JP4298617B2 - Treatment tool - Google Patents

Description

  The present invention relates to a processing tool.

  With the development of bio-related fields, many methods for detecting biological substances such as viruses, cells, genes, proteins, biochemical substances, etc. in samples have been developed.

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

  The in situ hybridization method is a method in which a chromosomal DNA (deoxyribonucleic acid) of a cell or mRNA (ribonucleic acid) in a tissue is targeted and hybridized with an RNA probe or a DNA probe, so that a specific gene on the chromosome is detected. This is a method of detecting the location or detecting the 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.

  The specific operation of such an in situ hybridization method includes, for example, a first step of fixing the biological sample so that the tissue structure of the biological sample does not collapse during the operation, and the probe easily reacts with the nucleic acid of the biological sample. In order to do so, a second step of removing an unnecessary part of the biological sample or a partial decomposition treatment with a proteolytic enzyme, a third step of reacting the probe with the biological sample, and a fourth step of washing and removing the excess probe A step, a fifth step of reacting with a reagent such as an enzyme-labeled antibody that binds to the label to detect the label in the probe hybridized with the nucleic acid of the biological sample, and a step of washing and removing excess antibody 6 steps, 7th step of reacting an enzyme with an enzyme substrate via an antibody, 8th step of washing away and removing the excess enzyme substrate, and taking an image of the enzyme substrate colored by the enzyme reaction It is performed by a ninth step of recording the like.

  Among these, in the second to eighth steps, after the biological sample is brought into contact with a treatment liquid (reaction liquid or washing liquid) containing a reagent, these liquids are replaced with the treatment liquid used in the next step. This is done by repeating the operation.

  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 a large number of samples at once.

  As described above, as a reaction apparatus (processing apparatus) capable of collectively replacing the processing liquid for a large number of samples, each of the biological samples and the processing liquid is accommodated in a plurality of capsule containers, and the processing liquid can be replaced. There has been proposed a method in which liquid is removed by mechanical suction and a new treatment liquid is supplied.

  However, in this apparatus, a soft biological sample may be damaged due to a suction operation of the processing liquid, and the apparatus itself is expensive and lacks versatility.

  In addition, a reaction vessel (processing tool) that can easily perform the in situ hybridization method as described above has also 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, A hollow plate 904 is provided at the tip of the hollow tube 904 and is composed of a pore plate 905.

  This reaction vessel 901 is used as follows. First, a liquid (sample solution) containing a biological sample 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 liquid) containing a biological sample 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 sample is processed with each processing liquid.

  However, in the reaction vessel 901, since the hollow tube 904 is inserted into the hollow tube 902, a gap is generated between them. The treatment liquid easily enters the gap, and when the treatment liquid enters the gap, the treatment liquid may be mixed into the treatment liquid to be used later, which may hinder accurate biological sample processing.

  In addition, since many processing solutions used in the in situ hybridization method are generally expensive, it is desirable to avoid the processing solutions being wasted as much as possible.

Japanese Patent Laid-Open No. 2003-169662

  An object of the present invention is to provide a processing tool that can easily and accurately process a sample such as a biological sample.

Such an object is achieved by the present inventions (1) to (30) below.
(1) a cylinder having a discharge port capable of discharging liquid at the tip;
A first filter fixed near the outlet in the cylinder;
The second filter is provided on the proximal 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
A stirrer housed in the space;
Is fixed to the center portion of the second filter, the operation member operated to move the second filter in the axial direction of the cylindrical body,
A gas injecting means for injecting gas into the liquid stored in the space while covering the front end opening in a state of being detachably attached to the outlet and attached to the outlet. Processing tool.

  (2) The processing tool according to (1), wherein the stirrer is configured to stir the liquid stored in the space in the vicinity of the second filter.

  (3) The processing tool according to (1) or (2), wherein the stirring bar floats in a liquid stored in the space.

  (4) The processing tool according to any one of (1) to (3), wherein the stirring bar has a specific gravity of 0.85 to 0.99.

  (5) The processing tool according to any one of (1) to (4), wherein the stirrer is granular.

  (6) The processing tool according to any one of (1) to (5), wherein the granular stirring bar has an average particle size of 0.5 to 5 mm.

  (7) The treatment according to any one of (1) to (6), wherein at least the vicinity of the surface of the stirrer is made of a material having poor reactivity with the liquid or a substance contained in the liquid. Ingredients.

