JP2006025767A - Reaction-treating device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a reaction-treating device by using in a plurality of reagents, capable of accurately treating with these reagents. <P>SOLUTION: This reaction-treating device 40 is a device for treating a biomaterial by supplying a plurality of kinds of all purpose reagents and a plurality of kinds of non-all purpose reagents to a treating tool 1 housing the biomaterial. The reaction-treating device 40 is equipped with a device main body 41, a first dispenser head 42 for sucking/discharging liquid in a disposable nozzle chip mounted as demountable, a second dispenser head 43 having a not disposable fixed nozzle, and a head-transferring mechanism 44 for transferring the first dispenser head 42 and the second dispenser head 43 relative to the device main body 41. The plurality of non-all purpose reagents are dispensed into the treating tool 1 by using the first dispenser head 42 while exchanging the nozzle chip for each of the kinds, and the plurality of all purpose reagents are dispensed into the treating tool 1 by the second dispenser head 43 commonly from the fixed type nozzle irrespective to their kinds. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to automation of a reaction processing apparatus.

  With the development of bio-related fields, there is a method for detecting biological substances such as viruses, cells, genes, proteins, glycoproteins, hormones, biochemical substances in samples (sample liquids) such as blood, lymph, saliva and urine. Many have been developed.

  For example, an in situ hybridization method is known as a method for detecting a gene.

  In situ hybridization is a method that targets chromosomal DNA (deoxyribonucleic acid) of cells or mRNA (ribonucleic acid) in tissues to form a hybrid with an RNA probe or DNA probe. This is a method of detecting the location or detecting a localized region of mRNA in the tissue.

  Here, an RNA probe or a DNA probe is a base sequence that specifically binds to a specific site such as chromosomal DNA of a cell or mRNA in a tissue, and is a radioisotope or fluorescence for detecting the presence of the base sequence. A label such as a substance is chemically bonded.

  A specific operation of such an in situ hybridization method includes, for example, a first step of fixing the cell so that the tissue structure of the cell (biological material) does not collapse during the operation, and the probe reacts with the nucleic acid in the cell. In order to make it easier to remove, the second step of removing unnecessary portions of the cell and the partial decomposition treatment with a proteolytic enzyme, the third step of reacting the probe with the nucleic acid in the cell, and removing the excess probe by washing 4th step to perform, 5th step of reacting with a reagent such as an enzyme-labeled antibody that binds to the labeled product in order to detect the labeled product in the probe hybridized with the nucleic acid in the cell, and washing the excess antibody or the like 6th step of removing, 7th step of reacting enzyme and enzyme substrate via antibody, 8th step of washing and removing excess enzyme substrate, and image of enzyme substrate colored by enzyme reaction Shoot It is performed by a ninth step of recording the like.

  Among these, the 2nd process-the 8th process, after making a cell (biological substance) contact the processing liquid (reaction liquid and washing liquid) containing a reagent, these liquids are used as the processing liquid used at the next process. This is done by repeating operations such as replacement.

  Therefore, in order to efficiently perform the in situ hybridization method, it is important that the replacement operation of these solutions can be performed for many biological substances at once.

  Thus, for a large number of biological materials, as a reaction device (processing device) that can collectively replace the treatment liquid, each of the biological materials and the treatment liquid is accommodated in a plurality of capsule containers, and the treatment liquid can be replaced. There has been proposed a method in which the processing liquid is removed by mechanical suction and a new processing liquid is supplied.

  However, in this apparatus, a weak biological material may be damaged by a suction operation of the treatment liquid, and the apparatus itself is expensive and lacks versatility.

  In addition, since this apparatus uses a large number of reagents, it has been strongly desired to appropriately control the temperature when processing these reagents while avoiding contamination and holding the reagents as necessary.

  For example, a reaction vessel (processing tool) that can easily perform the in situ hybridization method as described above when processing a biological material has been proposed (see, for example, Patent Document 1).

  As shown in FIG. 13, the reaction vessel 901 includes a hollow tube 902, a pore plate 903 provided inside the hollow tube 902, a hollow tube 904 inserted into the hollow tube 902, It consists of a pore plate 905 provided at the tip of the hollow tube 904.

  This reaction vessel 901 is used as follows. First, a liquid (sample solution) containing a biological material is stored in the hollow tube 902, and the hollow tube 904 is inserted into the hollow tube 902. As a result, a liquid (sample solution) containing a biological material is stored in a space defined by the pore plate 903 and the pore plate 905.

  Next, in this state, various processing liquids are sequentially supplied into the space through the pore plate 905. Thereby, the processing liquid in the space is sequentially replaced, and the biological material is processed by each processing liquid.

  The actual situation is that the series of processing operations as described above are performed manually and require time and effort.

Japanese Patent Laid-Open No. 2003-169662

  An object of the present invention is to provide a reaction processing apparatus capable of accurately processing these reagents using a plurality of reagents.

Such an object is achieved by the present inventions (1) to (7) below.
(1) A reaction processing apparatus capable of processing a biological material by supplying a plurality of types of general-purpose reagents and a plurality of types of non-general-purpose reagents to a processing tool containing a biological material,
The device body,
A first dispensing head that detachably mounts a disposable nozzle tip and sucks and discharges liquid into the nozzle tip;
A second dispensing head having a fixed nozzle capable of discharging liquid and used a plurality of times;
A head moving mechanism for moving the first dispensing head and the second dispensing head relative to the apparatus main body,
The plurality of types of non-general-purpose reagents are dispensed to the processing tool by the first dispensing head while exchanging the nozzle tips for each type. Regardless of the type, the reaction head is configured to dispense from the fixed nozzle to the processing tool in common by a dispensing head.

  (2) The reaction processing apparatus according to (1), further including a temperature adjustment unit that adjusts a temperature of at least one of the general-purpose reagent and the non-general-purpose reagent.

  (3) The reaction processing apparatus according to (2), wherein the temperature adjustment unit includes a cooling storage unit that stores the non-general-purpose reagent in a cooled state.

  (4) A storage unit that stores the general-purpose reagent for each type, a liquid feed pump that supplies the general-purpose reagent stored in the storage unit to the fixed nozzle, and a flow from the storage unit to the fixed nozzle A flow path switching unit that switches a path, and the type of the general-purpose reagent to be dispensed by the fixed nozzle is selected by switching the flow path switching unit. Reaction processing equipment.

  (5) The reaction processing apparatus according to (4), further including a heater for heating the general-purpose reagent supplied to the fixed nozzle.

  (6) Control for controlling the operation of the first dispensing head and the second dispensing head so that the non-general reagents and the general reagents are supplied to the processing tool in a predetermined order. The reaction processing apparatus according to any one of (1) to (5), comprising means.

(7) The treatment tool includes a cylindrical body including a discharge port capable of discharging a liquid at a tip portion;
A first filter fixed near the outlet in the cylinder;
The second filter is provided on the proximal end side of the first filter in the cylinder so as to be movable in the axial direction of the cylinder, and can hold a liquid in a space formed between the first filter and the first filter. And the filter
The reaction processing apparatus according to any one of (1) to (6), further including an operation member that moves the second filter in an axial direction of the cylindrical body.

  According to the present invention, when dispensing a non-general reagent into a treatment tool, since the disposable nozzle tip is exchanged between different types of non-general reagents, contamination between these reagents is performed. Can be reliably prevented.

  In addition, when the temperature adjusting unit is provided, the temperature of the non-general-purpose reagent that is easily decomposed at room temperature can be adjusted, and thus the non-general-purpose reagent can be appropriately managed.

  In addition, when dispensing multiple types of general-purpose reagents to the processing tool, since the fixed nozzle is used in common regardless of the type, the cost of the nozzle tip, which is a consumable item, is not incurred. Since the time for replacing the nozzle tip is not required, the processing efficiency can be improved.

  For this reason, according to the present invention, biological materials can be processed easily and accurately. Further, the processing cost is low.

  Hereinafter, the reaction processing apparatus of this invention is demonstrated in detail based on suitable embodiment shown to an accompanying drawing.

  FIG. 1 is a perspective view showing an embodiment of the reaction processing apparatus of the present invention, and FIG. 2 is a piping diagram in the reaction processing apparatus shown in FIG.

  The reaction processing apparatus 40 of the present invention shown in FIG. 1 includes an apparatus main body 41, a first dispensing head 42, a second dispensing head 43, a first dispensing head 42, and a second dispensing head. And a head moving mechanism 44 for moving 43 with respect to the apparatus main body 41.

The reaction processing apparatus 40 supplies a plurality of types of non-general-purpose reagents and a plurality of types of general-purpose reagents to the processing tool 1 containing a biological material, and processes the biological material by, for example, an in situ hybridization method. .
The processing tool 1 will be described in detail later.

