WO2013183614A1 - 電気泳動用カセットおよび電気泳動方法 - Google Patents
電気泳動用カセットおよび電気泳動方法 Download PDFInfo
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- WO2013183614A1 WO2013183614A1 PCT/JP2013/065422 JP2013065422W WO2013183614A1 WO 2013183614 A1 WO2013183614 A1 WO 2013183614A1 JP 2013065422 W JP2013065422 W JP 2013065422W WO 2013183614 A1 WO2013183614 A1 WO 2013183614A1
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- sample
- separation medium
- porous member
- sample separation
- electrophoresis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
- G01N27/44743—Introducing samples
Definitions
- the present invention relates to an electrophoresis cassette and an electrophoresis method.
- electrophoresis is known as a method for separating a biological sample.
- the electrophoresis method is a method for separating a sample to be separated such as protein or nucleic acid based on a difference in migration speed of electrophoresis.
- a method is generally used in which a sample is introduced into a gel containing an electrolyte and separated by applying a voltage to both ends of the gel.
- SDS-PAGE sodium dodecyl sulfate
- a protein forms a complex with a negatively charged SDS at a certain rate, and a voltage is applied to both ends of the gel so that the protein-SDS complex moves through the polyacrylamide gel toward the anode. Moving. At that time, proteins are separated according to molecular weight by the molecular sieving effect of polyacrylamide gel.
- Non-Patent Document 1 an acrylamide solution is poured into a space formed by two flat glass plates or resin plates and a spacer sandwiched therebetween, and a recess (well) for sample application is formed therein.
- a gel for SDS-PAGE is prepared by inserting a comb for polymerization and polymerizing in a container.
- Non-Patent Document 1 also includes the preparation of a separation gel for separating sample proteins and a concentration gel for concentrating sample proteins by adjusting acrylamide solutions having different concentrations and polymerizing them in a non-continuous manner. Are listed.
- Patent Document 1 discloses a novel method for preparing a support gel for electrophoresis and a support base / electrophoresis method, in which a portion on which a sample is arranged has a wedge-shaped recess, and the electrophoresis gel A container for making a gel is described.
- precast gel cassettes are filled with gel between two flat glass plates or two resin plates.
- a well for introducing the sample solution into the separation gel is formed in the gel.
- the user injects the sample solution to be separated into the well and starts an electrophoresis experiment.
- JP 2004-45107 A released on February 12, 2004
- Japanese Patent Application No. 2005-252755 released on August 31, 2005
- the shape of the well and the operation of applying the sample solution to the well are important parts because they affect the electrophoresis result.
- the gel is a soft elastic body, it is difficult to form wells with a uniform shape reproducibly on the gel, and the gel is easily deformed when the comb described above is removed. Currently, it takes time to work well.
- Patent Document 2 discloses a sample separated by first-dimensional electrophoresis without forming a well by bringing a gel subjected to first-dimensional electrophoresis into contact with a gel for second-dimensional electrophoresis. Is introduced into a gel for two-dimensional electrophoresis, but this is a technique for introducing a separated sample into a sample separation medium, not a technique for introducing an unseparated sample into a sample separation medium. .
- the present invention has been made in view of the above problems, and has as its main object to provide a technique for introducing an unseparated sample into a sample separation medium without forming a well in the sample separation medium.
- an electrophoresis cassette includes a porous member containing an unseparated sample, a sample separation medium for separating the sample, and the porous member is used for the sample separation. And a pushing tool for pushing the porous member or the sample separation medium so as to be pushed into the medium.
- the pusher device pushes the porous member toward the sample separation medium and pushes the porous member into the sample separation medium, or attaches the porous member to the pusher device and samples the entire pusher device.
- the porous member containing the unseparated sample can be inserted into the sample separation medium.
- the unseparated sample can be introduced into the sample separation medium from the position where the porous member is inserted. Therefore, an unseparated sample can be introduced into a sample separation medium without forming a well, and an unseparated sample can be introduced at a desired position at a desired timing.
- the unseparated sample does not diffuse into the sample separation medium, and high resolution can be obtained during electrophoresis.
- the electrophoresis cassette according to the present invention further includes an auxiliary device disposed on the sample separation medium and in close contact with the sample separation medium, and the porous member and the pushing device are provided on the auxiliary device. It is preferable that a through-hole to be inserted to the sample separation medium is provided.
- the porous member can be pushed into the sample separation medium through the through hole.
- the electrophoresis cassette according to the present invention includes an insulator for storing the sample separation medium with a part of an upper surface thereof exposed, and the auxiliary instrument is fitted to the insulator and the sample is stored. It is preferable to be in close contact with the exposed portion of the separation medium.
- the sample separation medium can be accommodated in the insulator, and electrophoresis can be performed like normal slab electrophoresis.
- the auxiliary device includes a first contact portion
- the pushing device includes a second contact portion, and the first contact portion and the second contact portion. It is preferable that the abutting portions abut against each other to stop the movement of the porous member when the porous member is pushed into the sample separation medium to a predetermined length.
- the porous member even when the porous member is strongly pushed into the sample separation medium, if the porous member is pushed into the sample separation medium to a predetermined length, the first contact portion and the second The contact portions come into contact with each other, and the movement of the porous member stops. Therefore, the pushing length when pushing the pushing tool and the porous member into the sample separation medium can always be made constant. Therefore, it is possible to prevent the pushing tool and the porous member from excessively entering the sample separation medium, to prevent the electrophoresis result of the unseparated sample from being disturbed, and to improve the analysis resolution.
