applying the adhesive liquid only to the position where the thin section is applied, the thin section exceeds the range where the adhesive liquid was applied. It can be prevented from moving. Moreover, when printing a sample number etc. on the edge part of a slide glass, it can not apply | coat an adhesive liquid to the printing area | region.

  In addition, as shown in FIG. 5, when the adhesive liquid is supplied while moving the slide glass 17 and the nozzle 53 relatively intermittently (hereinafter, sometimes referred to as “intermittent method”), the bonding is performed. The supply amount and position of the liquid can be controlled more precisely. According to the intermittent method, since the nozzle is temporarily stopped with respect to the slide glass, the supply position of the adhesive liquid can be determined more accurately. Furthermore, since the adhesive liquid is supplied in a state where the nozzle is stationary with respect to the slide glass, the adhesive liquid can be reliably supplied to the position. In the method of supplying the adhesive liquid while moving the slide glass and the nozzle relatively continuously (hereinafter sometimes referred to as “continuous method”), the adhesive liquid slides after the pump starts feeding the adhesive liquid. Since the slide glass moves while reaching the glass surface, the actual adhesive liquid supply position may deviate from a predetermined position, and fine adjustment may be necessary. On the other hand, in the intermittent method, it is easy to set the adhesive liquid supply position.

  Moreover, when supplying an adhesive liquid, moving the slide glass 17 and the nozzle 53 relatively intermittently, it is preferable to use a diaphragm pump as the pump 52, and it is more preferable to use a piezo pump. Since these pumps can adjust the supply amount of a small amount of adhesive liquid with high accuracy, it is possible to obtain a greater effect on the uniformity of adhesive liquid application by combining with the intermittent method that can accurately position the supply position. is there. As described above, the amount of adhesive liquid supplied on one slide glass is about several tens of μL. If this is divided and supplied in several places, the amount of adhesive liquid supplied to each position will be about 1-50 microliters. Thus, when the amount of the adhesive liquid supplied to one place is 1 to 50 μL, further 3 to 30 μL, particularly 5 to 20 μL, it is preferable to use a diaphragm pump as a pump, and use a piezo pump. More preferably.

  As the mechanical structure for realizing the moving means 54, various known structures for moving the slide glass and / or the nozzle can be used. In addition, the moving means 54 may be comprised as a part of slide glass conveyance part (70 of FIG. 1), and may be provided independently from the slide glass conveyance part.

  In the present embodiment, the position and supply amount for supplying the adhesive liquid 19 on the slide glass 17 can be set in advance. Since there is a preferable position for attaching the thin section 16 on the slide glass by an apparatus for analyzing and observing the sample later, the adhesive liquid supply position on the slide glass is appropriately set accordingly. Specifically, a desired set value is input from the input unit of the control unit 80 described above. When supplying the adhesive liquid while moving the slide glass and the nozzle relatively continuously, for example, when there are a supply start position, a supply end position, and a plurality of supply positions, the respective supply start positions and supply end positions. Input the relative movement speed of the slide glass and the nozzle, the adhesive liquid supply speed, and the like. When supplying the adhesive liquid while moving the slide glass and the nozzle relatively intermittently, for example, input a plurality of supply positions, the amount of adhesive liquid supplied at each supply position, etc.

  FIG. 6 shows an example of the setting screen in the case where the adhesive liquid is supplied while the slide glass and the nozzle are relatively intermittently moved on the touch panel screen which is an input unit of the control unit 80. The picture of the slide glass on the left side of FIG. 6 shows marks corresponding to 10 positions on the slide glass. By switching ON / OFF for each mark, the position where the adhesive liquid is actually supplied is shown. You can choose. Below the picture of the slide glass, the pitch of the mark (“dropping pitch” in the figure) and the distance from the right end of the slide glass to the right end mark (“dropping position” in the figure) can be entered. On the right side of FIG. 6, the amount of adhesive liquid supplied to each mark position on the slide glass (“drop amount” in the figure), the adhesive liquid purge amount during the initialization operation after turning on the apparatus power (“purge amount” in the figure) (Initial) "), an adhesive liquid purge amount (" purge amount "in the figure) to be performed every time one slide glass is processed can be input. Although FIG. 6 is an example, it is preferable that the adhesive liquid supply position and the supply amount on the slide glass can be easily set on the touch panel screen as described above.

  In the present embodiment, the adhesive liquid is supplied using a single nozzle, but the number of nozzles may be two or more. Even in that case, the adhesive liquid can be supplied while relatively moving the slide glass and the nozzle. However, if the piping from the pump is branched and connected to multiple nozzles, the amount of adhesive liquid supplied varies from nozzle to nozzle, so it is more uniform to sequentially supply to multiple adhesive liquid supply positions using a single nozzle. An adhesive liquid can be applied. Also, the use of a single nozzle is superior in terms of the simplicity of the device structure and the control. An advantage of using a plurality of nozzles is that processing time (cycle time) may be shortened.