  The apparatus main body 41 has an appearance almost like a rectangular parallelepiped table, and various devices are accommodated therein.

  On the apparatus main body 41, a holding unit 45 for holding the processing tool 1 is installed. In the present embodiment, six processing tools 1 having a multiple configuration (8-series configuration) can be installed in the holding unit 45, but the number of processing tools 1 to be installed is not limited to this.

  The first dispensing head 42 has a disposable nozzle tip 421 detachably attached thereto, and has a function of sucking and discharging a liquid into the nozzle tip 421.

  The term “disposable” as used in the present invention means that the nozzle tip 421 is replaced with a new one according to the type of reagent to be dispensed, and of course when the used nozzle tip 421 is discarded. This also includes the case where the collected used nozzle tip 421 is washed, dried and reused.

  The first dispensing head 42 is, for example, a rare and expensive reagent such as a probe solution, an antibody solution, an enzyme solution, a reagent that easily decomposes at room temperature (room temperature) (an unstable reagent), or contamination with other reagents. Is dispensed to the treatment tool 1 (referred to as “non-general purpose reagent” in the present invention). This non-general-purpose reagent is used for detection of a detection target contained in a biological substance, and is a treatment liquid (reaction liquid) containing a substance that can react with the detection target.

  On the apparatus main body 41, a cooling storage section (temperature adjustment section) 46 for storing a non-general purpose reagent is installed. In addition, this cooling storage part 46 may be filled with an inert gas. Thereby, a non-general purpose reagent can be preserve | saved in the atmosphere of an inert gas.

  The cooling storage section 46 is formed with small holes having a large capacity arranged in a matrix and large holes having a relatively large capacity arranged next to each other. Each small hole has a small amount of use. General-purpose reagents can be stored separately for each type, and non-general-purpose reagents with a relatively large amount of use can be stored separately for each type in each large hole.

  Moreover, the cooling storage part 46 has a built-in cooling device, and can store the non-general-purpose reagent in a cooled state. Thereby, alteration and deterioration of a non-general purpose reagent can be prevented. Although this cooling device is not specifically limited, For example, it can comprise with a Peltier device etc.

  When dispensing the non-generic reagent, first, the first dispensing head 42 is moved to the cooling reservoir 46 to suck the non-generic reagent into the nozzle tip 421, and then the first dispensing head 42. Is moved to the processing tool 1, and the aspirated non-general-purpose reagent is discharged into the processing tool 1.

  A chip remover 47 for removing the nozzle tip 421 from the first dispensing head 42 is installed on the apparatus main body 41.

  The chip remover 47 is formed with a circular hole 472 and a semicircular cutout 471. The notch 471 is formed integrally with the hole 472 in a part of the outer periphery of the hole 472.

  The diameter of the hole 472 is slightly larger than the outer diameter of the upper end of the nozzle tip 421. The nozzle tip 421 of the first dispensing head 42 moved downward by driving the head moving mechanism 44 (elevating mechanism 441) is inserted into the hole 472.

  The width of the notch 471 is smaller than the outer diameter of the upper end of the nozzle tip 421 and larger than the outer diameter of the tip mounting portion on which the nozzle tip 421 is mounted.

  Removal of the nozzle tip 421 from the first dispensing head 42 is performed as follows (hereinafter, this step is referred to as “nozzle tip removal step”).

  First, the first dispensing head 42 to which the nozzle tip 421 is attached is moved onto the tip remover 47 (hole 472).

  Next, the first dispensing head 42 is moved downward (lowered), and the nozzle tip 421 is inserted into the hole 472 to its upper end.

  Next, the first dispensing head 42 is moved in the depth direction of the reaction processing apparatus 40. As a result, the tip mounting portion of the first dispensing head 42 enters the notch 471.

  Next, the first dispensing head 42 is raised. Thereby, the edge of the notch 471 presses the upper end of the nozzle tip 421 downward, and the nozzle tip 421 is detached.

  A recovery container 49 for collecting (collecting) the removed nozzle tip 421 is installed below the tip remover 47 and inside the apparatus main body 41.

  The collection container 49 can be taken out from an opening 411 formed on the front surface of the apparatus main body 41. The used nozzle tip 421 removed by the tip remover 47 falls into the collection container 49 and is collected.

  Further, on the apparatus main body 41, a chip rack 48 that holds a large number of new nozzle chips 421 arranged in a matrix in a standing state is installed.

  When a new nozzle tip 421 is mounted after the nozzle tip 421 is removed from the first dispensing head 42 in the nozzle tip removal process, the first dispensing head 42 is moved above the tip rack 48, The first dispensing head 42 is lowered. As a result, the tip mounting portion of the first dispensing head 42 is fitted into the upper end opening of the new nozzle tip 421 so that the nozzle tip 421 can be mounted (hereinafter, this process is referred to as “nozzle chip mounting process”). Called).

  When dispensing a non-general reagent to the processing tool 1, the first dispensing head 42 replaces the nozzle tip 421 with a new one for each type of non-general reagent. Thereby, contamination between different types of non-general-purpose reagents can be reliably prevented, and biological substances can be processed accurately.

  Unlike the first dispensing head 42, the second dispensing head 43 has a fixed nozzle 431 that is not disposable, that is, used multiple times.

  The second dispensing head 43 may be subjected to contamination with generally widely used (general purpose) reagents such as physiological saline, ethanol aqueous solution, specific gravity adjusting liquid, and other reagents. A reagent, a reagent that is difficult to decompose at room temperature (room temperature) (stable reagent) (referred to as “general-purpose reagent” in the present invention) is dispensed (discharged) to the treatment tool 1. This general-purpose reagent is used, for example, for the purpose of improving the processing efficiency of a biological material with a non-general-purpose reagent, the purpose of improving the detection efficiency of a detection object, and a processing tool before and after processing the biological material with a non-general-purpose reagent (treatment liquid). 1 is used for the purpose of smoothly replacing liquids having different specific gravities, for the purpose of washing biological materials, and the like.

A shelf 50 is installed above a position near the rear of the apparatus main body 41.
On the shelf 50, a plurality of tanks (storage units) 51 for storing general-purpose reagents are placed. In the configuration shown in the drawing, a total of 17 tanks 51 are provided in three types, that is, three large-capacity tanks 511, four medium-capacity tanks, and ten small-capacity tanks 513.
A plurality of types of general-purpose reagents are stored for each type in each tank 51 according to the amount of use.

  The first dispensing head 42 and the second dispensing head 43 each move relative to the apparatus main body 41 by the operation of the head moving mechanism 44. Hereinafter, the configuration of each part of the head moving mechanism 44 will be sequentially described.

  The 1st dispensing head 42 and the 2nd dispensing head 43 move to an up-down direction by the raising / lowering mechanism 441, respectively.

  The elevating mechanism 441 moves together with the first dispensing head 42 and the second dispensing head 43 along the movable bridge 442 installed along the lateral direction of the apparatus main body 41.

  A drive belt 443 and a motor 444 for driving the drive belt 443 are installed on the movable bridge 442, and the lifting mechanism 441 moves along the movable bridge 442 by these drives.

  Both end portions of the movable bridge 442 are supported by rails 445 and 446 installed along the front-rear direction of the apparatus main body 41, and along the rails 445 and 446, the lifting mechanism 441 and the front-rear direction of the apparatus main body 41 are included. Move to.

  The rails 445, 446 are installed on walls 447, 448 erected near the left and right ends of the apparatus main body 41, and are positioned higher than the upper surface of the apparatus main body 41. It can pass over the holding unit 45, the cooling storage unit 46, the chip remover 47, the chip rack 48, and the like installed above.

  By the head moving mechanism 44 having the above configuration, the first dispensing head 42 and the second dispensing head 43 are integrally moved in the vertical direction, the horizontal direction, and the front-rear direction.

  In addition, unlike such a configuration, the first dispensing head 42 and the second dispensing head 43 may be movable separately.

  As shown in FIG. 2, a dispensing pump 53 configured by, for example, a syringe pump is connected to the tip mounting portion of the first dispensing head 42 via a tube pipe 52.

  The suction / discharge force generated by the operation of the dispensing pump 53 is transmitted to the first dispensing head 42 via the tube pipe 52, whereby the liquid can be sucked / discharged by the mounted nozzle tip 421.

  The flow paths 55 exiting from the tanks 51 merge to form a single flow path 56, and the single flow path 56 is connected to the fixed nozzle 431 of the second dispensing head 43.

  An electromagnetic valve (flow path switching means) 57 is installed in the middle of each flow path 55 before joining. In addition, a liquid feed pump 54 that feeds the general-purpose reagent in each tank 51 to the fixed nozzle 431 is installed in the middle of the merged one flow path 56.

