JP2019158520A - Micro flow passage chip and method for forming the same - Google Patents

Abstract

To make it easier to adjust the time for migration by a micro flow passage chip having a flow passage for migration.SOLUTION: A micro flow passage chip (1) includes: a plurality of reagent introduction parts; and a flow passage (15) for migration made of a passage groove (11). In at least one of the flow passage (15) for migration, there is a migration time adjusting region (18) having a groove with a different depth from those of the other parts in the flow passage (15) for migration.SELECTED DRAWING: Figure 9

Description

  The present invention relates to a microchannel chip and a manufacturing method thereof.

  In recent years, for example, in the fields of the pharmaceutical industry and the like, development of microchannel chips has been promoted. The microchannel chip includes a microchannel (microcavity) and a microchannel, and is used for performing chemical reaction, separation / analysis, and the like of a small amount of sample in a minute space. For example, one method for separating and analyzing a small amount of sample is capillary electrophoresis. A microchannel chip that can be used for such applications includes a plurality of reagent introduction parts (the above-described microchannels) and a migration channel that includes a channel groove and connects the plurality of reagent introduction parts (the above-described microchannels). With.

  International Publication No. 2012/060186 (Patent Document 1) is to manufacture a micro-channel chip by manufacturing a plastic substrate having a channel groove by a resin molding processing technique using a molding die and bonding a plastic film to it. Is disclosed. In such a microchannel chip, the depth of the channel groove is often constant regardless of the site (position) of the migration channel.

  When electrophoresis of a predetermined sample is performed in the electrophoresis channel of the microchannel chip, from the viewpoint of improving analysis accuracy, the electrophoresis time until the sample reaches a predetermined point in the electrophoresis channel is predetermined. It may be preferable to be within the range of the reference time. In such a case, if the actual migration time deviates from the reference time, it can be said that it is preferable to adjust the migration time so that it falls within the range of the reference time. -There was no suggestion in Patent Document 1.

International Publication No. 2012/060186

  In a microchannel chip having a migration channel, it is desired to easily adjust the migration time.

The microchannel chip according to the present invention is
A plurality of reagent introduction parts;
A migration channel comprising a channel groove and connecting a plurality of the reagent introduction parts,
An electrophoresis time adjustment region formed so as to have a groove depth different from other portions in the electrophoresis channel is provided in at least one location of the electrophoresis channel.

  According to this configuration, the planar view shape (the channel length and the channel width) of the migration channel is changed only by changing the groove depth of the channel groove at at least one site in the migration channel. Therefore, the migration time can be easily adjusted.

  Hereinafter, preferred embodiments of the present invention will be described. However, the scope of the present invention is not limited by the preferred embodiments described below.

As one aspect,
The plurality of reagent introduction parts each include a first electrode insertion part into which an electrode is inserted, a second electrode insertion part, and a third electrode insertion part,
The migration channel has an introduction zone, a sample concentration zone, and a sample separation zone in that order in the main channel connecting the first electrode insertion part and the second electrode insertion part, and the main channel in the main channel The third electrode insertion part is connected to the upstream side of the sample separation zone via a sub-flow channel,
It is preferable that the migration time adjustment region is provided in any one or more of the introduction zone, the sample concentration zone, the sample separation zone, and the sub-flow channel.

  According to this configuration, the sample flowing in the electrophoresis channel is concentrated in the sample concentration zone and then separated in the sample separation zone, so that even a very small amount of sample can be analyzed with high accuracy. In addition, the migration time can be appropriately adjusted by setting the migration time adjustment region at one or more of the introduction zone, the sample concentration zone, the sample separation zone, and the sub-flow path. In this case, the migration time adjustment region may be installed at two or more of the introduction zone, the sample concentration zone, the sample separation zone, and the sub-flow channel, and in this way, different areas in the migration channel. The migration time of each can be adjusted.

As one aspect,
It is preferable that the migration time adjustment region is provided in each of the introduction zone or the sample concentration zone and the sample separation zone or the sub flow channel.

  According to this configuration, the migration time until the sample reaches the sample separation zone and the migration time for the sample to migrate through the sample separation zone can be adjusted.

As one aspect,
The migration time adjustment region is preferably a region having a shallower groove depth than the other part.