- the electrophoresis cassette according to the present invention may be one in which a hollow portion is formed in the pushing device, and the porous member is loaded in the hollow portion.
- the porous member is loaded in the push-in device, even if time elapses before electrophoresis is performed, if the porous member is not pushed into the sample separation medium, both contact with each other. There is no. Therefore, it is possible to prevent the unseparated sample from diffusing into the sample separation medium before the porous member is pushed into the sample separation medium.
- the electrophoresis cassette according to the present invention may be one in which an acute angle portion protruding toward the sample separation medium is formed at the tip of the pushing instrument.
- the acute angle portion of the pushing tool comes into contact with the sample separation medium, so that a gas such as air enters the contact surface between the sample separation medium and the pushing tool. Can be prevented.
- the porous member may be attached to the tip of the pushing instrument.
- the porous member can be suitably pushed into the sample separation medium by the pushing tool.
- the porous member is preferably made of a material selected from the group consisting of resin, filter paper, agarose gel and glass filter.
- the porous member can suitably contain the unseparated sample, and the sample can be suitably introduced from the porous member into the sample separation medium.
- the electrophoresis is hardly affected.
- the porous member is preferably hydrophilized.
- the porous member can more suitably contain the unseparated sample, and the sample can be more suitably introduced from the porous member into the sample separation medium.
- the sample separation medium is preferably made of a gel.
- the porous member can be successfully pushed into the sample separation medium by the pushing member. Moreover, electrophoresis can be performed suitably.
- the electrophoresis cassette according to the present invention includes a plurality of the porous members, and the pushing instrument includes a plurality of pushing portions for pushing each of the plurality of porous members toward the sample separation medium. It may be.
- a plurality of porous members containing unseparated samples can be simultaneously pushed into the sample separation medium, and the samples in the plurality of porous members can be electrophoresed simultaneously.
- the porous member or the porous member containing the unseparated sample is pushed into the sample separation medium for separating the unseparated sample using the pushing tool.
- the method includes a pressing step of pressing the sample separation medium, and an electrophoresis step of electrophoresis of the unseparated sample moved from the pressed porous member to the sample separation medium.
- an auxiliary instrument provided with a through-hole that is in close contact with the sample separation medium and allows the porous member to pass through to the sample separation medium is disposed.
- the application step of applying the sample solution containing the unseparated sample to the through hole before the pushing step, and the penetration through which the sample solution has been applied after the applying step and before the pushing step It may further include a sample introduction step of inserting the porous member into the hole to cause the porous member to contain the unseparated sample.
- the sample solution containing the unseparated sample is applied to the through hole provided in the auxiliary instrument.
- an unseparated sample can be successfully contained in the porous member by inserting the porous member into the through hole to which the sample solution is applied. Accordingly, the sample can be easily introduced into the porous member without separately performing an operation for introducing the sample into the porous member.
- the electrophoresis method according to the present invention includes an injection step of putting a sample solution containing the unseparated sample into a sample introduction container before the pushing step, and after the injection step and before the pushing step, It may further include a sample introduction step of allowing the porous member to contain the unseparated sample by inserting the porous member into the sample introduction container containing the sample solution.
- an unseparated sample can be easily introduced into the porous member by inserting the porous member into the sample introduction container containing the sample solution.
- agarose in the injection step, agarose may be placed in the sample introduction container, and in the sample introduction step, the agarose in the sample introduction container may be heated and melted.
- agarose can be dissolved in the sample solution by heating and melting the agarose placed in the sample introduction container.
- agarose can be contained together. Since agarose is solidified in the porous member, it is possible to prevent the unseparated sample from moving from the porous member to the outside before the pushing step. Therefore, an unseparated sample can be efficiently introduced into the porous member.
- an electrophoresis cassette that can easily introduce a sample into a sample separation medium without forming a well in the sample separation medium and a related technique.
- FIG. 1 It is a perspective view which shows typically the structure of the cassette for electrophoresis which concerns on one Embodiment of this invention. It is sectional drawing which shows typically the structure of the cassette for electrophoresis which concerns on one Embodiment of this invention.
- A is a perspective view which shows typically the structure of the sample separation part which concerns on one Embodiment of this invention, (b) is when the auxiliary instrument of the sample separation part which concerns on one Embodiment of this invention is removed. It is a perspective view which shows a structure typically. It is a perspective view which shows typically the structure of the cassette for electrophoresis which concerns on other embodiment of this invention.
- FIG. 1 is a perspective view schematically showing a configuration of an electrophoresis cassette according to an embodiment of the present invention
- FIG. 2 is a cross-sectional view taken along line AA in FIG.
- the electrophoresis cassette 100 is attached to and detached from an electrophoresis apparatus that separates a sample to be separated such as protein and nucleic acid and / or a control sample thereof based on a difference in moving speed during electrophoresis. It can be installed.
- the electrophoresis cassette 100 includes a buffer cell 101, an electrophoresis buffer solution tank 102, sample separation medium support plates (insulators) 111 and 113, a sample separation medium 112, an auxiliary instrument 120, unseparated A porous member 132 containing a sample and a pushing tool 131 are provided.
