JP2016024021A - Electrophoretic analysis chip and electrophoretic analyzer - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an electrophoretic analysis chip capable of analyzing a specimen highly accurately.SOLUTION: An electrophoretic analysis chip includes a first reservoir 11 and a second reservoir 21 provided on a substrate 50, an electrophoretic passage 33 through which the first reservoir 11 and the second reservoir 21 are connected together and a specimen is electrophoresed, and a liquid level adjustment part 1 capable of adjusting the liquid level of at least one of a liquid held by the first reservoir 11 and a liquid held by the second reservoir 21.SELECTED DRAWING: Figure 3

Description

  The present invention relates to an electrophoresis analysis chip and an electrophoresis analysis apparatus.

  In the analysis of the specimen, for example, an analysis chip including two reservoirs each having an electrode disposed thereon and a microchannel connecting the two reservoirs is used. The analysis is performed, for example, by measuring a sample introduced into the microchannel via the reservoir.

US Pat. No. 5,482,608

However, the following problems exist in the conventional technology as described above.
If the liquid levels in the two reservoirs are different, flow may occur in the specimen in the microchannel due to the hydrostatic pressure flow. In sample measurement, if the sample in the microchannel moves before applying a voltage to the electrodes, there is a possibility that high-precision measurement may be hindered.

  The present invention has been made in consideration of the above points, and an object of the present invention is to provide an electrophoresis analysis chip and an electrophoresis analysis apparatus capable of analyzing a sample with high accuracy.

  According to the first aspect of the present invention, the first reservoir and the second reservoir provided on the substrate, the electrophoresis channel for connecting the first reservoir and the second reservoir, and the sample migrating, There is provided an electrophoresis analysis chip comprising: a liquid level adjustment unit capable of adjusting a liquid level of at least one of a liquid held in a first reservoir and a liquid held in the second reservoir.

  According to the second aspect of the present invention, the holding unit that holds the electrophoresis analysis chip according to the first aspect of the present invention and the liquid level adjustment unit of the electrophoresis analysis chip held in the holding unit are used. There is provided an electrophoretic analyzer comprising: a displacement portion that displaces at least one liquid level of the liquid held in the first reservoir and the liquid held in the second reservoir.

  According to the third aspect of the present invention, a load is applied to the holding unit that holds the electrophoresis analysis chip according to the first aspect of the present invention, and the liquid level adjustment unit of the electrophoresis analysis chip held in the holding unit. An electrophoretic analyzer including a load applying unit that can be applied is provided.

  In the present invention, the sample can be analyzed with high accuracy.

The perspective view which shows the basic structure of the extracellular endoplasmic reticulum analysis chip concerning an embodiment. II-II sectional view taken on the line of FIG. 1 is an external perspective view showing an electrophoresis analysis chip 300 according to a first embodiment. IV-IV sectional view taken on the line of FIG. The figure for demonstrating the liquid level adjustment process which concerns on embodiment. The figure which shows the relationship between the pushing distance to the load part 1 which concerns on embodiment, and the speed in a flow path. Sectional drawing of the electrophoresis analysis chip | tip 300A which concerns on 2nd Embodiment. The figure for demonstrating a liquid level adjustment process with the electrophoresis analysis chip | tip 300A which concerns on embodiment. FIG. 2 is a schematic configuration diagram of an electrophoresis analysis apparatus 400 that holds an electrophoresis analysis chip 300B according to an embodiment. The external appearance perspective view of the electrophoresis analysis chip 300B which concerns on embodiment. The top view in which the electrophoresis analysis chip 300B, the holding | maintenance part 410, and the displacement part 440 which concern on embodiment are shown. The schematic diagram which shows the electrophoresis analyzer 1500 which concerns on 2nd Embodiment. FIG. 14 is a schematic configuration diagram inside a housing 1510. Sectional drawing which shows the modification of an electrophoresis analysis chip | tip. The top view which shows the modification of an electrophoresis analysis chip | tip. The top view which shows the modification of an electrophoresis analysis chip | tip. Sectional drawing which shows the modification of a load provision member. Sectional drawing which shows the modification of a load provision member. Sectional drawing which shows the modification of a load provision member.

  In one embodiment, the present invention provides an electrophoresis analysis chip for use in analyzing a specimen. Examples of the sample include cells, extracellular vesicles, fine particles, latex particles (including latex particles modified with antibodies and further modified with cells), polymer micelles, and the like. In the present embodiment, a case will be described in which an extracellular endoplasmic reticulum analysis chip for analyzing extracellular endoplasmic reticulum is used as an electrophoresis analysis chip. In the present specification, an extracellular vesicle means a lipid vesicle including exosome (exosome), apoptotic body, microvesicle and the like. In the following, the extracellular endoplasmic reticulum analysis chip (electrophoresis analysis chip) according to the present embodiment will be described by taking as an example the case of analyzing exosomes.

[Exosome]
Exosomes (exosomes) are lipid vesicles having a diameter of about 30 to 100 nm. As a fusion of endosome and cell membrane, various cells such as tumor cells, dendritic cells, T cells, B cells, blood, urine, It is secreted into body fluids such as saliva.
Abnormal cells such as cancer cells existing in the body express a protein specific to the cell membrane. An exosome is a secreted product of a cell and expresses a cell-derived protein as a secretory source on its surface.

  Therefore, by analyzing the protein expressed on the surface of the exosome, abnormalities in the secretory cell can be detected. Here, the surface of the exosome is a membrane surface of a lipid vesicle secreted from a cell, and refers to a portion where the secreted exosome is in contact with the environment in the living body.

  Since exosomes are detected in blood circulating in the living body, abnormalities in the living body can be detected by analyzing exosomes without performing a biopsy test.

[Analysis of exosomes]
As an example, analysis of exosomes using an extracellular endoplasmic reticulum analysis chip can be performed as follows. First, the exosome to be detected is purified. Next, the exosome is brought into contact with the specific binding substance. Here, the specific binding substance means a substance that can specifically bind to a molecule present on the surface of the exosome, and will be described in detail later. Next, the zeta potential of exosome is measured and analyzed using an extracellular endoplasmic reticulum analysis chip. This analysis can be applied not only to exosomes but also to analyzes of the extracellular endoplasmic reticulum in general.

(Specific binding substance)
Specific binding substances include, for example, antibodies, modified antibodies, aptamers, ligand molecules, and the like. Examples of antibodies include IgG, IgA, IgD, IgE, IgM and the like. Examples of IgG include IgG1, IgG2, IgG3, and IgG4. Examples of IgA include IgA1 and IgA2. Examples of IgM include IgM1 and IgM2. Examples of the modified antibody include Fab, F (ab ′) ₂ , scFv and the like. Examples of aptamers include peptide aptamers and nucleic acid aptamers. Examples of the ligand molecule include a ligand of the receptor protein when the molecule to be detected present on the surface of the exosome is a receptor protein. For example, when the molecule present on the surface of the exosome is interleukin, examples of the ligand molecule include G protein.

  Further, the specific binding substance may be labeled with a labeling substance. Examples of the labeling substance include charged molecules such as biotin, avidin, streptavidin, neutravidin, glutathione-S-transferase, glutathione, fluorescent dye, polyethylene glycol, and melittic acid.

(Purification of exosomes)
Each step of this analysis will be described. First, the exosome is purified from a sample containing exosome. Examples of the sample include blood, urine, breast milk, bronchoalveolar lavage fluid, amniotic fluid, malignant exudate, saliva, cell culture fluid and the like depending on the purpose. Especially, it is easy to purify exosomes from blood and urine.

  Examples of the method for purifying exosomes include ultracentrifugation, ultrafiltration, continuous flow electrophoresis, chromatography, and a method using a μ-TAS (Micro-Total Analysis Systems) device.

(Reaction between exosome and specific binding substance)
Next, the exosome is brought into contact with a specific binding substance (antibody, aptamer, etc.). When the molecule to be detected is present on the surface of the exosome, a specific binding substance-exosome complex is formed. By appropriately selecting a specific binding substance, for example, an abnormality associated with a disease such as cancer, obesity, diabetes, or neurodegenerative disease can be detected. Details will be described later.

