JP2013092490A - Exchange product for reaction progress device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To suppress an increase in test costs of a biochemical test and in period of test time.SOLUTION: An exchange product 1010 comprises: a channel structure having a reaction channel 1163 on whose channel surface a reactant-containing body including reactants of a biochemical reaction is fixed and a nozzle insertion hole 1164 to which a nozzle for guiding test liquid to the reaction channel is inserted; and a sealing member 1160 having a first main surface 1162, and a second main surface 1161 which is adhered to the surface of the channel structure. The nozzle insertion hole has a first opening 1240 on the second surface 1161 and a second opening 1291 on a wall surface of the reaction channel. A first area on the second main surface of the sealing member seals the first opening of the nozzle insertion hole. A first water-repellent film 35a is formed on at least part of the first area of the sealing member.

Description

  The present invention relates to a replacement product for a reaction progressing device that advances a biochemical reaction.

  Biochemical reactions such as antigen-antibody reactions are used in biochemical tests. For example, the antibody is fixed inside the flow path, the sample liquid containing the antigen is supplied to the flow path, and the antigen contained in the sample liquid and the antibody fixed inside the flow path are combined. The presence or absence of antigen binding to the antibody, the amount of antigen binding to the antibody, and the like are measured by surface plasmon resonance (SPR), surface plasmon excitation fluorescence spectroscopy (SPFS), and the like. Before the sample liquid is supplied to the flow path, a cleaning liquid, a buffer liquid, or the like may be supplied to the flow path. In many cases, liquids such as a sample liquid, a cleaning liquid, and a buffer liquid are supplied to the flow path by the nozzle and collected from the flow path by the nozzle.

  In biochemical tests, disposable test chips incorporating biochemical reaction channels have become widespread for the purpose of facilitating the handling of sample solutions and the like. When the measurement is completed, the test chip is discarded as medical waste. However, the test chip after measurement has a risk of infection, contamination, etc. due to mixed antibodies remaining in the flow path. ing. For this reason, care must be taken when handling the inspection chip after measurement.

  For this reason, in the inspection apparatus of Patent Document 1, after completion of the measurement, a filling material having curability with respect to active energy such as ultraviolet rays is filled at the entrance / exit of the flow path of the inspection chip, and the filling material is irradiated with active energy and filled. The channel is blocked by curing the material.

JP 2008-139237 A

  However, in the technique of Patent Document 1, the cost of inspection increases due to the configuration of the apparatus for performing additional processing after the completion of the inspection such as filling of the filling material and irradiation of active energy, and the use of the filling material. There is a problem that inspection time is prolonged due to the additional processing.

  The present invention has been made to solve these problems, and can suppress an increase in inspection cost and an increase in inspection time due to risk reduction processing such as contamination in the inspection chip after the inspection is completed. The purpose is to provide technology.

  In order to solve the above-described problem, an exchange product according to the first aspect is a replacement product for a reaction progressing device for advancing a biochemical reaction, and includes a reactant containing a reaction product of the biochemical reaction on a road surface. A flow path structure having a reaction flow path in which is fixed, a nozzle insertion hole into which a nozzle for introducing a sample liquid into the reaction flow path is inserted, a first main surface, and a second main surface, And a seal member having the second main surface joined to the surface of the flow channel structure, and the nozzle insertion hole has a first opening on the surface of the flow channel structure. The reaction channel has a second opening on the wall surface, and the first region of the second main surface of the seal member closes the first opening of the nozzle insertion hole, and A first water repellent film is formed in at least a part of the first region.

  The replacement product according to the second aspect is the replacement product according to the first aspect, wherein the flow path structure has a third opening on the surface of the flow path structure and the reaction flow path. A wall opening having a fourth opening and storing the sample liquid overflowing from the reaction flow path, and formed on the surface of the flow path structure, with one end at a side surface of the liquid storage hole And a second region of the second main surface of the seal member closes at least a part of the groove and the third opening of the liquid reservoir hole, respectively. A gas vent hole capable of discharging the gas in the liquid reservoir hole to the outside of the exchange product when the sample liquid is introduced into the reaction channel is formed, and among the second region of the seal member, A second water repellent film is formed on at least a portion of the liquid reservoir hole that covers a part of the third opening. Together are formed, said portion of said third opening, characterized in that it is a part of the groove side of the third opening.

  The replacement product according to the third aspect is the replacement product according to the second aspect, and in the flow path structure, a third repellent is formed on a portion of the wall surface of the groove portion that constitutes the wall surface of the gas vent hole. A water film is formed.

  The replacement product according to the fourth aspect is the replacement product according to the second or third aspect, and in the seal member, a portion constituting the wall surface of the gas vent hole in the second main surface. A fourth water repellent film is formed.

  The exchange product according to the fifth aspect is an exchange product according to any one of the second to fourth aspects, and the size of the hole cross section of the gas vent hole is the sample liquid introduced into the reaction channel. Is a size that does not leak out of the replacement product through the vent hole regardless of the posture of the replacement product.

  A replacement product according to a sixth aspect is the replacement product according to any one of the first to fifth aspects, wherein the seal member is the first region in the first region of the second main surface. The portion where the water repellent film is formed has an introduction hole penetrating the seal member, and the introduction hole is sized so that the sample liquid introduced into the reaction channel is introduced regardless of the posture of the exchange product. It is a size that does not leak from the hole to the outside of the replacement product.

  A replacement product according to a seventh aspect is a replacement product according to any one of the first to sixth aspects, wherein the flow path structure has a third main surface and a fourth main surface. The reaction channel is formed, and the reaction channel has a fifth opening and a sixth opening on the third main surface and the fourth main surface, respectively, and the flow channel A lid member bonded to the third main surface of the member and having the nozzle insertion hole formed therein; and a fourth member surface of the flow path member bonded to the sixth opening of the flow channel member. It is characterized by comprising an obstruction to be obstructed.

  An exchange product according to an eighth aspect is the exchange product according to the seventh aspect, wherein the obstruction includes a light-transmitting dielectric medium having a light reflection surface, the flow path member, and the light reflection surface. And a conductor film interposed between them.

  According to any of the first to eighth aspects of the invention, the first opening of the nozzle insertion hole is formed by the first region of the second main surface of the seal member joined to the surface of the flow path structure of the replacement product. It is blocked. A first water repellent film is formed in at least a part of the first region. Therefore, even if an introduction hole penetrating the seal member is formed in a portion of the first region where the first water repellent film is formed, liquid leakage from the introduction hole is less likely to occur. For this reason, it is possible to suppress an increase in inspection cost due to risk reduction processing such as contamination in the inspection chip after completion of the inspection and an increase in inspection time.

It is a schematic diagram which shows an example of a structure of the measuring device using the test | inspection chip which concerns on embodiment. It is a perspective view showing typically an example of the appearance of the inspection chip concerning an embodiment. It is a figure showing typically an example of the section of the inspection chip concerning an embodiment. It is a figure showing typically an example of the section of the inspection chip concerning an embodiment. It is a figure which shows typically an example of the perspective view of the test | inspection chip which concerns on embodiment. It is a figure showing typically an example of the section of the inspection chip concerning an embodiment. It is a figure showing typically an example of the section of the inspection chip concerning an embodiment. It is a figure for demonstrating prevention of a liquid leak. It is a flowchart which shows an example of the manufacturing process of the test | inspection chip which concerns on embodiment. It is a flowchart which shows an example of the biopsy process using the test | inspection chip which concerns on embodiment.

  Hereinafter, an embodiment of the present invention will be described with reference to the drawings. In the drawings, parts having the same configuration and function are denoted by the same reference numerals, and redundant description is omitted in the following description. Each drawing is schematically shown. For example, the size and positional relationship of display objects in each drawing are not necessarily shown accurately.

<About embodiment>
<Configuration of Measuring Device 1000 and Inspection Chip 1010>
(Outline)
This desirable embodiment relates to a measuring device, a reaction progress device included in the measurement device, a replacement product for the reaction progress device, and a method for manufacturing the replacement product.

  FIG. 1 is a schematic diagram illustrating an example of a measuring apparatus 1000 that uses an inspection chip 1010 according to the embodiment. The schematic diagrams from FIG. 2 to FIG. 7 show an embodiment of the test chip 1010. FIG. 2 is a perspective view of the appearance, FIG. 3, FIG. 4 and FIG. 7 show cross sections, and FIG. 6 shows a cross section with the nozzle 1080 inserted. FIG. 7 shows a state where the nozzle 1080 shown in FIG. 6 is removed. FIG. 5 is an exploded perspective view.

  A measurement apparatus 1000 shown in FIG. 1 performs measurement by surface plasmon excitation fluorescence spectroscopy (SPFS). The measurement apparatus 1000 may perform measurement by surface plasmon resonance (SPR). The reaction progressing device 1215 included in the measuring device 1000 is used in addition to measurement by SPFS and SPR. For example, the reaction progressing device 1215 is also used for measurement using an ELISA, immunochromatography, or the like.