- the buffer cell 101 stores the sample separation unit 110.
- the electrophoresis buffer solution tanks 102 are respectively formed on both sides of the buffer cell 1-1.
- the two electrophoresis buffer tanks 102 formed on both sides are liquid tanks for filling a buffer used for electrophoresis, and sandwich the sample separation unit 110 in pairs.
- the electrophoresis cassette 100 fills the electrophoresis buffer solution tank 102 with a buffer solution, installs electrodes and the like in the buffer solution vessel, and applies a voltage between the two electrophoresis buffer solution tanks 102, thereby providing a sample separation medium.
- the unseparated sample introduced into 112 can be separated in the direction of arrow X in FIGS.
- An unseparated sample is a sample that has not been previously separated by electrophoresis or the like, and is separated by electrophoresis using the electrophoresis cassette according to the present embodiment. A method for introducing the unseparated sample into the porous member 132 will be described later.
- FIG. 3A is a perspective view schematically showing the configuration of a sample separation unit according to an embodiment of the present invention
- FIG. 3B is a schematic view of the configuration when an auxiliary instrument of the sample separation unit is removed. It is a perspective view shown in FIG.
- the sample separation unit 110 is formed by laminating a sample separation medium support plate 111, a sample separation medium 112, a sample separation medium support plate 113, and an auxiliary instrument 120 in this order.
- the auxiliary instrument 120 is provided with through holes 121 and 122, and the through holes are provided for allowing the porous member 132 and the pushing instrument 131 to pass through to the sample separation medium 112.
- the sample separation medium support plate 113 and the auxiliary instrument 120 can be bonded by using a known adhesive and bonding method.
- the sample separation medium 112 is a medium that separates various samples according to their properties by introducing an unseparated sample and performing electrophoresis.
- the sample separation medium 112 is sandwiched between sample separation medium support plates 111 and 113 described later. Further, the sample separation medium 112 may be produced in a space formed by the sample separation medium support plates 111 and 113 and the spacer, and the separately prepared sample separation medium 112 is moved and fixed in the space. Also good.
- the sample separation medium 112 is an acrylamide gel
- an acrylamide solution may be formed by pouring an acrylamide solution into the space and polymerizing the acrylamide, and the prepared acrylamide gel is moved to the space. It may be fixed.
- the sample separation medium 112 is not limited as long as it is a medium that can introduce and separate a sample.
- a gel is preferable.
- the gel include those gelled with a gelling agent selected from the group consisting of polyacrylamide, agarose, agar, and starch. As the gel, agarose gel or polyacrylamide gel is often used.
- sample separation medium support plate The sample separation medium support plates 111 and 113 are support plates that sandwich the sample separation medium 112 between the two support plates.
- the sample separation medium support plate 111 is disposed below the sample separation medium 112, and the sample separation medium support plate 113 is disposed above the sample separation medium 112. Since the sample separation medium support plate 113 is shorter than the sample separation medium support plate 111 in the direction of the arrow X in FIGS. 1 and 2, a part of the sample separation medium 112 is exposed from the sample separation medium support plate 113.
- the unexposed sample separation medium 112 is in contact with the sample separation medium support plate 113, and the exposed sample separation medium 112 is covered with an auxiliary instrument 120 described later.
- an insulator can be suitably used as the material of the sample separation medium support plates 111 and 113.
- the sample separation medium can be accommodated in an insulator, and electrophoresis can be performed like normal slab electrophoresis.
- the insulator for example, a resin such as acrylic resin, polystyrene, or polyethylene terephthalate, or glass, ceramic, or the like can be used to suitably form the support plate.
- the sample separation medium support plates 111 and 113 are bonded via spacers (not shown) arranged on both sides of the sample separation medium 112 so that the sample separation medium 112 can be accommodated.
- the auxiliary instrument 120 is fitted with the sample separation medium support plate 113 and covers the exposed portion of the sample separation medium 112 exposed from the sample separation medium support plate 113.
- the auxiliary instrument 120 has the through holes 121 and 122.
- the size of the through holes is such that the porous member 132 and the pushing instrument 131 can be inserted into the sample separation medium 112. There is no particular limitation on the number of through holes.
- the upper surface of the sample separation medium 112 where the through holes 121 and 122 are provided is exposed, and the through holes 121 and 122 of the auxiliary instrument 120 and the upper surface of the sample separation medium 112 form a groove.
- auxiliary device 120 examples include resins such as acrylic resin, polystyrene, and polyethylene terephthalate, glass, ceramic, and the like. By using these materials, the auxiliary instrument 120 used for electrophoresis can be suitably formed.
- the pushing tool 131 is a tool used to push the porous member 132 or the sample separation medium 112 so that the porous member 132 is pushed into the sample separation medium 112.
- the pushing tool 131 is preferably formed of a hard material. Examples of the material of the pushing tool 131 include resins such as acrylic resin, polystyrene, polyethylene terephthalate, and epoxy resin, or glass and ceramics. Can be mentioned.
- the porous member 132 is a porous member that contains an unseparated sample and moves the unseparated sample to the sample separation medium 112 by contacting the sample separation medium 112.