(Measurement of zeta potential)
As an example, the case where an antibody is used as a specific binding substance will be described. After reacting the exosome with the antibody, the zeta potential of the exosome reacted with the antibody is measured. The zeta potential is the surface charge of the fine particles in the solution. For example, exosomes are negatively charged while antibodies are positively charged. For this reason, the zeta potential of the antibody-exosome complex is positively shifted compared to the zeta potential of the exosome alone. Therefore, by measuring the zeta potential of the exosome reacted with the antibody, the expression of the antigen on the exosome membrane surface can be detected. This is true not only for antibodies but also for positively charged specific binding substances.

As an example, the exosome zeta potential ζ is obtained by performing exosome electrophoresis in the microchannel of an extracellular endoplasmic reticulum analysis chip, optically measuring the exosome electrophoresis speed S, and measuring the measured exosome electrophoresis. Based on the speed S, it can be calculated using the Smolkovsky equation shown in the following equation (1).
U = (ε / η) ζ (1)
In Equation (1), U is the electrophoretic mobility of the exosome to be measured, and ε and η are the dielectric constant and viscosity coefficient of the sample solution, respectively. The electrophoretic mobility U can be calculated by dividing the electrophoretic velocity S by the electric field strength in the microchannel.

  As an example, the exosome electrophoresis speed S is obtained by electrophoresing exosomes in a microchannel of an extracellular endoplasmic reticulum analysis chip, and as an example, irradiating a laser beam on exosomes flowing in the microchannel, Measurement can be performed by obtaining a particle image by scattered light. As an example of the laser beam, one having a wavelength of 488 nm and an intensity of 50 mW can be given.

[Basic structure of extracellular endoplasmic reticulum analysis chip]
FIG. 1 is a perspective view showing a basic structure of an extracellular endoplasmic reticulum analysis chip. 2 is a cross-sectional view taken along line II-II in FIG. The extracellular endoplasmic reticulum analysis chip 100 includes a first reservoir 110, a second reservoir 120, an electrophoresis channel 150 that connects the first reservoir 110 and the second reservoir 120, and a substrate 160. The migration channel 150 is, for example, a millimeter channel or a micro channel. For example, the migration channel 150 has a width of about 200 μm, a height of 50 μm, and a length of about 1000 μm. The electrophoresis channel 150 is a specific binding substance formed by the interaction between an extracellular vesicle or a specific binding substance that specifically binds to a molecule present on the surface of the extracellular vesicle and the extracellular vesicle. -Electrophoresis of an extracellular endoplasmic reticulum complex (for example, an antibody-exosome complex). An example of a specific binding substance includes an antibody, an aptamer, or a combination thereof. Aptamers are, for example, nucleic acid aptamers, peptide aptamers and the like. Examples of molecules recognized by the specific binding substance include antigens, membrane proteins, nucleic acids, sugar chains, glycolipids and the like.
The migration channel 150 has one end connected to the first reservoir and the other end connected to the second reservoir 120. In addition, the first reservoir 110 and the second reservoir 120 are provided on the base 160 and have an electrode 130 and an electrode 140, respectively. For example, the electrode 130 is provided at the bottom of the first reservoir 110, and the electrode 140 is provided at the bottom of the second reservoir 120. As shown in FIG. 2, the electrode 130 and the electrode 140 are each provided in the vicinity of the end of the migration channel 150. In addition, for example, a sample (eg, exosome to be analyzed) is introduced into the first reservoir 110, and a buffer solution is introduced into the second reservoir 120. The buffer solution may be introduced into the first reservoir 110.

  This extracellular endoplasmic reticulum analysis chip 100 is suitable for measuring the zeta potential of an extracellular endoplasmic reticulum. In the following, a method for measuring the zeta potential of exosome using this extracellular endoplasmic reticulum analysis chip will be described, taking as an example the case of analyzing exosomes as specimens or extracellular endoplasmic reticulum.

  First, a sample solution containing exosomes to be analyzed is introduced into the first reservoir 110. The exosome to be analyzed may have been reacted with a specific binding substance. The exosome is extracted from, for example, a culture supernatant or serum, and the sample solution is an exosome suspension in which the exosome is suspended in a buffer solution such as a phosphate buffer solution (PBS). Next, a sample solution containing exosomes is introduced into the migration channel 150. As an example, the exosome can be introduced into the electrophoresis channel 150 by connecting a syringe to the second reservoir 120 and sucking the sample solution. Next, the buffer solution is put into the first reservoir 110 and the second reservoir 120. By adjusting the liquid level (liquid level height) between the first reservoir 110 and the second reservoir 120 by the liquid level adjusting means described later, the generation of hydrostatic pressure flow generated in the migration channel 150 is prevented, and zeta potential measurement is performed. The accuracy can be improved. Subsequently, a voltage is applied between the electrodes 130 and 140 by a control unit (for example, a control unit CONT or a computer 1513 described later), and the exosome is electrophoresed. As an example, the control unit applies a voltage having an electric field strength of about 50 V / cm for about 10 seconds.

  During electrophoresis, the electrophoresis channel 150 is irradiated with laser light, and the scattered light passing through the exosome, which is emitted from the electrophoresis channel 150, is collected using an objective lens or the like, and a light receiving sensor (eg, high A sensitivity camera is used to image exosomes or specific binding substance-exosome complexes. The magnification of the objective lens is about 60 times as an example. The wavelength and intensity of the laser are, for example, a wavelength of 488 nm and an intensity of 50 mW.

  Subsequently, the control unit calculates the electrophoresis speed S of the exosome or the specific binding substance-exosome complex based on the photographed image. Then, the control unit calculates the electrophoretic mobility U by dividing the electrophoretic velocity S by the electric field strength. Subsequently, the control unit calculates the zeta potential of the exosome or the specific binding substance-exosome complex using the Smolkovsky equation described above.

  By using the extracellular endoplasmic reticulum analysis chip in this embodiment, not only the average value of the zeta potential of the specific binding substance-exosome complex but also the zeta potential of the specific binding substance-exosome complex is measured at the level of one particle. can do. Therefore, from the average value of the zeta potential, even if it seems that there is no exosome having a molecule (for example, antigen) recognized by the specific binding substance in the sample, it exists as a minor population. An exosome having the antigen can be detected.

[First Embodiment of Electrophoresis Analysis Chip]
FIG. 3 is an external perspective view showing an electrophoresis analysis chip 300 according to one embodiment. 4 is a cross-sectional view taken along line IV-IV in FIG. The electrophoresis analysis chip 300 of this embodiment includes a reservoir member 10, a flow path member 30, and a substrate 50 that are sequentially stacked. For example, the electrophoresis analysis chip 300 in the present embodiment has a laminated structure (laminated body) composed of at least the reservoir member 10, the flow path member 30, and the substrate 50. In this case, the laminated structure of the electrophoresis analysis chip is a three-layer structure. Further, for example, such a laminated structure of the electrophoresis analysis chip is formed by bonding the reservoir member 10, the flow path member 30, and the substrate 50 to each other.

  In the following description, the direction in which the reservoir member 10, the flow path member 30 and the substrate 50 are sequentially stacked is the Z direction (Z axis), and the length direction of the reservoir member 10, the flow path member 30 and the substrate 50 is the Y direction. The width direction of the reservoir member 10, the flow path member 30, and the substrate 50 orthogonal to the (Y axis), the Z direction, and the Y direction will be appropriately described as the X direction (X axis).

  The reservoir member 10 is made of a material that can be elastically deformed in at least one direction by an external force or the like. Examples of the material of the reservoir member 10 include elastomers such as silicone rubber and PDMS (polydimethylsiloxane). The reservoir member 10 includes a first reservoir 11 and a second reservoir 21 formed on a rectangular parallelepiped base material 15. The 1st reservoir | reserver 11 and the 2nd reservoir | reserver 21 are arrange | positioned at intervals in the Y direction. For example, the first reservoir 11 and the second reservoir 21 are arranged with an interval in the direction of the migration channel 33. The reservoir member 10 is provided with a through hole 16 having a rectangular shape in plan view that penetrates in the Z direction between the first reservoir 11 and the second reservoir 21.