  As shown in FIG. 1, the measurement apparatus 1000 includes a measurement mechanism 1002, a liquid feeding mechanism 1004, a control unit 1006, a display unit 1008, a test chip 1010, and a reagent chip 1012. Components other than these may be added to the measurement apparatus 1000.

  The measurement mechanism 1002 includes a laser diode 1100, a first band pass filter 1102, a linear polarization filter 1104, a neutral density (ND) filter 1106, a half-wave plate 1108, a half-wave plate driving mechanism 1110, a shaping optical system 1112, a mirror 1114, A mirror driving mechanism 1116, a condensing lens 1118, a second bandpass filter 1120, a bandpass filter driving mechanism 1122, an imaging lens 1124, a photomultiplier tube 1126, an inspection chip transport mechanism 1128, and a light absorber 1130 are provided. Components other than these may be added to the measurement mechanism 1002.

  The liquid feeding mechanism 1004 includes a liquid feeding pump 1140 and a liquid feeding pump transport mechanism 1142. Components other than these may be added to the liquid feeding mechanism 1004.

  The control unit 1006 controls the half-wave plate driving mechanism 1110, the mirror driving mechanism 1116, the band-pass filter driving mechanism 1122, the inspection chip transport mechanism 1128, the liquid feed pump 1140, and the liquid feed pump transport mechanism 1142, and the photomultiplier tube 1126. The light quantity measurement result is obtained from The control unit 1006 is a computer in which a program is installed. All or a part of the functions of the control unit 1006 may be carried by hardware without a program. The hardware may be an electronic circuit such as an operational amplifier or a comparator, or may be a mechanical mechanism.

  As shown in FIGS. 2 to 7, the inspection chip 1010 mainly includes, for example, a prism 1150, a conductor film 1152, a capturing body 1154, a flow path member 1156, a lid member 1158, a seal member 1160, and the like. The Other components may be added to the inspection chip 1010. The inspection chip 1010 is also called a sensor chip, an analysis chip, a sample cell, or the like. A flow path 1163 is formed in the flow path member 1156. The channel 1163 has a channel body 1168, a tip receiving hole 1170, and an end hole 1172. The lid member 1158 includes a nozzle insertion hole 1164 into which a nozzle 1180 for guiding the sample liquid 1202 and the like into the flow path 1163 is inserted, and a liquid reservoir hole 1166 that can store the sample liquid 1202 and the like overflowing from the flow path 1163. Is formed. Further, a groove 1249 is formed in the sealing surface 1228 of the lid member 1158, and one end thereof reaches the side surface of the liquid reservoir hole 1166.

  As shown in FIG. 1, the reagent chip 1012 includes a nozzle 1180, a sample container 1182, a dilution container 1184, a dilution liquid container 1186, a labeled antibody liquid container 1188, a cleaning liquid container 1190, and a holder 1192. Dilution liquid 1194, labeled antibody liquid 1196, and cleaning liquid 1198 are previously stored in dilution liquid container 1186, labeled antibody liquid container 1188, and cleaning liquid container 1190, respectively. The specimen 1200 is accommodated in the specimen container 1182 before the reagent chip 1012 is attached to the measuring apparatus 1000. The dilution container 1184 stores the sample solution 1202 after the reagent chip 1012 is attached to the measuring apparatus 1000.

  The inspection chip 1010 and the reagent chip 1012 are exchange products 1216 that are exchanged for each specimen 1200.

  The liquid feeding mechanism 1004, the inspection chip 1010, and the reagent chip 1012 constitute a reaction progressing device 1215 that progresses a biochemical reaction.

  The first band pass filter 1102, the linear polarization filter 1104, the neutral density filter 1106, the half-wave plate 1108, the shaping optical system 1112, and the mirror 1114 constitute an excitation optical system 1212. The condensing lens 1118, the second band pass filter 1120, and the imaging lens 1124 constitute a detection optical system 1214.

(Summary of biochemical reaction progress)
When the biochemical reaction is allowed to proceed, the cleaning liquid 1198 is supplied to the flow path 1163 by the liquid feeding mechanism 1004, the flow path 1163 is washed, and the cleaning liquid 1198 is recovered from the flow path 1163 by the liquid feeding mechanism 1004. Subsequently, the sample liquid 1202 is supplied to the flow path 1163 by the liquid feeding mechanism 1004, the antigen contained in the sample liquid 1202 and the antibody contained in the capturing body 1154 are combined, and the sample liquid 1202 is fed by the liquid feeding mechanism 1004. It is recovered from the flow path 1163. Subsequently, the labeled antibody solution 1196 is supplied to the flow path 1163 by the liquid feeding mechanism 1004, and the antigen bound to the antibody contained in the capturing body 1154 and the labeled antibody contained in the labeled antibody solution 1196 are combined. When the cleaning liquid 1198 is collected from the flow path 1163, the nozzle 1180 is inserted deeply into the nozzle insertion hole 1164 until the tip 1252 of the nozzle 1180 reaches the tip accommodation hole 1170.

(Outline of measurement)
When measurement is performed, the excitation light 1210 is applied to the prism 1150. The irradiated excitation light 1210 is reflected at the interface between the prism 1150 and the conductor film 1152. When the excitation light 1210 is reflected at the interface between the prism 1150 and the conductor film 1152, an evanescent wave leaks from the interface between the prism 1150 and the conductor film 1152 toward the conductor film 1152, and the conductor film 1152. The surface plasmon and evanescent wave interfere with each other. When the plasmon and the evanescent wave resonate, the electric field of the evanescent wave is remarkably enhanced. The enhanced electric field excites the labeled antibody, and surface plasmon excitation fluorescence 1218 is emitted from the labeled antibody. The amount of surface plasmon excitation fluorescence 1218 is measured by a photomultiplier tube 1126. The presence / absence of the antigen, the amount of antigen captured, and the like are calculated by the control unit 1006 from the measurement result of the light amount of the surface plasmon excitation fluorescence 1218, and the calculation result is displayed on the display unit 1008.

(Inspection chip)
As shown in FIGS. 2 to 7, in the inspection chip 1010, a prism 1150, a conductor film 1152, a capturing body 1154, a flow path member 1156, a lid member 1158 and a seal member 1160 are stacked.

○ Prism 1150:
The prism 1150 is a trapezoidal column and has an incident surface 1220, a reflecting surface 1222, and an exit surface 1224 for the excitation light 1210. One inclined side surface of the trapezoidal column is the incident surface 1220, the wide parallel side surface of the trapezoidal column is the reflecting surface 1222, and the other inclined side surface of the trapezoidal column is the exit surface 1224. The incident surface 1220, the reflecting surface 1222, and the emitting surface 1224 are arranged in a mutual relationship such that the excitation light 1210 incident from the incident surface 1220 is reflected by the reflecting surface 1222 and is emitted from the emitting surface 1224. The external shape of the prism 1150 may be other than the trapezoidal column, and the prism 1150 may be replaced with a shape that is not included in the category of “prism”. For example, the outer shape of the prism 1150 may be a cylindrical body, and the prism 1150 may be replaced with a plate.

  The prism 1150 is a dielectric medium made of a material that is transparent to the excitation light 1210. That is, the prism 1150 is a dielectric medium that has a reflection surface 1222 that is a light reflection surface and has optical transparency to the excitation light 1210. The prism 1150 is made of glass, resin, or the like. The prism 1150 is preferably made of a resin having a refractive index of 1.4 to 1.6 and a small birefringence. Thereby, the prism 1150 is manufactured at low cost. When the prism 1150 is made of resin, the prism 1150 is preferably manufactured by injection molding. However, the prism 1150 may be manufactured by other methods.

○ Conductor film 1152:
The conductor film 1152 is made of a conductor that generates surface plasmon resonance. The conductor film 1152 is preferably made of gold. However, the conductor film 1152 may be replaced with a film made of a metal such as silver, copper, or aluminum, or an alloy containing these metals. The film thickness of the conductor film 1152 is desirably 100 nm or less, and more desirably 30 to 70 nm. However, the thickness of the conductor film 1152 may be outside this range. The conductor film 1152 is formed on the reflective surface 1222 by sputtering, vapor deposition, plating, or the like. However, the conductor film 1152 may be formed by other methods. The prism 1150 provided with the conductor film 1152 is also referred to as a “composite 1148”.

  The conductor film 1152 has a main surface 1230 (“sixth main surface”) and a main surface 1232 (“fifth main surface”). One main surface 1230 is in close contact with the reflecting surface 1222 of the prism 1150, and the other main surface 1232 is bonded to the main surface 1234 of the flow path member 1156 to block the opening 1235 of the flow path 1163 in the flow path member 1156. To do. That is, the conductor film 1152 is interposed between the flow path member 1156 and the reflection surface 1222 (light reflection surface), and the complex 1148 operates as an obstruction that blocks the opening 1235 of the flow path 1163. . On the road surface of the flow path 1163 in which the opening 1235 is blocked by the composite 1148, a capturing body 1154 (“reactant-containing body”) containing a reaction product of a biochemical reaction is fixed (“fixed”).