- the porous member 132 is not limited as long as it is a member that can contain an unseparated sample and can introduce the contained sample into the sample separation medium 112, but is not limited to resin, filter paper, agarose gel, Mention may be made of substances such as glass filters. Although it is not limited to these as resin, For example, an epoxy resin, an acrylic resin, etc. can be used suitably. Moreover, as a form of resin, the form of the filter which has a through-hole can be used suitably, for example. That is, the resin used as the porous member 132 can be, for example, a resin filter. If these substances are used, the sample can be suitably introduced into the sample separation medium 112, and even if inserted into the sample separation medium 112, the electrophoresis is hardly affected.
- porous member 132 is more preferably hydrophilized. By making the porous member 132 hydrophilic, a sample solution containing an unseparated sample can be easily introduced into the porous member 132.
- the porous member 132 made of an epoxy resin, an acrylic resin, a glass filter or the like can be hydrophilized by a method such as corona discharge treatment, ozone irradiation, plasma polymerization treatment, or primer treatment.
- the porous member 132 made of filter paper, agarose gel, or the like is hydrophilic and does not need to be hydrophilized.
- porous member 132 containing the unseparated sample and the pushing tool 131 may be separate members, or both may be bonded.
- the porous member 132 and the pushing tool 131 can be bonded by using a known bonding method such as a double-sided tape.
- FIG. 5 and 6 are enlarged views showing variations of the pushing device according to one embodiment of the present invention and a porous member containing an unseparated sample. More specifically, FIGS. 5 (b), (g) and (l), and FIGS. 6 (b) and (h) are shown in FIGS. 5 (a), (f) and (k), and FIG. 6 (a) and (g), and FIGS. 5 (c), (h) and (m) and FIGS. 6 (c, d) and (i) are respectively the same as FIGS. ) And (k) and FIGS. 6A and 6G are side views.
- FIGS. 6 (e) and (j) are cross-sectional views before the porous member 132 is pushed into the sample separation medium 112, respectively.
- FIGS. 6 (f) and (k) are cross-sectional views after the porous member 132 has been pushed into the sample separation medium 112, respectively.
- FIGS. 5A to 5E show a basic pushing tool 131 and a porous member 132 containing an unseparated sample.
- the porous member 132 does not need to be bonded to the pushing tool 131. However, as shown in the figure, when the porous member 132 and the pushing tool are bonded, these are inserted into the through holes 121 and 122, respectively. It is easier to insert.
- FIGS. 5 (i) and 5 (j) show modified examples of the basic pushing tool 131 and the porous member 132.
- the pushing instrument 131 has an abutting portion (second abutting portion) 143 having a width wider than that of the through-hole 121 at the end on the side where the porous member 132 is not bonded. Therefore, as shown in FIGS. 5 (i) and 5 (j), when the pushing tool 131 is inserted into the through-hole 121 and the porous member 132 is pushed into the sample separation medium 112, the contact portion 143 becomes the auxiliary tool 120. It contacts with the upper surface (first contact part).
- the pushing instrument 131 is strongly pushed into the sample separation medium 112
- the contact portion 143 and the upper surface of the auxiliary instrument 120 are brought into contact with each other, and the movement of the porous member 132 within the sample separation medium 112 is performed. Stop. Therefore, the pushing length when the pushing tool 131 and the porous member 132 are pushed into the sample separation medium 112 can be made constant at all times. Therefore, it is possible to prevent the pushing tool 131 and the porous member 132 from excessively entering the sample separation medium 112 and to prevent the electrophoresis result of the unseparated sample from being disturbed.
- the indentation length is determined in advance by appropriately adjusting the depth of the through-hole 121 and the length of the sample separation medium, and the shapes of the indenter 131 and the porous member 132 in the direction in which the porous member 132 is indented. Can do.
- the pushing instrument 131 includes a concave shape (second contact portion) 146 on its side surface, and the through hole 121 has a convex shape (first contact surface) on its side surface. Part) 147 may be provided. Accordingly, as shown in FIGS. 6 (j) and (k), when the pushing tool 131 is inserted into the through hole 121 and the porous member 132 is pushed into the sample separation medium 112, the concave shape 146 becomes the convex shape 147. And the movement of the porous member 132 in the sample separation medium 112 stops.
- the pushing length when the pushing tool 131 and the porous member 132 are pushed into the sample separation medium 112 can be made constant at all times. Therefore, it is possible to prevent the pushing tool 131 and the porous member 132 from excessively entering the sample separation medium 112 and to prevent the electrophoresis result of the unseparated sample from being disturbed.
- a recess (hollow portion) 144 is formed on the bottom surface of the pusher 131 (the tip of the pusher 131 on the side of the sample separation medium 112), and in the recess 144, A porous member 132 containing a sample for separation may be loaded.
- the porous member 132 is not brought into contact with the sample separation medium 112 unless it is pushed into the sample separation medium 112. Therefore, it is possible to prevent the unseparated sample from diffusing into the sample separation medium 112 before the porous member 132 is pushed into the sample separation medium 112.
- the pusher 131 when the pusher 131 is pushed into the sample separation medium 112, as shown in FIG. 5 (o), the pusher 131 made of a hard material and the sample separation medium support plate 111 are not the porous member 132. Contact. Therefore, the porous member 132 containing the unseparated sample can be always inserted at the same position of the sample separation medium 112.
- a penetrating portion (recessed portion) 145 is formed on the side surface of the pushing instrument 131, and a porous member 132 containing an unseparated sample is loaded in the penetrating portion 145. May be. Thus, even if time elapses before electrophoresis is performed, the porous member 132 is not brought into contact with the sample separation medium 112 unless it is pushed into the sample separation medium 112.