  The first reservoir 11 includes a first holding space 12 that has a circular cross section in a plane parallel to the XY plane, extends in the Z direction, and penetrates the base material. The first holding space 12 is formed surrounded by the first inner wall surface 13. The first holding space 12 is open to the upper surface (first surface) 10 a of the reservoir member 10. On the first surface 10a, as an example, a first protrusion 14 that surrounds and divides the periphery of the opening of the first holding space 12 is projected with a height of several hundreds of micrometers. As shown in FIG. 3, the first protrusion 14 has a chord-like shape that connects between the arc-shaped arc portion 14 a formed around the same axis as the first holding space 12 and the end of the arc-shaped portion 14 a. And a straight portion 14b. The arc portion 14 a is disposed on the −Y side of the first holding space 12. The straight line portion 14 b is disposed on the + Y side of the first holding space 12. As shown in FIG. 4, a part of the inner peripheral surface of the straight portion 14 b constitutes a first inner wall surface 13 that forms the first holding space 12. As an example, a chamfer is formed at a portion of the first holding space 12 that opens to the upper surface 10a except for a part of the inner peripheral surface of the linear portion 14b that forms the first holding space 12 at a height of 1 mm. .

  The second reservoir 21 includes a second holding space 22 that has a circular cross section in a plane parallel to the XY plane, extends in the Z direction, and penetrates the base material. The second holding space 22 is formed surrounded by the second inner wall surface 23. The second holding space 22 opens on the upper surface 10 a of the reservoir member 10. A second ridge 24 is provided on the first surface 10 a so as to surround and divide the periphery of the opening of the second holding space 22. As shown in FIG. 3, the second protrusion 24 has a chord-like shape connecting the arc portion 24 a having a circular arc shape in plan view formed around the same axis as the second holding space 22 and the end of the arc portion 24 a. And a straight portion 24b. The arc portion 24 a is disposed on the + Y side of the second holding space 22. The straight portion 24 b is disposed on the −Y side of the second holding space 22. As shown in FIG. 4, a part of the inner peripheral surface of the straight portion 24 b constitutes a second inner wall surface 23 that forms the second holding space 22. As an example, a chamfer is formed at a portion of the second holding space 22 that opens to the upper surface 10 a except for a part of the inner peripheral surface of the straight portion 24 b that forms the second holding space 22. .

  On the + Y side of the base 15 with respect to the first reservoir 11, an upper surface 10c that is parallel to the upper surface 10a and located on the −Z side of the upper surface 10a is exposed. The + Y side edge of the upper surface 10a and the −Y side edge of the upper surface 10c are connected by a side surface (second surface) 10d parallel to the ZX plane. At least the position of the side surface 10d in the Y direction (eg, the flow path direction) is set so that the first inner wall surface 13 facing the side surface 10d has a thickness capable of elastic deformation when a load is applied to the side surface 10d. (As an example, about 1 mm). As shown in FIG. 3, the side surface 10d is provided with a straight portion 14b and a notch 17 for separating a portion other than the straight portion 14b so that the first inner wall surface 13 can be easily elastically deformed. The notch 17 has a thickness reaching the upper surface 10c from the upper surface 10a, and is formed in a wedge shape whose width gradually decreases from the side surface 10d toward the -Y side.

  The flow path member 30 includes a first holding space 31 (first holding space on the flow path member side), a second holding space 32 (second holding space on the flow path member side), and an electrophoresis flow path 33. The first holding space 31 is provided through the Z holding direction at a position communicating with the first holding space 12. The second holding space 32 is provided so as to penetrate in the Z direction at a position communicating with the second holding space 22. The migration channel 33 is provided so as to connect the first holding space 31 and the second holding space 32 to the surface 30 a facing the substrate 50. For example, the migration channel 33 is formed in a size of about 200 μm in width, 50 μm in height, and about 1000 μm in length. As an example, the flow path member 30 is formed of PDMS as in the case of the reservoir member 10, but, as will be described later, the flow path member 30 is not subjected to a load for directly adjusting the liquid level. May be formed of a large plastic material.

  The substrate 50 has a length that extends outward in the Y direction from the reservoir member 10 and the flow path member 30. An electrode 51 and an electrode 52 are exposed from the channel member 30 on the surface 50 a of the substrate 50 facing the channel member 30. The electrode 51 is provided so that one end faces the first holding space 31 and the other end extends to the + Y side from the reservoir member 10 and the flow path member 30 and is exposed. The electrode 52 is formed such that one end faces the second holding space 32 and the other end extends and is exposed to the −Y side from the reservoir member 10 and the flow path member 30. Examples of the material of the electrode 51 and the electrode 52 include gold, platinum, and carbon.

  As an example, the reservoir member 10 may be prepared by mixing PDMS (SILPOT 184 manufactured by Toray Dow Corning Co., Ltd.) and a cross-linking agent in a ratio of 10: 1 and degassing PDMS into a resin mold. And by solidifying by heating at 80 ° C. for 7 hours. As an example, the flow path member 30 is prepared by pouring PDMS having the same composition as the PDMS used in the preparation of the reservoir member 10 into a glass material having a flow path mold made of a cube having a width of 200 μm and a length of 40 mm, and being cured by heating. And can be produced by peeling from the flow path mold after curing. As an example, the substrate 50 is manufactured by forming the electrode 51 and the electrode 52 on a glass substrate by sputtering or the like. As an example, the reservoir member 10, the flow path member 30 and the substrate 50 are each plasma-irradiated and bonded to each other, whereby the first holding spaces 12 and 31 of the first reservoir 11, the second holding space 22 of the second reservoir 21, Thus, an electrophoresis analysis chip 300 is prepared in which 32 is connected via the electrophoresis channel 33.

Although not shown in FIGS. 3 and 4, the electrophoresis analysis chip 300 may be provided with a cover that covers the opening of the first holding space 12 and the opening of the second holding space 22. However, the cover does not seal the first holding space 12 and the second holding space 22, but covers the first holding space 12 and the second holding space 22 in a state where the air is open to the atmosphere. By covering the opening of the first holding space 12 and the opening of the second holding space 22 with the cover, it is possible to prevent contamination of the sample by the sample or the sample by the surrounding environment.
Further, as an example, the cover is provided with a recess that fits with the first protrusion 14 and the second protrusion 24, and the recess is fitted with the first protrusion 14 and the second protrusion 24, so that the cover is It is possible to cover the opening of the first holding space 12 and the opening of the second holding space 22 while being positioned on the electrophoresis analysis chip 300.

Next, a procedure for adjusting the liquid levels in the first reservoir 11 and the second reservoir 21 before measuring a sample using the electrophoresis analysis chip 300 will be described.
First, a liquid containing a specimen (hereinafter referred to as a specimen liquid as appropriate) is dropped into the second holding space 22 of the second reservoir 21. At this time, the sample liquid is introduced into the second holding space 22 through the opening (sample inlet) of the second holding space 22 on the upper surface 10a. Next, the sample liquid is introduced into the migration channel 33 by inserting, for example, a syringe into the first holding space 12 of the first reservoir 11 to aspirate the sample liquid. Next, the same sample liquid as the sample liquid dropped into the second holding space 22 of the second reservoir 21 is dropped into the first holding space 12 of the first reservoir 11, and the first holding space is sandwiched between the migration flow paths 33. 12 and 31 and the second holding spaces 22 and 32 are connected by the sample liquid.

  At this time, if the liquid level of the sample liquid held in the second holding spaces 22 and 32 is higher than the liquid level of the sample liquid held in the first holding spaces 12 and 31, the hydrostatic pressure is generated in the migration channel 33. The sample liquid containing the sample flows from the second holding spaces 22 and 32 toward the first holding spaces 12 and 31. The flow of the specimen (particle) in the migration channel 33 can be confirmed by the particle image described above. In this case, of the side surface 10d of the reservoir member 10, the position where the thickness between the first inner wall surface 13 and the Y axis including the axis of the first holding space 12 intersects with the side surface 10d is the liquid surface. A load portion 1 (see FIGS. 3 and 4) serving as a position adjusting portion is used, and a load is applied to the load portion 1 in the −Y direction by a stick-like load applying member Sf. For example, the load applying member Sf is provided in an electrophoretic analyzer described later.

