JP6350541B2 - Surface plasmon resonance fluorescence analysis method, surface plasmon resonance fluorescence analyzer, and surface plasmon resonance fluorescence analysis system - Google Patents

Description

  The present invention relates to a surface plasmon resonance fluorescence analysis method for measuring a substance to be detected by emitting a fluorescent substance labeled with a substance to be detected using an electric field of an evanescent wave generated by surface plasmon resonance, and detecting the fluorescence, More particularly, the present invention relates to a surface plasmon resonance fluorescence analysis apparatus and a surface plasmon resonance fluorescence analysis system, and more specifically, a surface plasmon resonance fluorescence analysis method and a surface plasmon resonance fluorescence useful for automatically inspecting a plurality of specimen chips in a short time continuously. The present invention relates to an analyzer and a surface plasmon resonance fluorescence analysis system.

  Conventionally, as a surface plasmon resonance fluorescence analyzer (hereinafter, also simply referred to as “fluorescence analyzer”), the one described in Patent Document 1 is known. In this analyzer, an enhanced electric field (enhanced electric field) is formed in the vicinity of the surface of the conductor film by causing surface plasmon resonance in the conductor film formed on the prism. Sensitive and highly accurate specimen inspection is performed.

  For example, in the fluorescence analyzer 100 shown in FIGS. 7 and 8, the prism portion 110 on which the conductive film 112 is formed, the light source 120 that emits the light α toward the prism portion 110, and the prism portion 110 reflects the light. A light receiving unit 130 for measuring the light ray α and a fluorescence detection unit 140 for detecting light (fluorescence) based on the enhanced electric field in the vicinity of the conductor film 112.

  The prism unit 110 includes a triangular prism (hereinafter simply referred to as “prism”) 114, a conductor film 112 formed on a predetermined surface 114 b facing the apex angle of the prism 114, and the conductor film. An antibody solid phase film 113 formed on a surface 112a (surface opposite to the prism 114) 112a and having a capturing body (antibody) capturing a specific antigen in a specimen (sample solution) fixed on the surface; A flow path member 117 having a flow path 116 through which a specimen can flow while in contact with the surface of the antibody solid phase film 113.

  In the prism unit 110, the prism 114 causes the light beam α emitted from the light source 120 to enter the prism 114 from one inclined surface (incident surface) 114a, and is formed by the conductor film 112 provided on the predetermined surface 114b. The reflected light ray α is emitted from the other inclined surface (exit surface) 114c to the outside. Specifically, the light beam α incident on the prism 114 is totally reflected by the surface 112a of the conductor film 112 from the back surface (surface on the prism 114 side) 112b side of the conductor film 112, and is emitted from the exit surface 114c to the outside of the prism 114. It is injected.

  The light source 120 emits a light ray α toward the incident surface 114 a of the prism 114. The light source 120 is configured to be able to change the incident angle θ of the light ray α to the conductor film 112. The light receiving unit 130 receives the light beam α reflected from the conductive film 112 and emitted from the exit surface 114c of the prism 114 to the outside of the prism 114, and measures the intensity thereof. The fluorescence detection unit 140 is disposed at a position facing the conductor film 112 with the flow path 116 interposed therebetween, and detects fluorescence of the fluorescent material excited by an enhanced electric field formed near the surface of the conductor film 112. .

  Usually, in such a fluorescence analyzer 100, the detection target substance is inspected by the following steps (A) to (D) (see, for example, Patent Document 2).

  (A) The sample solution is caused to flow through the flow path 116 and contact the capturing body. When the specimen flows through the flow path 116, the target substance (specific antigen) in the specimen is captured by the antibody immobilized on the antibody solid phase film 113 by the antigen-antibody reaction (this process is referred to as a primary reaction process).

  (B) When inspecting the specimen (signal measurement step), the incident angle θ of the light ray α to the conductor film 112 for forming an enhanced electric field near the surface of the conductor film 112 (specifically, the conductor film 112 The incident angle θ of the light ray α incident on the front surface 112a from the back surface 112b side, that is, the measurement angle θ3 is determined.

  Specifically, the light α is emitted from the light source 120 while changing the incident angle θ of the light α to the conductor film 112. At this time, the light beam α reflected by the conductor film 112 is received by the light receiving unit 130, and the intensity thereof is measured. As a result, a resonance angle (SPR angle) θ1, which is an incident angle to the conductive film 112 at which surface plasmon resonance occurs in the conductive film 112, is obtained (see FIG. 9). Specifically, the incident angle to the conductor film 112 where the intensity of the reflected light is the smallest (that is, the incident angle where the reflectance is the smallest) becomes the resonance angle θ1. Here, there is a difference between the resonance angle θ1 at which surface plasmon resonance occurs in the conductor film 112 and the incident angle of the light beam α (the maximum angle of enhanced electric field θ2) to the conductor film 112 where the enhancement electric field is the largest. Therefore, the fluorescence analyzer 100 adjusts the predetermined angle from the obtained resonance angle θ1 (usually ± 0.5 °), and determines the measurement angle θ3 so as to substantially coincide with the enhanced electric field maximum angle θ2. To do.

  The emission direction of the light source 120 is adjusted so that the incident angle of the light ray α on the conductor film 112 becomes the measurement angle θ3. As a result, the enhanced electric field formed in the vicinity of the surface 112a of the conductor film 112 (near the channel 116 side of the conductor film 112) is substantially maximized (until the measurement angle θ3 is obtained from the incident angle θ using the optical system). This process is referred to as a resonance angle adjustment process).