  In such a configuration, by opening only the electromagnetic valve 57 to one tank 51 and closing the electromagnetic valve 57 to the other tank 51, the general-purpose reagent in the one tank 51 is transferred to the fixed nozzle 431. Can be delivered and dispensed. Then, by switching the electromagnetic valve 57 to be opened, the general-purpose reagent can be sent from the desired tank 51 to the fixed nozzle 431 and dispensed.

  As described above, the plurality of types of general-purpose reagents are dispensed from the fixed nozzle 431 to the processing tool 1 in common regardless of the type of the general-purpose reagent using the second dispensing head 43. Thereby, since the cost of the nozzle tip which is a consumable item is not required for the second dispensing head 43, the cost can be reduced. Further, since the second dispensing head 43 does not require time for nozzle tip replacement, the processing efficiency can be improved.

  The second dispensing head 43 is provided with a heater 58 for heating the general-purpose reagent to be supplied around the flow path 56 before the fixed nozzle 431. By heating the general-purpose reagent dispensed to the processing tool 1 by the heater 58, the reaction in the processing tool 1 can be promoted.

  Inside the holding part 45 that holds the processing tool 1, a liquid receiving part 59 that receives the liquid discharged from the processing tool 1 is installed. The drainage liquid collected in the liquid receiving portion 59 passes through the flow path 60 and is collected and stored in the drainage tank 61.

  The drainage tank 61 is hermetically sealed, and a suction pump 62 that decompresses the inside of the drainage tank 61 is connected to the drainage tank 61. When the electromagnetic valve 63 installed between the liquid receiver 59 and the flow path 60 is opened, the drainage liquid accumulated in the liquid receiver 59 is transferred to the drainage tank 61 by the suction force of the suction pump 62.

  A shaking device (not shown) that shakes (vibrates) the holding unit 45 is provided inside the apparatus main body 41 below the holding unit 45. By the operation of this shaking device, a floating stirrer 8 described later can be moved in the liquid stored in the processing tool 1 to stir the liquid.

  In the present embodiment, the liquid receiving part 59, the flow path 60, the drainage tank 61, the suction pump 62 and the electromagnetic valve 63 constitute a drainage recovery means.

  A nozzle cleaning tank 64 is connected to the flow path 60 to the drainage tank 61 via a flow path 65.

  For example, when the type of general-purpose reagent supplied to the fixed nozzle 431 of the second dispensing head 43 is changed, the fixed nozzle 431 is moved to the nozzle cleaning tank 64 before dispensing to the processing tool 1. Then, the liquid feeding pump 54 is operated, and a certain amount of the general-purpose reagent is discharged into the nozzle cleaning tank 64. As a result, it is possible to wash away the pre-change general-purpose reagent remaining in the fixed nozzle 431 and the flow path 56 and replace it with the post-change general-purpose reagent. Therefore, when the treatment tool 1 is subsequently dispensed, the modified general-purpose reagent can be immediately discharged from the fixed nozzle 431.

  When the electromagnetic valve 66 installed in the flow path 65 is opened, the drainage liquid accumulated in the nozzle cleaning tank 64 is transferred to the drainage tank 61 by the suction force of the suction pump 62.

  The reaction processing apparatus 40 includes a dispensing pump 53, a liquid feed pump 54, a head moving mechanism 44 (first dispensing head 42, second dispensing head 43), a Peltier element in the cooling reservoir 46, a heater 58, an electromagnetic wave. The control means 70 for controlling the operation of each part of the reaction processing apparatus 40 such as the valves 57, 63 and 66 has a processing protocol (each non-general reagent, each general purpose reagent and The supply (processing) to the processing tool 1 in a predetermined order is executed based on the control program.

  Moreover, the reaction processing apparatus 40 is provided with an input unit 71 having buttons such as a keyboard. Thereby, for example, the processing protocol, the processing speed, and the like can be set as appropriate.

Next, the processing tool 1 used for this reaction processing apparatus 40 is demonstrated.
FIG. 3 is a partial cross-sectional perspective view showing a configuration of a processing tool used in the present invention, and FIGS. 4 to 11 are diagrams (longitudinal cross-sectional views) for explaining how to use the reaction processing apparatus of the present invention, respectively. FIG. 12 is a graph showing a change in the specific gravity of the liquid stored in the space defined between the first filter and the second filter. In the following description, the upper side in FIG. 3 (the same applies to FIGS. 4 to 11) is referred to as the “base end”, and the lower side is referred to as the “tip”. Also, in FIGS. 4 to 11, in order to avoid complication of the drawings, some of them are omitted.

  The processing tool 1 shown in FIG. 3 has a column main body 2, and a first filter 5 and a second filter 6 provided in the column main body 2.

  The processing tool 1 stores a liquid (sample solution) containing a biological material in a space 20 defined by the first filter 5 and the second filter 6 in the column body 2, and in this state, the space By supplying the treatment liquid as described above into the inside 20 through the second filter 6, the treatment liquid is brought into contact with the biological material to perform the treatment.

  Further, as will be described later, the processing tool 1 includes an extension cup 3, a cap 4, and a positioning unit 100 that are detachably attached to the column main body 2.

  In this embodiment, the plurality of column main bodies 2, the plurality of extension cups 3, and the main body portions 110 of the plurality of positioning means 100 are integrally formed (connected side by side).

Hereinafter, the structure of each part of the processing tool 1 is demonstrated sequentially.
The column body (tubular body) 2 has a substantially cylindrical shape as a whole.

  The column main body 2 has a main body portion 21 and a discharge port 22 whose diameter is reduced with respect to the main body portion 21 at the distal end thereof, and these are integrally formed.

  A liquid such as a processing liquid or a specific gravity adjusting liquid is supplied into the column main body 2 (main body portion 21) from the base end opening 210. Then, the biological material is treated with the treatment liquid. Further, the liquid after processing the biological material is discharged out of the main body 21 from the discharge port 22.

  The cross-sectional shape of the column body 2 is preferably circular, but may be any other shape such as an ellipse, a rectangle, a hexagon, and the like.

  Examples of the constituent material of the column body 2 include various resin materials, various glass materials, various ceramic materials, various metal materials, and the like, but it is preferable to use a substantially transparent or translucent material. Thereby, it is possible to easily observe the biological material stored in the main body 21.

  In addition, when the biological substance treatment process includes a heating (heating) process, it is preferable to use a material having excellent heat resistance as the constituent material of the column body 2. Examples of the heat-resistant resin material include polypropylene.

  The column main body 2 (main body portion 21) preferably has a length from the base end face of the first filter 5 provided therein to the base end of the column main body 2 of about 5 to 30 mm, and 10 to 20 mm. More preferred is the degree. Thereby, for example, when the biological substance after processing is observed from the proximal end side of the column main body 2 using an optical microscope or the like, the distance between the biological substance and the objective lens can be made relatively short. The state of the substance can be confirmed (observed) more accurately.

  The outer diameter of the main body 21 (maximum outer diameter of the column main body 2) is preferably about 5 to 20 mm, and more preferably about 8 to 15 mm.

  The inner diameter of the main body 21 (the maximum inner diameter of the column main body 2) is preferably about 3 to 18 mm, and more preferably about 6 to 13 mm.

  Moreover, it is preferable that the length of the discharge port 22 is about 3-15 mm, and it is more preferable that it is about 6-10 mm.

  The outer diameter of the discharge port 22 is preferably about 2.5 to 7 mm, and more preferably about 3 to 5 mm.

  The inner diameter of the discharge port 22 is preferably about 0.5 to 5 mm, and more preferably about 1 to 3 mm.

  In the column main body 2, the first filter 5 is fixed (adhered) in the vicinity of the discharge port 22, and the second filter 6 is disposed on the proximal end side of the first filter 5 in the axial direction of the column main body 2 ( In FIG. 3, it is provided to be movable in the vertical direction.

  Each of the first filter 5 and the second filter 6 is composed of a member having a large number of relatively small-diameter continuous (through) pores.

  A space 20 is defined in the column main body 2 (main body portion 21) by the first filter 5 and the second filter 6. The biological material 10 is accommodated in the space 20, and the biological material is processed by the processing liquid supplied in the space 20.

  The volume of the space 20 can be adjusted by setting the axial position of the column body 2 of the second filter 6.

  As the first filter 5, a biological material stored in the space 20 or a material having pores with a diameter such that a floating stirrer 8 described later cannot pass is suitably used.

  The average pore diameter of the pores of the first filter 5 is appropriately set depending on the biological material, the size of the floating stirrer 8 and the like, and is not particularly limited, but is preferably about 10 to 150 μm, and 20 to 100 μm. More preferred is the degree.

  Moreover, it is preferable that the average thickness of the 1st filter 5 is about 0.5-5 mm, and it is more preferable that it is about 1-3 mm.