- a depression for loading the porous member 132 may be formed on the side surface of the pushing device instead of the through portion.
- the unseparated sample can be prevented from diffusing into the sample separation medium 112 before the porous member 132 containing the unseparated sample is pushed into the sample separation medium 112, and the sample separation medium can be prevented.
- the porous member 132 can always be inserted at the same position 112.
- an acute angle portion that protrudes toward the sample separation medium 112 is formed on the bottom surface of the pusher 131 (the tip of the pusher 131 on the sample separation medium 112 side). May be.
- the acute angle portion of the pusher 131 comes into contact with the sample separation medium 112, so that a gas such as air is brought into contact with the sample separation medium 112 and the pusher 131. Can be prevented from entering.
- the porous member 132 or the sample is inserted so that the porous member 132 containing the unseparated sample is pushed into the sample separation medium 112 for separating the sample.
- the method includes a pressing process of pressing the separation medium 112 and an electrophoresis process of electrophoresis of an unseparated sample moved from the pressed porous member 132 to the sample separation medium 112.
- the electrophoresis method according to the present embodiment includes an applying step for applying a sample solution to the through-hole 121 and a sample introduction for allowing the porous member 133 not containing a sample to contain an unseparated sample before the pushing step. And a process.
- each said process is demonstrated using figures.
- FIG. 8 is a schematic view showing a method for introducing an unseparated sample into a porous member.
- FIGS. 8A and 8B are diagrams showing a basic sample introduction method.
- the sample solution 135 is put into a well installed in the sample loading chip (sample introduction container) 136 (injection step).
- the sample solution 135 contains an unseparated sample.
- a porous member 133 such as a filter paper not containing a sample is attached to the tip of the pushing instrument 131, and the pushing instrument 131 and the porous member 133 are inserted into the well into which the sample solution 135 has been introduced, and the porous member 133 is inserted. Is immersed in the sample solution 135. Thereby, an unseparated sample can be easily introduced into the porous member 133.
- the porous member 133 and the pushing tool 131 can be bonded by a known bonding method, for example, using a double-sided tape.
- FIGS. 8 (c) and 8 (d) are diagrams showing a modification of the sample introduction method.
- agarose containing an SDS-PAGE loading buffer is placed in advance in the sample loading chip 136, and then the sample solution 135 is placed in the sample loading chip 136.
- the sample loading chip 136 is placed on the heat block 137 and heated to melt the agarose and dissolve it in the sample solution 135.
- tip of the pushing tool 131 is immersed in the sample solution 135 in which the agarose melt
- an unseparated sample and dissolved agarose can be introduced into the porous member.
- agarose solidifies in the porous member 133 it can prevent that an unseparated sample moves outside from the porous member 133 before the pushing process demonstrated later. Therefore, an unseparated sample can be efficiently introduced into the porous member 133.
- FIGS. 8 (e) and 8 (f) are diagrams showing another modification of the sample introduction method.
- the sample solution 135 containing an unseparated sample is added to the through-hole 121 formed in the auxiliary instrument 120 (apply process).
- a porous member 133 attached to the tip of the pushing tool 131 is inserted there, and the porous member 133 is immersed in the sample solution 135. Accordingly, the porous material 133 can be easily made porous without providing a mechanism (for example, the sample loading tip 136) for introducing an unseparated sample into the porous member 133 as shown in FIGS. 8 (a) to 8 (d).
- An unseparated sample can be introduced into the member 133.
- FIG. 9 is a schematic diagram illustrating a method of pushing a porous member containing an unseparated sample into a sample separation medium according to an embodiment of the present invention
- FIG. 10 includes an unseparated sample into the sample separation medium. It is the schematic which shows another method of pushing in the porous member to do.
- the pushing tool 131 and the porous member 132 containing the unseparated sample are inserted into the through-hole 121 of the auxiliary tool 120. Then, the pressing member 141 that presses the pushing tool 131 and the porous member 132 is moved onto the pushing tool 131.
- the pressing member 141 may have a configuration for transporting the pressing instrument 131 and the porous member 132 to the through hole 121 in a state where the pressing instrument 131 is supported.
- the porous member 132 and the pushing instrument 131 containing the unseparated sample are transported and inserted into the through hole 121. Also good. Then, as shown in FIG. 10C, the pushing tool 131 and the porous member 132 may be pressed to push the porous member 132 containing the unseparated sample into the sample separation medium 112.
- the operation of pushing the porous member 132 containing the unseparated sample into the sample separation medium 112 may be an automatic operation or a manual operation.
- an unseparated sample moves to the sample separation medium. Therefore, an unseparated sample can be introduced at the position of the sample separation medium 112 in which the porous member 132 is inserted, and the sample can be prevented from diffusing into the sample separation medium 112. Therefore, an unseparated sample can be introduced at an optimum position of the sample separation medium 112.
- an unseparated sample can be easily introduced into the sample separation medium 112 without forming a well in the sample separation medium 112.
- the unseparated sample moved from the pushed porous member 132 to the sample separation medium 112 is electrophoresed (electrophoresis step).