  When a load in the −Y direction is applied to the load portion 1 by the load applying member Sf, as shown in FIG. 5, the load portion 1 is mainly thinner than other portions of the first inner wall surface 13. The first inner wall surface 13 at a position opposite to is elastically deformed as the first flexible portion 13 a and bulges into the first holding space 12. In the side surface 10d, since the straight portion 14b and a portion other than the straight portion 14b are separated by the notch 17, when the load is applied to the load portion 1, substantially all of the applied load is the first inner wall surface. It contributes as a force for elastically deforming 13 and the first inner wall surface 13 can be elastically deformed with a minimum necessary load. Since the load applied by the load applying member Sf is consumed by elastic deformation of the load portion 1 thinner than other portions, deformation of other portions of the reservoir member 10 and deformation of the migration flow path 33 of the flow path member 30 are performed. Is suppressed.

  The volume of the first holding space 12 decreases as the first flexible portion 13 a bulges into the first holding space 12. That is, the volume of the first holding space 12 is adjusted by applying a load in the −Y direction to the load portion 1 by the load applying member Sf. The liquid level of the sample liquid held in the first holding space 12 rises (displaces) in accordance with the volume change of the first holding space 12. Therefore, by adjusting the load in the −Y direction on the load portion 1 by the load applying member Sf and adjusting the volume of the first holding space 12, the liquid of the sample liquid held in the first holding space 12 is adjusted. The position can be adjusted. The liquid level of the sample liquid is adjusted so that the liquid level of the sample liquid in the first holding space 12 is flush with the liquid level of the sample liquid in the first holding space 12 and the liquid level of the sample liquid in the second holding space 22. This can be confirmed by the above-described particle image that the flow of the specimen (particles) in the migration channel 33 has stopped.

  Although the liquid level of the sample liquid held in the first holding space 12 may rise, the sample liquid may overflow from the first holding space 12, but the first around the opening of the first holding space 12. Since the protrusion 14 is provided in a protruding manner, the sample liquid can be prevented from leaking from the first reservoir 11. Further, the electrophoresis analysis chip of the present embodiment adjusts the load on the load portion 1 from the outside of the first holding space 12 by the load applying member Sf, and adjusts the volume of the first holding space 12 to thereby adjust the first holding space 12. The liquid level of the sample liquid held in the holding space 12 can be adjusted.

[Example]
After a sample liquid containing 10 ¹⁰ latex particles having an average particle diameter of about 100 nm / ml was dropped into the second holding space 22 of the second reservoir 21, the sample liquid was introduced into the electrophoresis channel 33 using, for example, a syringe. Next, the same sample liquid as the sample liquid dropped into the second holding space 22 of the second reservoir 21 was dropped into the first holding space 12 of the first reservoir 11. Here, for example, the electrophoresis channel 33 was irradiated with laser light through the through-hole 16, and the scattered light of the specimen (latex particles) in the migration channel 33 was observed with a microscope (a dark field microscope as an example). An infinite number of specimens (latex particles) flowing in the direction from the second reservoir 21 toward the first reservoir 11 with Brownian motion were observed in the electrophoresis channel 33.

  When the load application member Sf is used to apply a load in the −Y direction and push it in while observing the flow of the sample with a microscope, the liquid level of the sample liquid held in the first holding space 12 rises. The flow rate of the sample in the electrophoresis channel 33 changed. FIG. 6 shows the pushing distance to the load unit 1 when the electrophoresis analysis chip 300 having the first holding spaces 12 and 31 and the second holding spaces 22 and 32 having a diameter of 4 mm and a height (depth) of 3 mm is used. It is a figure which shows the relationship between a flow path speed. As shown in FIG. 6, it was confirmed that the flow velocity of the specimen in the electrophoresis channel 33 decreases as the pushing distance to the load unit 1 increases. In this case, the pushing distance to the load unit 1 is a change in position in the Y direction when a load is applied to the load unit 1 and pushed.

  Further, when the amount of pushing into the load section 1 is increased, the flow of the specimen in the migration channel 33 in the direction from the second reservoir 21 to the first reservoir 11 is stopped, and only the Brownian motion of the specimen is observed. It was. The time required to apply the load to the load unit 1 and stop the flow of the specimen is, for example, adding or aspirating the specimen liquid to the first reservoir 11 or the second reservoir 21 using, for example, a pipette as in the past. When it was repeated as appropriate, it took about 5 minutes, but when the electrophoresis analysis chip 300 of this embodiment was used, it took about 1 minute.

  Thus, when the flow of the sample in the migration channel 33 stops, as described above, the electrophoresis speed S of the sample in the migration channel 33 is optically measured to measure the zeta potential with high accuracy. Is possible.

As described above, in the present embodiment, the load is held in the first holding space 12 by a simple operation of applying a load to the load portion 1 and elastically deforming the first inner wall surface 13 (first flexible portion 13a). The liquid level of the sample liquid can be easily adjusted. Therefore, in this embodiment, the flow of the sample in the migration channel 33 can be stopped or reduced without increasing the size of the apparatus, and the analysis of the sample can be performed with high accuracy with a simple configuration. Moreover, even if it is not a skilled engineer, the volume of the holding space 12 is changed only by a simple operation of applying a load to the holding space 12 to deform it, and the liquid level in the first holding space and the liquid in the second holding space are changed. Accordingly, the flow of the sample in the electrophoresis channel 33 can be stopped or reduced.
In the present embodiment, since the side surface 10d is separated by the notch 17, the first inner wall surface 13 can be efficiently elastically deformed with the minimum necessary load when applying a load to the load portion 1. In the present embodiment, the first protrusion 14 surrounds the opening of the first holding space 12, and the second protrusion 24 surrounds the opening of the second holding space 22. It is possible to suppress. In the present embodiment, the load applied by the load applying member Sf is mainly consumed by the elastic deformation of the load portion 1 in a portion thinner than other portions, and therefore the reservoir is applied by the load applied by the load applying member Sf. It can suppress that the other location of the member 10, and the migration flow path 33 of the flow path member 30 deform | transform.

  The liquid level adjustment of the sample liquid described in the first embodiment is an adjustment in the direction in which the liquid level of the sample liquid held in the first holding space 12 of the first reservoir 11 is raised. When dropping the sample liquid into the space 12, it is preferable that the liquid level be lower than the liquid level held in the second holding space 22 of the second reservoir 21. Further, the side surface 10d (and the upper surface 10c) serving as the load portion 1 at the time of liquid level adjustment can also be provided on the second reservoir 21 side. In the case where the load unit 1 is provided in both the first reservoir 11 and the second reservoir 21, which of the first holding space 12 of the first reservoir 11 or the second holding space 22 of the second reservoir 21 is the liquid level of the sample liquid. Even when the liquid level is high, by applying a load to the load unit 1 having a lower liquid level, the flow of the sample in the electrophoresis channel 33 can be stopped by adjusting the liquid level of the sample liquid. When the liquid level of the sample liquid held in the second holding space 22 is adjusted (when raised), the sample liquid may overflow from the opening of the second holding space 22, but the second holding space Since the second protrusion 24 protrudes around the opening 22, the sample liquid can be prevented from leaking from the second reservoir 21.

[Second Embodiment of Electrophoresis Analysis Chip]
Next, a second embodiment of the electrophoresis analysis chip 300A will be described with reference to FIGS. In these drawings, the same components as those of the first embodiment shown in FIGS. 1 to 6 are denoted by the same reference numerals, and the description thereof is omitted.

7 and 8 are cross-sectional views of the electrophoresis analysis chip 300A according to the second embodiment.
The first reservoir 11 of the electrophoresis analysis chip 300A shown in FIG. 7 includes a first cylinder portion 18 provided on the upper surface 10a. The first cylinder portion 18 is provided so that the first holding space 12 and the axis line coincide with each other. An inner peripheral surface 18 a of the first cylindrical portion 18 is formed flush with the first inner wall surface 13. The inner peripheral surface 18 a of the first cylindrical portion 18 forms a part of the first inner wall surface 13. The thickness of the first cylindrical portion 18 is set to a thickness that allows the inner peripheral surface 18a to elastically deform and bulge into the first holding space 12 when a load is applied to the outer peripheral surface 18b.