  (C) A fluorescent substance is labeled on the antigen captured by the antibody immobilized on the antibody solid phase film 113 by flowing the fluorescently labeled antibody through the flow path 116 (this step is referred to as a secondary reaction step). .

  (D) This labeled fluorescent substance is excited by an enhanced electric field formed in the vicinity of the surface of the conductor film 112 to emit light. When the light receiving unit 130 measures the fluorescence, the fluorescence analyzer 100 measures the reaction amount of the antigen with high sensitivity and high accuracy (this step is referred to as a signal measurement step).

JP 2006-208069 A International Publication No. 2012/108323 Pamphlet

  By the way, as shown in FIG. 10A, such a conventional fluorescence analyzer is incorporated in a casing 82 that is easy to carry, and is used with the sample chip 1 attached thereto. Usually, one specimen chip insertion port 83 for inserting the specimen chip 1 containing the substance to be detected is formed in the housing of the fluorescence analyzer. The sample chip insertion port 83 has a height sufficiently higher than the thickness of the sample chip 1 so that the sample chip 1 does not contact the insertion port 83 when the sample chip 1 is inserted. In FIG. 10A, reference numeral 85 denotes a memory card slot, 86 denotes a print output port, 87 denotes an operation panel, and 88 denotes an input / output terminal.

  On the other hand, in order to improve the processing capacity of the test, it is desired to provide a plurality of sample chip insertion ports 83 and 83 so that analysis of a plurality of sample chips can be performed at a time. For that purpose, for example, as shown in FIG. 10B, it is conceivable to form two specimen chip insertion ports 83 in the housing 82. For example, the sample chips 1a and 1b are simultaneously inserted into the sample chip insertion openings 83 and 83, both tests are completed, and then the sample chips 1a and 1b are simultaneously removed from the housing 82 to obtain the next sample. The inspection can be performed by inserting the chips 1c and 1d in the same manner. In such a system, one sample chip insertion port 83 is formed as in the fluorescence analyzer 100 shown in FIG. 10A, and the sample chips 1a, 1b, 1c, 1d,. Compared with the embodiment in which the ・ is sequentially inserted, the time required for exchanging the sample chip can be omitted.

  Note that the number of the sample chip insertion ports 83 can be three or more.

  Here, the enhanced electric field maximum angle θ2 varies from chip to chip, in other words, the degree of electric field enhancement at a certain incident angle θ varies from chip to chip, so that the appropriate measurement angle θ3 varies from chip to chip. Therefore, even when the specimens are sequentially tested in the housing 82, each time a second specimen chip 1b is examined or a third specimen chip is examined, a new one is newly provided. It is necessary to perform the resonance angle adjustment step (B). If the next sample chip is tested with the measurement angle θ3 of the first sample chip 1a without newly measuring the appropriate measurement angle θ3, the same amount of fluorescent material should be emitted. Since the intensity of the signal to be obtained is different for each of the sample chips 1a, 1b,..., The quantitative property (measurement accuracy) of a specific antigen in the sample is deteriorated.

  As shown in FIG. 10 (A), when testing a plurality of sample chips 1a, 1b,... In the fluorescence analyzer 100 according to the embodiment having one sample chip insertion port 83, for example, one sample chip is inserted. The primary reaction process described above for the inspection of the sample chip 1a is (A), the resonance angle adjustment process is (B), the secondary reaction process is (C), and the signal measurement process is (D). ) Are performed in this order, and after the inspection of one sample chip 1a is completely completed, the next inspection of the sample chip 1b is performed in the order of (A) to (D) described above. As shown in B), the inspection time becomes extremely long.

  For example, in one embodiment, for one sample chip, the time required for the primary reaction step (A) is about 120 seconds, the time required for the resonance angle adjustment step (B) is about 20 seconds, and the time required for the secondary reaction step (C). The time required is about 50 seconds, and the time required for the signal measurement step (D) is about 20 seconds. Therefore, it takes about 210 seconds (3 minutes 30 seconds) to inspect one sample chip 1a. Further, it takes 2 × 210 seconds (about 7 minutes) to inspect the second sample chip 1b. When inspecting n specimen chips, a time of n × 210 seconds is the minimum required.

  In the case where there are a plurality of sample chip insertion ports 83 as shown in FIG. 10B, since the time occupied by the measurement using the optical system is small in the inspection time, the insertion port / liquid feeding is performed to reduce the cost of the apparatus. It is possible to reduce the number of optical systems relative to the number of systems. In that case, it is necessary to prevent at least the measurement timings at which the optical system is operated from overlapping between the insertion openings.

  10B, when there are a plurality of (for example, two) sample chip insertion openings 83, the inspection of the sample chips 1a, 1b... Inserted into the insertion openings 83, 83 is started simultaneously. Instead, as shown in FIG. 12, it is also possible to start at random with a time interval.

  Even when starting at random in this way, the resonance angle adjustment step (B) and the signal measurement step (D) overlap more than the number of optical systems (for example, the signal measurement step (D) for the sample chip 1a). And the resonance angle adjustment step (B) with respect to the sample chip 1b, etc.), the time must be adjusted, resulting in a time loss.

  Furthermore, it is assumed that the number of specimen chip insertion ports 83 is three, the number of optical systems is one, and the number of liquid feeding systems is three. In such a case, as shown in FIG. 13, the primary reaction step (A) is performed on the first sample chip, the second sample chip, and the third sample chip using a plurality of liquid feeding systems. Can be performed simultaneously. However, when the resonance angle adjustment step (B) of the first sample chip is performed, the second sample chip and the third sample chip must stand by. Therefore, time loss also occurs in such a case. In addition, there is a concern that antigen dissociation occurs during the waiting time.