○ Capture body 1154:
The capturing body 1154 (reactant-containing body) includes an antibody that reacts with an antigen to be detected. More generally, the trap 1154 includes a second reactant that reacts with the first reactant of the biochemical reaction. When the sample liquid 1202 containing the antigen to be detected comes into contact with the capturing body 1154, the antigen to be detected contained in the sample liquid 1202 binds to the antibody contained in the capturing body 1154 and is detected in the sample liquid 1202. The target antigen is captured by the capturing body 1154. The trap 1154 provides a reaction field for biochemical reactions. The capturing body 1154 is preferably fixed to the other main surface 1232 of the conductor film 1152 by surface treatment.

  When the capture body 1154 includes an antibody, the capture body 1154 includes a protein. When the capturing body 1154 contains a protein, a storage reagent is preferably applied to the capturing body 1154. Thereby, the capturing performance of the capturing body 1154 is maintained for a long time. The storage reagent is desirably a humectant that does not affect the capture performance of the capture body 1154. For example, the storage reagent is a liquid mainly composed of a sucrose aqueous solution. The inspection chip 1010 is stored in a state where a storage reagent is applied to the capturing body 1154 and dried. Therefore, the capturing body 1154 is washed with the cleaning liquid 1198 before the sample liquid 1202 is brought into contact with the capturing body 1154.

○ Channel member 1156:
A channel 1163 (“reaction channel”) is formed in the channel member 1156. The flow path member 1156 is preferably made of an adhesive sheet, and also serves as a bonding medium for bonding the composite 1148 and the lid member 1158. The flow path member 1156 has main surfaces 1234 and 1236, and a flow path 1163 is formed. However, the flow path member 1156 may be made of an elastic body other than the adhesive sheet. For example, the flow path member 1156 may be made of an elastic sheet, an O-ring, or the like. When the flow path member 1156 is not a joining medium, the composite 1148 and the lid member 1158 are joined by adhesion, laser welding, ultrasonic welding, clamp pressure bonding, or the like. The flow path member 1156 is preferably made of resin.

  The flow path 1163 has openings 1235 (“sixth opening”) and 1237 (“fifth opening”) on one main surface 1234 and the other main surface 1236 of the flow path member 1156, respectively. 1156 extending in parallel. One opening 1235 is blocked by the other main surface 1232 of the conductor film 1152 formed on the reflection surface 1222 of the prism 1150. The other opening 1237 is closed by the joint surface 1226 of the lid member 1158 except for the tip accommodation hole 1170 and the end hole 1172.

  The tip accommodation hole 1170 in the flow channel 1163 is at one end of the flow channel 1163, and the end hole 1172 is at the other end of the flow channel 1163. The flow path main body 1168 extends from the tip accommodation hole 1170 to the end hole 1172.

  The flow path member 1156, the lid member 1158, and the complex 1148 constitute a flow path structure 1050 (FIG. 5). One main surface 1161 of the seal member 1160 is joined to the seal surface 1228 of the lid member 1158 that forms the surface of the flow path structure 1050. Note that when the reaction progressing device 1215 is used in addition to the measurement by SPR, one opening 1235 may be blocked by a plug other than the complex 1148. In addition, when the reaction progressing device 1215 is used in addition to the measurement by SPFS or SPR and the opening 1235 is closed by an obstruction other than the complex 1148, preferably all or a part of the obstruction is made of resin, and more desirably. All the obstructions are made of resin. The other main surface 1236 of the flow path member 1156 is bonded to the bonding surface 1226 of the lid member 1158.

○ Lid member 1158:
The lid member 1158 has, for example, a rectangular parallelepiped shape, and is joined to the main surface 1236 of the flow path member 1156. The lid member 1158 is made of a material transparent to the surface plasmon excitation fluorescence 1218 and scattered light. The lid member 1158 is preferably made of, for example, resin and manufactured by injection molding or the like.

  The lid member 1158 has a joint surface 1226 and a seal surface 1228, and a nozzle insertion hole 1164 and a liquid reservoir hole 1166 are formed. The seal surface 1228 constitutes the surface of the flow path structure 1050.

  One end of the nozzle insertion hole 1164 is an opening 1291 (“second opening”) provided in the joint surface 1226, and the other end is a nozzle insertion port 1240 (“first opening”) provided in the seal surface 1228. is there. The nozzle insertion hole 1164 extends from the nozzle insertion opening 1240 to the tip receiving hole 1170. That is, the opening 1291 is formed in the wall surface of the flow path 1163, and the nozzle insertion hole 1164 passes through the lid member 1158 and communicates with one end side of the flow path 1163. Further, the nozzle insertion hole 1164 extends perpendicular to the flow path 1163 of the flow path member 1156. However, the flow path 1163 and the nozzle insertion hole 1164 need only extend in different directions and do not necessarily extend perpendicular to each other. Further, the nozzle insertion opening 1240 is blocked by one main surface 1161 of the seal member 1160.

  One end of the liquid reservoir hole 1166 is an opening 1292 (“fourth opening”) provided in the joint surface 1226, and the other end is an opening 1242 (“third opening”) provided in the seal surface 1228. The liquid reservoir hole 1166 extends from the liquid reservoir port 1166 to the end hole 1172. That is, the liquid reservoir hole 1166 passes through the lid member 1158 and communicates with the end hole 1172 side of the flow path 1163. As shown in FIGS. 3 and 4, for example, a liquid reservoir hole 1166 that is wider on the opening 1242 side than on the opening 1292 side is employed. A liquid reservoir hole 1166 having a uniform width (diameter) may be employed from the opening 1292 side to the opening 1242 side, such as a cylindrical or quadrangular cylindrical hole. The opening 1242 is blocked by one main surface 1161 of the seal member 1160.

  Further, a groove 1249 is formed on the seal surface 1228. The lid member 1158 and the main surface 1161 form a gas vent hole 1248 by at least a part of the opening in the seal surface 1228 of the groove portion 1249 being blocked by one main surface 1161 of the seal member 1160. The gas vent hole 1248 is configured such that when the sample liquid 1202 or the like is introduced into the flow path 1163, the gas in the flow path 1163 can be discharged to the outside of the inspection chip 1010 through the liquid reservoir hole 1166.

  One end of the groove 1249 reaches the side surface of the liquid reservoir hole 1166 and forms one opening 1251 of the gas vent hole 1248. In other words, one end of the groove 1249 communicates with the side surface of the liquid reservoir hole 1166. Further, the other end of the groove portion 1249 desirably reaches the side surface 1229 of the lid member 1158 and forms the other opening 1250 of the gas vent hole 1248. Even if the other end of the groove portion 1249 does not reach the side surface 1229, the opening 1250 can be formed in the seal surface 1228 by blocking a part of the opening of the groove portion 1249 in the seal surface 1228 with the main surface 1161. Therefore, even if the other end of the groove 1249 does not reach the side surface 1229, the usefulness of the present invention is not impaired.

  When the lid member 1158 is joined to the main surface 1236 of the flow path member 1156, the sample liquid 1202 containing the antigen to be detected flows through the nozzle insertion hole 1164, the flow path 1163, and the liquid reservoir hole 1166. It will be in a state to get. Therefore, normally, after the lid member 1158 is joined to the main surface 1236 of the flow path member 1156, the nozzle insertion hole 1164, the flow path 1163, and the liquid reservoir hole 1166 are subjected to blocking processing.

  The blocking process is a phenomenon in which an antigen to be detected is adsorbed at a place other than the capturing body 1154 among the places where the sample liquid 1202 in the nozzle insertion hole 1164, the flow path 1163, and the liquid reservoir hole 1166 can come into contact (“non-specific adsorption”). It is also a surface treatment for preventing. In the blocking process, for example, a liquid containing a protein such as skim milk or albumin is flowed into the nozzle insertion hole 1164, the flow path 1163, and the liquid reservoir hole 1166, and then the liquid is collected and the portion where the liquid is in contact It is performed by drying.

  Occurrence of non-specific adsorption is almost prevented by the blocking treatment. However, since the protein has hydrophilicity, the part subjected to the blocking treatment is hydrophilized, and the influence of the hydrophilization extends to the groove 1249 communicating with the liquid reservoir hole 1166. And the water repellency of the part which comprises the wall surface of the vent hole 1248 among the wall surfaces of the groove part 1249 is impaired, and it becomes inadequate water repellency. For this reason, in the inspection chip 1010, in order to increase the water repellency of the wall surface of the gas vent hole 1248, the portion constituting the wall surface of the gas vent hole 1248 out of the wall surface of the groove 1249 is preferably subjected to the blocking process. A water repellent film 35c is formed. The water repellent film 35c will be described later. Further, a water repellent film may be formed on at least a portion including the opening 1251 of the gas vent hole 1248 in the wall surface of the liquid reservoir hole 1166.