The second reservoir 21 of the electrophoresis analysis chip 300A includes a second cylinder portion 28 provided on the upper surface 10a. The second cylindrical portion 28 is provided so that the second holding space 22 and the axis coincide with each other. An inner peripheral surface 28 a of the second cylindrical portion 28 is formed flush with the second inner wall surface 23. The inner peripheral surface 28 a of the second cylindrical portion 28 forms a part of the second inner wall surface 23. The thickness of the second cylindrical portion 28 is set to a thickness that allows the inner peripheral surface 28a to elastically deform and bulge into the second holding space 22 when a load is applied to the outer peripheral surface 28b.
Other configurations are the same as those of the first embodiment.

A procedure for adjusting the liquid level of the sample liquid in the electrophoresis analysis chip 300A having the above-described configuration will be described.
As an example, when the liquid level of the sample liquid held in the second holding space 22 of the second reservoir 21 is higher than the liquid level of the sample liquid held in the first holding space 12 of the first reservoir 11, the hydrostatic pressure Due to the flow, the sample flows in the migration channel 33 in the direction from the second reservoir 21 toward the first reservoir 11. When the specimen flows in the direction from the second reservoir 21 toward the first reservoir 11, a load is applied to the first cylinder portion 18 of the first reservoir 11 as shown in FIG. 8. For example, a part of the outer peripheral surface 18a of the first cylindrical portion 18 is set as the load portion 1 and a load is applied by the load applying member Sf.

  When a load is applied to the outer peripheral surface 18b as the load portion 1, the first cylindrical portion 18 is elastic so that the inner peripheral surface 18a at a position facing the load portion 1 falls into the first holding space 12 as a first flexible portion. Deform. When the inner peripheral surface 18a is elastically deformed into the first holding space 12, the volume of the first holding space 12 is reduced. As the volume of the first holding space 12 decreases, the liquid level of the sample liquid held in the first holding space 12 rises. Accordingly, in the same manner as described above, the first holding space 12 and the second holding space 12 are adjusted by adjusting the load applied amount to the load portion 1, that is, the push amount applied to the outer peripheral surface 18b by the load applying member Sf while observing the microscope. The liquid level difference of the sample liquid between the holding spaces 22 is eliminated. As a result, the sample flow in the migration channel 33 caused by the difference in the level of the sample liquid between the first holding space 12 and the second holding space 22 is suppressed.

  On the contrary, the liquid level of the sample liquid held in the first holding space 12 of the first reservoir 11 is higher than the liquid level of the sample liquid held in the second holding space 22 of the second reservoir 21. Due to the hydrostatic pressure flow, the specimen flows in the migration channel 33 in the direction from the first reservoir 11 to the second reservoir 21. In this case, a load is applied as the load portion 1 to the outer peripheral surface 28b of the second cylindrical portion 28 of the second reservoir 21, and the inner peripheral surface 28a is used as the first flexible portion in the second holding space 22. By elastically deforming, similarly to the above, the liquid level difference of the sample liquid between the first holding space 12 and the second holding space 22 is eliminated, and the sample liquid between the first holding space 12 and the second holding space 22 is eliminated. It is possible to suppress the specimen flow in the electrophoresis channel 33 caused by the liquid level difference between the two.

  In the electrophoresis analysis chip 300A of the present embodiment, in addition to obtaining the same functions and effects as those of the first embodiment, the level of the sample liquid in either the first holding space 12 or the second holding space 22 is determined. Even if it is high, the liquid level can be easily adjusted. Furthermore, in the present embodiment, since the first cylindrical portion 18 and the second cylindrical portion 28 which are the load portions 1 are cylindrical and are formed with a uniform thickness, the thickness is the same as in the first embodiment. Compared with the case where the load portion 1 with a small thickness is located at a specific location, it is possible to stably apply the load without variation in the amount of load applied depending on the position where the load is applied.

[First Embodiment of Electrophoresis Analyzer]
Next, a first embodiment of an electrophoresis analyzer that performs electrophoresis analysis while holding an electrophoresis analysis chip will be described with reference to FIGS. 9 to 11. In these drawings, the same components as those of the first embodiment of the electrophoresis analysis chip 300 shown in FIGS. 1 to 6 are denoted by the same reference numerals, and the description thereof is omitted.
FIG. 9 is a schematic configuration diagram of an electrophoresis analyzer 400 that holds the electrophoresis analysis chip 300B. FIG. 10 is an external perspective view of the electrophoresis analysis chip 300B used in the present embodiment.

The electrophoretic analyzer 400 includes a holding unit 410, measurement units 420 and 430, a displacement unit 440, and a control unit CONT.
The holding unit 410 holds the periphery of the substrate 50 of the electrophoresis analysis chip 300B from the −Z side, and holds the side surfaces of the substrate 50 from both sides in the Y direction. The holding unit 410 holds the electrophoresis analysis chip 300B and can move at least in the X direction.

  As shown in FIG. 10, the electrophoresis analysis chip 300 </ b> B has a first reservoir 11, a second reservoir 21, an electrophoresis channel 33, and electrodes 51, 52 compared to the electrophoresis analysis chip 300 described in the first embodiment. A plurality of lanes each having the first protrusion 14 and the second protrusion 24 are arranged in the X direction (four lanes in FIG. 10). That is, in the electrophoresis analysis chip 300B, a plurality (four in FIG. 10) of electrophoresis channels 33 extending in the Y direction are provided in parallel to each other and spaced in the X direction. A cut 17 is provided on the side surface 10d of the electrophoresis analysis chip 300B so as to separate the first reservoirs 11 adjacent in the X direction. The 1st protrusion 14 is protrudingly provided around the said opening part so that the opening part of the 1st holding space 12 in each lane may be divided independently. The 1st protrusion 24 is protrudingly provided around the said opening part so that the opening part of the 2nd holding space 22 in each lane may be divided independently. The through hole 16 is formed between the first reservoir 11 and the second reservoir 21 in a size that spans multiple (five) lanes.

  The measurement unit 420 includes a light irradiation unit 421, a detection unit 422, and a mobility calculation unit (not shown). The light irradiation unit 421 irradiates the migration channel 33 with light from the + Z side through the through hole 16. The detection unit 422 detects the sample in the migration channel 33. The mobility calculation unit calculates the zeta potential from the mobility of the specimen from the detection result of the detection unit 422.

  The measurement unit (second measurement unit) 430 includes liquid level sensors 431 and 432. For example, the liquid level sensor 431 is held in the first holding space 12 by projecting detection light onto the sample liquid held in the first holding space 12 and receiving light reflected by the liquid surface. Information on the liquid level of the sample liquid is detected. The liquid level sensor 431 outputs information on the detected liquid level of the sample liquid to the control unit CONT. For example, the liquid level sensor 432 is held in the second holding space 22 by projecting detection light onto the sample liquid held in the second holding space 22 and receiving light reflected by the liquid surface. Information on the liquid level of the sample liquid is detected. The liquid level sensor 432 outputs information related to the detected liquid level of the sample liquid to the control unit CONT.

  The control unit CONT detects information about the liquid level of the sample liquid held in the first holding space 12 detected by the liquid level sensor 431 and the sample liquid held in the second holding space 22 detected by the liquid level sensor 432. Is displayed on the display device DP.

FIG. 11 is a plan view showing the electrophoresis analysis chip 300 </ b> B, the holding unit 410, and the displacement unit 440.
The displacement unit 440 can displace the liquid level of the sample liquid held in the first holding space 12 for each lane using the load units 1 of a plurality of lanes. The displacement part 440 includes a holding frame 441, push sesame 442 and 443, a displacement adjustment screw 444 and an urging member 445. The push sesame 442, 443, the displacement adjusting screw 444, and the urging member 445 are provided independently for each lane.

  The holding frame 441 has a rectangular tube shape extending in the X direction. The holding frame 441 is disposed on the + Y side of the first reservoir 11 in the electrophoresis analysis chip 300B. The holding frame 441 is provided integrally with the holding unit 410. A hole 441b penetrating in the Y direction is formed in the side wall 441a of the holding frame 441 facing the electrophoresis analysis chip 300B in the same position as the load portions 1 of the plurality of lanes in the X direction. A female threaded portion 441d is formed through the side wall 441c of the holding frame 441 facing the side wall 441a so as to be coaxial with the hole 441b.