  As described above, in the conventional fluorescence analyzer 100 or the fluorescence analyzer 110 that is assumed to have a plurality of sample chip insertion ports, in general, it takes a certain amount of time to test a plurality of sample chips 1. As a result, there is a problem that the cost required for the inspection becomes high.

  Further, when the number of liquid feeding systems 90 and the number of optical systems 150 are the same number and a plurality of sets are prepared, each of them is independent. You can also start. For example, if three sets of the liquid feeding system and the optical system are prepared, it is possible to simultaneously start the inspection for the first sample chip, the second sample chip, and the third sample chip.

  However, in such a case, there is a problem that the apparatus becomes large. Further, for example, the optical system 150 including the light source 120, the light receiving unit 130, and the fluorescence detection means 140 shown in FIG. 7 is expensive, and therefore the number of the optical systems 150 is matched to the number of the liquid feeding systems 90. Will greatly increase the cost of the fluorescence analyzer 100.

  In view of such circumstances, the present invention can reduce the time required for inspection as much as possible when inspecting a plurality of sample chips, and can inspect at low cost without increasing the size of the apparatus. It is an object of the present invention to provide a surface plasmon resonance fluorescence analysis method, a surface plasmon resonance fluorescence analysis apparatus, and a surface plasmon resonance fluorescence analysis system.

In order to achieve at least one of the above-described objects, a surface plasmon resonance fluorescence analysis method reflecting one aspect of the present invention includes:
The sample chip for holding a substance to be detected with a plurality, wherein the target substance is held respectively by the plurality of sample chips, continuously detecting surface plasmon resonance fluorescence analysis by surface plasmon resonance fluorescence analysis A method,
The sample chip has a prism and a conductor film formed on one surface of the prism,
When the plurality of sample chips are respectively inserted into fluorescence analyzers having a plurality of (n) sample chip insertion ports,
(B) an initial measurement step in which initial measurement is performed by irradiating the conductive film from the light source via the prism with light from the light source to the sample chip inserted into the sample chip insertion port of the fluorescence analyzer;
(C) injecting a sample solution containing the substance to be detected to the sample chip, and the sample solution is brought into contact with the catching捉体on the conductive film, the capturing member with the substance to be detected contained in the sample solution A primary reaction step of binding
(D) a secondary reaction step of labeling the substance to be detected with a fluorescent substance;
(E) irradiating the conductive film from the prism side with excitation light at a measurement angle, and measuring the amount of fluorescence emitted from the fluorescent substance that labels the substance to be detected, and a signal measurement step ,
The step (c) or the step (d) is performed after the step (b), and the step (e) is performed after the step (c) and the step (d) are completed.
Wherein among the plurality of sample chips, the 1 th of said step (c) or the step for the analyte chip (d) is completed Then simultaneously, the step (e) and two eyes for said one th sample chips of the started process (d) or the step (c) for the analyte chip, the (n-1) -th said step for the analyte chip (c) or said step (d) completing then simultaneously, the (n -1) The step (e) for the first sample chip and the step (d) or the step (c) for the nth sample chip are started .

In order to achieve at least one of the above-described objects, a surface plasmon resonance fluorescence analyzer that reflects one aspect of the present invention is provided.
In a surface plasmon resonance fluorescence analyzer for injecting a fluid into a guide part of a sample chip and reacting a reagent and a substance to be detected and measuring a reaction result from the detection part of the sample chip using an optical system,
A plurality (n) of sample chip insertion ports for inserting the sample chips into a housing of the surface plasmon resonance fluorescence analyzer;
A liquid feeding system for injecting fluid into the specimen chip;
The inserted the specimen tip from the specimen tip insertion port, and a drive member for liquid-based feed said a flow body guide portion is fixed at a predetermined position for communicating,
An optical system for optically detecting the substance to be detected;
The number of the optical system is set to a number smaller than the number of the sample chip insertion ports ,
When a plurality of sample chips are inserted into each of the plurality of sample chip insertion ports,
(B) an initial measurement step of performing initial measurement by irradiating the conductor film with light from the light source through the prism to the sample chip inserted into the sample chip insertion port of the surface plasmon resonance fluorescence analyzer;
(C) Injecting a sample solution containing the substance to be detected into the sample chip, bringing the sample solution into contact with a capturing body on the conductor film, and detecting the substance and the capturing body contained in the sample solution A primary reaction step of binding
(D) a secondary reaction step of labeling the substance to be detected with a fluorescent substance;
(E) performing a signal measurement step of irradiating the conductive film from the prism side with excitation light at a measurement angle and measuring the amount of fluorescence emitted from the fluorescent substance that labels the detection target substance. Composed,
The step (c) or the step (d) is performed after the step (b), and the step (e) is performed after the step (c) and the step (d) are completed.
At the same time when the step (c) or the step (d) for the first sample chip is completed among the plurality of sample chips, the step (e) and the second step for the first sample chip are completed. The step (d) or the step (c) for the sample chip is started, and at the same time the step (c) or the step (d) for the (n-1) -th sample chip is completed, the (n-1) ) Ru is configured to initiate said step (d) or the step (c) with respect to th sample chip to said step (e) and the n-th sample chips.