  On the other hand, the second filter 6 is configured to hold a liquid (sample liquid, processing liquid, or specific gravity adjusting liquid) in the space 20.

  Specifically, when the second filter 6 supplies liquid from the base end side, the space 20 is filled with the liquid, and the liquid substantially does not exist on the base end side, the pores The passage of the liquid is stopped by the capillary phenomenon generated in the inside, and when the liquid is again supplied to the proximal end side from this state, the supplied liquid passes through the pores and flows into the space 20 (the distal end side). Has been.

  Such characteristics of the second filter 6 are adjusted to a desired one by combining conditions such as the thickness of the second filter 6, the number of pores, the shape of the pores, and the pore diameter. can do.

  The average pore diameter of the pores of the second filter 6 is not particularly limited, but is preferably about 10 to 150 μm, and more preferably about 20 to 100 μm. If the average pore diameter of the pores of the second filter 6 is too small, depending on the viscosity of the liquid or the like, the liquid passing speed may be slow, and the processing efficiency of the biological material 10 may be reduced, while the average pore diameter is large. If too much, the capillary phenomenon does not occur sufficiently, and it may be difficult to hold the liquid in the space 20.

  Moreover, it is preferable that the average thickness of the 2nd filter 6 is about 0.5-5 mm, and it is more preferable that it is about 1-3 mm.

  Such fine pores of the first filter 5 and the second filter 6 may be linear, for example, an indefinite shape whose inclination angle changes, continuous pores communicating with each other, or the like. .

  That is, each of the first filter 5 and the second filter 6 is, for example, a porous plate having a plurality of through pores penetrating in the thickness direction, a fibrous porous body such as a woven fabric or a nonwoven fabric, and the like. The non-fibrous porous material can be used. Among these, it is particularly preferable to use a pore plate. Thereby, the passage resistance of the liquid of the 1st filter 5 and the 2nd filter 6 can be reduced more.

  As the constituent materials of the first filter 5 and the second filter 6, the same materials as those mentioned in the column main body 2 can be used, respectively, but a material that can ensure sufficient strength for the filter. In addition, a liquid (a treatment liquid or a specific gravity adjusting liquid) or a material having a low reactivity with a substance (for example, a labeling reagent) contained in these liquids (a material that does not substantially react) is preferable.

  In this specification, “reaction” is a concept including electroadsorption, chemical bonding, and the like.

  Examples of such a material include various resin materials such as polyolefin (for example, high density polyethylene, polypropylene), polyamide, fluorine resin (for example, polytetrafluoroethylene), and silicone resin, stainless steel, titanium, or titanium alloy. Such various metal materials are mentioned.

  Moreover, the 1st filter 5 and the 2nd filter 6 may be formed (covered) with a surface layer made of the above material on the surface of the base material.

  In this way, by configuring at least the vicinity of the surface of the first filter 5 and the second filter 6 with a liquid (treatment liquid or specific gravity adjusting liquid) or a material having poor reactivity with substances contained in these liquids. It is possible to more reliably prevent the processing accuracy of the biological material from being lowered.

  Further, the first filter 5 and the second filter 6 are preferably hydrophilic. Thereby, the liquid passes through the filters 5 and 6 more smoothly.

  Examples of the hydrophilization method include plasma treatment, ultraviolet irradiation, and treatment with a surfactant.

  A rod-like operation member 7 for moving the second filter 6 in the axial direction of the column main body 2 is fixed (adhered) to the center of the second filter 6.

  The operation member 7 may be formed integrally with the second filter 6, and as shown in FIG. 3, a member different from the second filter 6 is bonded by fusion or an adhesive. For example, it may be fixed to the second filter 6 by, for example.

  The second filter 6 can be moved closer to or away from the first filter 5 by moving the operating member 7 in the axial direction of the column body 2. Thereby, the volume of the space 20 can be adjusted.

  In addition, by fixing the operation member 7 to the central portion of the second filter 6, the second filter 6 is moved by the first filter 5 when the second filter 6 is moved in the axial direction of the column body 2. It can prevent suitably that it inclines with respect to.

  In addition, the operation member 7 is configured to be positioned substantially at the center of the column body 2. Thereby, when attaching the extension (extension cylinder) 3 mentioned later to the column main body 2, it can prevent that the base end part of the operation member 7 becomes obstructive.

The cross-sectional area of the operating member 7 is preferably about 0.5 to ¹⁰ mm ² , and more preferably about 1 to 5 mm ² .

  Moreover, it is preferable that the length of the operation member 7 is about 5-20 mm, and it is more preferable that it is about 10-15 mm.

A floating stirrer (stirrer) 8 is accommodated in the space 20.
The floating stirrer 8 is configured to stir the liquid in the vicinity of the second filter 6 by floating in the liquid stored in the space 20.

  As a result, the liquid stored in the space 20 and the liquid flowing into the space 20 through the second filter 6 from the base end side of the second filter 6 are moved to the vicinity of the second filter 6. And the concentration (specific gravity) in each part of the liquid existing in the space 20 can be made more uniform.

  The specific gravity of the floating stirrer 8 is not particularly limited as long as it floats in various liquids (particularly water), but is preferably about 0.85 to 0.99, 0.9 to 0.99. More preferably, it is about 0.9 to 0.96. If the specific gravity of the floating stirrer 8 is too small, the buoyancy of the floating stirrer 8 will increase, and the contact friction with the second filter 6 will increase, possibly resulting in insufficient stirring. On the other hand, if the specific gravity of the floating stirrer 8 is too large, the floating stirrer 8 is likely to sink into the liquid stored in the space 20, and the liquid may be insufficiently stirred and mixed in the vicinity of the second filter 6. is there. In general, the biological material generally exists in contact with the first filter 5, but the floating stirrer 8 is submerged in the liquid (sample liquid, processing liquid, specific gravity adjusting liquid) stored in the space 20. Therefore, there is a possibility that the biological material is likely to be damaged (broken).

  Such a floating stirrer 8 has a material (substantially reacts) at least near the surface thereof with a liquid (treatment liquid or specific gravity adjusting liquid) or a material having a poor reactivity with a substance (for example, a labeling reagent) contained in these liquids. The material is preferably made of a material that does not generate Thereby, it can prevent more reliably that the process accuracy of the biological material 10 falls.

  As such a material, the same materials as those mentioned for the first filter 5 and the second filter 6 can be used, but it is preferable to use a material having a relatively low specific gravity.

  The floating stirring bar 8 is preferably hydrophobic. As a result, the floating stirring bar 8 moves more smoothly in the liquid, and the liquid can be mixed more uniformly.

Examples of the hydrophobizing method include treatment with a surfactant.
The form of the floating stirring bar 8 may be any of a solid body, a hollow body, and a porous body.

  When the floating stirrer 8 is formed of a solid body, the constituent material of the floating stirrer 8 may be selected in consideration of the specific gravity of the material itself. When the floating stirrer 8 is composed of a hollow body or a porous body, Since the specific gravity of the floating stirrer 8 can be adjusted by setting the hollow portion, the porosity, etc., a material having a relatively large specific gravity can also be used.

  Moreover, the shape of the floating stirrer 8 is not particularly limited, and may be any shape such as granular, flat plate, and rod, but is preferably granular (particularly spherical). Thereby, the contact friction with the 2nd filter 6 can be weakened, and the stirring effect by free floating is acquired. Moreover, even if the floating stirrer 8 collides with the biological material, the impact can be reduced, so that the biological material can be suitably prevented from being damaged.

  When the floating stirrer 8 is granular, the average particle size is preferably about 0.5 to 5 mm, more preferably about 1 to 3 mm. Thereby, the impact on the biological material can be reduced while suitably maintaining the ability to stir the liquid. Therefore, for example, it is possible to suitably prevent the biological material from being damaged.

  Such floating stirrer 8 can be moved more smoothly and randomly by allowing bubbles to coexist in the liquid. Thereby, stirring and mixing of a liquid can be performed more reliably.

  As shown in FIG. 3, a cap 4, which is almost entirely flexible, is detachably attached to the distal end portion (discharge port 22) of the column main body 2.

  The cap 4 can cover the tip opening 221 in a state of being attached to the discharge port 22, and supply internal air (gas) to the liquid stored in the space 20 when being removed from the discharge port 22. (Injection) and has the function of generating bubbles as described above in the liquid.

  Specifically, the cap 4 is attached to the tip of the column body 2 so as to cover the tip opening 221 of the discharge port 22. In this state, the liquid is stored in the space 20, the cap 4 is held (clamped) with a finger or the like to be deformed, the internal volume is reduced, and the air in the cap 4 is discharged into the space 20 through the discharge port 22. To send. Thereby, air is supplied to the liquid stored in the space 20, and bubbles are generated in the liquid.

  Examples of the constituent material of the cap 4 include silicone rubber and various thermoplastic elastomers.