- electrophoresis step By performing electrophoresis, the sample is separated according to the property of the sample. As described above, since unseparated samples can be prevented from diffusing into the sample separation medium 112, high resolution can be obtained during electrophoresis.
- FIG. 4 is a perspective view schematically showing a configuration of an electrophoresis cassette according to another embodiment of the present invention
- FIG. 7 shows a pushing instrument and an unseparated sample according to another embodiment of the present invention. It is an enlarged view which shows the variation with the porous member to contain.
- (B), (e), and (h) of FIG. 7 are front views of (a), (d), and (g) of FIG. 7, and (c), (f), and (i) of FIG. ) Are side views of FIGS. 7A, 7D and 7G, respectively.
- the pusher 131 includes a plurality of pushers 142, and each pusher 142 is bonded to the porous member 132.
- the auxiliary instrument 120 is provided with a through-hole 122 through which the pushing instrument 131 and the plurality of porous members 132 are inserted to the sample separation medium 112.
- the plurality of porous members 132 containing the unseparated sample can be simultaneously pushed into the sample separation medium 112, and the samples in the plurality of porous members 132 can be electrophoresed simultaneously.
- FIGS. 7 (d) to 7 (f) show modified examples of the pusher 131 and the porous member 132 shown in FIGS. 7 (a) to 7 (c).
- porous members 132 containing unseparated samples are respectively loaded in the recesses 144 formed on the bottom surfaces of the plurality of pushing portions 142 (tip portions on the sample separation medium 112 side of the pushing portions 142). Yes. This has the same effect as the configuration shown in FIGS. 5K to 5O according to the first embodiment.
- FIGS. 7 (g) to 7 (i) show other modified examples of the pushing tool 131 and the porous member 132 shown in FIGS. 7 (a) to 7 (c).
- porous members 132 containing unseparated samples are respectively loaded in through portions (cut-out portions) 145 formed on the side surfaces of a plurality of push-in portions 142, respectively. This has the same effect as the structure shown in FIGS. 7 (d) to 7 (f).
- FIGS. 7G to 7I acute angles projecting toward the sample separation medium 112 respectively on the bottom surfaces of the plurality of push-in portions 142 (tip portions on the sample separation medium 112 side of the push-in portions 142).
- the part is formed. This has the same effect as the structure shown in FIGS. 6A to 6C according to the first embodiment.
- the electrophoresis cassette according to the present invention can be used in the field of manufacturing analyzers for various samples (particularly biological samples).
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Abstract
Description
まず、本発明の第1実施形態を、図1および2を用いて詳細に説明する。図1は、本発明の一実施形態に係る電気泳動用カセットの構成を模式的に示す斜視図であり、図2は、図1においてA-Aで示す矢印断面図である。
本発明の一実施形態に係る電気泳動用カセット100は、タンパク質、核酸などの分離対象サンプルおよび/またはその対照サンプルを、電気泳動時の移動速度等の違いに基づいて分離する電気泳動装置に着脱可能に設置されるものである。
未分離のサンプルは、予め電気泳動等により分離されていないサンプルであり、本実施形態に係る電気泳動用カセットを用いた電気泳動によって分離される。なお、未分離のサンプルを、多孔質部材132に導入する方法については後述する。