  Further, the groove 1249 is not formed, and a gas vent hole 1248 is formed by making a hole in a part of the seal member 1160 that closes the opening 1242 of the liquid reservoir hole 1166 at the time of measurement, similarly to the introduction hole 1241 described later. Even if done, it does not impair the usefulness of the present invention. In this case, the gas vent hole 1248 is formed in a portion where a water repellent film 35 b described later is formed on the main surface 1161 in the portion of the seal member 1160 that closes the liquid reservoir hole 1166.

  In addition, the size of the opening 1251 of the gas vent hole 1248, that is, the size of the hole cross section, is such that the sample liquid 1202 or the like introduced into the flow path 1163 is from the gas vent hole 1248 to the test chip 1010 regardless of the posture of the test chip 1010. It is formed in a size that does not leak to the outside. The size of the opening 1251 will be described later.

  The inspection chip 1010 is preferably held so that the flow path 1163 extends in the horizontal direction, the nozzle insertion hole 1164 and the liquid reservoir hole 1166 extend in the vertical direction, and the seal surface 1228 faces upward in the vertical direction. Is done. This makes it difficult for liquid to spill from the flow path 1163. The liquid reservoir hole 1166 and the end hole 1172 may be a nozzle insertion hole and a tip accommodation hole different from the nozzle insertion hole 1164 and the tip accommodation hole 1170, respectively.

  In addition, the flow path member 1156 and the lid member 1158, which are separate from each other, are joined to each other, so that a structure in which the flow path 1163 and the nozzle insertion hole 1164 are formed can be easily manufactured. However, the flow path member 1156 and the lid member 1158 may be replaced with an integrated structure in which the flow path 1163, the nozzle insertion hole 1164, and the groove 1249 are formed.

○ Sealing member 1160:
The seal member 1160 has one main surface 1161 (“second main surface”) and the other main surface 1162 (“first main surface”). One main surface 1161 of the seal member 1160 is attached to the seal surface 1228 of the lid member 1158. The seal member 1160 is bonded to the seal surface 1228 of the lid member 1158 with an adhesive or the like. The seal member 1160 closes the nozzle insertion opening 1240 and the liquid reservoir hole 1166 with one main surface 1161 and blocks the groove 1249 except for the opening 1250. That is, the seal member 1160 closes the flow path 1163 except for the opening 1250.

  The seal member 1160 is preferably made of resin. The sealing member 1160 may be a laminate of a sheet made of resin and a sheet made of aluminum. The aluminum sheet has an anti-drying action. When the sealing member 1160 is constituted by a resin sheet and an aluminum sheet, a structure in which an aluminum sheet is inserted between two resin sheets is desirably employed. Further, when a resin sheet is included in the constituent members of the seal member 1160, the resin sheet is bonded to the seal surface 1228 of the lid member 1158.

  A first region 1310 (FIG. 5) of the main surface 1161 of the seal member 1160 is a region that blocks the nozzle insertion port 1240 of the nozzle insertion hole 1164. A water repellent film 35 a (“first water repellent film”) is formed in at least a part of the first region 1310.

  At the time of measurement, the nozzle 1180 breaks the seal member 1160 and is inserted into the nozzle insertion hole 1164 at the part of the first region 1310, that is, the portion of the first region 1310 where the water repellent film 35a is formed. An introduction hole 1241 (FIG. 6) is formed in the trace where the nozzle 1180 is removed from the seal member 1160. That is, the introduction hole 1241 is formed in a portion of the first region 1310 where the water repellent film 35 a is formed, and penetrates the seal member 1160. The size of the introduction hole 1241 is a size such that the sample liquid 1202 or the like introduced into the flow path 1163 does not leak out of the inspection chip 1010 from the introduction hole 1241 regardless of the posture of the inspection chip 1010. The size of the introduction hole 1241 will be described later.

  The second region 1320 (FIG. 5) of the main surface 1161 of the seal member 1160 is a region that blocks at least a part of the groove 1249 and the opening 1242 of the liquid reservoir hole 1166. The second region 1320 closes at least a part of the groove 1249 and the opening 1242, so that the gas in the liquid reservoir hole 1166 is transferred to the outside of the inspection chip 1010 when the sample liquid 1202 is introduced into the flow path 1163. A vent hole 1248 that can be discharged is formed.

  A water repellent film 35 b (“second water repellent film”) is formed in at least a portion of the second region 1320 of the seal member 1160 that covers a part of the opening 1242 of the liquid reservoir hole 1166. The part of the opening 1242 is a part of the opening 1242 on the groove 1249 side.

  Further, a water repellent film 35 d (“fourth water repellent film”) is desirably formed on a portion of the main surface 1161 of the seal member 1160 that forms the wall surface of the gas vent hole 1248. That is, the water repellent film 35d is desirably formed on a portion of the second region 1320 that constitutes the wall surface of the gas vent hole 1248. However, even if the water-repellent film 35d is not formed on the portion, the usefulness of the present invention of the present invention is not impaired. When the water repellent film 35d is formed in the portion, it is possible to further prevent liquid leakage from the gas vent hole 1248 as compared with the case where the water repellent film 35d is not formed.

  Further, as described above, the water repellent film 35c (“third water repellent film”) is preferably formed on the portion of the wall surface of the groove 1249 formed in the lid member 1158 that constitutes the wall surface of the gas vent hole 1248. Is formed. If the water repellent film 35c is formed, liquid leakage from the gas vent hole 1248 can be further prevented as compared with the case where the water repellent film 35c is not formed. However, even if the water repellent film 35c is not formed, the present invention. It does not impair the usefulness.

  The water repellent films 35a to 35d are indicated by thick broken lines in FIGS. The water-repellent films 35a to 35d are water-repellent films having a substantially uniform film thickness, and the film thickness is desirably set in the range of 1 nm to 1 μm. However, the film thickness of the water repellent films 35a to 35d may be a film thickness outside this range. Further, the water repellent films 35a to 35d are formed on the main surface 1161 by various methods such as coating with a coater, brush coating, ink jet coating, and the like. It is formed by coating and drying. As the water repellent, for example, a fluorine water repellent or a silicone water repellent is used.

  In the inspection chip 1010, a water repellent film using a water repellent is formed on one main surface 1161 of the seal member 1160, and the water repellency of the main surface 1161 is enhanced. Incidentally, the resin (plastic or the like) constituting the main surface 1161 of the seal member 1160 has a certain degree of water repellency, and can also be given water repellency by appropriately selecting the surface structure of the main surface 1161. Can do. However, if a water repellent film using a water repellent is formed on the main surface 1161, the water repellency of the main surface 1161 can be further enhanced as compared to the case where the water repellent film is not formed. Further, due to the improved water repellency, the liquid inside the test chip 1010 is less likely to leak.

  That is, according to the inspection chip 1010, the nozzle insertion port 1240 of the nozzle insertion hole 1164 is formed by the first region 1310 in one main surface 1161 of the seal member joined to the surface (seal surface 1228) of the flow path structure 1050. It is blocked. A water repellent film 35 a is formed in at least a part of the first region 1310. Accordingly, even if the introduction hole 1241 penetrating the seal member 1160 is formed in the portion of the first region 1310 where the water repellent film 35a is formed, liquid leakage from the introduction hole 1241 is less likely to occur. For this reason, it is possible to suppress an increase in inspection cost due to risk reduction processing such as contamination in the inspection chip after completion of the inspection and an increase in inspection time.

  The seal member 1160 is constituted by a plurality of seal members, for example, a seal member that closes the nozzle insertion port 1240 and a seal member that closes the opening 1242 of the liquid reservoir hole 1166 and the groove 1249. Even if it is made, it does not impair the usefulness of the present invention.

(Size of introduction hole 1241 and size of opening 1251 of vent hole 1248)
FIG. 8 is a view for explaining prevention of liquid leakage of the liquid L1 from the hole H1 of the container C1. The liquid L1 in the container C1 has, for example, a liquid surface shape as indicated by the liquid surface W1 due to the surface tension in the hole H1.

  The greater the surface tension of the liquid L1, the greater the contact angle θ1. The pressure P1 applied to the liquid L1 by the surface tension is expressed by the equation (1). Further, the pressure P2 applied to the liquid L1 by gravity is given by the equation (2). In this case, if the pressures P1 and P2 satisfy the expression (3), the liquid L1 in the container C1 does not leak from the hole H1 to the outside of the container. The expressions (1) to (3) do not depend on the inner diameter Φ2 of the container C1.

  Therefore, in the formation of the opening 1251 of the introduction hole 1241 and the gas vent hole 1248, for example, first, using the formulas (1) to (3), an approximate hole size Φ1 (hole diameter for preventing liquid leakage) is used. ) Is required. Next, the hole size for preventing liquid leakage is determined by experimentally adjusting the size from the determined approximate hole size Φ1. Then, based on the determined hole size Φ1, the diameter of the nozzle 1180 and the mold size for forming the groove 1249 are set, and the nozzle and the mold that match the set size are respectively employed. Note that the groove 1249 may be formed to have a size set by, for example, cutting instead of injection molding. If the groove portion 1249 is formed in the lid member 1158 in advance by injection molding or the like, for example, the gas vent hole 1248 of the gas vent hole 1248 is compared with a case where a gas vent hole penetrating the second region 1320 of the seal member 1160 is formed at the time of inspection. It becomes easier to form the water repellent film 35 c on the wall surface. Therefore, if the groove 1249 is formed in the lid member 1158 in advance, liquid leakage from the test chip 1010 is less likely to occur.