  Note that the cap 4 may be configured to have a portion having flexibility in a part thereof. In this case, air can be supplied into the space 20 in the same manner as described above by deforming the portion having such flexibility.

  Moreover, it is preferable that the internal volume of the cap 4 is set to be smaller than the volume on the tip side of the first filter 5 in the column body 2. The effect obtained by such a configuration will be described later.

Specific values of the internal volume of the cap 4 is preferably about 10 to 100 mm ^3, more preferably about 20 to 50 mm ^3.

  Further, as shown in FIG. 3, an extension cup (extension cylinder) 3 is detachably attached to the base end portion of the column main body 2.

The extension cup 3 has a substantially cylindrical shape as a whole.
The extension cup 3 has a main body portion 31 and a mounting portion (reduced diameter portion) 32 whose inner diameter and outer diameter are reduced with respect to the main body portion 31 at the distal end thereof, and these are integrally formed. ing.

  The outer diameter of the mounting portion 32 is set to be approximately equal to or slightly larger than the inner diameter of the main body portion 21 of the column main body 2. The extension cup 3 is attached (fixed) to the column main body 2 by the mounting portion 32 being inserted and fitted inside the base end portion of the main body portion 21.

  In addition, the extension cup 3 of the present embodiment has such a length that the base end of the operation member 7 does not protrude from the base end of the extension cup 3 attached (connected) to the column main body 2.

  Therefore, by attaching the extension cup 3 to the column main body 2, the base end portion of the operation member 7 is accommodated in the extension cup 3. For this reason, for example, when the liquid (processing liquid or specific gravity adjusting liquid) is supplied into the column main body 2 using the first dispensing head 42 and the second dispensing head 43, they are attached to the front ends thereof or attached thereto. The tip of the chip (the member that supplies the liquid) is prevented from hitting (contacting) the operation member 7, and the liquid (processing liquid and specific gravity adjusting liquid) is supplied more accurately into the column body 2. Can do.

  Further, the tip end of the tip can be guided into the column main body 2 along the inner surface of the extension cup 3.

  As described above, the extension cup 3 has a function of guiding the liquid or a member supplying the liquid into the column main body 2 when supplying the liquid (processing liquid or specific gravity adjusting liquid) into the column main body 2. is there.

  In addition, since the capacity of the entire processing tool 1 can be increased by mounting the extension cup 3 on the column main body 2, a relatively large amount of liquid (processing liquid or specific gravity adjusting liquid) can be placed in the column main body 2 at one time. Will be able to supply. As a result, the processing efficiency of the biological material 10 can be improved.

Such a length of the extension cup 3 (full-length) _{L 2} is preferably in the range of about 1 / 10-3 / 4 column length of the body 2 (the entire length) _{L 1,} about 3 / 10-1 / 2 It is more preferable that As a result, the effects as described above are more remarkably exhibited.

  The main body 31 of the extension cup 3 preferably has a cross-sectional shape similar to that of the main body 21 of the column main body 2. In particular, the outer diameter (maximum outer diameter of the extension cup 3) is the same as that of the column main body 2. The outer diameter of the main body 21 (the maximum outer diameter of the column main body) is preferably substantially equal. When the processing tool 1 is not used, a plurality of the processing tools 1 are stored in a storage container or the like with the extension cup 3 attached to the column body 2. Can be stored well.

  In addition, in a state where the extension cup 3 is attached to the column main body 2, it is preferable that liquid tightness (particularly, air tightness) is secured between them. Thereby, for example, it is possible to more reliably prevent liquid from leaking out from a connecting portion (connecting portion) between the extension cup 3 and the column main body 2.

  As the constituent material of the extension cup 3, the same materials as those mentioned in the column main body 2 can be used.

  The cross-sectional shape of the extension cup 3 is also preferably circular, but may be any other shape such as an ellipse, a rectangle, a hexagon, and the like, similar to the column main body 2.

  In addition, a sheet material (sheet-like sealing member) 9 is detachably attached (provided) to the extension cup 3 so as to close the proximal end opening 310 on the proximal end surface.

  As a constituent material of the sheet material (sealing member) 9, for example, one kind or two or more kinds of various resin materials such as polyethylene terephthalate, polypropylene, polyethylene, silicone-based resin, silicone rubber, and various metal materials are used. They can be used in combination.

  The average thickness of the sheet material 9 is preferably about 0.05 to 10 mm, and more preferably about 0.1 to 5 mm.

  This sheet material 9 is removed immediately before the operation of supplying the processing liquid and specific gravity adjusting liquid into the column main body 2 is started. Thereby, the inside of the column main body 2 (processing tool 1) is sealed until the processing liquid and the specific gravity adjusting liquid are supplied, and foreign matters such as dust can be prevented from entering the column main body 2 (processing tool 1). As a result, the biological material 10 can be processed more accurately.

  In addition, in a state where the sheet material 9 is adhered to the extension cup 3, it is preferable that airtightness is secured between them.

  Note that the sealing member attached to the base end portion of the extension cup 3 is not limited to a sheet-like member, and it is sufficient that the base end opening 310 can be sealed, for example, the base end portion outside the extension cup 3. It may be a cap-like thing fixed by fitting, screwing or the like, or a plug-like thing fixed by fitting, screwing, or the like inside the base end part.

  Further, as in the present embodiment, the column main body 2 and the extension cup 3 have a multiple configuration, which makes it possible to process a plurality of biological materials at a time, which is convenient.

  The positioning means 100 is used by being attached to the base end portion of the column main body 2 as described above, and performs positioning of the second filter 6 with respect to the column main body 2.

  As shown in FIG. 1, the positioning means 100 is provided with a main body portion 110 that is detachably attached to a base end portion of the column main body 2 (main body portion 21), and penetrates the main body portion 110 in the thickness direction. And a hole (concave portion) 120.

  The main body 110 has a substantially columnar shape (block shape). The inner diameter of the main body 110 is set to be approximately equal to or slightly smaller than the outer diameter of the main body 21 of the column main body 2. Thus, the positioning means 100 is attached (fixed) to the column main body 2 by inserting and fitting the distal end side of the main body 110 into the inner side of the base end of the column main body 2.

A ring-shaped stepped portion 112 is formed at the base end portion of the main body 110.
When the positioning means 100 is mounted on the column main body 2 (hereinafter referred to as “the mounting state of the positioning means 100”), the base end of the column main body 2 abuts on the stepped portion 112, whereby the positioning means 100 (main body section). 110) positioning of the column body 2 in the axial direction is performed.

  The operation member 7 is inserted into the hole 120 with the positioning means 100 mounted. The hole 120 is formed in the central portion of the main body 110, and is configured such that the central axis of the hole 120 and the central axis of the column main body 2 substantially coincide with each other when the positioning means 100 is mounted.

  Thereby, in the mounted state of the positioning means 100, the central axis of the operation member 7 inserted into the hole 120 and the central axis of the column main body 2 substantially coincide with each other.

  Then, the center axis of the operation member 7 and the center axis of the column body 2 substantially coincide with each other, whereby the second filter 6 is positioned in the horizontal direction, so that the first filter 5 and the second filter 6 are substantially aligned. It comes to be parallel.

  Further, a guide hole 121 that communicates with the hole 120 and guides the operation member 7 to the hole 120 is formed at the distal end of the main body 110.

  The guide hole 121 is open at the tip of the main body 110 and has a transverse area gradually decreasing toward the hole 120. The inner surface of the guide hole 121 has a mortar shape. By providing such a guide hole 121, when the positioning means 100 is mounted on the column main body 2, the central axis of the operation member 7 is slightly shifted (tilted) with respect to the central axis of the column main body 2. However, the operating member 7 can be guided into the hole 120 and inserted reliably.

  The angle θ formed between the inner surface of the guide hole 121 and the front end surface of the main body 110 is not particularly limited, but is preferably about 30 to 60 °, more preferably about 35 to 45 °. Thereby, the operation member 7 can be inserted into the hole 120 more smoothly and reliably.

  From this viewpoint, it is preferable that the diameter (transverse area) of the base end portion of the operation member 7 is gradually reduced toward the base end direction. Thereby, the operation member 7 can be inserted into the hole 120 more smoothly and reliably.

  In this embodiment, the cross-sectional area of the guide hole 121 is substantially constant at the tip, and the step 112 is formed at the boundary with the tapered portion. In the mounted state of the positioning means 100, the base end of the column main body 2 abuts on the stepped portion 112, thereby positioning the column main body 2 in the axial direction of the positioning means 100.

  The cross-sectional shapes of the hole 120 and the operation member 7 may be different from each other, but are preferably similar to each other. Each of these cross-sectional shapes is preferably circular, but may be any other shape such as an ellipse, a rectangle, a hexagon, and the like.