  The present invention is not limited to the above-described embodiment, and various modifications can be made within the scope of the technical idea.

  For example, in FIG. 4, without stopping the movement of the slide glass 17, the supply of the adhesive liquid 19 was started (FIG. 4B) and stopped (FIG. 4D). On the other hand, the slide glass is temporarily stopped at the supply start position and supply stop position of the adhesive liquid, and the supply of the adhesive liquid is started or stopped. Between the positions, the slide glass is continuously moved while being bonded. You may make it continue supply of a liquid.

  Further, for example, the thin section temporary holding means is not limited to the one using a carrier tape, and the carrier member may be in the form of a sheet or film. The material of the carrier member is not particularly limited, although resin or paper is generally used.

  In addition, for example, the method and apparatus configuration relating to surface determination and temperature determination can be variously changed including the presence or absence thereof.

10 Sample block 11 Sample (subject)
12 Embedding agent 13 Sample block base 14 Rough cutting height 15 Discarded or main cutting height 16 Thin section 17 Slide glass 18 Slide glass with thin section 19 Adhesive liquid 20 Sample block transport section 21 Sample block storage section 22 Height detection section 23 Imaging unit 30 Cutting unit 31 Cutter 32 Supply reel 33 Carrier tape (temporary holding means)
34 to 39, 61, 62 Guide roller 40 Take-up reel 50 Adhesive liquid application part 51 Tank 52 Pump 53 Nozzle (adhesive liquid supply port)
54 Moving means 60 Thin section pasting section 65 Extending section 70 Slide glass transport section 80 Control section 100 Thin section sample preparation device

Claims (12)

Cutting means for cutting a sample block in which a sample is embedded in an embedding agent to produce a thin slice;
Temporary holding means for temporarily holding the thin section;
Supply means for supplying an adhesive liquid onto the slide glass;
Moving means for relatively moving the slide glass and the adhesive liquid supply port of the supply means;
Position setting means for setting the supply position of the adhesive liquid to be supplied onto the slide glass;
A transposing means for transferring the thin section from the temporary holding means onto a slide glass supplied with the adhesive liquid;
A thin-section sample preparation apparatus. The supply means is operable in synchronism with the moving means;
The thin-section sample preparation apparatus according to claim 1. The moving means is capable of relatively intermittently moving the slide glass and the adhesive liquid supply port,
The supply means can supply the adhesive liquid in a state where the slide glass is stationary with respect to the adhesive liquid supply port.
The thin-section sample preparation apparatus according to claim 2. The moving means is capable of relatively continuously moving the slide glass and the adhesive liquid supply port,
The supply means can supply the adhesive liquid in a state where the slide glass is moved relative to the adhesive liquid supply port.
The thin-section sample preparation apparatus according to claim 2. The supply means has a positive displacement pump;
The thin-section sample preparation apparatus as described in any one of Claims 1-4. The supply means is configured such that the adhesive liquid supply port is a single nozzle.
The thin-section sample preparation device according to any one of claims 1 to 5. Cutting a sample block in which a sample is embedded in an embedding agent to produce a thin section;
Holding the thin section in a temporary holding means;
A moving step of relatively moving the slide glass and the adhesive liquid supply port;
In synchronization with the moving step, based on a preset position on the slide glass, supplying an adhesive liquid from the adhesive liquid supply port on the slide glass;
Transposing the thin section from the temporary holding means onto the glass slide;
A method for preparing a sliced piece sample. In the step of supplying the adhesive liquid, when the adhesive liquid supply port is on the slide glass, the slide glass and the adhesive liquid supply port are relatively intermittently moved so that the slide glass is bonded to the slide glass. Supplying the adhesive liquid in a stationary state with respect to the liquid supply port;
The thin-section sample preparation method according to claim 7. In the step of supplying the adhesive liquid, when the adhesive liquid supply port is on the slide glass, the slide glass and the adhesive liquid supply port are relatively continuously moved so that the slide glass is bonded to the slide glass. Supplying the adhesive liquid in a state of moving relative to the liquid supply port;
The thin-section sample preparation method according to claim 7. Cutting the sample block in which the sample is embedded in the embedding agent with a cutting means;
Holding the sliced sample in a temporary holding means;
Sequentially supplying the adhesive liquid to a plurality of positions on the slide glass;
Transferring the sliced sample from the temporary holding means onto the glass slide;
A method for preparing a sliced piece sample. The step of supplying the adhesive liquid is performed using a positive displacement pump.
The thin-section sample preparation method as described in any one of Claims 7-10. The step of supplying the adhesive liquid is performed using a single nozzle.
The thin-section sample preparation method as described in any one of Claims 7-11.

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