  (8) The processing tool according to any one of (1) to (7), wherein the stirring bar has hydrophobicity.

  (9) The processing tool according to any one of (1) to (8), wherein the stirrer includes a solid body, a hollow body, or a porous body.

(10) a cylinder having a discharge port capable of discharging liquid at the tip;
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
Is fixed to the center portion of the second filter, the operation member operated to move the second filter in the axial direction of the cylindrical body,
A gas injecting means for removably attaching to the discharge port and covering the opening of the tip in a state of being attached to the discharge port and injecting a gas into the liquid stored in the space is provided. Processing tool.

  (11) The processing tool according to any one of (1) to (10), wherein the gas injection unit includes a cap that is detachably attached to the discharge port.

  (12) The cap has a portion having flexibility in at least a part of the cap, the internal volume is reduced by deforming the portion, and the internal air as the gas passes through the discharge port. The processing tool according to (11), wherein the processing tool is configured to be fed into the inside.

  (13) The processing tool according to (11) or (12), wherein an internal volume of the cap is smaller than a volume on a tip side of the first filter in the cylindrical body.

  (14) The processing tool according to any one of (1) to (13), wherein a length from a base end surface of the first filter to a base end of the cylindrical body is 5 to 30 mm.

  (15) The processing tool according to any one of (1) to (14), wherein the first filter includes a pore plate having a plurality of through pores penetrating in the thickness direction.

  (16) The first filter according to any one of (1) to (15), wherein at least the vicinity of the surface is made of a material having poor reactivity with the liquid or a substance contained in the liquid. Processing tools.

  (17) The processing tool according to any one of (1) to (16), wherein the first filter has hydrophilicity.

  (18) The processing tool according to any one of (1) to (17), wherein the second filter is configured by a pore plate having a plurality of through pores penetrating in the thickness direction.

  (19) The processing tool according to any one of (1) to (18), wherein the second filter is made of a material having poor reactivity with the liquid or a substance contained in the liquid.

  (20) The processing tool according to any one of (1) to (19), wherein the second filter has hydrophilicity.

  (21) The processing tool according to any one of (1) to (20), wherein the operation member has a rod shape.

  (22) The processing tool according to any one of (1) to (21), wherein the operation member is positioned substantially at a center of the cylindrical body.

  (23) The processing tool according to any one of (1) to (22), wherein the operation member is fixed to a central portion of the second filter.

  (24) The processing tool according to any one of (21) to (23), wherein the processing tool includes at least one extension cylinder that is detachably attached to a base end portion of the cylinder.

  (25) The processing tool according to (24), wherein the extension cylinder has a function of guiding the liquid or a member that supplies the liquid into the cylinder when supplying the liquid into the cylinder.

  (26) A plurality of the extension cylinders, and when the extension cylinder is attached to the cylinder body, by appropriately selecting and using the extension cylinder, the total length of the processing tool can be adjusted. 24) The processing tool according to (25).

  (27) The processing tool according to (26), wherein the plurality of extension cylinders include ones having different lengths.

  (28) The processing tool according to any one of (24) to (27), wherein a plurality of the extension cylinders are connected side by side.

  (29) The processing tool according to any one of (1) to (28), wherein a plurality of the cylindrical bodies are connected side by side.

  (30) The processing tool according to any one of (1) to (29), which is used for storing a sample in the space and processing the sample with a processing liquid in this state.

  According to the present invention, by providing the gas injection means, the gas is supplied (injected) into the liquid stored in the space defined by the first filter and the second filter to generate bubbles. Can be made. Thereby, for example, when the liquid is stirred with a stirrer, the stirrer can be moved smoothly, and the liquid can be stirred more reliably. As a result, a sample such as a biological sample can be processed easily and accurately.

Hereinafter, the processing tool of the present invention will be described in detail with reference to the accompanying drawings.
Although the processing tool of the present invention can be used for various purposes, a case where the processing tool is applied to a sample processing tool used to process a sample using various processing solutions will be described below as a representative.

  FIG. 1 is a longitudinal sectional view showing an embodiment when the processing tool of the present invention is applied to a sample processing tool. In the following description, the upper side in FIG. 1 is referred to as a “base end” and the lower side is referred to as a “tip”.

  A sample processing tool 1 shown in FIG. 1 is detachably attached to a column main body 2 and a first filter 5 and a second filter 6 provided in the column main body 2. An extension cup 3 and a cap 4 are provided.