  When the height h of the liquid level introduced into the flow path 1163, the nozzle insertion hole 1164, and the liquid reservoir hole 1166 is assumed to be, for example, 5 mm, the diameter of the introduction hole 1241 is usually 2 to 3 mm. The hole diameter of the opening 1251 of the gas vent hole 1248 is set to about 1 mm. Note that even if the hole diameter of the opening 1251 is set to be approximately the same as the hole diameter of the introduction hole 1241, the usefulness of the present invention is not impaired.

  Further, when the modeling expressing the liquid leakage from the introduction hole 1241 in the nozzle insertion hole 1164 and the gas vent hole 1248 in the liquid reservoir hole 1166 is realized more accurately than the required specifications are satisfied, the expression (1) Even if the pore size is determined only by a calculation formula corresponding to the formula (3) or the like, the usefulness of the present invention is not impaired. Moreover, even if the pore size is obtained by experiments alone without using the equations (1) to (3), the usefulness of the present invention is not impaired.

  If the hole diameters of the opening 1251 of the introduction hole 1241 and the gas vent hole 1248 are set as described above, for example, even if the inspection chip 1010 is turned over so that the main surface 1162 of the seal member 1160 faces the ground, The liquid inside the inspection chip can be prevented from leaking outside.

(Reagent chip)
As shown in FIG. 1, a nozzle 1180, a sample container 1182, a dilution container 1184, a dilution liquid container 1186, a labeled antibody liquid container 1188 and a washing liquid container 1190 are coupled by a holder 1192. These may be provided separately.

(Nozzle and liquid pump)
In the nozzle 1180, a liquid storage space for storing a liquid is formed inside. The liquid storage space has a liquid inlet / outlet at the tip 1252 and an opening to which the liquid feed pump 1140 is attached at the other end. The diameter of the nozzle 1180 becomes thinner as it is closer to the tip 1252 side. The larger the diameter of the nozzle 1180 is on the tip 1252 side, the thicker it may be. The diameter of the nozzle 1180 may be uniform. The diameter of the nozzle 1180 may change irregularly. The nozzle 1180 is preferably made of resin. Thereby, the nozzle 1180 is manufactured at low cost. The material of the nozzle 1180 may be other than resin such as glass.

(Mounting the nozzle to the pump)
As shown in FIG. 6, when the nozzle 1180 is attached to the liquid feed pump 1140, the pump tip is press-fitted into an opening at an end different from the tip 1252 of the nozzle 1180. The nozzle 1180 may be attached to the liquid feed pump 1140 by other methods.

(Liquid ejection and inhalation)
The liquid feed pump 1140 makes the internal liquid storage space positive or negative. When the liquid storage space is set to a positive pressure, the liquid is discharged from the liquid storage space via the liquid inlet / outlet of the tip 1252. When the liquid storage space is set to a negative pressure, the liquid is sucked into the liquid storage space via the liquid inlet / outlet.

  When the liquid is supplied to the flow path 1163, as shown in FIG. 6, the seal member 1160 is broken to insert the nozzle 1180 into the nozzle insertion hole 1164, and the liquid is discharged from the liquid storage space via the liquid inlet / outlet. The When the liquid is recovered from the flow path 1163, the liquid is sucked into the liquid storage space via the liquid inlet / outlet in a state where the nozzle 1180 is inserted into the nozzle insertion hole 1164. When the measurement is completed and the nozzle 1180 is removed from the nozzle insertion hole 1164, an introduction hole 1241 is formed in the nozzle insertion port 1240 as shown in FIG.

(Tip position)
When the nozzle 1180 is inserted into the nozzle insertion hole 1164, the tip 1252 crosses the boundary between the nozzle insertion hole 1164 and the flow path 1163, and the tip 1252 is accommodated in the tip accommodation hole 1170. Thus, the tip 1252 is disposed inside the flow path 1163 and the liquid is easily recovered from the flow path 1163. Therefore, when the sample liquid 1202 is supplied to the flow path 1163 after the cleaning liquid 1198, the cleaning liquid 1198 is sufficiently recovered from the flow path 1163, and the sample liquid 1202 is not diluted with the cleaning liquid 1198 and is contained in the sample liquid 1202. And the biochemical reaction (interaction) with the antibody contained in the capturing body 1154 is less affected by the cleaning liquid 1198. Two or more nozzle insertion holes and a tip accommodation hole may be provided, and the nozzle insertion hole and the tip accommodation hole into which the nozzle 1180 is inserted when the liquid is supplied to the flow channel 1163, the liquid is supplied from the flow channel 1163. It may be different from the nozzle insertion hole and the tip accommodation hole into which the nozzle 1180 is inserted when it is collected.

  Desirably, the tip 1252 is accommodated in the tip accommodation hole 1170 both when the liquid is supplied to the channel 1163 and when it is recovered from the channel 1163. Thereby, supply and collection | recovery of a liquid are performed in the same position, and conveyance of the liquid feeding pump 1140 reduces. However, it is necessary that the tip 1252 be accommodated in the tip accommodation hole 1170 when the liquid is collected, but it is not always necessary that the tip 1252 is accommodated in the tip accommodation hole 1170 when the liquid is supplied. Alternatively, the tip 1252 may be housed away from the tip housing hole 1170.

(Optical path of excitation light and reflected light)
As shown in FIG. 1, the excitation light 1210 is guided from the laser diode 1100 to the incident surface 1220 by the excitation optical system 1212. The excitation light 1210 sequentially passes through the first band pass filter 1102, the linear polarization filter 1104, the neutral density filter 1106, the half-wave plate 1108, and the shaping optical system 1112, and is reflected by the mirror 1114. The reflected light 1280 emitted from the emission surface 1224 is absorbed by the light absorber 1130.

(Laser diode)
The laser diode 1100 emits a beam of substantially monochromatic light and linearly polarized excitation light 1210. The light quantity and wavelength of the laser diode 1100 are stabilized. The laser diode 1100 may be replaced with other types of light sources. For example, the laser diode 1100 may be replaced with a light emitting diode, a mercury lamp, a laser other than the laser diode, or the like. When the light emitted from the light source is not a beam, the light is converted into a beam by a lens, a mirror, a slit, or the like. When the light emitted from the light source is not monochromatic light, the light is converted into monochromatic light by a diffraction grating or the like. When the light emitted from the light source is not linearly polarized light, the light is converted into linearly polarized light by a polarizer or the like.

(First bandpass filter)
When the excitation light 1210 passes through the first bandpass filter 1102, the wavelength distribution of the excitation light 1210 is narrowed. Thereby, even if the wavelength distribution of the excitation light 1210 emitted from the laser diode 1100 is wide, the excitation light 1210 having a narrow wavelength distribution can be obtained. When the wavelength distribution of the excitation light 1210 emitted from the laser diode 1100 is sufficiently narrow, the first band pass filter 1102 may be omitted.

(Linear polarization filter)
When the excitation light 1210 passes through the linear polarization filter 1104, the distribution of the polarization direction of the excitation light 1210 is narrowed. Thereby, even if the distribution of the polarization direction of the excitation light 1210 emitted from the laser diode 1100 is wide, the excitation light 1210 having a narrow distribution of the polarization direction can be obtained. When the distribution of the polarization direction of the excitation light 1210 emitted from the laser diode 1100 is sufficiently narrow, the linear polarization filter 1104 may be omitted.

(Nettenuation filter)
When the excitation light 1210 passes through the neutral density filter 1106, the light amount of the excitation light 1210 is decreased. When the amount of excitation light 1210 emitted from the laser diode 1100 is appropriate, the neutral density filter 1106 may be omitted.

(Half wave plate and half wave plate drive mechanism)
When the excitation light 1210 passes through the half-wave plate 1108, the polarization direction of the excitation light 1210 is adjusted. The half-wave plate 1108 is rotated around a rotation axis perpendicular to the half-wave plate 1108 by a half-wave plate driving mechanism 1110. The half-wave plate driving mechanism 1110 rotates the half-wave plate 1108 by a rotary stepping motor or the like. The polarization direction of the excitation light 1210 passing through the half-wave plate 1108 is adjusted by the rotation angle of the half-wave plate 1108. The polarization direction of the excitation light 1210 can be adjusted between the polarization direction in which the oozing of the evanescent wave from the reflecting surface 1222 is maximized and the polarization direction in which the oozing of the evanescent wave from the reflecting surface 1222 is eliminated. In the initial state, the rotation angle of the half-wave plate 1108 is set to the rotation angle at which the p-polarized component is expected to be incident on the reflecting surface 1222.