次に、サンプル分離部110の個々の構成物について図3を用いて説明する。図3は、(a)は本発明の一実施形態に係るサンプル分離部の構成を模式的に示す斜視図であり、(b)はサンプル分離部の補助器具を取り除いたときの構成を模式的に示す斜視図である。
サンプル分離媒体112は、未分離のサンプルを導入し、電気泳動することで種々のサンプルをその性質に応じて分離する媒体である。
サンプル分離媒体支持板111、113は両支持板の間にサンプル分離媒体112を挟持する支持板である。サンプル分離媒体支持板111は、サンプル分離媒体112の下側に配置され、サンプル分離媒体支持板113は、サンプル分離媒体112の上側に配置される。サンプル分離媒体支持板113は、サンプル分離媒体支持板111よりも図1および2の矢印X方向において短いため、サンプル分離媒体112の一部は、サンプル分離媒体支持板113から露出している。そして、露出していないサンプル分離媒体112はサンプル分離媒体支持板113と接触しているとともに、露出したサンプル分離媒体112は後述する補助器具120に覆われている。
補助器具120は、サンプル分離媒体支持板113と嵌合しており、サンプル分離媒体支持板113から露出しているサンプル分離媒体112の露出部分を覆っている。また、補助器具120は、貫通孔121、122を有しているが、貫通孔の大きさについては、多孔質部材132および押し込み器具131をサンプル分離媒体112まで挿通させることができる程度の大きさがあればよく、貫通孔の数については、特に限定されない。
押し込み器具131は、多孔質部材132がサンプル分離媒体112に押し込まれるように、多孔質部材132またはサンプル分離媒体112を押圧するために用いる器具である。なお、押し込み器具131は、硬質の素材から形成されることが好ましく、押し込み器具131の材料としては、例えば、アクリル樹脂、ポリスチレン、ポリエチレンテレフタラート、エポキシ樹脂等の樹脂、あるいは、ガラス、セラミック等が挙げられる。
多孔質部材132は、未分離のサンプルを含有し、サンプル分離媒体112と接触することにより、未分離のサンプルをサンプル分離媒体112に移動させる多孔質性の部材である。
以下では、押し込み器具131と多孔質部材132とのバリエーションについて、図5および6を用いて説明する。図5および6は、本発明の一実施形態に係る押し込み器具と未分離のサンプルを含有する多孔質部材とのバリエーションを示す拡大図である。より詳細には、図5(b)、(g)および(l)、ならびに、図6(b)および(h)は、それぞれ図5(a)、(f)および(k)、ならびに、図6(a)および(g)の正面図であり、図5(c)、(h)および(m)ならびに図6(c、d)および(i)は、それぞれ図5(a)、(f)および(k)ならびに図6(a)および(g)の側面図である。また、図5(d)、(i)および(n)ならびに図6(e)および(j)は、それぞれ多孔質部材132をサンプル分離媒体112に押し込む前の断面図であり、図5(e)、(j)および(o)ならびに図6(f)および(k)は、それぞれ多孔質部材132をサンプル分離媒体112に押し込んだ後の断面図である。
以下、本発明の一実施形態に係る電気泳動方法について説明する。まず、本発明の一実施形態に係る電気泳動方法は、未分離のサンプルを含有する多孔質部材132が、上記サンプルを分離するためのサンプル分離媒体112に押し込まれるように多孔質部材132またはサンプル分離媒体112を押圧する押し込み工程と、押し込んだ多孔質部材132からサンプル分離媒体112に移動した未分離のサンプルを電気泳動する電気泳動工程とを包含していることを特徴としている。
まず、図8を用いて、サンプルを含有しない多孔質部材(以下、「多孔質部材」ともいう。)133に未分離のサンプルを導入するサンプル導入工程について説明する。図8は、多孔質部材に未分離のサンプルを導入する方法を示す概略図である。
次に、未分離のサンプルを含有する多孔質部材132が、サンプル分離媒体112に押し込まれるように、多孔質部材132またはサンプル分離媒体112を押圧する押し込み工程について、図9および10を用いて説明する。図9は、本発明の一実施形態に係るサンプル分離媒体へ未分離のサンプルを含有する多孔質部材を押し込む方法を示す概略図であり、図10は、サンプル分離媒体へ未分離のサンプルを含有する多孔質部材を押し込む別の方法を示す概略図である。
上記押し込み工程後に、押し込んだ多孔質部材132からサンプル分離媒体112に移動した未分離のサンプルを電気泳動する(電気泳動工程)。電気泳動を行うことにより、サンプルの性質に応じてサンプルが分離される。上述の通り、未分離のサンプルがサンプル分離媒体112に拡散することを防止できるため、電気泳動時には高い分解能が得られる。
第1実施形態では、押し込み器具131が一つの多孔質部材132を押圧する構成について説明しているが、本実施形態においては、押し込み器具131が、複数の多孔質部材132の各々をそれぞれ押し込む複数の押し込み部142を備えている構成について、図4および7を用いて説明する。なお、第1実施形態と同一の部材については、同一の番号を付し、その説明を省略し、第1実施形態と同様の方法等(電気泳動方法、サンプル導入工程等)については、その説明を省略する。
101 バッファーセル
102 電気泳動緩衝液槽
110 サンプル分離部
111、113 サンプル分離媒体支持板(絶縁物)
112 サンプル分離媒体
120 補助器具
121、122 貫通孔
131 押し込み器具
132 多孔質部材
133 サンプルを含有しない多孔質部材
135 サンプル溶液
136 サンプルローディングチップ(サンプル導入用容器)
137 ヒートブロック
141 押圧部材
142 押し込み部
143 当接部(第二当接部)
144 凹部(くりぬき部)
145 貫通部(くりぬき部)
146 凹形状(第二当接部)
147 凸形状(第一当接部)
Claims (15)
- 未分離のサンプルを含有する多孔質部材と、
上記サンプルを分離するためのサンプル分離媒体と、
上記多孔質部材が上記サンプル分離媒体に押し込まれるように上記多孔質部材または上記サンプル分離媒体を押圧する押し込み器具とを備えていることを特徴とする電気泳動用カセット。 - 上記サンプル分離媒体上に配置され、上記サンプル分離媒体に密着している補助器具をさらに備えており、
上記補助器具に、上記多孔質部材および上記押し込み器具を上記サンプル分離媒体まで挿通させる貫通孔が設けられていることを特徴とする請求項1に記載の電気泳動用カセット。 - 上記サンプル分離媒体を、その上面の一部を露出させて収納する絶縁物を備えており、
上記補助器具は、上記絶縁物に嵌合するとともに、上記サンプル分離媒体の露出部分に密着していることを特徴とする請求項2に記載の電気泳動用カセット。 - 上記補助器具は、第一当接部を備えており、
上記押し込み器具は、第二当接部を備えており、
上記第一当接部および上記第二当接部は、上記多孔質部材が上記サンプル分離媒体に予め定められた長さまで押し込まれたとき、互いに当接して上記多孔質部材の移動を停止させるようになっていることを特徴とする請求項2または3に記載の電気泳動用カセット。 - 上記押し込み器具の側面に、くりぬき部が形成されており、
上記くりぬき部内に、上記多孔質部材が装填されていることを特徴とする請求項1~4の何れか一項に記載の電気泳動用カセット。 - 上記押し込み器具の先端部に、上記サンプル分離媒体に向かって突出する鋭角部が形成されていることを特徴とする請求項1~5の何れか一項に記載の電気泳動用カセット。