  The sample processing tool 1 stores a liquid (sample liquid) containing a sample 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 Various processing solutions are supplied into the sample 20 through the second filter 6 so that various processing solutions are brought into contact with the sample to perform various processing.

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.

  Various processing liquids are supplied into the column main body 2 (main body portion 21) through the base end opening 210, and various processings are performed on the sample by the processing liquid. Further, the processing liquid after processing the sample is discharged out of the main body portion 21 (column main body 2) through 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, observation of the sample etc. accommodated in the main-body part 21 can be performed easily.

  When the sample processing step includes a heating (heating) step, 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) is preferably about 5 to 30 mm in length from the base end surface to the base end of the first filter 5 provided therein, and is about 10 to 20 mm. Is more preferable. Thus, for example, when the sample after processing is observed from the proximal end side of the column body 2 using an optical microscope or the like, the distance between the sample and the objective lens can be made relatively short, and the state of the sample Can be confirmed 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. 1, 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. A sample is stored in the space 20, and the sample is processed with various processing liquids supplied into 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 sample housed in the space 20 or a filter having a pore diameter that does not allow passage of a floating stirrer 8 described later is suitably used.

  The average pore diameter of the pores of the first filter 5 is appropriately set depending on the size of the sample and the floating stirrer 8 and is not particularly limited, but is preferably about 10 to 150 μm, and about 20 to 100 μm. It is more preferable that

  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 and the like, the liquid passing speed may be slow, and the processing efficiency of the sample may be reduced. On the other hand, if the average pore diameter is too large. There is a possibility that the capillary phenomenon does not occur sufficiently and it is 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, tetrafluoroethylene), and silicone resin, stainless steel, titanium, or a titanium alloy. Various metal materials.

  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 sample processing accuracy 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. 1, a member different from the second filter 6 may be 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 and 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, which may prevent sufficient 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. Further, when a biological sample is handled as a sample, the biological sample generally exists in contact with the first filter 5, but a liquid (sample liquid, processing liquid) in which the floating stirrer 8 is accommodated in the space 20. Or the specific gravity adjustment liquid), the biological sample may be easily 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 low reactivity with a substance (eg, 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 precision of a sample 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. Further, when a biological sample is handled as a sample, even if the floating stirrer 8 collides with the biological sample, the impact can be reduced, so that the biological sample 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 sample can be reduced while suitably maintaining the ability to stir the liquid. Therefore, for example, even when a biological sample is handled as a sample, the biological sample can be suitably prevented from being damaged.

  Such a 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.

  A cap 4, which is substantially flexible as a whole, 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 grasped with a finger or the like and deformed to reduce the internal volume, and the air in the cap 4 is sent into the space 20 through the discharge port 22. 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.

  An extension cup (extension cylinder) 3 is detachably attached to the base end portion of the column 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 a liquid (treatment liquid or specific gravity adjusting liquid) is supplied into the column main body 2 using a pipette, a dispensing pump, or the like, the tip of a tip (a member that supplies liquid) attached thereto However, it is prevented from hitting (contacting) the operation member 7, and the liquid (processing liquid or specific gravity adjusting liquid) can be supplied into the column body 2 more accurately.

  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.

  Further, by attaching the extension cup 3 to the column main body 2, the capacity of the entire sample processing tool 1 can be increased, so that a relatively large amount of liquid (processing liquid and specific gravity adjusting liquid) can be put in the column main body 2 at a time. Will be able to supply. As a result, the processing efficiency of the sample 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 sample processing tool 1 is not used, a plurality of sample processing tools 1 are stacked and stored in a storage container or the like. In this case, the dead space is reduced and the plurality of sample processing tools 1 are efficiently stored. Can do.

  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 quadrangle, a hexagon, etc., as with the column body 2.

  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 or more of polyethylene terephthalate, polypropylene, polyethylene, silicone resin, various resin materials such as silicone rubber, various metal materials, etc. 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.

  The sheet material 9 is removed immediately before the operation of supplying the processing liquid and the specific gravity adjusting liquid into the column main body 2 is started. Thus, the inside of the column main body 2 (sample 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 inside of the column main body 2 (sample processing tool 1). As a result, more accurate processing of the sample can be performed.

  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.