  As shown in FIG. 11, the push sesame 442 is provided with an acute edge portion at the −Y side end facing the load portion 1. The + Y side of the push sesame 442 is integrally fixed to the push sesame 443. The push sesame 443 includes a shaft portion 443a and an engaging portion 443b. The shaft portion 443a is held in the hole portion 441b so as to be movable in the Y direction. The shaft portion 443a is formed in such a length that the tip projects from the side wall 441a when the engaging portion 443b is engaged with the side wall 441c. The engaging portion 443b is formed with a larger diameter than the shaft portion 443a. The engaging portion 443b is disposed in the internal space of the holding frame 441. The urging member 445 is configured by a coil spring, for example, disposed between the side wall 441a and the engaging portion 443b. The urging member 445 urges the engaging portion 443b in a direction (+ Y side) in which the engaging portion 443b is separated from the side wall 441a. The displacement adjustment screw 444 includes a male screw portion 444a and a head portion 444b. The male screw portion 444a is screwed with the female screw portion 441d. The head portion 444b is disposed outside the + Y side of the side wall 441c.

  In the electrophoretic analyzer 400 having the above configuration, when adjusting the liquid level of the first reservoir 11, first, the displacement adjusting screw 444 corresponding to the lane in which the liquid level is adjusted is rotated. When the displacement adjustment screw 444 is rotated via the head portion 444b, the male screw portion 444a screwed with the female screw portion 441d is rotated against the biasing force of the biasing member 445 and the pitch and rotation of the female screw portion 441d and the male screw portion 444a. It moves in the -Y direction with a movement amount corresponding to the number.

  When the displacement adjustment screw 444 moves in the −Y direction, the push sesame 442 and the push sesame 443 move in the −Y direction. When the push sesame 443 moves in the −Y direction, the tip of the push sesame 443 pushes the load portion 1 of the adjustment target lane. When the load unit 1 is pushed in and elastically deformed, as described above, the volume of the first holding space 12 is reduced, so that the liquid level of the specimen liquid held in the first holding space 12 is increased.

  Information relating to the liquid level of the sample liquid held in the first holding space 12 is detected by the liquid level sensor 431. Information relating to the liquid level of the sample liquid held in the second holding space 22 is detected by the liquid level sensor 432. Information on the liquid level of the sample liquid held in the first holding space 12 and information on the liquid level of the sample liquid held in the second holding space 22 are displayed on the display unit DP via the control unit CONT. Therefore, the operator (operator) is held in the first holding space 12 by adjusting the rotation speed and rotation direction of the displacement adjustment screw 444 in accordance with the liquid level information of the sample liquid displayed on the display unit DP. The liquid level of the sample liquid and the liquid level of the sample liquid held in the second holding space 12 can be made the same.

  Similarly, with respect to the other lanes, by adjusting the rotation speed and the rotation direction of the displacement adjusting screw 444 corresponding to each lane, the liquid level of the sample liquid held in the first holding space 12 for all the lanes can be determined. The liquid level of the sample liquid held in the second holding space 12 can be made the same. In this way, when the difference between the liquid level of the sample liquid held in the first holding space 12 and the liquid level of the sample liquid held in the second holding space 22 is adjusted, the detection is performed as described above. The detection of the sample in the electrophoresis channel 33 by the unit 422 and the zeta potential calculation process by the mobility calculation unit are performed.

  As described above, in the electrophoresis analysis apparatus 400 of the present embodiment, the sample liquid held in the first holding space 12 independently for each lane with respect to the electrophoresis analysis chip 300B provided with a plurality of lanes. The liquid level can be adjusted. Therefore, in the present embodiment, it is possible to suppress the flow of the specimen that occurs due to the liquid level difference between the specimen liquids held in the first holding space 12 and the second holding space 22 for each lane. As a result, in this embodiment, even when a plurality of lanes are provided, it is possible to detect the electrophoresis speed of the specimen for each lane with a reduced error factor and calculate the zeta potential of the specimen with high accuracy. It becomes. Moreover, in this embodiment, since the liquid level is adjusted by pressurization from one direction (+ Y side) for a plurality of lanes arranged in parallel, the apparatus configuration can be simplified and reduced in size and weight. it can. Moreover, in this embodiment, since the notch 17 is provided between adjacent lanes and the load part 1 is separated, the influence of the load (elastic deformation) applied to the load part 1 in each lane is affected by the load of other lanes. It can suppress reaching part 1. In the present embodiment, the first ridge 14 divides the periphery of the opening of the first holding space 12 for each lane, and the second ridge 24 surrounds the opening of the second holding space 22 for each lane. Therefore, even if at least one of the sample liquid held in the first holding space 12 and the sample liquid held in the second holding space 22 overflows from the opening, the sample liquid remains in the other. It is possible to suppress the occurrence of so-called contamination mixed with the sample liquid used in the lane. In the present embodiment, the holding unit 410 and the displacement unit 440 on which the electrophoresis analysis chip 300B is arranged and held may be integrally configured such that the holding unit 410 is provided with the displacement unit 440. In this case, the holding unit 410 is provided in advance with a positioning unit that indicates a position where the electrophoresis analysis chip 300 </ b> B is disposed and a position where the push block 442 of the displacement unit 440 can contact the load unit 1.

In the present embodiment, information related to the liquid level of the sample liquid held in the first holding space 12 and information related to the liquid level of the sample liquid held in the second holding space 22 are displayed on the liquid level display unit DP. Therefore, the displacement unit 440 can be operated and finely adjusted while referring to the displayed liquid level information. Therefore, in this embodiment, even when the liquid level is adjusted manually, it is possible to easily suppress the flow of the specimen caused by the liquid level difference of the specimen liquid.
In addition, the electrophoresis analysis chip in each of the embodiments described above includes a reservoir member having a first reservoir and a second reservoir, and a flow having an electrophoresis channel that connects the first reservoir and the second reservoir to migrate a sample. A laminated structure including a path member and a substrate, and the reservoir member is a liquid capable of adjusting a liquid level of at least one of a liquid held in the first reservoir and a liquid held in the second reservoir. A position adjustment unit is provided. The first reservoir includes a first holding space, and the liquid level adjustment unit can adjust the volume of the first holding space. Since the electrophoresis analysis chip is manufactured by sequentially stacking and adhering the reservoir member 10, the flow path member 30, and the substrate 50, it can be easily manufactured and the manufacturing cost can be reduced. Become. Further, when the members are stacked, an alignment mark (positioning reference portion) may be provided on any one or more of the reservoir member 10, the flow path member 30, and the substrate 50.

[Second Embodiment of Electrophoresis Analyzer]
Next, a second embodiment of the electrophoresis analyzer will be described with reference to FIGS. In these drawings, the same components as those of the first embodiment of the electrophoresis analyzer 400 shown in FIGS. 9 to 11 are denoted by the same reference numerals, and the description thereof is omitted.

FIG. 12 is a schematic diagram showing an electrophoretic analyzer 1500 according to one embodiment.
The electrophoretic analyzer 1500 includes a housing 1510, an electrophoretic analysis chip introduction port 1520, a holding unit 410 that holds the electrophoretic analysis chip 300B, a signal cable 1511, a computer 1513, and an operation screen 1515.

  As shown in FIG. 13, a displacement portion 440 is provided in the housing 1510. The displacement part 440 in the present embodiment is different from the electrophoretic analyzer 400 in that the structure of the holding frame 441 and the push block 443 and the actuator that drives the push block 443 are added.

  A hole 441e is formed through the side wall 441c of the holding frame 441 so as to be coaxial with the hole 441b. The push sesame 443 includes a shaft portion 443a, an engaging portion 443b, and a shaft portion 443c. The shaft portion 443c is held in the hole portion 441e so as to be movable in the Y direction. The shaft portion 443c is formed to have a length that protrudes in the + Y direction from the side wall 441c when the engaging portion 443b comes into contact with the side wall 441c by being biased by the biasing member 445.

  An actuator 445 is provided at a position facing the tip of the shaft portion 443c. The actuator 445 applies a force that moves the pushing sesame 443 in the −Y direction against the biasing force of the biasing member 445 to the shaft portion 443c under the control of the control unit CONT.