  A screw (insertion member) 130 is inserted and fixed to the base end side of the hole 120 by screwing. In this state, when the positioning means 100 (main body 110) is attached to the column main body 2, the proximal end of the operation member 7 inserted into the hole 120 comes into contact with the distal end of the screw 130, and the operation member 7 is Pressed in the direction. Thereby, the operation member 7 moves (lowers) toward the distal direction.

  As the operating member 7 moves in the distal direction, the second filter 6 also moves (lowers) in the distal direction. Therefore, by adjusting the insertion depth of the screw 130 into the hole 120, the moving distance of the operation member 7 in the distal direction can be adjusted. Thereby, the position in the axial direction (vertical direction) of the column main body 2 of the second filter 6 can be adjusted, that is, the positioning of the second filter 6 in the axial direction of the column main body 2 can be performed.

  Note that the insertion member is not limited to the screw 130, and may be any member that can be inserted and fixed in the hole 120, for example, a pin. In addition, when the positioning member 100 is mounted on the column main body 2 by configuring the insertion member with a screw, even when the proximal end of the operation member 7 abuts against the distal end of the operation member with a relatively strong force, the operation member Can be prevented from unintentionally moving, falling off (disengaging) from the hole 120, and the like.

  Moreover, you may make it provide the scale which shows an insertion depth along an axial direction in the surrounding surface of the part (this embodiment screw part of the screw 130) inserted in the hole 120 of an insertion member. This is convenient because the insertion depth of the insertion member into the hole 120 can be confirmed.

  In the present invention, as the insertion member, a plurality of insertion members having different lengths to be inserted into the hole 120 are prepared, and by appropriately selecting and using them, the insertion member can be inserted into the hole 120 of the insertion member. The insertion depth (insertion amount) can be adjusted.

  Examples of the constituent material of the positioning means 100 (the main body 110 and the screw 130) include various resin materials, various glass materials, various ceramic materials, various metal materials, and the like.

  In addition, as in the present embodiment, the positioning means 100 has a multiple configuration, which makes it possible to position the plurality of second filters 6 with respect to the column main body 2 at once, which is convenient.

  Then, by setting whether or not the screw 130 is inserted into the hole 120, or by setting the insertion depth, the position in the vertical direction (the axial direction of the column body 2) is desired for each second filter 6. Can be adjusted.

  In the illustrated configuration, the case where all of the column body 2, the extension cup 3 and the positioning means 100 have a multiple configuration is shown. However, one or two of these are configured in a multiple configuration. There may be.

  In the present embodiment, a configuration having only one extension cup 3 (one stage) is shown. However, in the present invention, a plurality of (multi-stage) extension cups 3 are provided and combined (connected). It can also be set as the structure to be used.

  In this case, the length of one extension cup 3 only needs to satisfy the relationship described above with respect to the length of the column body 2.

  Further, in this case, the processing tool 1 is used in a state where the extension cup 3 is attached to the column main body 2 in multiple stages so that the base end of the operation member 7 does not protrude from the base end.

  In this case, it is preferable to prepare extension cups 3 having different lengths. Thus, by appropriately selecting and using the number and / or type of extension cups 3, the overall length of the processing tool 1 can be adjusted to a desired one according to the purpose and application.

Next, the usage method (action) of the reaction processing apparatus 40 will be described.
[1] First, as shown in FIG. 4A, the second filter 6 is removed from the column body 2 together with the extension cup 3 and the operation member 7.

  Then, the column main body 2 is installed so that the cap 4 is downward (front end side) and the first filter 5 is substantially horizontal.

  Next, as shown in FIG. 4B, a liquid (sample solution) 11 containing the biological material 10 is supplied into the column body 2 from the proximal end opening 210 of the column body 2.

  Examples of the sample solution 11 include those obtained by suspending or dissolving the biological material 10 in physiological saline or a buffer solution, blood, lymph, saliva, urine, spinal fluid, ascites, pleural effusion, amniotic fluid, and the like.

  Here, examples of the biological substance 10 include various cells, tissue sections, viruses, microorganisms such as bacteria, genes (DNA, RNA), proteins, glycoproteins, hormones, biochemical substances, and the like.

  Further, when handling a block material such as a biological tissue section as the biological material 10, for example, after placing the biological material 10 on the first filter 5, a physiological saline, a buffer solution or the like is added to the column main body. 2 to supply.

[2] Next, the second filter 6 is mounted at a predetermined position in the axial direction of the column body 2.
The mounting operation of the second filter 6 is performed as follows using the positioning means 100 as described above.

  First, as shown in FIG. 1 and FIG. 4C, the screw 130 is inserted into the base end side of the hole 130 by screwing so as to have a predetermined insertion depth.

  Next, as shown in FIG. 5 (d), the base end portion of the operation member 7 is held with a finger or the like, and the second filter 6 is inserted into the column main body 2 from the base end opening 210.

  Next, as shown in FIGS. 5 (d) and 5 (e), the positioning means 100 is made substantially horizontal to approach (lower) the column body 2 so that the operation member 7 is inserted into the hole 120. ). Accordingly, the base end portion of the operation member 7 is guided by the hole 120 while being in sliding contact with the inner surface of the guide hole 121 and is inserted into the hole 120. Then, the proximal end of the operation member 7 comes into contact with the distal end of the screw 130.

  Further, when the positioning means 100 is moved closer to the column main body 2, the operation member 7 is moved (lowered) toward the distal end by being pressed by the screw 130, and accordingly, the second filter 6 is also moved to the distal end. Move (down) in the direction.

  Then, as shown in FIG. 5 (f), when the base end portion of the column main body 2 is inserted into the guide hole 121 and comes into contact with the stepped portion 112, further movement of the positioning means 100 in the distal end direction is prevented. Is done. As a result, the movement of the operating member 7 and the second filter 6 in the distal direction is stopped, and the second filter 6 is positioned in the vertical direction (the axial direction of the column body 2).

  At this time, the central axis of the hole 120 and the central axis of the column body 2 substantially coincide. As a result, the central axis of the operation member 7 and the central axis of the column main body 2 substantially coincide with each other, the second filter 6 is positioned in the horizontal direction, and the first filter 5 and the second filter 6 are substantially parallel. Will come to be located.

  In this way, by using the positioning means 100, it is possible to simultaneously position the second filter 6 both in the horizontal direction and in the vertical direction by one operation of mounting it on the column main body 2. Thereby, it becomes possible to perform the various processes of the biological material 10 by the processing tool 1 more efficiently.

  Next, as shown in FIG. 6G, the positioning means 100 is removed (removed) from the column body 2.

  [3] Next, as shown in FIG. 6 (h), the extension cup 3 is attached to the base end of the column body 2.

  At this time, the sheet material 9 is adhered to the base end face of the extension cup 3 so as to close the base end opening 310.

  [4] Next, as shown in FIG. 6 (i), the cap 4 is grasped with a finger or the like and crushed (deformed). As a result, the air in the cap 4 passes through the first filter 5 and is supplied into the space 20. In the sample solution 11, air bubbles (bubbles) of approximately the same amount as the cap 4 is crushed. ) 111 occurs.

  At this time, the air in the extension cup 3 is compressed by the volume of the bubbles 111 (air supplied from within the cap 4).

  [5] Next, as shown in FIG. 7 (j), the cap 4 is removed from the discharge port 22 while being crushed.

  The sample liquid 11 is pressed (pressurized) toward the distal end by the compressed air in the extension cup 3, and the passage of the first filter 5 is started by removing the cap 4.

  Here, as described above, in the present embodiment, the internal volume of the cap 4 is set to be smaller than the volume on the tip side of the first filter 5 in the column main body 2, so that the sample solution 11 is discharged from the discharge port 22. Outflow from the inside is prevented. Thereby, it can prevent that the sample liquid 11 disperses outside, and does not pollute a hand, the surrounding environment, etc.

  Therefore, for example, when a microorganism or the like is handled as the biological material 10, the processing operation can be performed more safely.

  [6] Next, in this state, after the processing tool 1 is installed on the holding unit 45 of the reaction processing apparatus 40, as shown in FIG. 7 (k), the sheet material adhered to the base end of the extension cup 3. 9 is removed (removed).

  When the sheet material 9 is removed, the sample solution 11 stored in the space 20 passes through the first filter 5 and is discharged from the discharge port 22, and the sample solution 11 on the proximal end side of the second filter 6. Passes through the second filter 6 and flows into the space 20.

  As described above, when the sample liquid 11 is not substantially present on the base end side of the second filter 6, the outflow of the sample liquid 11 from the discharge port 22 is stopped.

  The sample liquid 11 discharged from the discharge port 22 is collected and stored in the drainage tank 61 via the liquid receiving part 59 and the flow path 60.