Next, the usage method of such a sample processing tool 1 is demonstrated.
2 to 9 are diagrams (longitudinal sectional views) for explaining how to use the sample processing tool shown in FIG. 1, and FIG. 10 is a diagram between the first filter and the second filter. It is a graph which shows the change of specific gravity of the liquid accommodated in the space formed.

  In the following description, the upper side in FIGS. 2 to 9 is referred to as “base end”, and the lower side is referred to as “tip”. Moreover, in FIG. 6 to FIG. 9, some parts are omitted in order to avoid complicated drawings.

  [1] First, as shown in FIG. 2A, the second filter 6 together with the extension cup 3 and the operation member 7 is removed from the column body 2.

  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. 2B, the liquid (sample liquid) 11 containing the sample 10 is supplied into the column main body 2 from the base end opening 210 of the column main body 2.

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

  Here, as the sample 10, for example, microorganisms such as viruses and bacteria, nucleic acids such as DNA and RNA, antigens and antibodies, proteins such as markers used for diagnosis of various diseases, glycoproteins, hormones, amino acids, Examples include biochemical substances such as peptides and nucleotides.

  When the sample 10 is handled in the form of a block such as a biological tissue section, for example, after the sample 10 is placed on the first filter 5, physiological saline, buffer solution, or the like is placed in the column 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, for example, an injection guide 12 and an injector 13 as shown in FIG.

  The injection guide 12 is composed of a flat plate member, and a through hole 121 penetrating in the thickness direction is formed at the center thereof. On the other hand, the injector 13 is composed of a block-shaped member.

  First, as shown in FIG. 3D, the base end portion of the operation member 7 is grasped 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 FIG. 3 (e), the operation member 7 is inserted into the through hole 121 of the injection guide 12, and the injection guide 12 is placed on the base end surface of the column main body 2.

  At this time, the central axis of the column main body 2 and the central axis of the through-hole 121 of the injection guide 12 are substantially matched. 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, and the first filter 5 and the second filter 6 are positioned substantially in parallel. That is, the second filter 6 is positioned in the horizontal direction.

  Next, the distal end surface of the injector 13 is brought into contact with the proximal end of the operation member 7, and the injector 13 is moved (lowered) toward the distal end in this state. Thereby, the operation member 7 moves toward the distal end direction by being pressed by the injector 13, and accordingly, the second filter 6 also moves toward the distal end direction.

  Then, when the distal end surface of the injector 13 comes into contact with the proximal end surface of the injection guide 12 as shown in FIG. 3 (f), the further movement in the distal end direction is prevented. 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).

  Note that the insertion depth of the second filter 6 (the separation distance from the first filter 5) can be adjusted by setting the thickness of the injection guide 12.

  Next, the injection guide 12 is removed from the column body 2 as shown in FIG.

  [3] Next, as shown in FIG. 4 (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. 4 (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. 5 (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 the sample 10 labeled with a microorganism or radioactive substance is handled, the processing operation can be performed more safely.

  [6] Next, as shown in FIG. 5 (k), the sheet material 9 adhered to the base end of the extension cup 3 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.

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

  Here, the treatment liquid 14 is not particularly limited, and examples thereof include a reaction solution containing a reagent that reacts with the sample 10 and a cleaning liquid.

  In addition, the processing liquid 14 may have a specific gravity smaller, larger, or substantially equal to the specific gravity of the sample liquid 11, but depending on the specific gravity relationship between the processing liquid 14 and the sample liquid 11, an appropriate method may be used. Substitution is preferred.

[7A] When the specific gravity of the treatment liquid 14 is smaller than the specific gravity of the sample liquid 11 First, as shown in FIG. 6A, the treatment liquid 14 is introduced into the extension cup 3 from the proximal end opening 310 using a pipette or the like. Supply.

  Thereby, 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 processing liquid 14 passes through the second filter 6 and enters the space 20. Inflow.

  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. 6C, the space 20 can be almost completely replaced with the processing 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 sample processing tool 1 is started. Thereby, the floating stirrer 8 moves relative to the column body 2 and the sample liquid 11 is stirred.

  The direction in which the sample processing 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 (the front-rear direction, the left-right direction, the rotation in the 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. 7. 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, as shown in FIG. 8A, 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 from the proximal end opening 310 using a pipette or the like. .