- 上記押し込み器具の先端部に、上記多孔質部材が取り付けられていることを特徴とする請求項1~4の何れか一項に記載の電気泳動用カセット。
- 上記多孔質部材が、樹脂、ろ紙、アガロースゲルおよびガラスフィルターからなる群より選ばれる物質からなることを特徴とする請求項1~7の何れか一項に記載の電気泳動用カセット。
- 上記多孔質部材が、親水化されていることを特徴とする請求項1~8の何れか一項に記載の電気泳動用カセット。
- 上記サンプル分離媒体が、ゲルからなることを特徴とする請求項1~9の何れか一項に記載の電気泳動用カセット。
- 複数の上記多孔質部材を備え、上記押し込み器具は、上記複数の多孔質部材の各々をそれぞれ上記サンプル分離媒体側に押し込む複数の押し込み部を備えていることを特徴とする請求項1~10の何れか一項に記載の電気泳動用カセット。
- 未分離のサンプルを含有する多孔質部材が、上記未分離のサンプルを分離するためのサンプル分離媒体に押し込まれるように、押し込み器具を用いて上記多孔質部材または上記サンプル分離媒体を押圧する押し込み工程と、
押し込んだ上記多孔質部材から上記サンプル分離媒体に移動した上記未分離のサンプルを電気泳動する電気泳動工程とを包含していることを特徴とする電気泳動方法。 - 上記サンプル分離媒体上に、上記サンプル分離媒体に密着するとともに、上記多孔質部材を上記サンプル分離媒体まで挿通させる貫通孔が設けられた補助器具が配置されており、
上記押し込み工程の前に、上記貫通孔に上記未分離のサンプルを含むサンプル溶液をアプライするアプライ工程と、
上記アプライ工程の後、上記押し込み工程の前に、上記サンプル溶液がアプライされた上記貫通孔に上記多孔質部材を挿入することで、上記多孔質部材に上記未分離のサンプルを含有させるサンプル導入工程とをさらに包含することを特徴とする請求項12に記載の電気泳動方法。 - 上記押し込み工程の前に、上記未分離のサンプルを含むサンプル溶液をサンプル導入用容器に入れる注入工程と、
上記注入工程の後、上記押し込み工程の前に、上記サンプル溶液が入った上記サンプル導入用容器に上記多孔質部材を挿入することで、上記多孔質部材に上記未分離のサンプルを含有させるサンプル導入工程とをさらに包含することを特徴とする請求項12に記載の電気泳動方法。 - 上記注入工程では、アガロースを上記サンプル導入用容器に入れ、
上記サンプル導入工程では、上記サンプル導入用容器内の該アガロースを加熱して溶融させることを特徴とする請求項14に記載の電気泳動方法。
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Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS55161238U (ja) * | 1979-05-04 | 1980-11-19 | ||
JP2003254940A (ja) * | 2002-02-25 | 2003-09-10 | Alfa Wassermann Spa | 基板上に流体サンプルを供給するためのアプリケータ |
JP2010025584A (ja) * | 2008-07-15 | 2010-02-04 | Toppan Printing Co Ltd | 電気泳動器具および電気泳動方法 |
JP2011022061A (ja) * | 2009-07-17 | 2011-02-03 | Toppan Printing Co Ltd | 電気泳動用カセット封止材 |
JP2011133311A (ja) * | 2009-12-24 | 2011-07-07 | Sharp Corp | 電気泳動用器具および電気泳動装置 |
JP2012073078A (ja) * | 2010-09-28 | 2012-04-12 | Toppan Printing Co Ltd | 電気泳動用ゲルカセット及びその製造方法 |
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Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2671290B1 (fr) * | 1991-01-04 | 1993-04-16 | Sebia Sa | Dispositif pour l'application d'echantillons biologiques sur une plaque d'electrophorese. |
IL112147A (en) * | 1994-01-19 | 1999-12-22 | Du Pont | Sample holder and method for automated electrophoresis |
US5759375A (en) * | 1996-05-17 | 1998-06-02 | Purdue Research Foundation | Miniaturized disposable gels for DNA analysis |
US6617111B2 (en) * | 2001-05-10 | 2003-09-09 | Invitrogen Corporation | Method for measuring unit activity of an enzyme |
US6905585B2 (en) * | 2002-09-11 | 2005-06-14 | Temple University Of The Commonwealth System Of Higher Education | Automated system for high-throughput electrophoretic separations |
-
2013
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Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS55161238U (ja) * | 1979-05-04 | 1980-11-19 | ||
JP2003254940A (ja) * | 2002-02-25 | 2003-09-10 | Alfa Wassermann Spa | 基板上に流体サンプルを供給するためのアプリケータ |
JP2010025584A (ja) * | 2008-07-15 | 2010-02-04 | Toppan Printing Co Ltd | 電気泳動器具および電気泳動方法 |
JP2011022061A (ja) * | 2009-07-17 | 2011-02-03 | Toppan Printing Co Ltd | 電気泳動用カセット封止材 |
JP2011133311A (ja) * | 2009-12-24 | 2011-07-07 | Sharp Corp | 電気泳動用器具および電気泳動装置 |
JP2012073078A (ja) * | 2010-09-28 | 2012-04-12 | Toppan Printing Co Ltd | 電気泳動用ゲルカセット及びその製造方法 |
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