Furthermore, in order to realize at least one of the above-described objects, a surface plasmon resonance fluorescence analysis system reflecting one aspect of the present invention is provided.
A surface composed of a surface plasmon resonance fluorescence analyzer for injecting a fluid into the guide part of the sample chip, reacting the reagent and the substance to be detected, and measuring the reaction result from the detection part of the sample chip using an optical system, and a control unit A plasmon resonance fluorescence analysis system comprising:
The sample chip has a prism and a conductor film formed on one surface of the prism,
The surface plasmon resonance fluorescence analyzer is
A plurality (n) of sample chip insertion ports for inserting the sample chips into a housing of the surface plasmon resonance fluorescence analyzer;
A liquid feeding system for injecting fluid into the specimen chip;
A driving member for fixing the sample chip inserted from the sample chip insertion port at a predetermined position where the fluid guide unit and the liquid feeding system communicate;
An optical system for optically detecting the substance to be detected;
The number of the optical system is set to a number smaller than the number of the sample chip insertion ports,
When a plurality of the sample chips are inserted into each of the plurality of sample chip insertion ports, the control unit
(B) an initial measurement step in which initial measurement is performed by irradiating the conductor film with light from the light source through the prism to the sample chip inserted into the sample chip insertion port of the surface plasmon resonance fluorescence analyzer;
(C) Injecting a sample solution containing the substance to be detected into the specimen chip, bringing the sample solution into contact with the capturing body on the conductor film, and detecting the substance and the capturing body contained in the sample solution A primary reaction step of binding
(D) a secondary reaction step of labeling the substance to be detected with a fluorescent substance;
(E) performing a signal measurement step of irradiating the conductive film from the prism side with excitation light at a measurement angle and measuring the amount of fluorescence emitted from the fluorescent substance that labels the detection target substance. Composed,
The step (c) or the step (d) is performed after the step (b), and the step (e) is performed after the step (c) and the step (d) are completed.
At the same time when the step (c) or the step (d) for the first sample chip is completed among the plurality of sample chips, the step (e) and the second step for the first sample chip are completed. The step (d) or the step (c) for the sample chip is started, and at the same time the step (c) or the step (d) for the (n-1) -th sample chip is completed, the (n-1) ) Ru is configured to initiate said step; (d) step (c) with respect to th sample chip to said step (e) and the n-th sample chips.

  According to the surface plasmon resonance fluorescence analysis method, the surface plasmon resonance fluorescence analysis apparatus, and the surface plasmon resonance fluorescence analysis system according to the present invention, a plurality of specimen chips can be efficiently and continuously provided in a short time while suppressing the increase in size of the apparatus. It can be inspected and can contribute to a reduction in cost required for biochemical inspection.

FIG. 1 is a schematic configuration diagram of a surface plasmon resonance fluorescence analyzer according to a preferred embodiment of the present invention. FIG. 2 is a schematic configuration diagram of a state in which the positions of the first optical filter and the second optical filter are switched in the surface plasmon resonance fluorescence analyzer of FIG. FIG. 3 is a diagram showing the relationship between the intensity of plasmon scattered light generated near the surface of the conductor film formed on the prism and the electric field enhancement in the surface plasmon resonance fluorescence analyzer of FIG. FIG. 4 is an enlarged perspective view for explaining a reagent chip used in the liquid feeding system. FIG. 5 is an external view of the surface plasmon resonance fluorescence analyzer according to the present embodiment. FIG. 6A is a flow chart showing the relationship with the elapsed time when the sample chip is inspected by a so-called random method in the surface plasmon resonance fluorescence analyzer according to this embodiment, and FIG. 6B is this embodiment. 6 is a flowchart showing a relationship with an elapsed time when a sample chip is inspected by a so-called batch method using the surface plasmon resonance fluorescence analyzer according to FIG. FIG. 7 is a schematic diagram of a conventional surface plasmon resonance fluorescence analyzer. FIG. 8 is an enlarged vertical sectional view showing the structure of the prism portion of the surface plasmon resonance fluorescence analyzer shown in FIG. FIG. 9 is a diagram showing the relationship between the reflectance of light rays in the conductor film of the prism portion and the intensity of plasmon scattered light generated near the surface of the conductor film. FIG. 10A is an external view of a fluorescence analyzer having one sample chip insertion port, and FIG. 10B is an external view of a fluorescence analyzer having two sample chip insertion ports. FIG. 11A is a flowchart showing the relationship between the order of each process and the elapsed time when testing a plurality of sample chips in a conventional fluorescence analyzer, and FIG. It is a flowchart which shows the elapsed time in the case of test | inspecting the specimen chip | tip. FIG. 12 is a flowchart showing the relationship with the elapsed time in the case where a plurality of sample chips are randomly measured in a fluorescence analyzer provided with a plurality of sample chip insertion ports. FIG. 13 is a flowchart showing the relationship between the inspection method and the inspection time when there are three sample chip insertion ports, one optical system, and three liquid delivery systems in the fluorescence analyzer.

  Hereinafter, preferred embodiments of the present invention will be described with reference to the accompanying drawings.

  As shown in FIGS. 1 and 2, the surface plasmon resonance fluorescence analyzer (fluorescence analyzer) 10 according to the present embodiment includes a light source 40 that emits light toward the sample chip 20 and light generated in the sample chip 20. A light measuring unit 50 that measures the sample chip 20 and a holding unit 12 that detachably holds the sample chip 20. The fluorescence analyzer 10 also includes a calculation unit 14 for analyzing the light measured by the light measurement unit 50 and a display unit 16 for displaying various information such as calculation results in the calculation unit 14. 1 and 2 show a state in which the sample chip 20 is attached to the fluorescence analyzer 10. FIG.