  The specific gravity adjusting liquid 15 is preferably one that does not affect the sample 10 (one that does not substantially react with the sample 10). For example, the specific gravity is adjusted in a solvent or dispersion medium used to adjust the treatment liquid 14. The thing etc. which added the substance for use 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 main body 2 in the tip direction, so that it is 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 due to the discharge 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. 8 (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 thus moves (lowers) 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. 9C, 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 center 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 is continued 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. 9C.

  Next, the above operation is repeated by using the same specific gravity adjusting liquid 15 as the specific gravity adjusting liquid 15 or the specific gravity adjusting liquid 15 having a larger specific gravity than the specific gravity adjusting liquid 15. 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.

  Here, for example, a case where the sample liquid 11 having a specific gravity of 1.0 stored 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. 10, 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 sample processing tool 1 is stopped, and the processing liquid 14 is supplied into the column body 2.
Here, since the specific gravity of the mixed liquid 16 stored in the space 20 is larger than that of the processing liquid 14, as shown in FIG. 6B, 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.

  As described above, in the sample processing tool 1, the liquid stored in the space 20 is removed by the weight of the liquid supplied to the base end side from the second filter 6 without using a machine such as a suction device. Can be replaced. For this reason, when handling a biological sample as the sample 10, the damage etc. can be prevented suitably. In addition, since the configuration is simple, the cost can be kept low.

  In addition, the sample processing tool 1 can be made into the structure (multiple structure) by which many things were united. Hereinafter, the sample processing tool 1 having a multiple configuration will be described.

  FIG. 11 is a partial cross-sectional perspective view showing a sample processing tool having a multiple configuration. In the following description, the upper side in FIG. 11 is referred to as “base end”, and the lower side is referred to as “tip”.

  Hereinafter, the sample processing tool 1 shown in FIG. 11 will be described focusing on the differences from the sample processing tool 1 shown in FIG. 1 and the like, and the description of the same matters will be omitted.

  The sample processing tool 1 shown in FIG. 11 is the same as the sample processing tool 1 except that a plurality of column main bodies 2 and a plurality of extension cups 3 are integrally formed (connected side by side).

  As described above, the sample processing tool 1 having a multiple configuration is convenient because a plurality of samples can be processed at a time.

  In this case, the number of connections of the column main body 2 is limited to some extent. However, when the mounting portion 32 of the extension cup 3 is fitted inside the base end portion of the column main body 2, The inner diameter can be set relatively large. For this reason, the internal volume of the column main body 2 can be increased without increasing the length of the column main body 2 more than necessary.

  In addition, by adopting a configuration in which the mounting portion 32 of the extension cup 3 is fitted inside the base end portion of the column main body 2, the liquid can be reliably supplied into the column main body 2.

  FIG. 11 shows the case where both the column body 2 and the extension cup 3 have a multiple configuration, but only one of the column body 2 and the extension cup 3 has a multiple configuration. May be.

  Here, another embodiment of a method for attaching the extension cup (extension cylinder) 3 to the column main body (cylinder body) 2 will be described.

  FIG. 12 is a view (longitudinal sectional perspective view) for explaining another embodiment of a method of mounting the extension cylinder to the cylinder. In the following description, the upper side in FIG. 12 is referred to as “base end” and the lower side is referred to as “tip”.

  In the configuration shown in FIG. 12A, the mounting portion 32 of the extension cup 3 is constituted by an enlarged diameter portion in which the inner diameter of the main body portion 31 is increased, and is fitted to the outer side (outer circumference) of the base end portion of the column main body 2. Configured to get.

  In this case, since the inner diameter of the main body 31 of the extension cup 3 and the inner diameter of the main body 21 of the column main body 2 are substantially equal, the second filter 6 can be used by being positioned in the extension cup 3. become. That is, the volume of the space 20 can be set larger. This allows processing of larger quantities of samples or larger size samples.

  Further, in the configuration shown in FIG. 12B, the mounting portion 32 of the extension cup 3 is constituted by a reduced diameter portion in which the outer diameter of the main body portion 31 is reduced, while the proximal end portion of the column main body 2 is A diameter-enlarged portion is formed in which the inner diameter of the main body portion 21 is increased, and the mounting portion 32 of the extension cup 3 can be fitted into the diameter-expanded portion.

  In this case, the extension cup 3 can have the advantages of both the configuration shown in FIGS. 1 and 11 and the configuration shown in FIG.

  In the above embodiment, a configuration having only one extension cup 3 (one stage) has been described. However, in the present invention, a plurality of (multi-stage) extension cups 3 are provided and used in combination (connected). It can also be configured.