  According to this configuration, for example, when a microchannel chip having a channel groove is manufactured by a resin molding processing technique using a molding die, a groove depth of the channel groove is obtained by cutting or polishing a part of the molding die. Thus, the migration time can be easily adjusted.

The manufacturing method of the microchannel chip according to the present invention is as follows.
A chip manufacturing process for manufacturing a micro-channel chip having the above-mentioned channel for migration composed of a channel groove having a predetermined depth using a mold having a linear projection having a shape corresponding to the shape of a desired channel for electrophoresis. When,
Electrophoresis of a predetermined sample in the migration channel using the microchannel chip produced in the chip production step, and a migration time measurement step of measuring the migration time;
When the actual migration time acquired in the migration time measurement step is out of a predetermined reference time, a projection height adjustment step for adjusting the height in at least a partial region of the linear projection,
including.

  According to this configuration, in the chip manufacturing process, it is possible to manufacture a microchannel chip having a migration channel having a desired shape by using a mold having linear protrusions. Thereafter, when the migration time measurement step is executed and the actual migration time is out of the predetermined reference time, the height in at least a part of the linear projection of the mold is adjusted in the projection height adjustment step. Adjust. Then, in the next chip manufacturing process, a region having a groove depth different from that of other portions in the migration channel can be provided in at least one location of the migration channel including the channel groove. Then, the migration time of the specimen is adjusted in the regions having different groove depths. As described above, the migration time can be easily adjusted only by finely adjusting the protruding height of the linear protrusion while using an existing mold. Therefore, it is possible to easily obtain a homogeneous microchannel chip in which the migration time is within a predetermined reference time range.

  Further features and advantages of the present invention will become more apparent from the following description of exemplary and non-limiting embodiments described with reference to the drawings.

Sectional drawing of the microchannel chip concerning an embodiment Schematic diagram of flow path for electrophoresis Flow chart showing the micro-channel chip manufacturing procedure Schematic diagram showing one aspect of the chip manufacturing process Schematic diagram showing one aspect of the chip manufacturing process Cross-sectional view of microchannel chip obtained after the first chip manufacturing process Schematic diagram showing one aspect of the protrusion height adjustment process Schematic showing one aspect of the chip manufacturing process again Cross-sectional view of microchannel chip obtained after another chip manufacturing process

  Embodiments of a microchannel chip and a manufacturing method thereof will be described with reference to the drawings. As shown in FIG. 1, the microchannel chip 1 of this embodiment includes a resin substrate 10 and a resin film 20 bonded to the resin substrate 10. A flow path groove 11 is formed on the joint surface of the resin substrate 10 with the resin film 20, and a migration flow path 15 is formed by the flow path groove 11 covered with the resin film 20.

  As the resin constituting the resin substrate 10, a resin excellent in heat resistance and transparency can be selected as appropriate. The resin substrate 10 can be made of, for example, a resin selected from the group consisting of polycarbonate, cycloolefin copolymer, cycloolefin polymer, polymethylpentene, polystyrene, polymethyl (meth) acrylate, and polyethylene terephthalate.

  The resin substrate 10 has a flow channel 11 on one surface. In the present embodiment, the flow channel 11 is configured to have a branch. The channel groove 11 may have a width of 1 mm or less and a depth of 0.01 mm or more and 0.5 mm or less, for example. In this way, experiments and the like on a minute scale can be performed. A part of the channel groove 11 passes through the resin substrate 10 and opens to the other surface, and communicates with the reagent introduction unit 12 and the waste liquid storage unit 14 (see FIG. 2) through the opening. .

  The outer shape and size of the resin substrate 10 can be appropriately set in consideration of handling properties, analysis suitability (analysis method and suitability for an analysis apparatus), and the like. For example, in the case of a quadrangle (square or rectangular), for example, the side is preferably 10 mm or more and 200 mm or less, more preferably 10 mm or more and 100 mm or less. The outer shape of the resin substrate 10 may be another polygonal shape, a circular shape, an elliptical shape, or the like.

  The resin substrate 10 can be manufactured by a resin molding processing technique using a molding die 5 (see FIG. 4 and the like). The resin substrate 10 can be produced by, for example, injection molding, transfer molding, or extrusion molding. Preferably, the resin substrate 10 can be produced by injection molding.