[7] Next, the sample liquid 11 stored in the space 20 is replaced with the processing liquid 14.
In the space 20, the biological material 10 is processed with the replaced processing liquid 14.

  The processing liquid 14 has a specific gravity smaller than, larger or substantially equal to the specific gravity of the sample liquid 11, but is replaced by an appropriate method depending on the specific gravity relationship between the processing liquid 14 and the sample liquid 11. Preferably it is done.

[7A] When the specific gravity of the processing liquid 14 is smaller than the specific gravity of the sample liquid 11 First, the first dispensing head 42 is moved above the tip rack 48 by the operation of the head moving mechanism 44 and then lowered. As a result, the nozzle tip 421 is attached to the tip attachment portion of the first dispensing head 42.

  Next, after moving the first dispensing head 42 above the cooling reservoir 46, the first dispensing head 42 is lowered. Then, the processing liquid (non-general-purpose reagent) 14 is sucked into the nozzle tip 421.

  Next, after the first dispensing head 42 is moved above the predetermined processing tool 1, the first dispensing head 42 is lowered and the tip of the nozzle tip 421 is positioned in the extension cup 3. Then, as shown in FIG. 8A, the processing liquid 14 is supplied into the extension cup 3.

  Next, a chip remover removal process is performed. That is, after the first dispensing head 42 is moved above the tip remover 47, it is lowered. Then, the first dispensing head 42 is raised with the tip mounting portion inserted into the notch 471, and the nozzle tip 421 is removed from the tip mounting portion.

  The above operation is repeated for the number (predetermined number) of column bodies 2 (processing tools 1) to be processed.

  When the processing liquid 14 is supplied into the column main body 2, the sample liquid 11 passes through the first filter 5 and is discharged out of the column main body 2 from the discharge port 22, and the processing liquid 14 is supplied to the second filter 2. 6 and flows into the space 20.

  At this time, since the specific gravity of the processing liquid 14 is smaller than the specific gravity of the sample liquid 11, the processing liquid 14 does not mix with the sample liquid 11 in the space 20 as shown in FIG. While forming a clear interface 141 at the boundary with the liquid 11, it descends uniformly. As a result, as shown in FIG. 8C, the space 20 can be almost completely replaced with the treatment liquid 14.

  In this case, the formation of a clear interface 141 at the boundary between the treatment liquid 14 and the sample liquid 11 is important in terms of improving the replacement efficiency, and thus the stirring operation by the floating stirring bar 8 is omitted. .

[7B] When the specific gravity of the treatment liquid 14 is greater than or substantially equal to the specific gravity of the sample liquid 11 In this case, prior to supplying the treatment liquid 14 into the column body 2, the specific gravity of the liquid stored in the space 20 is Adjustment is made to be greater than the specific gravity of the treatment liquid 14.

  [7B-1] First, shaking (vibration) of the processing tool 1 (holding unit 45) is started by a shaking device provided inside the apparatus main body 41 below the holding unit 45. Thereby, the floating stirrer 8 moves relative to the column body 2 and the sample liquid 11 is stirred.

  The direction in which the treatment tool 1 is shaken may be the vertical direction, the front-rear direction, the left-right direction, the rotation direction, or a combination thereof, but is the horizontal direction (front-rear direction, left-right direction, rotation in a horizontal plane, or a combination thereof). Is preferred. Thereby, the sample liquid 11 and the liquid (specific gravity adjusting liquid 15) flowing into the space 20 in the vicinity of the second filter 6 can be efficiently stirred and mixed.

  Here, by forming the bubbles 111 in the sample liquid 11, the floating stirrer 8 collides with the bubbles 111 as shown in FIG. 9. Thereby, it can prevent that the floating stirring element 8 contacts the 2nd filter 6, and cannot move with the frictional force between them.

  Further, the floating stirrer 8 and the bubbles 111 repeatedly collide, whereby the floating stirrer 8 can be moved at random, and the sample liquid 11 and the liquid flowing into the space 20 can be more efficiently stirred. Can be mixed.

  [7B-2] Next, after the second dispensing head 43 is moved above the predetermined processing tool 1, the second dispensing head 43 is lowered and the tip of the fixed nozzle 431 is positioned in the extension cup 3.

  Next, the predetermined electromagnetic valve 57 is opened, and the liquid feed pump 54 is operated. As a result, the specific gravity adjusting liquid (general purpose reagent) 15 is transferred from the general purpose reagent tank 51 to the second dispensing head 43 (fixed nozzle 431) via the flow paths 55 and 56. Then, as shown in FIG. 10A, a specific gravity adjusting liquid 15 having a specific gravity larger than that of the sample liquid 11 is supplied into the extension cup 3.

  The specific gravity adjusting liquid 15 is preferably one that does not affect the biological material 10 (one that does not substantially react with the biological material 10). For example, in the solvent or dispersion medium used to adjust the treatment liquid 14, What added the substance for adjusting specific gravity etc. can be used.

  The specific gravity of the specific gravity adjusting solution 15 is preferably 3 times or less, more preferably 2 times or less that of the sample solution 11. If the specific gravity of the specific gravity adjusting liquid 15 is too small, the specific gravity of the sample liquid 11 may not be effectively increased. On the other hand, if the specific gravity of the specific gravity adjusting liquid 15 is too large, the specific gravity adjusting liquid 15 rapidly moves (lowers) along the wall surface of the column body 2 to the front end, and is therefore mixed with the sample liquid 11 by the floating stirrer 8. There is a possibility that the specific gravity of the sample liquid 11 is difficult to increase effectively because it is discharged from the outlet 22 to the outside of the column body 2 before.

  By supplying the specific gravity adjusting liquid 15, the sample liquid 11 passes through the first filter 5 and is discharged out of the column body 2 from the discharge port 22, and the specific gravity adjusting liquid 15 is discharged from the second filter 6. And flows into the space 20.

  Here, as shown in FIG. 10 (b), the specific gravity adjusting liquid 15 that has flowed into the space 20 has a specific gravity greater than that of the sample liquid 11, and therefore moves (down) along the wall surface of the column body 2 in the distal direction. Although starting, the floating stirring bar 8 is stirring the sample liquid 11 in the vicinity of the second filter 6, whereby the sample liquid 11 and the specific gravity adjusting liquid 15 are stirred and mixed.

  Then, the mixed liquid 16 obtained by mixing the sample liquid 11 and the specific gravity adjusting liquid 15 has a specific gravity almost in the middle of these, and the specific gravity is still larger than that of the sample liquid 11, so that it moves (descends) toward the tip. )

  Thereafter, as shown in FIG. 11 (c), the space 20 can be almost completely replaced with the mixed solution 16.

  The mixed liquid 16 may move (down) in the tip direction along the wall surface of the column body 2 in some cases. At this time, the sample liquid 11 having a small specific gravity moves (rises) in the proximal direction in the central portion of the column body 2 and is mixed with the specific gravity adjusting liquid 15 flowing into the space 20 to obtain a mixed liquid 16. It will be. This cycle continues until the supply of the specific gravity adjusting liquid 15 is stopped, and the space 20 can be almost completely replaced with the mixed liquid 16 as shown in FIG.

  Next, the specific gravity adjusting liquid 15 identical to the specific gravity adjusting liquid 15 or the solenoid valve 57 to be opened is changed, and the specific gravity adjusting liquid 15 having a larger specific gravity than the specific gravity adjusting liquid 15 is supplied, and the above operation is repeated. Thereby, the specific gravity of the liquid in the space 20 is adjusted to be larger than the specific gravity of the processing liquid 14.

  After this adjustment, as shown in FIG. 11D, as described above, the processing liquid 14 is supplied (discharged) from the nozzle tip 421 of the first dispensing head 42 to the column main body 2 (main body portion 21). To do.

  Here, for example, a case where the sample liquid 11 having a specific gravity of 1.0 housed in the space 20 is replaced with the processing liquid 14 having a specific gravity of 2.0 will be described as an example.

  In this case, as shown in FIG. 12, first, the specific gravity adjusting liquid 15 having a specific gravity of 2.0 is placed in the column body 2 in which the sample liquid 11 having a specific gravity of 1.0 is stored, and the volume of the space 20 (sample liquid 11). Is supplied in the space 20, the mixed liquid 16 having a specific gravity of about 1.5 is obtained.

  Next, when the specific gravity adjusting liquid 15 having a specific gravity of 2.0 is supplied in an amount substantially equal to the volume of the space 20, a mixed liquid 16 having a specific gravity of about 1.75 is obtained in the space 20.

  Next, when the specific gravity adjusting liquid 15 having a specific gravity of 2.0 is supplied in an amount substantially equal to the volume of the space 20, a mixed liquid 16 having a specific gravity of about 1.9 is obtained in the space 20.