  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.

  In this case, the sample processing tool 1 is used in a state where the extension cup 3 is mounted on 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. Accordingly, by appropriately selecting and using the number and / or type of extension cups 3, the total length of the sample processing tool 1 can be adjusted to a desired one according to the purpose and application.

  The treatment tool of the present invention has been described above. However, the present invention is not limited to these, and the configuration of each part can be replaced with any one that can exhibit the same function, or any The thing of the structure of can also be added.

  For example, as the stirrer, a suspended stirrer supported in the vicinity of the second filter 6 by being suspended from the upper part in addition to the one suspended in the liquid as described above may be used. Good. The stirrer 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. Furthermore, the operation member is not limited to a rod-shaped member, and may be, for example, a tubular member.

  In the embodiment, the cap has a function of supplying (injecting) air (gas) into a space formed between the first filter and the second filter, and other functions. It may be.

  In the above embodiment, the cap has been described as an example of the gas injection means (gas supply means). However, as the gas injection means, for example, a tube that is detachably attached to the discharge port, It can also be set as the structure which has the gas (gas) supply source connected to the edge part side opposite to being connected to a discharge port.

  Moreover, in the said embodiment, although the case where the processing tool of this invention was applied to the sample processing tool used for processing a sample using various process liquids was demonstrated as a representative, the processing tool of this invention is, for example, The present invention can also be applied to an agarose gel having a density gradient (gradient), an apparatus for adjusting sucrose, and the like.

It is a longitudinal cross-sectional view which shows embodiment at the time of applying the processing tool of this invention to a sample processing tool. It is a figure (longitudinal sectional view) for demonstrating the usage method of the sample processing tool shown in FIG. It is a figure (longitudinal sectional view) for demonstrating the usage method of the sample processing tool shown in FIG. It is a figure (longitudinal sectional view) for demonstrating the usage method of the sample processing tool shown in FIG. It is a figure (longitudinal sectional view) for demonstrating the usage method of the sample processing tool shown in FIG. It is a figure (longitudinal sectional view) for demonstrating the usage method of the sample processing tool shown in FIG. It is a figure (longitudinal sectional view) for demonstrating the usage method of the sample processing tool shown in FIG. It is a figure (longitudinal sectional view) for demonstrating the usage method of the sample processing tool shown in FIG. It is a figure (longitudinal sectional view) for demonstrating the usage method of the sample processing tool shown in FIG. 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 fragmentary sectional perspective view which shows the sample processing tool of a multiple structure. It is a figure (longitudinal section perspective view) for demonstrating the other form of the mounting method to the cylinder of an extension cylinder. 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 Sample processing tool 2 Column main body 20 Space 21 Main body part 210 Base end opening 22 Outlet 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 Sample 11 Sample liquid 111 Bubble 12 Injection guide 121 Through hole 13 Injector 14 Treatment liquid 141 Interface 15 Specific gravity adjusting liquid 16 Mixed liquid 901 Reaction vessel 902 Hollow tube 903 Porous plate 904 Hollow tube 905 Pore plate

Claims (6)

A cylinder having a discharge port capable of discharging liquid at the tip;
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
A stirrer housed in the space;
Is fixed to the center portion of the second filter, the operation member operated to move the second filter in the axial direction of the cylindrical body,
A gas injecting means for removably attaching to the discharge port and covering the opening of the tip in a state of being attached to the discharge port and injecting a gas into the liquid stored in the space is provided. Processing tool. A cylinder having a discharge port capable of discharging liquid at the tip;
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
Is fixed to the center portion of the second filter, the operation member operated to move the second filter in the axial direction of the cylindrical body,
A gas injecting means for removably attaching to the discharge port and covering the opening of the tip in a state of being attached to the discharge port and injecting a gas into the liquid stored in the space is provided. Processing tool.   The processing tool according to claim 1, wherein the gas injection unit is configured by a cap that is detachably attached to the discharge port.   The cap has a portion having flexibility at least at a part thereof, the inner volume is reduced by deforming the portion, and the internal air is sent out as the gas into the space through the discharge port. The processing tool according to claim 3, which is configured as described above.   The processing tool according to any one of claims 1 to 4, wherein a plurality of the cylindrical bodies are connected side by side.   The processing tool according to claim 1, wherein the processing tool is used for storing a sample in the space and processing the sample with a processing liquid in this state.

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