  As the resin constituting the resin film 20, a resin excellent in heat resistance and transparency can be appropriately selected. The resin film 20 can be made of, for example, a resin selected from the group consisting of polycarbonate, cycloolefin copolymer, cycloolefin polymer, polymethylpentene, polystyrene, polymethyl (meth) acrylate, and polyethylene terephthalate. The resin constituting the resin film 20 may be the same resin as the resin constituting the resin substrate 10 or may be a different resin.

  Although the thickness of the resin film 20 is not specifically limited, For example, it can be 0.01 mm or more and 1 mm or less. By being 0.01 mm or more, wrinkles and the like are hardly generated at the time of joining, and the flow path groove 11 can be sufficiently sealed. Moreover, the favorable followability to the unevenness | corrugation of the resin substrate 10 can be acquired because it is 1 mm or less.

  The resin substrate 10 and the resin film 20 are laminated such that the surface of the resin substrate 10 on which the flow channel 11 is formed and one surface of the resin film 20 are in contact with each other. The resin film 20 is bonded to the resin substrate 10 so as to cover the flow path groove 11. Thus, a micro flow path (electrophoretic flow path 15) including the flow path grooves 11 is formed between the resin substrate 10 and the resin film 20. Such a microchannel chip 1 can be suitably used as a microanalysis chip for chemical analysis such as capillary electrophoresis.

  As an example, FIG. 2 shows a schematic diagram of the migration channel 15 in the microchannel chip 1 for analyzing a tumor marker. As shown in these drawings, the microchannel chip 1 includes a plurality of reagent introduction units 12 and a migration channel 15 that connects the plurality of reagent introduction units 12 to each other. In the present embodiment, the microchannel chip 1 includes six reagent introduction parts 12. In the following, when referring to them without distinction, they are collectively referred to as “reagent introduction unit 12”, and when referring to them with particular distinction, reference numerals “12A” to “12F” correspond to the display of FIG. "Will be described.

  The reagent introduction unit 12 is configured as a cylindrical (for example, cylindrical) storage tank. The reagent introduction part 12 is provided so as to protrude from the surface of the resin substrate 10 opposite to the joint surface with the resin film 20. In addition, the plurality of reagent introduction portions 12 are aligned so as to be arranged in a line at equal intervals along the long side of the rectangular resin substrate 10 in plan view.

  The reagent introduction parts 12A, 12B, and 12F are parts for introducing a buffer solution for electrophoresis. The reagent introduction part 12D is a part for introducing a first labeled antibody solution (for example, a solution containing a DNA-labeled antibody for tumor marker). Reagent introduction parts 12C and 12E are sites for introducing an immune reaction solution between a tumor marker as a specimen and a second labeled antibody solution (for example, a solution containing a fluorescently labeled tumor marker antibody).

  Some of the plurality of reagent introduction parts 12 (in this example, a total of three of the reagent introduction part 12A, the reagent introduction part 12B, and the reagent introduction part 12F into which a buffer solution is introduced) are inserted with respective electrodes. The electrode insertion portion 13 is formed. In this embodiment, the reagent introduction part 12B constitutes the first electrode insertion part 13A, the reagent introduction part 12F constitutes the second electrode insertion part 13B, and the reagent introduction part 12A constitutes the third electrode insertion part 13C. Yes. In the present embodiment, a cathode is inserted into the first electrode insertion portion 13A and the third electrode insertion portion 13C, and an anode is inserted into the second electrode insertion portion 13B. An electrophoresis channel 15 is provided so as to connect the electrode insertion portions 13 to each other. The migration channel 15 includes a main channel 15A and a sub-channel 15B connected to the main channel 15A.

  The main channel 15A is a channel that connects the reagent introduction part 12B, which is the first electrode insertion part 13A, and the reagent introduction part 12F, which is the second electrode insertion part 13B. The sub flow path 15B is a flow path that connects the reagent introduction part 12A, which is the third electrode insertion part 13C, and the reagent introduction part 12F, which is the main flow path 15A and then the second electrode insertion part 13B. Reagent introduction parts 12C, 12D, and 12E are connected to main flow path 15A via a connection flow path, respectively. When the first electrode insertion portion 13A side (reagent introduction portion 12B side) in the main flow path 15A is defined as “upstream side” and the second electrode insertion portion 13B side (reagent introduction portion 12F side) is defined as “downstream side”, Reagent introduction parts 12D, 12E, 12C, and 12A are connected in this order from the upstream side toward the downstream side.