  The half-wave plate driving mechanism 1110 may be replaced with a mechanism that rotates the laser diode 1100 around the optical axis. When the optimization of the polarization direction is omitted, the half-wave plate driving mechanism 1110 may be omitted and the half-wave plate 1108 may be fixed. When the optimization of the polarization direction is omitted and the laser diode 1100 is held in a posture in which the p-polarized component is mainly incident on the reflection surface 1222, the half-wave plate 1108 and the half-wave plate driving mechanism 1110 are omitted. Good.

(Shaping optics)
When the excitation light 1210 passes through the shaping optical system 1112, the beam size, cross-sectional shape, etc. of the excitation light 1210 are shaped by a slit, a zoom optical system, or the like.

(Mirror and mirror drive mechanism)
When the excitation light 1210 is reflected by the mirror 1114, the traveling direction of the excitation light 1210 is bent in the direction from the optical axis direction of the laser diode 1100 toward the incident surface 1220. The mirror 1114 is rotated in the direction perpendicular to the optical path of the excitation light 1210 by the mirror drive mechanism 1116 and moved in the optical axis direction of the laser diode 1100.

  When the incident angle θ of the excitation light 1210 on the reflecting surface 1222 is adjusted by the mirror 1114 and the mirror driving mechanism 1116, the irradiation position is moved by changing the incident angle θ while the mirror angle is adjusted by the mirror driving mechanism 1116. The mirror position is adjusted by the mirror drive mechanism 1116 so as to cancel. Thereby, only incident angle (theta) is adjusted and the irradiation position of the excitation light 1210 in the reflective surface 1222 is maintained.

  The mirror drive mechanism 1116 rotates the mirror 1114 around a rotation axis parallel to the mirror surface by a rotary stepping motor or the like, and adjusts the mirror angle. The mirror drive mechanism 1116 moves the mirror 1114 along the optical axis direction of the laser diode 1100 by a linear stepping motor or the like on a linear stage or the like, and adjusts the mirror position.

  The mirror 1114 and the mirror driving mechanism 1116 have an advantage that the incident angle θ can be adjusted by a simple mechanism. However, the incident angle θ may be adjusted by a mechanism that adjusts the positions and postures of the laser diode 1100 and the excitation optical system 1212. Alternatively, the incident angle θ may be adjusted by a mechanism that adjusts the posture of the inspection chip 1010.

(Optical path of surface plasmon excitation fluorescence)
The surface plasmon excitation fluorescence 1218 is guided from the capturing body 1154 to the photomultiplier tube 1126 by the detection optical system 1214. The surface plasmon excitation fluorescence 1218 sequentially passes through the condenser lens 1118, the second band pass filter 1120, and the imaging lens 1124.

(Condensing lens and imaging lens)
The condensing lens 1118 condenses the surface plasmon excitation fluorescence 1218 and converts it into parallel light. The imaging lens 1124 images the surface plasmon excitation fluorescence 1218 onto the photomultiplier tube 1126. The condensing lens 1118 and the imaging lens 1124 constitute a conjugate optical system. Thereby, the influence of stray light is suppressed.

(Second bandpass filter and bandpass filter drive mechanism)
The second band pass filter 1120 is inserted into and extracted from the optical path of the measurement light between the condenser lens 1118 and the imaging lens 1124 by the band pass filter driving mechanism 1122. “Measurement light” is scattered light and surface plasmon excitation fluorescence 1218.

  When the amount of the surface plasmon excitation fluorescence 1218 is measured, a second bandpass filter 1120 is inserted into the optical path of the measurement light, and the second bandpass filter 1120 selectively emits light having the same wavelength as the excitation light 1210. Attenuated. Thereby, the scattered light is attenuated, and the surface plasmon excitation fluorescence 1218 is mainly guided to the photomultiplier tube 1126, and the sensitivity and accuracy of measurement are improved. When the amount of scattered light is measured, the second bandpass filter 1120 is removed from the optical path of the measurement light.

  A neutral density filter may be inserted into and removed from the optical path of the measurement light. When the amount of surface plasmon excitation fluorescence 1218 is measured, the neutral density filter is removed from the optical path of the measurement light. When the amount of scattered light is measured, a neutral density filter is inserted into the optical path of the measurement light. As a result, the difference in the amount of light between the surface plasmon excitation fluorescence 1218 guided to the photomultiplier tube 1126 and the scattered light becomes small, and the measurement of the amount of light becomes easy. The amount of light of the surface plasmon excitation fluorescence 1218 may be measured by a relatively high sensitivity photomultiplier tube 1126, and the amount of scattered light may be measured by a relatively low sensitivity photodiode, phototransistor, photoresistor, or the like.

(Photomultiplier tube)
The amount of measurement light is measured by a photomultiplier tube 1126. The photomultiplier tube 1126 has the advantage of good sensitivity and signal-to-noise (S / N) ratio. However, the photomultiplier tube 1126 may be replaced with another type of light quantity sensor. For example, the photomultiplier tube 1126 may be replaced with a cooled charge coupled device (CCD) camera or the like.

<Production of inspection chip 1010>
(Preparation of each component)
The flowchart of FIG. 9 shows an example of the manufacturing procedure of the inspection chip 1010. In the assembly of the inspection chip 1010, each component and material constituting the inspection chip 1010 are prepared (step S11). Specifically, a prism 1150, a material for the conductor film 1152, a material for the capturing body 1154, a flow path member 1156, a lid member 1158, a seal member 1160, a water repellent, and the like are prepared.

(Formation of water repellent film 1)
When the respective components constituting the inspection chip 1010 are prepared, the water repellent film 35a, 35b, and 35d is then applied by applying a water repellent agent to a predetermined portion of the main surface 1161 of the seal member 1160. Is formed (step S12).

(Production of complex)
When the water repellent film is formed, a composite 1148 is manufactured (step S13). Specifically, a conductive film 1152 is formed by fixing a material such as gold on the reflecting surface 1222 of the prism 1150 by sputtering, vapor deposition, plating, or the like, and the prism 1150 and the conductive film 1152 are formed. A composite 1148 is produced.

(Fixing the capturing body)
When the composite 1148 is manufactured, the capturing body 1154 is fixed (“fixed”) to a part of the main surface 1232 of the conductor film 1152 that forms the road surface of the flow path 1163 (step S14). When the flow path member 1156 is joined to the composite body 1148, the portion becomes a part surrounded by the opening 1237 of the flow path 1163. The capture body 1154 is fixed by fixing a material of the capture body 1154 including an antibody which is a reaction product of a biochemical reaction to the main surface 1232 by a surface treatment.

(Join the flow path member)
When the capturing body 1154 is fixed to the composite body 1148, the flow path member 1156 is bonded to the main surface 1232 of the conductor film 1152 by adhesion or the like (step S15). Through the bonding of the flow path member 1156, a flow path 1163 having an opening 1237 is formed on the conductor film 1152.

(Covering the lid member)
When the capturing body 1154 is fixed, the lid member 1158 is joined to the main surface 1236 of the flow path member 1156 by joining the joining surface 1226 of the lid member 1158 by adhesion or the like (step S16). By joining the lid member 1158, a nozzle insertion hole 1164, a liquid reservoir hole 1166, a groove portion 1249, and the like are formed.

(Blocking process)
When the lid member is joined, the nozzle insertion hole 1164, the flow path 1163, and the liquid reservoir hole 1166 are subjected to a blocking process using a liquid containing a protein such as skim milk or albumin (step S17), and the sample liquid Nonspecific adsorption by the antigen to be detected contained in 1202 is prevented.

(Formation of water repellent film 2)
When the blocking process is performed, the water-repellent film 35c is formed on the portion of the wall surface of the groove 1249 of the lid member 1158 that forms the wall surface of the gas vent hole 1248 (step S18). Although the water repellency of the groove portion 1249 is impaired by the blocking treatment, the water repellency of the groove portion 1249 is sufficient to prevent liquid leakage from the gas vent hole 1248 by forming the water repellent film 35c after the blocking treatment. Enhanced.

(Join seal member)
When the water repellent film 35c is formed in the groove 1249, the seal member 1160 is joined to the seal surface 1228 of the lid member 1158 (step S19). The joining is performed by, for example, joining one main surface 1161 of the seal member 1160 to the seal surface 1228 after an adhesive is applied to the seal surface 1228. By joining the seal member 1160, the nozzle insertion port 1240 and the liquid reservoir hole 1166 are closed. Further, except for the opening 1250, the groove 1249 is also closed to form a gas vent hole 1248, and the manufacture of the inspection chip 1010 is completed.

<Measurement procedure>
(Preparation and mounting of inspection chip and reagent chip)
The flowchart of FIG. 10 shows a measurement procedure. In the measurement, the inspection chip 1010 and the reagent chip 1012 are prepared and attached to the measurement apparatus 1000 (step S101). The inspection chip 1010 and the reagent chip 1012 attached to the measuring apparatus 1000 are arranged in the pretreatment chamber 1800. When the reagent chip 1012 is prepared, the sample 1200 is stored in the sample container 1182. The specimen 1200 is typically a collected material from a human such as blood, but may be a collected material from a non-human organism or a non-living material.