  The electrophoretic analyzer 1500 having the above-described configuration operates as follows. First, the electrophoresis analysis chip 300 </ b> B into which the specimen to be examined is introduced is set in the holding unit 410. Subsequently, when the holding unit 410 moves in the direction of the arrow β, the electrophoresis analysis chip 300B moves to the inside of the electrophoresis analysis apparatus 1500.

The liquid level of the first reservoir 11 is adjusted for the electrophoresis analysis chip 300B that has moved into the housing 1510.
First, the control unit CONT detects information about the liquid level of the sample liquid held in the first holding space 12 detected by the liquid level sensor 431 and the liquid of the sample liquid held in the second holding space 22 detected by the liquid level sensor 432. The driving amount of the actuator 445 is obtained based on the information regarding the position. The drive amount of the actuator 445 is, for example, a table that holds in advance a relationship between the push amount of the push sesame 442 and the liquid level change amount of the sample liquid held in the first holding space 12, or a function derived based on the relationship. And so on. The control unit CONT drives the actuator 445 and applies a load to the load unit 1 via the push sesame 442, confirms the detection results of the liquid level sensors 431 and 432, and holds the sample held in the first holding space 12. Until the difference between the liquid level and the liquid level of the specimen liquid held in the second holding space 22 falls within a predetermined value, the actuator 445 is driven and the detection results of the liquid level sensors 431 and 432 are confirmed. Repeat (feedback control).

  In this way, when the difference between the liquid level of the sample liquid held in the first holding space 12 and the liquid level of the sample liquid held in the second holding space 22 is adjusted, the detection is performed as described above. The detection of the sample in the electrophoresis channel 33 by the unit 422 and the zeta potential calculation process by the mobility calculation unit are performed.

  As described above, the electrophoretic analyzer 1500 according to the present embodiment can be held in the first holding space 12 in addition to obtaining the same operation and effect as the electrophoretic analyzer 400 according to the first embodiment. Since the control unit CONT controls the displacement unit 440 according to the information related to the liquid level of the sample liquid and the information related to the liquid level of the sample liquid held in the second holding space 22, the liquid level of the sample liquid can be easily and quickly. It becomes possible to suppress the flow of the specimen caused by the difference.

  As described above, the preferred embodiments according to the present invention have been described with reference to the accompanying drawings, but the present invention is not limited to the examples. Various shapes, combinations, and the like of the constituent members shown in the above-described examples are examples, and various modifications can be made based on design requirements and the like without departing from the gist of the present invention.

  For example, in the electrophoretic analyzers 400 and 1500 of the above-described embodiment, the configuration for adjusting the liquid level of the sample liquid with respect to the first reservoir 11 is illustrated, but the configuration is not limited to this configuration. A displacement part 440 may be provided in both of the second reservoirs 21 and the liquid level of one or both of the first reservoir 11 and the second reservoir 21 may be adjusted according to the detected liquid level of the sample liquid.

  In the above-described embodiment, the push sesame 442 is brought into contact with a part of the electrophoresis analysis chips 300, 300A, and 300B to apply a load. However, the present invention is not limited to this configuration. For example, as shown in FIG. 14, the first outer peripheral surface 19 (or the second outer peripheral surface) of the first inner wall surface 13 (or the second inner wall surface 23) that forms the first holding space 12 (or the second holding space 22). It is also possible to adopt a configuration in which a flexible balloon BL that forms a gas reservoir 19a (or a gas reservoir 29a) with the liquid level adjusting unit is provided as a liquid level adjusting unit. The balloon BL can be provided on the entire circumference or a part of the first outer circumferential surface 19 (or the second outer circumferential surface 29). By operating (pushing in) the balloon BL and changing the volume of the gas reservoir 19a (or the gas reservoir 29a), the pressure of the gas reservoir 19a (or the gas reservoir 29a) can be increased. When the pressure of the gas reservoir 19a (or the gas reservoir 29a) increases, the first outer peripheral surface 19 (or the second outer peripheral surface 29) and the first inner wall surface 13 (or the first inner wall surface 13) facing the gas reservoir 19a (or the gas reservoir 29a). 2 inner wall surface 23) is elastically deformed into the first holding space 12 (or the second holding space 22). Therefore, in the electrophoretic analysis chips 300, 300A, 300B provided with the balloon BL, the volume of the first holding space 12 (or the second holding space 22) changes according to the above volume change, and according to the volume change. Thus, it is possible to adjust the liquid level of the sample liquid held in the first holding space 12 (or the second holding space 22).

  In addition, although the electrophoresis analysis chips 300, 300A, and 300B in the above embodiment are formed of a flexible material and are elastically deformed when a load is applied, an example is shown in FIG. In addition, only the load portion 1 may be formed of a flexible material that can be elastically deformed, and other portions may be formed of a material having rigidity. Further, the electrophoresis analysis chips 300, 300A, and 300B are formed of a material having rigidity, and as shown in FIG. 16, the first inner wall surface 13 (or the second holding space 22) that forms the first holding space 12 (or the second holding space 22). By providing the notch groove 13b in a part of the second inner wall surface 23) and forming a plurality of thin portions (two in FIG. 16), the region sandwiched by the notch grooves 13b is made the load portion 1 that can be elastically deformed. It is also possible. In the case of adopting this configuration, the notch groove 13b can be formed in the same process when the electrophoresis analysis chips 300, 300A, 300B are formed (manufactured), so that the electrophoresis analysis chips 300, 300A, 300B can be formed. It is possible to form the load portion 1 that can be elastically deformed while ensuring the strength (rigidity), which can contribute to a reduction in manufacturing cost.

  In the above-described embodiment, the electrophoretic analysis chips 300, 300A, 300B are configured to apply a load along the Y direction that intersects the extending direction (Z direction) of the first holding space 12 and the second holding space 22. Although illustrated, it cannot be overemphasized that the structure which provides a load along a X direction may be sufficient. In the present invention, a configuration in which a load is applied along the extending direction of the first holding space 12 and the second holding space 22 is also applicable. FIG. 17 shows an example of the configuration of the load applying member Sv that applies a load along the extending direction of the first holding space 12 and the second holding space 22. As shown in FIG. 17, the load applying member Sv includes a lid Sv1 that closes an opening of the first holding space 12 (or the second holding space 22), and the first holding space 12 (or the second holding space 12 of the lid Sv1). And a protrusion Sv2 provided on the surface facing the holding space 22). In the load applying member Sv, when the lid portion Sv1 moves downward, the cylindrical wall portion is elastically deformed in the compression direction, and the amount of protrusion Sv2 immersed in the sample liquid increases. As a result, the volume of the first holding space 12 (or the second holding space 22) can be reduced, and the liquid level of the sample liquid can be raised.

  FIG. 18 shows an example of the configuration of the load applying member Sw that applies a load along the extending direction of the first holding space 12 and the second holding space 22. The load applying member Sw has an outer diameter at the tip portion smaller than the diameter of the opening of the first holding space 12 (or the second holding space 22). The load applying member Sw has a truncated cone part that gradually increases in diameter as it is separated from the tip part. The truncated cone portion has a region having an outer diameter larger than the diameter of the opening of the first holding space 12 (or the second holding space 22). Using the load applying member Sw having the above-described configuration, after the tip portion is inserted into the opening of the first holding space 12 (or the second holding space 22), the load applying member Sw is moved downward to thereby perform the first holding. The opening of the space 12 (or the second holding space 22) is elastically deformed in the direction in which the diameter increases. As a result, the volume of the first holding space 12 (or the second holding space 22) is increased, and the liquid level of the sample liquid can be lowered.

  In the above embodiment, the configuration in which the electrophoretic analysis chips 300, 300A, and 300B are elastically deformed mechanically (physically) is exemplified. However, the electrophoretic analysis chips 300, 300A, and 300B can be elastically deformed thermally. It is. As an example, the electrophoretic analysis chips 300, 300A, and 300B can be elastically deformed by applying heat using a deformable polymer such as a shape memory polymer. The shape memory polymer memorizes the shape formed by molding or machining by the stationary phase in the resin, and transforms the memorized shape into a free shape at a temperature within the temperature range above the shape recovery temperature and below the melting point. Can do. And shape memory polymer can fix this deformation | transformation by cooling to the temperature below shape recovery temperature, maintaining this deformed state. The shape formed by molding or machining is restored by heating the fixed deformation after cooling to a temperature below the shape recovery temperature to a temperature above the shape recovery temperature and below the melting point.