  The microchannel chip 1 includes a plurality of waste liquid storage units 14. In this embodiment, the microchannel chip 1 includes four waste liquid storage units 14. In the following, when referring to them without distinction, they are collectively referred to as “waste liquid storage unit 14”, and when referring to them with particular distinction, reference numerals “14A” to “14D” are used in accordance with the display of FIG. "Will be described.

  The waste liquid storage unit 14 is configured as a cylindrical (for example, cylindrical) storage tank. The waste liquid storage part 14 is provided so as to protrude from the surface of the resin substrate 10 opposite to the joint surface with the resin film 20. In addition, the plurality of waste liquid storage units 14 are arranged so as to be arranged in one row at equal intervals, and are arranged in parallel with the rows of the plurality of reagent introduction units 12.

  The waste liquid reservoirs 14A and 14B are parts (waste liquid reservoirs) for storing surplus portions of the immune reaction liquid introduced from the reagent introduction parts 12C and 12E, respectively. The waste liquid storage unit 14C is a part (waste liquid reservoir) for storing an excess of the buffer solution introduced from the reagent introduction unit 12B. The waste liquid storage part 14D is a part (a waste liquid reservoir) for storing an excess of the first labeled antibody liquid introduced from the reagent introduction part 12D.

  The waste liquid reservoirs 14A to 14D are each connected to the main flow path 15A via a connection flow path. These are connected in the order of the waste liquid reservoirs 14C, 14D, 14B, and 14A from the upstream side toward the downstream side. More specifically, the waste liquid storage unit 14C is connected to the main channel 15A at a position downstream of the reagent introduction unit 12B and upstream of the reagent introduction unit 12D. The waste liquid storage unit 14D is connected to the main flow path 15A at a position downstream of the reagent introduction unit 12D and upstream of the reagent introduction unit 12E. The waste liquid storage part 14B is connected to the main flow path 15A at a position downstream of the reagent introduction part 12E and upstream of the reagent introduction part 12C. The waste liquid storage unit 14A is connected to the main flow path 15A at a position downstream of the reagent introduction unit 12C and upstream of the reagent introduction unit 12A.

  The migration channel 15 of the present embodiment includes an introduction zone Z1, a sample concentration zone Z2, and a sample separation zone Z3 in the main channel 15A. These are provided in the order of the introduction zone Z1, the sample concentration zone Z2, and the sample separation zone Z3 from the upstream side to the downstream side. The introduction zone Z1 is a zone for introducing a buffer solution for electrophoresis from the upstream side of various reagents, and is provided from the reagent introduction part 12B to the waste liquid storage part 14C. The specimen concentration zone Z2 is a zone that concentrates a tumor marker, which is a specimen, using an immune reaction and isotachophoresis (ITP), and is provided from the reagent introduction part 12E to the waste liquid storage part 14A. . When performing isotachophoresis, a voltage is applied between the first electrode insertion portion 13A (reagent introduction portion 12B) and the second electrode insertion portion 13B (reagent introduction portion 12F). The specimen separation zone Z3 is a zone for separating a tumor marker as a specimen from other components using capillary zone electrophoresis (CZE), and is provided from the reagent introduction part 12A to the reagent introduction part 12F. Yes. When performing capillary zone electrophoresis, a voltage is applied between the third electrode insertion portion 13C (reagent introduction portion 12A) and the second electrode insertion portion 13B (reagent introduction portion 12F).

  The migration channel 15 further includes a detection zone Z4 on the downstream side of the sample separation zone Z3 in the main channel 15A. The detection zone Z4 is a zone for detecting the specimen that has migrated through the main flow path 15A, and is provided so as to face the light detection unit provided in the analysis apparatus in a state of being loaded in the analysis apparatus. The light detection unit of the present embodiment is configured to be able to detect fluorescence.

  The migration channel 15 further includes a sub-channel zone Z5 in the sub-channel 15B.