(Mounting the nozzle to the pump)
After the inspection chip 1010 and the reagent chip 1012 are arranged in the pretreatment chamber 1800, the nozzle 1180 is attached to the liquid feed pump 1140 (step S102). At this time, the liquid feed pump transport mechanism 1142 is controlled by the control unit 1006, so that the liquid feed pump 1140 is transported toward the nozzle 1180, and the pump tip is press-fitted into the opening at the end different from the tip 1252 of the nozzle 1180. The press-fitting amount is adjusted by the press-fitting torque or the transport distance of the liquid feed pump 1140.

(Detection of tip)
After the nozzle 1180 is attached, the tip 1252 is detected (step S103). At this time, the liquid feed pump transport mechanism 1142 is controlled by the control unit 1006, the tip 1252 is pressed against the upper surface of the inspection chip 1010, and it is detected that the front end 1252 is in contact with the upper surface of the inspection chip 1010 due to a change in torque or the like. . Thereby, the relative positional relationship between the inspection chip 1010 and the tip 1252 is specified, and the tip 1252 can be accommodated in the tip accommodation hole 1170. The tip 1252 may be detected from the projection image of the nozzle 1180.

(Inspection chip cleaning)
After the tip 1252 is detected, the inspection chip 1010 is washed (step S104). At this time, the liquid feed pump transport mechanism 1142 is controlled by the control unit 1006, the nozzle 1180 is inserted into the cleaning liquid solution 1190, and the tip 1252 is immersed in the cleaning liquid 1198. In a state where the tip 1252 is immersed in the cleaning liquid 1198, the liquid feed pump 1140 is controlled by the control unit 1006, and the cleaning liquid 1198 is sucked into the liquid storage space in the nozzle 1180.

  After the cleaning liquid 1198 is sucked into the liquid storage space, the liquid feed pump transport mechanism 1142 is controlled by the control unit 1006, the nozzle 1180 breaks the seal member 1160 and is inserted into the nozzle insertion hole 1164, and the tip 1252 is the tip storage hole. The cleaning liquid 1198 is supplied to the inspection chip 1010 by being stored in 1170. In a state where the tip 1252 is housed in the tip housing hole 1170, the liquid feed pump 1140 is controlled by the control unit 1006, and the cleaning liquid 1198 is supplied from the liquid housing space to the flow path 1163 via the liquid inlet / outlet, which is necessary for cleaning. After a lapse of time, the cleaning liquid 1198 is recovered from the flow path 1163 to the liquid storage space via the liquid inlet / outlet. At this time, since the tip 1252 is inserted deeply beyond the boundary between the nozzle insertion hole 1164 and the flow path 1163, the cleaning liquid 1198 is almost completely recovered from the flow path 1163. The recovered cleaning liquid 1198 is discarded into the cleaning liquid container 1190 or a separately provided waste liquid container. The supply and recovery of the liquid may be repeated twice or more.

  The feeding of the cleaning liquid 1198 from the cleaning liquid container 1190 to the inspection chip 1010 may be repeated twice or more. By cleaning the inspection chip 1010, biochemical reaction inhibitors such as a storage reagent are removed, and the biochemical reaction can proceed appropriately. Instead of the cleaning liquid 1198 or in addition to the cleaning liquid 1198, a liquid such as a buffer liquid or a processing liquid may be supplied to the flow path 1163 and recovered from the flow path 1163.

  When cleaning of the inspection chip 1010 is completed, the liquid feed pump transport mechanism 1142 is controlled by the control unit 1006, whereby the nozzle 1180 is removed from the nozzle insertion port 1240 of the nozzle insertion hole 1164. When the nozzle 1180 is removed from the nozzle insertion hole 1164, an introduction hole 1241 is formed in the seal member 1160.

(Immune reaction between antigen and antibody)
After the test chip 1010 is washed, an immune reaction between the antibody and the antigen is allowed to proceed (step S105).

  At this time, the liquid feed pump transport mechanism 1142 and the liquid feed pump 1140 are controlled by the control unit 1006, the specimen 1200 is fed from the specimen container 1182 to the dilution container 1184, and the diluent 1194 is fed from the dilution liquid container 1186 to the dilution container 1184. The liquid is fed and the specimen 1200 is diluted. The diluted container 1184 contains the prepared sample solution 1202. Before the reagent chip 1012 is attached to the measuring apparatus 1000, the sample solution 1202 may be prepared outside the measuring apparatus 1000. The specimen 1200 may be used as the sample liquid 1202 as it is. Instead of dilution of the specimen 1200, or in addition to dilution of the specimen 1200, pretreatment such as blood cell separation and mixing of reagents may be performed.

  After the sample liquid 1202 is prepared, the liquid feed pump transport mechanism 1142 is controlled by the control unit 1006, the nozzle 1180 is inserted into the dilution container 1184, and the tip 1252 is immersed in the sample liquid 1202. In a state where the tip 1252 is immersed in the sample liquid 1202, the liquid feed pump 1140 is controlled by the control unit 1006, and the sample liquid 1202 is sucked into the liquid storage space.

  After the sample liquid 1202 is sucked into the liquid storage space, the liquid feed pump transport mechanism 1142 is controlled by the control unit 1006, and the nozzle 1180 is inserted into the introduction hole 1241 formed in the seal member 1160, so that the nozzle again It is inserted into the insertion hole 1164. Then, the tip 1252 is accommodated in the tip accommodation hole 1170, and the sample liquid 1202 is sent to the inspection chip 1010. In a state where the tip 1252 is housed in the tip housing hole 1170, the liquid feeding pump 1140 is controlled by the control unit 1006, and the sample solution 1202 is supplied from the liquid housing space to the flow path 1163 via the liquid inlet / outlet, thereby causing an immune reaction. After the necessary time has elapsed, the sample liquid 1202 is recovered from the flow path 1163 to the liquid storage space via the liquid inlet / outlet. As a result, the sample liquid 1202 comes into contact with the capturing body 1154, and the antibody contained in the capturing body 1154 and the antigen contained in the sample liquid 1202 are combined. Since the washing solution 1198 is almost completely recovered, the sample solution 1202 is not diluted in the washing solution 1198, and the immune reaction between the antibody and the antigen is not affected by the washing solution 1198. Since the tip 1252 is inserted deeply beyond the boundary between the nozzle insertion hole 1164 and the channel 1163, the sample solution 1202 is almost completely recovered from the channel 1163. The collected sample liquid 1202 is discarded into a dilution container 1184 or a separate waste liquid container.

(Setting incident angle to measurement angle)
After the immune reaction between the antibody and the antigen, the incident angle is set to the measurement angle (step S106).

  At this time, the inspection chip transport mechanism 1128 is controlled by the control unit 1006, and the inspection chip 1010 is loaded from the pretreatment chamber 1800 to the measurement position 1802. Further, the band-pass filter driving mechanism 1122 is controlled by the control unit 1006, and the second band-pass filter 1120 is removed from the optical path of the measurement light. Thereby, the scattered light is not shielded, and the amount of the scattered light can be measured by the photomultiplier tube 1126.

  In a state where the inspection chip 1010 is loaded at the measurement position 1802 and the second bandpass filter 1120 is removed, the mirror driving mechanism 1116 is controlled by the control unit 1006 and the incident angle θ is scanned in the vicinity of the expected resonance angle. The The incident angle θ satisfies the total reflection condition. While the incident angle θ is being scanned, the control unit 1006 acquires the measurement result of the amount of scattered light from the photomultiplier tube 1126. Thereby, the relationship between the incident angle θ and the amount of scattered light is obtained. The control unit 1006 specifies the resonance angle θr, which is the incident angle θ at which the amount of scattered light is maximized, from the relationship between the incident angle θ and the amount of scattered light, and is the incident angle θ that is measured from the resonance angle θr. The measurement angle θm is determined. In general, the measurement angle θm coincides with the resonance angle θr, but some correction may be performed. Instead of the incident angle θ at which the amount of scattered light is maximized, the incident angle θ at which the amount of reflected light 1280 is minimized may be the resonance angle θr. However, since the incident angle θ at which the amount of reflected light 1280 is minimized and the incident angle θ at which the electric field enhancement is maximized are slightly shifted, the incident angle θ at which the amount of reflected light is minimized is defined as the resonance angle θr. In this case, the measurement angle θm is determined by adding or subtracting a small angle to the incident angle θ at which the amount of reflected light is minimized. After the measurement angle θm is determined, the mirror driving mechanism 1116 is controlled by the control unit 1006, and the incident angle θ is set to the measurement angle θm. Desirably, the half-wave plate driving mechanism 1110 is controlled by the control unit 1006, and the polarization direction of the excitation light 1210 is optimized.

(Immune reaction between antigen and labeled antibody)
After the incident angle θ is set to the measurement angle θm, the immune reaction between the antigen and the labeled antibody is allowed to proceed (step S107).