  Further, the surface plasmon resonance fluorescence analysis system according to the present embodiment is an analysis system in which continuous operation is enabled by the apparatus 10 and the control unit 58 of FIG.

  The sample chip 20 includes a prism 21, a conductor film 25 formed on the surface of the prism 21, and a flow path through which a solution such as a sample, a reagent, and a cleaning solution flows while contacting the conductor film 25 on the conductor film 25. And a flow path member 30 that forms 31.

  The prism 21 is a film-forming surface (predetermined surface) on which an incident surface 22 for allowing the light α from the light source 40 to enter the prism 21 and a conductor film 25 for reflecting the light α incident on the prism 21 are formed. ) 23 and an emission surface 24 on which the light beam α reflected by the conductor film 25 is emitted to the outside of the prism 21 are formed on the surface thereof, and is formed of transparent glass or resin. The prism 21 of the present embodiment is made of transparent glass or resin having a refractive index of about 1.40 to 1.75. In addition, the prism may have a shape in which the apex angle portion of the triangular prism is cut off as in a side view as in the present embodiment, or may have a triangular shape in a side view (see the dotted line portion in FIG. 1). That is, the prism includes an incident surface, a film formation surface, and an emission surface on its surface, and a light beam α incident on the prism from the incident surface is totally reflected by the conductor film on the film formation surface. Any shape that exits from the exit surface to the outside without being irregularly reflected inside may be used.

  The conductor film 25 is a metal thin film formed (formed) on the film formation surface 23 of the prism 21. The conductor film 25 of this embodiment is made of gold. The conductor film 25 is a member for amplifying an evanescent wave generated by totally reflecting the light beam α. That is, by providing the conductor film 25 on the film formation surface 23 to cause surface plasmon resonance, the light α is totally reflected by the surface without the conductor film 25 (film formation surface 23) to generate an evanescent wave. As compared with the case, the electric field formed can be enhanced. The electric field enhancement (electric field enhancement) in this embodiment is about 10 times that of the case without the conductor film 25 (see FIG. 3). The conductor film 25 is formed by various film forming methods such as a sputtering method, a vapor deposition method, and a plating method so that the film thickness becomes 30 to 70 nm. The material of the conductor film 25 is not limited to gold, but may be any metal that generates surface plasmons, and may be, for example, silver, copper, aluminum, or the like (including an alloy).

  A capturing body 26 for capturing a specific antigen is fixed on the surface (surface opposite to the prism) 25a of the conductive film 25. The capturing body 26 is fixed to the surface 25a of the conductor film 25 by surface treatment, for example.

  The flow path member 30 is provided on the film formation surface 23 of the prism 21 and has a flow path 31 through which a specimen flows. The flow path member 30 is formed of a transparent resin. The flow path member 30 of the present embodiment is a plate-like member that expands in the horizontal direction. The flow channel 31 guides the sample from the outside of the sample chip 20 to the detection unit 32 and the detection unit 32 in which the antigen-antibody reaction is performed, or from the detection unit 32 to the external surface (the lower surface in FIG. 1) ) And the conductor film 25 on the prism 21 are surrounded by the groove provided in 30b. That is, in the detection unit 32, the specimen flows while being in contact with the surface of the conductive film 25 (the surface on which the capturing body 26 is fixed) 25a. As for each guide part 33, one edge part opens in the surface (upper surface in FIG. 1) 30a of the flow-path member 30, and the other edge part (edge part on the opposite side to the said one edge part) is a detection part. 32. As described above, the guide portion 33, the detection portion 32, and the guide portion 33 are sequentially connected to form a single flow path 31.

  The flow path member 30 is bonded (bonded) to the prism 21 with an adhesive. In the present embodiment, a seal member 35 made of an elastic body is provided at a position surrounding the detection unit 32 from the horizontal direction and between the flow path member 30 and the prism 21. Thereby, the leakage of the sample from the joint portion between the flow path member 30 and the prism 21 is prevented. In addition, joining of the flow path member 30 and the prism 21 is not limited to adhesion, and may be laser welding, ultrasonic welding, pressure bonding using a clamp member, or the like. Further, as long as the flow path member 30 and the prism 21 are joined in a liquid-tight manner, the seal member 35 surrounding the detection unit 32 may be omitted.

  The light source 40 is a light source device that emits light α toward the specimen chip 20, specifically, the incident surface 22 of the prism 21. The light source 40 includes a light emitting element 41 such as a semiconductor laser or an LED, an angle changing unit 42 that changes the emission direction of the light α by the light emitting element 41, and a polarizing plate (not shown) that polarizes the light α from the light emitting element 41. And have.

  The angle changing unit 42 is in a state where the light beam α emitted from the light emitting element 41 enters the prism 21 from the incident surface 22 and is totally reflected by the conductor film 25 (specifically, the back surface of the conductor film 25). In the state of passing through the conductor film 25 from the side (surface opposite to the surface 25a and being totally reflected by the surface 25a), the conductor film 25 of this light ray α (specifically, the conductor film) The emission direction of the light ray α by the light emitting element 41 is changed so as to change the incident angle θ with respect to the surface 25a of 25. Specifically, the angle changing unit 42 changes the emission direction of the light emitting element 41 so as to change the incident angle θ of the light ray α to the conductor film 25 without changing the position where the light ray α is reflected on the conductor film 25. It is configured to be changeable. The angle changing unit 42 of the present embodiment is connected to the control unit 58 of the light measuring unit 50 and changes the emission direction of the light beam α by the light emitting element 41 in accordance with an instruction signal from the control unit 58.