  In the microchannel chip 1 having such a configuration, a tumor marker as a specimen can be analyzed as follows. When a voltage is applied between the first electrode insertion portion 13A and the second electrode insertion portion 13B after the introduction of each reagent, the first labeled antibody (DNA labeled) is detected in the specimen concentration zone Z2 according to the principle of isotachophoresis. The tumor marker antibody) moves toward the first electrode insertion portion 13A. Thereafter, an immune complex of the tumor marker, the first labeled antibody, and the second labeled antibody (fluorescently labeled tumor marker antibody) is formed, and the complex moves in the direction of the second electrode insertion portion 13B. The separation zone Z3 is reached. In the present embodiment, the time from when the voltage is applied until the immune complex reaches the specimen separation zone Z3 is referred to as “handoff time”, which is an example of “electrophoresis time”. Since the unreacted (free) second labeled antibody has no charge in the molecule, it remains at that position and does not reach the sample separation zone Z3.

  When the immune complex of the tumor marker, the first labeled antibody, and the second labeled antibody reaches the specimen separation zone Z3, the electrodes are switched and a voltage is applied between the third electrode insertion portion 13C and the second electrode insertion portion 13B. Apply. The immune complex and the unreacted (free) first labeled antibody move in the direction of the second electrode insertion portion 13B in the specimen separation zone Z3 at respective moving speeds according to the charge and the molecular size. Then, the concentration of the tumor marker can be measured from the peak area of the fluorescence intensity when the immune complex including the second labeled antibody having the tumor marker as the core and having the fluorescent dye reaches the detection zone Z4. Since the unreacted first labeled antibody does not have a fluorescent dye in the molecule, even if it reaches the detection zone Z4, it does not affect the fluorescence intensity and does not affect the measurement of the tumor marker concentration. In the present embodiment, the time from when the immune complex reaches the specimen separation zone Z3 until it reaches the detection zone Z4 is referred to as “zone migration time”, which is also an example of “migration time”.

  From the viewpoint of improving analysis accuracy, it may be preferable that the handoff time and the zone migration time described above are within a predetermined reference time range. The reference time for the handoff time and the zone migration time is a time having a predetermined range defined by a lower limit value and an upper limit value, respectively. In order to keep at least one of the handoff time and the zone migration time within the range of the reference time, the microchannel chip 1 of the present embodiment has the migration time adjustment region 18 (see FIG. 9). This migration time adjustment region 18 is a region formed so as to have a groove depth different from other portions in the migration channel 15, and more specifically than the other sites in the migration channel 15. This is a region having a shallow groove depth. In the present embodiment, the migration time adjustment region 18 is formed so as to have the same groove width as other parts in the migration channel 15 and a shallower groove depth than the other parts.

  The migration time adjustment region 18 may be provided at any one of the migration channels 15, but may be provided at two or more locations in the migration channel 15. As described above, the migration channel 15 includes the main channel 15A including the introduction zone Z1, the sample concentration zone Z2, the sample separation zone Z3, and the detection zone Z4, and the subchannel 15B including the subchannel zone Z5. Contains. As an example, the migration time adjustment region 18 may be provided in any two locations of the introduction zone Z1, the sample concentration zone Z2, the sample separation zone Z3, and the sub-flow channel 15B (sub-flow channel zone Z5).

  As one aspect, it is mentioned that the migration time adjustment region 18 is provided in each of the introduction zone Z1 or the sample concentration zone Z2 and the sample separation zone Z3 or the sub-flow channel zone Z5.

  For example, when the handoff time is shorter than a predetermined reference time, the handoff time can be adjusted to be longer by providing the shallow groove migration time adjustment region 18 in the introduction zone Z1. By reducing the groove depth of the introduction zone Z1 and reducing its cross-sectional area, the resistance of the introduction zone Z1 increases and the voltage distribution increases. Accordingly, the voltage distribution in the sample concentration zone Z2 becomes relatively small, so that the flow in the sample concentration zone Z2 becomes slow, and as a result, the handoff time can be lengthened. The handoff time can be adjusted to be longer as the groove depth in the migration time adjustment region 18 of the introduction zone Z1 is made shallower. Thus, the handoff time can be kept within the range of the reference time. The migration time adjustment region 18 may be provided in the entire introduction zone Z1, or may be provided only in a part of the introduction zone Z1.