  As an example, as shown in FIG. 19A, the recess 5 formed in the first inner wall surface 13 (or the second inner wall surface 23) has a shape at a temperature lower than the shape recovery temperature. Under a temperature within the temperature range of the shape recovery temperature or higher and lower than the melting point, as shown in FIG. 19B, the shape can be stored as a shape in which the recess 5 is not formed. Accordingly, the first inner wall surface 13 (or the second inner wall surface 23) is heated to the state shown in FIG. 19B where the recess 5 is not formed by heating from the state shown in FIG. 19A where the recess 5 is formed. By deforming, the volume of the first holding space 12 (or the second holding space 22) can be reduced. Contrary to the above configuration, the shape of the recess 5 formed in the first inner wall surface 13 (or the second inner wall surface 23) is a shape under a temperature within the temperature range of the shape recovery temperature or higher and lower than the melting point. The shape when the temperature is lower than the temperature may be a state in which the depression 5 is not formed. In this configuration, the first inner wall surface 13 (or the second inner wall surface is heated to the state shown in FIG. 19A where the recess 5 is formed by heating from the state shown in FIG. 19B where the recess 5 is not formed. By deforming 23), the volume of the first holding space 12 (or the second holding space 22) can be increased.

  Accordingly, both the first region where the dent 5 is formed after heating and the second region where the dent 5 is not formed after heating are provided, and the liquid level of the sample liquid held in the first holding space 12 and the second When the liquid level is lowered according to the liquid level of the sample liquid held in the holding space 22, the first region is heated, and when the liquid level is raised, the second region is heated. Good. Further, the correlation between the temperature and the deformation amount when gradually heating from the temperature lower than the shape recovery temperature to the temperature higher than the shape recovery temperature is obtained in advance, and the first holding space 12 (or the second holding space 22). It is good also as a structure which can adjust the volume change of every minute amount.

Such a shape memory polymer is not particularly limited, and examples thereof include polymer materials such as elastomers having shape memory properties.
Specific examples of the shape memory elastomer include polyurethane, polyisoprene, polyethylene, polynorbornene, styrene-butadiene copolymer; epoxy resin, phenol resin, acrylic resin, polyester, melanin resin; polycaprolactone, polyvinyl chloride, Polymers such as polystyrene, polybutylene succinate, polyethylene terephthalate, polybutylene terephthalate, polyphenylene sulfide, etc., which are crosslinked by a thermal chemical crosslinking method using a peroxide such as an organic peroxide or benzoyl peroxide, for example. Can be mentioned.

  DESCRIPTION OF SYMBOLS 1 ... Load part (liquid level adjustment part), 10a ... Upper surface (1st surface), 10d ... Upper surface (2nd surface), 11 ... 1st reservoir, 12 ... 1st holding space, 13 ... 1st inner wall surface, 13a ... 1st flexible part, 14 ... 1st protrusion, 18 ... 1st cylinder part, 21 ... 2nd reservoir, 22 ... 2nd holding space, 24 ... 2nd protrusion, 28 ... 2nd cylinder part, 33 ... Electrophoretic flow path, 110 ... first reservoir, 120 ... second reservoir, 130, 140 ... electrode, 150 ... electrophoretic flow path, 300, 300A, 300B ... extracellular endoplasmic reticulum analysis chip (electrophoresis analysis chip), 400, 1500 DESCRIPTION OF SYMBOLS ... Electrophoresis analyzer 410 ... Holding | maintenance part 420 ... Measurement part 430 ... Measurement part (2nd measurement part), 400 ... Displacement part, BL ... Balloon (liquid level adjustment part, volume change part), CONT ... Control part , Sf, Sv ... Load Wood

Claims (19)

A first reservoir and a second reservoir provided on the substrate;
An electrophoresis channel that connects the first reservoir and the second reservoir, and the sample migrates;
A liquid level adjusting unit capable of adjusting the liquid level of at least one of the liquid held in the first reservoir and the liquid held in the second reservoir;
An electrophoresis analysis chip comprising: The first reservoir includes a first holding space;
The electrophoresis analysis chip according to claim 1, wherein the liquid level adjustment unit is capable of adjusting a volume of the first holding space. The electrophoresis analysis chip according to claim 1, wherein the second reservoir has a sample introduction port for introducing the sample. The electrophoresis analysis chip according to claim 2, wherein the first holding space is formed by an inner wall surface having a first flexible portion at least in part. 5. The electrophoresis analysis chip according to claim 4, wherein the liquid level adjustment unit includes a volume changing unit that is arranged in the first reservoir and can adjust the volume of the first holding unit by changing the volume. The electrophoretic analysis chip according to claim 4, wherein the liquid level adjustment unit includes a load unit that elastically deforms the first flexible unit in accordance with the load when a load is applied. The base material includes a first tube portion that protrudes from the first surface, and an inner peripheral surface forms a part of the first inner wall surface,
The electrophoresis analysis chip according to claim 6, wherein the load portion is at least a part of an outer peripheral surface of the first tube portion. The base material has a first surface in which the first holding space is opened,
A first protrusion that divides the periphery of the opening of the first holding space;
The electrophoresis analysis chip according to claim 6, wherein the load portion is the first protrusion. The base material includes a first surface in which the first holding space is opened, and a second surface that intersects with the first surface and is separated from the first inner wall surface by a thickness allowing the elastic deformation. Prepared,
The electrophoresis analysis chip according to claim 6, wherein the load portion is the second surface. A plurality of the first reservoir, the second reservoir, and the migration channel;
The second surface is provided to be separated from the plurality of first inner wall surfaces by a thickness that allows the elastic deformation, and is separated by a notch provided between the adjacent first reservoirs. Item 10. The electrophoresis analysis chip according to Item 9. The specimen is an extracellular vesicle, or a specific binding substance formed by the interaction between a specific binding substance that specifically binds to a molecule present on the surface of the extracellular vesicle and the extracellular vesicle. The electrophoretic analysis chip according to any one of claims 1 to 10, comprising an extracellular endoplasmic reticulum complex. An electrophoresis analysis chip for analyzing a sample,
A reservoir member having a first reservoir and a second reservoir;
A flow path member that connects the first reservoir and the second reservoir and has an electrophoresis flow path in which a sample migrates;
A laminated structure composed of a substrate,
The electrophoretic analysis chip, wherein the reservoir member includes a liquid level adjustment unit capable of adjusting a liquid level of at least one of a liquid held in the first reservoir and a liquid held in the second reservoir. A holding unit for holding the electrophoresis analysis chip according to any one of claims 1 to 12,
Using the liquid level adjustment unit of the electrophoresis analysis chip held in the holding unit, at least one liquid level of the liquid held in the first reservoir and the liquid held in the second reservoir is determined. A displacement part to be displaced;
An electrophoretic analyzer comprising:   The electrophoretic analyzer according to claim 13, further comprising a measurement unit that measures a zeta potential of the specimen. A second measuring unit for measuring at least one liquid level of the liquid held in the first reservoir and the liquid held in the second reservoir;
A control unit that controls the displacement unit using a measurement result of the second measurement unit;
The electrophoretic analyzer according to claim 13 or 14, comprising: A holding unit for holding the electrophoresis analysis chip according to any one of claims 6 to 10,
A load applying unit capable of applying a load to the liquid level adjusting unit of the electrophoresis analysis chip held in the holding unit;
An electrophoretic analyzer comprising: A plurality of the first reservoir, the second reservoir, and the migration channel are provided in a direction crossing the migration channel,
The electrophoretic analyzer according to claim 16, wherein the load applying unit can apply a load independently of each other for each of the plurality of liquid level adjusting units.   The electrophoretic analyzer according to claim 16 or 17, further comprising a measurement unit that measures a zeta potential of the specimen. A second measuring unit for measuring at least one liquid level of the liquid held in the first reservoir and the liquid held in the second reservoir;
A control unit for controlling the load applying unit using a measurement result of the second measurement unit;
The electrophoretic analyzer according to claim 16, comprising:

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