  The polarizing plate polarizes the light α emitted from the light emitting element 41 so as to be P-polarized with respect to the conductor film 25 provided on the film formation surface 23 of the prism 21. When a semiconductor laser is used as the light emitting element 41, a polarizing plate is not provided if the semiconductor laser is disposed so that the polarization plane of the semiconductor laser itself coincides with the polarization plane of P-polarized light with respect to the conductor film 25. Good.

  The light measuring unit 50 includes a light receiving unit 51 that receives light generated on the surface 25a side of the conductor film 25, a first optical filter 54, a second optical filter 55, and the first optical filter 54. A position switching device (position switching unit) 56 that switches the position with the second optical filter 55 and a control unit 58 that controls each component of the light measurement unit 50, the angle changing unit 42 of the light source 40, and the like. Prepare.

  The light receiving unit 51 includes a light receiving element 52 that detects light, and an optical system 53 that includes a lens or the like and guides light generated on the surface 25 a side of the conductor film 25 to the light receiving element 52. In this embodiment, a photomultiplier tube (so-called photomultiplier tube) is used as the light receiving element 52 in order to detect weak light (fluorescence generated by exciting a fluorescent substance labeled with an antigen in a specimen). ing.

  The first optical filter 54 is an optical filter that attenuates incident light and emits it. The first optical filter 54 of the present embodiment is a neutral density filter (ND filter). The first optical filter 54 attenuates the plasmon scattered light generated in the conductor film 25 of the sample chip 20, so that the plasmon by the light receiving element (photomal in this embodiment) 52 for detecting weak light. The scattered light can be measured.

  The second optical filter 55 is an optical filter for blocking light of a predetermined wavelength. The second optical filter 55 of the present embodiment is a long pass filter. The second optical filter 55 is not limited to a long pass filter, and may be a band pass filter or the like.

  The second optical filter 55 has light of a wavelength other than fluorescence (light generated by exciting a fluorescent substance labeled with an antigen in the specimen) in the light receiving unit 51 (for example, light from the light source 40 (leakage light)). And plasmon scattered light) are prevented from entering. That is, the second optical filter 55 removes noise components from the light incident on the light receiving unit 51, thereby improving the detection accuracy and detection sensitivity of weak fluorescence in the light receiving unit 51.

  The position switching device 56 is a device that switches the positions of the first optical filter 54 and the second optical filter 55 between an intermediate position and a retracted position. The intermediate position is a position between the conductor film 25 and the light receiving unit 51. Specifically, the intermediate position is a position between the sample chip 20 and the light receiving unit 51 (the position of the first optical filter 54 in FIG. 1). Further, the retracted position is a position (position of the second optical filter 55 in FIG. 1) farther from the conductor film 25 and the light receiving unit 51 than the intermediate position. Specifically, the retracted position is a position where light generated on the surface 25 a side of the conductor film 25 can directly enter the light receiving unit 51.

  Specifically, in the position switching device 56, when the first optical filter 54 is in the intermediate position, the second optical filter 55 is in the retracted position (see FIG. 1), and when the first optical filter 54 is in the retracted position. The positions of the first optical filter 54 and the second optical filter 55 are switched so that the second optical filter 55 is in the intermediate position (see FIG. 2). The position switching device 56 is connected to the control unit 58 and switches the positions of the first optical filter 54 and the second optical filter 55 based on an instruction signal from the control unit 58.

  The control unit 58 controls each component of the light source 40 and the light measurement unit 50. Specifically, the control unit 58 adjusts the emission direction of the light beam α by the light source 40 based on the light intensity measured by the light receiving unit 51. The control unit 58 of the present embodiment adjusts the emission direction of the light beam α of the light source 40 based on the intensity of the plasmon scattered light generated on the surface 25a side of the conductor film 25 of the sample chip 20. Specifically, the control unit 58 controls the light emitting element 41 and the angle changing unit 42 of the light source 40 when the sample chip 20 is held by the holding unit 12, so that the light beam α is on the back surface side (prisms) of the conductor film 25. 21) from the light source 40 so as to be incident on the conductor film 25 while changing the incident angle θ to the conductor film 25. At this time, the light receiving unit 51 detects the intensity of the plasmon scattered light generated on the surface 25 a side of the conductor film 25, and the control unit 58 controls the angle changing unit 42 based on the intensity change, and the light beam α by the light source 40. Adjust the injection direction. Specifically, when the light beam α is incident on the conductor film 25 while changing the incident angle θ, the intensity of the plasmon scattered light detected by the light receiving unit 51 changes as shown in FIG. The control unit 58 obtains the emission direction of the light beam α by the light source 40 when the light beam α is incident on the conductor film 25 at the incident angle θ5 when the intensity change reaches a peak (maximum value). Then, the control unit 58 adjusts the direction of the light emitting element 41 and outputs an instruction signal to the angle changing unit 42 so that the light source 40 can emit the light beam α in the emission direction.

  The control unit 58 instructs the position switching device 56 to switch the positions of the first optical filter 54 and the second optical filter 55. Specifically, when the control unit 58 adjusts the emission direction of the light beam α by the light source 40 using the plasmon scattered light, the first optical filter 54 is in the intermediate position and the second optical filter 55 is retracted. An instruction signal is output to the position switching device 56 so that the position is reached. On the other hand, the control unit 58 instructs the position switching device 56 so that the first optical filter 54 is in the retracted position and the second optical filter 55 is in the intermediate position when the fluorescence analyzer 10 performs the specimen inspection. Is output.