  For example, when the handoff time is longer than a predetermined reference time, the handoff time can be adjusted to be shortened by providing the shallow groove migration time adjustment region 18 in the sample concentration zone Z2. By reducing the groove depth of the sample concentration zone Z2 and reducing its cross-sectional area, the resistance of the sample concentration zone Z2 increases and voltage distribution increases. Thereby, the flow in the specimen concentration zone Z2 becomes faster, and as a result, the handoff time can be shortened. The handoff time can be adjusted to be shorter as the groove depth in the migration time adjustment region 18 of the specimen concentration zone Z2 is made shallower. Thus, the handoff time can be kept within the range of the reference time. The migration time adjustment region 18 may be provided in the entire sample concentration zone Z2, or may be provided only in a partial region of the sample concentration zone Z2.

  For example, when the zone migration time is longer than a predetermined reference time, the zone migration time can be adjusted to be shortened by providing a shallow groove migration time adjustment region 18 in the sample separation zone Z3. it can. By reducing the groove depth of the sample separation zone Z3 and reducing its cross-sectional area, the resistance of the sample separation zone Z3 increases and voltage distribution increases. As a result, the flow in the specimen separation zone Z3 becomes faster, and as a result, the zone migration time can be shortened. The zone migration time can be adjusted to be shorter as the groove depth in the migration time adjustment region 18 of the sample separation zone Z3 is made shallower. Thus, the zone migration time can be kept within the reference time range. The migration time adjustment region 18 may be provided in the entire sample separation zone Z3, or may be provided only in a partial region of the sample separation zone Z3.

  For example, when the zone migration time is shorter than a predetermined reference time, the zone migration time can be adjusted to be longer by providing the shallow groove migration time adjustment region 18 in the sub-flow channel zone Z5. Can do. By reducing the groove depth of the sub-flow zone Z5 and reducing its cross-sectional area, the resistance of the sub-flow zone Z5 increases and voltage distribution increases. Accordingly, the voltage distribution in the sample separation zone Z3 becomes relatively small, so that the flow in the sample separation zone Z3 becomes slow, and as a result, the zone migration time can be lengthened. The zone migration time can be adjusted to be longer as the groove depth in the migration time adjustment region 18 of the sub-flow zone Z5 is made shallower. Thus, the zone migration time can be kept within the reference time range. The migration time adjustment region 18 may be provided in the entire sub-flow channel zone Z5, or may be provided only in a partial region of the sub-flow channel zone Z5.

  For example, when the handoff time is shorter than a predetermined reference time and the zone migration time is longer than a predetermined reference time, the migration time adjustment region 18 of the shallow groove is provided in the introduction zone Z1 and the sample separation zone Z3. Provided. By doing in this way, both handoff time and zone migration time can be settled in the range of reference time. Further, for example, when the handoff time is longer than a predetermined reference time and the zone migration time is shorter than the predetermined reference time, the shallow groove migration time adjustment region 18 is set to the sample concentration zone Z2 and the secondary concentration time. It is provided in the flow path zone Z5. By doing in this way, both handoff time and zone migration time can be settled in the range of reference time.

  For example, when both the handoff time and the zone migration time are shorter than a predetermined reference time, the shallow groove migration time adjustment region 18 is provided in the introduction zone Z1 and the sub-flow channel zone Z5. By doing in this way, both handoff time and zone migration time can be settled in the range of reference time. For example, when both the handoff time and the zone migration time are longer than the predetermined reference time, the shallow groove migration time adjustment region 18 is provided in the sample concentration zone Z2 and the sample separation zone Z3. By doing in this way, both handoff time and zone migration time can be settled in the range of reference time.

  As shown in FIG. 3, the manufacturing method of the microchannel chip 1 includes a chip manufacturing process, a migration time measuring process, and a protrusion height adjusting process. The chip manufacturing process and the migration time measurement process are always executed in this order, and the protrusion height adjustment process is selectively executed after the migration time measurement process depending on the result obtained in the migration time measurement process.

  The chip manufacturing process is a process of manufacturing the microchannel chip 1 including the migration channel 15 including the channel groove 11. As shown in FIG. 4, in the chip manufacturing process, the microchannel chip 1 is manufactured by a resin molding processing technique using the mold 5 (in this example, injection molding). The mold 5 of the present embodiment includes a first mold 51 having a linear protrusion 52 and a second mold 54 having a recess 55. The recess 55 is formed in a shape corresponding to the desired outer shape of the resin substrate 10. The linear protrusion 52 is formed in a shape corresponding to a desired shape in plan view of the migration flow path 15. In the initial stage, the linear protrusion 52 has a constant height regardless of the part. In addition, the structure of the shaping | molding die 5 shown here is an example to the last, The specific structure can be changed suitably.