  Next, when the specific gravity adjusting liquid 15 having a specific gravity of 2.5 is supplied in an amount substantially equal to the volume of the space 20, a mixed liquid 16 having a specific gravity of about 2.2 is obtained in the space 20.

  Next, the shaking of the processing tool 1 is stopped, and the processing liquid 14 is supplied into the column body 2 as described in [7A] above.

  Here, since the mixed liquid 16 stored in the space 20 has a higher specific gravity than the processing liquid 14, as shown in FIG. 8B, the processing liquid 14 is While forming a clear interface 141 at the boundary with the mixed liquid 16, the liquid descends uniformly. Thereby, the inside of the space 20 can be almost completely replaced with the treatment liquid 14.

  Next, the nozzle tip 421 that has been used is removed through the nozzle tip removal step described above.

  Next, after the nozzle tip mounting process described above, the first dispensing head 42 is replaced with a new (different) nozzle tip 421, and different column main bodies 2 (a predetermined number of column main bodies 2) The above [7] is executed (repeated).

  [7] is not limited to being repeated a plurality of times, and may be executed only once.

  As described above, the reaction processing apparatus 40 (processing tool 1) does not use a machine such as a suction device, and the weight of the liquid supplied to the base end side from the second filter 6 allows the reaction processing apparatus 40 (processing tool 1) to enter the space 20. The stored liquid can be replaced. For this reason, damage to the biological material 10 can be suitably prevented. In addition, since the configuration is simple, the cost can be kept low.

  When dispensing the treatment liquid 14 (non-general-purpose reagent) to the treatment tool 1, the first dispensing head 42 replaces the nozzle tip 421 with a new one for each kind of treatment liquid 14 and dispenses it. I do. Thereby, contamination between the different types of processing liquids 14 can be reliably prevented, and the biological material 10 can be processed accurately.

  Further, a plurality of types of specific gravity adjusting liquids 15 (general-purpose reagents) are dispensed from the fixed nozzle 431 to the processing tool 1 in common regardless of the type using the second dispensing head 43. Thereby, since the cost of the nozzle tip which is a consumable item is not required for the second dispensing head 43, the cost can be reduced. Further, since the second dispensing head 43 does not require time for nozzle tip replacement, the processing efficiency can be improved.

  As mentioned above, although the reaction processing apparatus of the present invention has been described, the present invention is not limited thereto, and the configuration of each part can be replaced with any one that can exhibit the same function, or Arbitrary configurations can also be added.

  For example, the processing tools are not limited to those having a multiple configuration, and may be individual one by one.

  Moreover, as a stirrer used for the treatment tool, a suspended stirrer supported in the vicinity of the second filter by being suspended from the upper part in addition to the one suspended in the liquid as described above is used. It may be. Further, the stirring bar can be omitted as necessary.

  Further, the operation member is not limited to a linear one, and may be, for example, one that branches off at the tip side to form a Y-shape, and the number of installation members is not limited to one. It may be.

  In the above embodiment, the cap has been described as having a function of supplying (injecting) air (gas) into the space formed between the first filter and the second filter. It may have a function, and may have a function which supplies air (gas), and other functions.

  In the above embodiment, the cap is representatively described as an example of the gas injection means (gas supply means). However, as the gas injection means, for example, it is built in the apparatus main body, and the processing tool is installed in the holding unit. In this state, it is also possible to have a configuration including a tube attached to the discharge port and a gas (gas) supply source connected to the end side opposite to that connected to the tube discharge port.

  Moreover, a processing tool is not limited to what has two filters (double filter), For example, a test tube, a microplate, a slide glass etc. may be sufficient.

  Further, the reaction processing apparatus is not limited to the temperature adjustment performed so that the non-general-purpose reagent is cooled, but may be configured such that the temperature adjustment is performed so that the general-purpose reagent is cooled. Good.

  Moreover, this temperature adjustment is not limited to cooling, For example, constant temperature and heating (heating) may be sufficient.

It is a perspective view which shows embodiment of the reaction processing apparatus of this invention. It is a piping diagram in the reaction processing apparatus shown in FIG. It is a fragmentary sectional perspective view which shows the structure of the processing tool used by this invention. It is a figure for demonstrating the usage method of the reaction processing apparatus of this invention. It is a figure for demonstrating the usage method of the reaction processing apparatus of this invention. It is a figure for demonstrating the usage method of the reaction processing apparatus of this invention. It is a figure for demonstrating the usage method of the reaction processing apparatus of this invention. It is a figure for demonstrating the usage method of the reaction processing apparatus of this invention. It is a figure for demonstrating the usage method of the reaction processing apparatus of this invention. It is a figure for demonstrating the usage method of the reaction processing apparatus of this invention. It is a figure for demonstrating the usage method of the reaction processing apparatus of this invention. It is a graph which shows the change of specific gravity of the liquid stored in the space defined between the 1st filter and the 2nd filter. It is a longitudinal cross-sectional view which shows the structure of the conventional processing tool (reaction container).

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Processing tool 2 Column main body 20 Space 21 Main body part 210 Base end opening 22 Discharge port 221 Front end opening 3 Extension cup 31 Main body part 310 Base end opening 32 Mounting part 4 Cap 5 First filter 6 Second filter 7 Operation member 8 Floating stirrer 9 Sheet material 10 Biological substance 11 Sample liquid 111 Bubble 14 Processing liquid 15 Specific gravity adjusting liquid 16 Mixed liquid 40 Reaction processing apparatus 41 Apparatus main body 411 Opening 42 First dispensing head 421 Nozzle tip 43 Second dispensing head 431 Fixed nozzle 44 Head moving mechanism 441 Lifting mechanism 442 Movable bridge 443 Drive belt 444 Motor 445 446 Rail 447 448 Wall 45 Holding part 46 Cooling storage part 47 Tip remover 471 Notch 472 Hole 48 Tip rack 49 Recovery container 50 Shelf 51 General purpose reagent 511 Large capacity tank 512 Medium capacity tank 513 Small capacity tank 52 Tube piping 53 Dispensing pump 54 Liquid feed pump 55, 56 Channel 57 Solenoid valve 58 Heater 59 Liquid receiver 60 Channel 61 Drain tank 62 Suction pump 63 Electromagnetic Valve 64 Nozzle cleaning tank 65 Flow path 66 Solenoid valve 66 Solenoid valve 70 Control means 71 Input part 100 Positioning means 110 Main body part 112 Step part 120 Hole 121 Guide hole 130 Screw 141 Interface 901 Reaction vessel 902 Hollow tube 903 Porous plate 904 Hollow tube 905 pore plate

Claims (7)

A reaction processing apparatus capable of processing a biological material by supplying a plurality of types of general-purpose reagents and a plurality of types of non-general-purpose reagents to a treatment tool containing a biological material,
The device body,
A first dispensing head that detachably mounts a disposable nozzle tip and sucks and discharges liquid into the nozzle tip;
A second dispensing head having a fixed nozzle capable of discharging liquid and used a plurality of times;
A head moving mechanism for moving the first dispensing head and the second dispensing head relative to the apparatus main body,
The plurality of types of non-general-purpose reagents are dispensed to the processing tool by the first dispensing head while exchanging the nozzle tips for each type. Regardless of the type, the reaction head is configured to dispense from the fixed nozzle to the processing tool in common by a dispensing head.   The reaction processing apparatus according to claim 1, further comprising a temperature adjustment unit that adjusts a temperature of at least one of the general-purpose reagent and the non-general-purpose reagent.   The reaction processing apparatus according to claim 2, wherein the temperature adjustment unit includes a cooling storage unit that stores the non-general-purpose reagent in a cooled state.   A storage unit that stores the general-purpose reagent for each type, a liquid feed pump that supplies the general-purpose reagent stored in the storage unit to the fixed nozzle, and a flow path from the storage unit to the fixed nozzle is switched. The reaction processing apparatus according to any one of claims 1 to 3, further comprising: a channel switching unit, wherein the type of the general-purpose reagent dispensed by the fixed nozzle is selected by switching the channel switching unit.   The reaction processing apparatus of Claim 4 which has a heater which heats the general purpose reagent supplied to the said fixed nozzle.   Control means for controlling the operation of the first dispensing head and the second dispensing head so as to supply each non-general reagent and each general-purpose reagent to the processing tool in a predetermined order. The reaction treatment apparatus according to claim 1. The treatment tool includes a cylindrical body having a discharge port capable of discharging a liquid at a tip portion;
A first filter fixed near the outlet in the cylinder;
The second filter is provided on the proximal end side of the first filter in the cylinder so as to be movable in the axial direction of the cylinder, and can hold a liquid in a space formed between the first filter and the first filter. And the filter
The reaction processing apparatus according to claim 1, further comprising: an operation member that moves the second filter in an axial direction of the cylindrical body.

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