  The holding unit 12 is a part that holds the sample chip 20 when the sample is examined. Specifically, the holding unit 12 detachably holds the sample chip 20 so that the sample chip 20 is in a predetermined posture with respect to the light source 40. The predetermined attitude is an attitude in which the light beam α emitted from the light source 40 enters the prism 21 from the incident surface 22 and the light beam α incident on the prism 21 is totally reflected by the conductor film 25. is there.

  The reagent chip 60 shown in FIG. 4 is a member that can be mounted in the liquid feeding system. A specimen such as blood to be examined is pretreated (blood cell separation, dilution, mixing, etc.), and then the specimen is injected into the specimen chip 20. The reagent chip 60 is provided with a plurality of storage units 61, and each storage unit 61 is individually sealed with a reagent, a diluent, a cleaning solution, and the like in addition to a specimen such as blood.

  The calculation unit 14 is a part for calculating an output signal transmitted from the light measurement unit 50 (light receiving unit 51) and performing an analysis on the fluorescence measured by the light measurement unit 50 at the time of examination of the specimen. . Specifically, for example, the calculation unit 14 counts the number of fluorescences per unit area detected by the light measurement unit 50, calculates the amount of increase in fluorescence over time, and the like. The calculation result in the calculation unit 14 is output to the display unit 16 connected to the calculation unit 14.

  The display unit 16 displays the calculation result based on the output signal from the calculation unit 14. The display unit 16 may display a result on a screen such as a monitor, or may print out the result like a printer. The display unit 16 may be a combination of these.

  For example, as shown in FIG. 5, the fluorescence analyzer 10 configured as described above is used by being incorporated in one casing 820 that is easy to carry.

  For example, three specimen chip insertion ports 830 for inserting three specimen chips 20, 20, 20 are formed in the housing 820 that accommodates the fluorescence analyzer 10. Note that the number of specimen chip insertion ports 830 formed in the housing 820 is not limited to three, and may be two or more. In FIG. 5, reference numeral 85 is a memory card slot, 86 is a print output port, 87 is an operation panel, and 88 is an input / output terminal.

  Further, in the housing 820 of the present embodiment, a liquid feeding system 90 and an optical system 150 are provided as in the case shown in FIG. 10B, and the liquid feeding system 90 and the optical system 150 are provided. Between them, a transport mechanism capable of reciprocating both is provided. The transport mechanism allows the sample chip 20 to be transported from the liquid feeding system 90 to the optical system 150 or from the optical system 150 to the liquid feeding system 90. Furthermore, in the present embodiment, the number of optical systems 150 is smaller than the number of liquid feeding systems 90.

  However, the number of optical systems 150 and the number of liquid feeding systems 90 may be the same.

  Here, the liquid feeding system in this specification refers to blood, reagents, diluents, washing liquids, and the like stored in the storage units 61 of the reagent chip 60 in the flow channel 31 of the sample chip 20 shown in FIG. A system consisting of the components necessary for flow.

  Specifically, in addition to the reagent chip 60 of FIG. 4, a liquid feed pump, a liquid feed pump transport mechanism, and the like for injecting fluid from the reagent chip 60 to the flow path 31 of the sample chip 20 are mentioned.

  Further, the optical system means a system composed of members necessary for optically detecting a substance to be detected, such as the light source 40, the light measuring unit 50, and the position switching device 56, as described with reference to FIG. .

  In the present embodiment, since the number of these optical systems 150 is smaller than the number of liquid feeding systems 90, it contributes to downsizing the housing 820 and reducing the unit price of the fluorescence analyzer 100.

  In the fluorescence analyzer 10 configured as described above, the inspection is performed as follows.

(A) Preparation Step As shown in FIG. 5, a plurality of prepared sample chips 20 are inserted into the fluorescence analyzer 10 having three sample chip insertion ports 830, 830, and 830, respectively.

  Here, the three sample chips 20, 20, and 20 are a first sample chip, a second sample chip, and a third sample chip, respectively.

(B) Initial measurement process When the resonance angle is adjusted in the initial measurement process, first, the first sample chip 20 is placed at the examination position as shown in FIG. A measurement angle that is an incident angle of the light ray α to the conductive film 25 when the sample chip 20 is inspected is determined.

  Note that the initial measurement process broadly refers to a measurement process using an optical system that is a process other than a signal measurement process, specifically, either “resonance angle adjustment” or “blank measurement”, or It includes both. In addition, the initial measurement process includes acquisition of position information and the like other than “resonance angle adjustment” and “blank measurement”.

  Specifically, the control unit 58 outputs an instruction signal to the position switching device 56 so that the first optical filter 54 is in the intermediate position and the second optical filter 55 is in the retracted position. The position of the second optical filter 55 is switched (see FIG. 1).

  The control unit 58 changes the incident angle θ of the light beam α incident on the conductor film 25 of the prism 21 by the angle changing unit 42 while causing the light source 40 to emit the light beam α. At this time, the light ray α enters the prism 21, and the light ray α passes through the conductive film 25 from the back surface side of the conductive film 25 and is totally reflected on the surface 25 a, whereby the evanescent wave is converted into the conductive film 25. Ooze out to the surface 25a side of the surface, thereby generating plasmon scattered light. The light measuring unit 50 (specifically, the light receiving element (photomal) 52) measures the intensity of the plasmon scattered light. At this time, since the first optical filter 54 is disposed at the intermediate position, the plasmon scattered light generated on the surface 25a side of the conductor film 25 is attenuated by passing through the first optical filter 54. It reaches the light receiving element 52 in a state. Therefore, failure of the light receiving element 52 due to entering strong light (specifically, light that is stronger than fluorescence measured when examining the specimen) is prevented.