  In the chip manufacturing process, as shown in FIG. 5, molten resin is injected from the gate into a cavity space 58 formed by clamping the first mold 51 and the second mold 54. After cooling, the mold is opened and the resin substrate 10 is taken out. The resin substrate 10 has a shape corresponding to the shape of the linear protrusion 52 in plan view, and includes a migration channel 15 including a channel groove 11 having a predetermined depth. Thereafter, as shown in FIG. 6, the microchannel chip 1 including the migration channel 15 is manufactured by bonding the resin film 20 to the surface of the resin substrate 10 on which the channel groove 11 is formed. At this time, the migration channel 15 of the microchannel chip 1 has a certain groove depth regardless of the part.

  The migration time measurement step is a step of measuring the migration time by performing electrophoresis (capillary electrophoresis) of a predetermined sample in the migration channel 15 using the microchannel chip 1 produced in the chip production step. In the present embodiment, both the handoff time and the zone migration time are measured using the above-described tumor marker as a specimen.

  Thereafter, the hand-off time actually measured in the migration time measurement step is compared with a reference time predetermined for the hand-off time. Further, the zone migration time actually measured in the migration time measurement step is compared with a reference time predetermined for the zone migration time. Then, when at least one of the handoff time and the zone migration time is out of the reference time for each, the protrusion height adjusting step is executed.

  The protrusion height adjusting step is a step of adjusting the height in at least a partial region of the linear protrusion 52 of the mold 5 (first mold 51 in this example). In the present embodiment, the height of the linear protrusion 52 in the region corresponding to at least one of the introduction zone Z1, the sample concentration zone Z2, the sample separation zone Z3, and the sub-flow channel zone Z5 is adjusted in the protrusion height adjustment step. To do. When the handoff time deviates in the short direction from the reference time, the height of the linear protrusion 52 in the region corresponding to the introduction zone Z1 is adjusted, and when the handoff time deviates in the long direction, it corresponds to the sample concentration zone Z2. The height of the linear protrusion 52 in the region is adjusted. Further, when the zone migration time deviates in the short direction from the reference time, the height of the linear protrusion 52 in the region corresponding to the sub-flow zone Z5 is adjusted, and when it deviates in the long direction, the sample separation is performed. The height of the linear protrusion 52 in the region corresponding to the zone Z3 is adjusted.

  In the present embodiment, the height adjustment of the linear protrusion 52 is performed by cutting or polishing the protruding tip side of the linear protrusion 52 as shown in FIG. Thereby, in the projection height adjustment step, the linear projection 52 is adjusted so that the projection height is lower than other portions in at least a part of the region.

  When the protrusion height adjusting process is completed, the chip manufacturing process is performed again using the mold 5 as shown in FIGS. As shown in FIG. 9, the microchannel chip 1 obtained at this time has an adjustment of migration time having a groove depth shallower than other portions of the migration channel 15 in at least one location of the migration channel 15. The region 18 is provided. Thereafter, the migration time measurement step is performed again, and the handoff time and the zone migration time are reconfirmed. If at least one of them is out of the reference time, the protrusion height adjusting step, the chip manufacturing step, and the migration time measuring step are repeated again. If both the handoff time and the zone migration time are within the reference time range, the microchannel chip 1 is finally completed.

  According to the manufacturing method as described above (also referred to as “migration time adjustment method”), handoff can be performed simply by finely adjusting the protruding height of the linear protrusion 52 while using the existing mold 5. Time and zone migration time can be easily adjusted. There is no need to remake the mold 5 at high cost, which is very advantageous in terms of cost. Therefore, the homogeneous microchannel chip 1 in which the handoff time and the zone migration time are within a predetermined reference time range can be easily manufactured without incurring an increase in cost.

[Other Embodiments]
(1) In the above embodiment, the example in which the migration time adjustment region 18 is configured as a region having a groove depth shallower than other portions in the migration channel 15 has been described. However, without being limited to such a configuration, for example, the migration time adjustment region 18 may be configured as a region having a deeper groove depth than other portions in the migration channel 15.