  At this time, the liquid feed pump transport mechanism 1142 is controlled by the control unit 1006, the nozzle 1180 is inserted into the labeled antibody liquid container 1188, and the tip 1252 is immersed in the labeled antibody liquid 1196. In a state where the tip 1252 is immersed in the labeled antibody solution 1196, the liquid feeding pump 1140 is controlled by the control unit 1006, and the labeled antibody solution 1196 is sucked into the liquid storage space.

  After the labeled antibody solution 1196 is sucked into the liquid storage space, the liquid feed pump transport mechanism 1142 is controlled by the control unit 1006, and the nozzle 1180 is inserted into the introduction hole 1241 formed in the seal member 1160. It is inserted into the nozzle insertion hole 1164. Then, the distal end 1252 is accommodated in the distal end accommodation hole 1170, and the labeled antibody solution 1196 is sent to the test chip 1010. In a state where the tip 1252 is housed in the tip housing hole 1170, the liquid feed pump 1140 is controlled by the control unit 1006, and the labeled antibody solution 1196 is supplied from the liquid housing space to the flow path 1163 via the liquid inlet / outlet. Thereby, the labeled antibody solution 1196 comes into contact with the capturing body 1154, and the labeled antibody contained in the labeled antibody solution 1196 and the antibody contained in the capturing body 1154 are bound. Since the washing solution 1198 and the sample solution 1202 are almost completely recovered, the labeled antibody solution 1196 is not diluted in the washing solution 1198 and the sample solution 1202, and the immune reaction between the antigen and the labeled antibody is influenced by the washing solution 1198 and the sample solution 1202. I do not receive it. The immune reaction between the antigen and the labeled antibody may be allowed to proceed before the incident angle θ is set to the measurement angle θm.

(Measurement of light intensity of surface plasmon excitation fluorescence)
After the immune reaction between the antigen and the labeled antibody, the light amount of the surface plasmon excitation fluorescence 1218 is measured (step S108). At this time, the band pass filter drive mechanism 1122 is controlled by the control unit 1006, and the second band pass filter 1120 is inserted into the optical path of the measurement light. Further, the control unit 1006 acquires the measurement result of the light amount of the surface plasmon excitation fluorescence 1218 from the photomultiplier tube 1126. The measurement result of the light amount of the surface plasmon excitation fluorescence 1218 is converted into the presence / absence of an antigen, the capture amount of the antigen, etc., and displayed on the display unit 1008.

(Removal and disposal of inspection chip)
When the light amount of the surface plasmon excitation fluorescence 1218 is measured, the inspection chip 1010 is removed from the measuring apparatus 1000 and discarded (step S109). In the inspection chip 1010, the nozzle insertion port 1240 of the nozzle insertion hole 1164 is closed by the first region 1310 of one main surface 1161 of the seal member joined to the seal surface 1228 of the flow path structure 1050. A water repellent film 35 a is formed in at least a portion including the introduction hole 1241 in the first region 1310. Therefore, it is difficult for the liquid inside the inspection chip 1010 to leak from the introduction hole 1241. Similarly, the formation of the water-repellent films 35b to 35d makes it difficult for liquid leakage from the gas vent holes 1248 to occur.

  Here, when the inspection chip 1010 is removed and discarded, the posture of the inspection chip 1010 may be such that, for example, the main surface 1162 of the seal member 1160 faces the ground. However, in the test chip 1010, the formation of the above-described water-repellent film makes it difficult for liquid leakage of the sample liquid 1202 or the like from the introduction hole 1241 or the gas vent hole 1248 to the outside of the test chip 1010. Therefore, an increase in inspection cost and an increase in inspection time due to risk reduction processing such as contamination in the inspection chip 1010 after completion of the inspection can be suppressed.

  Further, in the inspection chip 1010, the sizes of the opening 1251 of the introduction hole 1241 and the gas vent hole 1248 are such that the sample liquid 1202 and the like inside the inspection chip 1010 can pass through these holes regardless of the posture of the inspection chip 1010. It is formed in a size that does not leak to the outside. Therefore, increase in inspection cost and prolongation of inspection time can be further suppressed.

(Manual transfer of liquid pump, etc.)
The liquid feed pump 1140 is desirably automatically transported by the liquid feed pump transport mechanism 1142 controlled by the control unit 1006, but all or part of the transport of the liquid feed pump 1140 may be performed manually. Even when the liquid feed pump 1140 is manually conveyed, the structure of the inspection chip 1010 described above is useful for substantially complete recovery of the cleaning liquid 1198.

  It is not essential that the liquid feeding pump 1140 is reciprocally conveyed between the liquid feeding source and the liquid feeding destination. That is, the liquid may be supplied from the liquid supply source to the liquid supply pump 1140 through a tube or the like, and the liquid supply pump 1140 may be lifted up and down exclusively.

  Although the invention has been shown and described in detail, the above description is illustrative in all aspects and not restrictive. Therefore, embodiments of the present invention can be modified or omitted as appropriate within the scope of the invention.

1000 Measuring device 1010 Inspection chip (replacement product)
1215 Reaction progressing device 1163 flow path (reaction flow path)
1237 opening (5th opening)
1235 opening (6th opening)
1154 Trapping body (reactant-containing body)
1152 Conductor film 1150 Prism (dielectric medium)
1148 Complex (occlusion)
1230 Main surface (6th main surface)
1232 principal surface (fifth principal surface)
1156 Channel member 1158 Lid member 1160 Seal member 1161 Main surface (second main surface)
1162 Main surface (first main surface)
1234 Main surface (4th main surface)
1236 Main surface (third main surface)
1164 Nozzle insertion hole 1240 Nozzle insertion opening (first opening)
1241 Introduction hole 1291 Opening (second opening)
1166 Liquid reservoir hole 1242 Opening (third opening)
1248 Vent hole 1292 Opening (fourth opening)
35a to 35d Water repellent film 1180 Nozzle 1310 First region 1320 Second region

Claims (8)

It is a replacement product for a reaction progress device that advances a biochemical reaction,
A flow path structure having a reaction flow path in which a reactant containing body containing the reaction product of the biochemical reaction is fixed on a road surface, and a nozzle insertion hole into which a nozzle for introducing a sample liquid is inserted into the reaction flow path Body,
A seal member having a first main surface and a second main surface, wherein the second main surface is joined to the surface of the flow path structure;
With
The nozzle insertion hole has a first opening on the surface of the flow channel structure and a second opening on the wall surface of the reaction flow channel,
A first region of the second main surface of the seal member closes the first opening of the nozzle insertion hole;
A replacement product, wherein a first water repellent film is formed in at least a part of the first region of the seal member. A replacement product according to claim 1,
The channel structure is
A liquid reservoir hole having a third opening on the surface of the flow path structure and a fourth opening on a wall surface of the reaction flow path, and capable of storing the sample liquid overflowing from the reaction flow path;
A groove formed on the surface of the flow channel structure and having one end reaching the side surface of the liquid reservoir hole;
Further comprising
The second region of the second main surface of the seal member closes at least a part of the groove and the third opening of the liquid reservoir hole, so that the sample liquid flows into the reaction channel. A gas vent hole is formed that can discharge the gas in the liquid reservoir hole to the outside of the replacement product when introduced,
A second water repellent film is formed on a portion of the second region of the seal member that covers at least a part of the third opening of the liquid reservoir hole, and
The replacement part, wherein the part of the third opening is a part of the third opening on the groove side. A replacement product according to claim 2,
In the flow channel structure,
A replacement product, wherein a third water-repellent film is formed on a portion of the wall surface of the groove portion constituting the wall surface of the gas vent hole. A replacement product according to claim 2 or claim 3,
In the sealing member,
A replacement product, wherein a fourth water repellent film is formed on a portion of the second main surface that constitutes the wall surface of the gas vent hole. A replacement product as claimed in any one of claims 2 to 4,
The size of the hole cross section of the vent hole is
A replacement product characterized in that the sample liquid introduced into the reaction channel has a size that does not leak out of the replacement product from the gas vent hole regardless of the posture of the replacement product. A replacement product as claimed in any one of claims 1 to 5,
The sealing member is
In the portion of the first main surface of the second main surface where the first water repellent film is formed, there is an introduction hole that penetrates the seal member;
The size of the introduction hole is
A replacement product characterized in that the sample liquid introduced into the reaction channel has a size that does not leak out of the replacement product from the introduction hole regardless of the posture of the replacement product. A replacement product as claimed in any one of claims 1 to 6,
The channel structure is
A third main surface and a fourth main surface, wherein the reaction channel is formed, and the reaction channel has a fifth opening and a sixth in the third main surface and the fourth main surface; A flow path member each having an opening of
A lid member joined to the third main surface of the flow path member and formed with the nozzle insertion hole;
An obstruction that is bonded to the fourth main surface of the flow path member and closes the sixth opening of the flow path member;
An exchange product characterized by comprising: A replacement product according to claim 7,
The obstruction is
A light transmissive dielectric medium having a light reflecting surface;
A conductor film interposed between the flow path member and the light reflecting surface;
An exchange product characterized by comprising:

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