(2) In the above-described embodiment, the configuration in which the migration time adjustment region 18 is provided in each of the introduction zone Z1 or the sample concentration zone Z2 and the sample separation zone Z3 or the sub-flow channel zone Z5 has been described as an example. . However, without being limited to such a configuration, for example, the migration time adjustment region 18 may be provided in each of the introduction zone Z1, the sample concentration zone Z2, the sample separation zone Z3, and the sub-flow channel zone Z5. Alternatively, the migration time adjustment region 18 may be provided in any one of the introduction zone Z1, the sample concentration zone Z2, the sample separation zone Z3, and the sub-flow channel zone Z5.

(3) In the above embodiment, the configuration in which the migration channel 15 includes the main channel 15A and the sub-channel 15B has been described as an example. However, the present invention is not limited to such a configuration, and for example, the migration channel 15 may be configured in a linear type including only the main channel 15A. In this case, the main flow path 15A is mainly composed of the sample separation zone Z3 without the sample concentration zone Z2. The migration time adjustment region 18 is preferably provided in the sample separation zone Z3.

(4) In the above embodiment, the configuration in which the microchannel chip 1 is manufactured by injection molding in the chip manufacturing process has been described as an example. However, without being limited to such a configuration, the chip manufacturing process may be executed by, for example, transfer molding or extrusion molding.

(5) In the above embodiment, the microchannel chip 1 for analyzing a tumor marker has been described as an example. However, the present invention is not limited to such a configuration, and the microchannel chip 1 may be used for analysis of any other specimen. The analysis conditions in that case may be appropriately set according to the type of specimen.

(6) The configurations disclosed in each of the above-described embodiments (including the above-described embodiments and other embodiments; the same applies hereinafter) are applied in combination with the configurations disclosed in the other embodiments unless a contradiction arises. It is also possible to do. Regarding other configurations as well, the embodiments disclosed in the present specification are examples in all respects, and can be appropriately modified without departing from the gist of the present disclosure.

DESCRIPTION OF SYMBOLS 1 Microchannel chip 5 Mold 11 Channel groove 12 Reagent introduction part 13A First electrode insertion part 13B Second electrode insertion part 13C Third electrode insertion part 15 Migration channel 15A Main channel 15B Subchannel 18 Adjustment of migration time Region 52 Linear projection Z1 Introduction zone Z2 Sample concentration zone Z3 Sample separation zone

Claims (5)

A plurality of reagent introduction parts;
A migration channel comprising a channel groove and connecting a plurality of the reagent introduction parts,
A microchannel chip in which a migration time adjustment region formed so as to have a groove depth different from other portions in the migration channel is provided in at least one location of the migration channel. The plurality of reagent introduction parts each include a first electrode insertion part into which an electrode is inserted, a second electrode insertion part, and a third electrode insertion part,
The migration channel has an introduction zone, a sample concentration zone, and a sample separation zone in that order in the main channel connecting the first electrode insertion part and the second electrode insertion part, and the main channel in the main channel The third electrode insertion part is connected to the upstream side of the sample separation zone via a sub-flow channel,
The microchannel chip according to claim 1, wherein the migration time adjustment region is provided in any one or more of the introduction zone, the sample concentration zone, the sample separation zone, and the subchannel.   The microchannel chip according to claim 2, wherein the migration time adjustment region is provided in each of the introduction zone or the sample concentration zone and the sample separation zone or the subchannel.   4. The microchannel chip according to claim 1, wherein the migration time adjustment region is a region having a groove depth shallower than the other part. 5. A chip manufacturing process for manufacturing a micro-channel chip having the above-mentioned channel for migration composed of a channel groove having a predetermined depth using a mold having a linear projection having a shape corresponding to the shape of a desired channel for electrophoresis. When,
Electrophoresis of a predetermined sample in the migration channel using the microchannel chip produced in the chip production step, and a migration time measurement step of measuring the migration time;
When the actual migration time acquired in the migration time measurement step is out of a predetermined reference time, a projection height adjustment step for adjusting the height in at least a partial region of the linear projection,
The manufacturing method of the microchannel chip | tip containing this.

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