  The control unit 58 is based on the angle-dependent data in the plasmon scattered light measured by the light measurement unit 50 (data representing the relationship between the incident angle θ of the light ray α on the conductor film 25 and the light intensity: see FIG. 3). An incident angle θ5 at which the intensity of plasmon scattered light is maximized is obtained. If the incident angle θ5 of the ray α to the conductor film 25 when the intensity of the plasmon scattered light is maximum is known, the control unit 58 makes the incident of the ray α to the conductor film 25 having the maximum electric field enhancement intensity. The angle θ can be derived with high accuracy. This is because the intensity of the plasmon scattered light is maximum when the magnitude of the electric field (electric field enhancement intensity) formed in the vicinity of the surface 25a of the conductor film 25 due to surface plasmon resonance is maximized in the conductor film 25. Based on becoming.

  The control unit 58 controls the angle changing unit 42 so that the incident angle at which the light beam α is incident on the conductor film 25 is the incident angle θ5 when the intensity of the plasmon scattered light is maximum (that is, the electric field enhancement is the maximum). The direction of the light beam α emitted from the light source 40 is adjusted so that the incident angle θ5) becomes.

  Next, the control unit 58 switches the positions of the optical filters 54 and 55. That is, according to the instruction signal of the control unit 58, the position switching device 56 causes the first optical filter 54 and the second light so that the first optical filter 54 is in the retracted position and the second optical filter 55 is in the intermediate position. The position of the filter 55 is switched (see FIG. 2).

  In the initial measurement process, when blank measurement is performed, the sample chip 20 is filled with the measurement liquid, and measurement is performed by applying light from the light source 40.

  Note that blank measurement refers to self-fluorescence and excitation light of the chip itself as noise that should not be detected light by measuring fluorescence in advance in a state where nothing is captured by a capture body such as a capture antibody. The fluorescence intensity such as leakage of light is determined by a blank measurement process, and this value is adjusted to be subtracted from the fluorescence intensity measured in the signal measurement process performed later.

(C) Primary reaction step A sample solution is caused to flow through the flow path 31 of the sample chip 20, and the sample solution is brought into contact with the capturing body 26 to cause an antigen-antibody reaction or the like. The substance to be detected (specific antigen) in the specimen and the capturing body 26 are bound by the antigen-antibody reaction.

(D) Secondary reaction step By flowing an antibody labeled with a fluorescent substance through the flow path 31, the target substance captured by the capturing body 26 is labeled with the fluorescent substance. At this time, since an enhanced electric field is formed in the vicinity of the surface of the conductive film 25 of the detection unit 32, the fluorescent substance labeled with the antigen is excited by this electric field and shines.

(E) Signal Measurement Step The excitation light is irradiated from the light source 40 to the conductor film 25 at a measurement angle, and the amount of fluorescence emitted from the fluorescent substance that labels the substance to be detected is measured.

  When the fluorescence passes through the second optical filter 55, the noise component is removed and reaches the light receiving unit 51, and the light receiving element 52 of the light receiving unit 51 detects the fluorescence from which the noise component has been removed. The light receiving unit 51 outputs the detection result as an output signal to the calculation unit 14 via the control unit 58. The calculation unit 14 receives this output signal and performs a quantitative analysis of the antigen by calculating the output signal.

  And the calculating part 14 sends a calculation result to the display part 16, and the said display part 16 displays a calculation result.

  In the fluorescence analyzer 10 of the present embodiment, three sample chips 20 are inserted into the housing 820, and the first sample chip 20 is disposed at the inspection position in FIG. At this time, after the second sample chip 20 and the third sample chip 20 are inserted, the second sample chip 20 and the third sample chip 20 are arranged not at the examination position but at the standby position.

  From this state, the inspection of the first sample chip 20, the second sample chip 20, and the third sample chip 20 is performed as follows.

  First, focusing on the first sample chip 20, as shown in FIG. 6A, the initial measurement b, the primary reaction step c, the secondary reaction step d, and the signal measurement step e described above are performed. Done in order.

  In the present embodiment, the initial measurement process b for the second sample chip 20 is performed immediately after the initial measurement process b for the first sample chip 20 is completed.

  Furthermore, in this embodiment, after the initial measurement process b for the second sample chip 20 is completed, the initial measurement process b for the third sample chip 20 is performed.

  In this way, the initial measurement step b is performed on the n-th sample chip 20 after the initial measurement step b for the (n-1) -th sample chip 20 is completed. At this time, in the sample chip set in the apparatus, when the initial measurement step b or the signal measurement step e is performed on the nth sample chip, the initial measurement step b is also performed on the other sample chips. The signal measurement step e is also not performed. In addition, while the primary reaction step c is being performed on the n-1st sample chip, the initial measurement step b is performed on the nth sample chip. The initial measurement step b for the nth sample chip is performed before the signal measurement step e for the n−1th sample chip, for example, during the secondary reaction step d. Alternatively, it may be performed while the primary reaction step c to the secondary reaction step d are performed on the (n-1) th sample chip.

  On the other hand, the initial measurement step b and the signal measurement step e for each sample chip are sequentially performed for each sample chip in each step, and the primary reaction step c or the secondary reaction step d is simultaneously performed simultaneously on a plurality of sample chips. You may carry out.

  If the initial measurement step b is performed in this order, the expensive optical system 150 can be shared and effectively used, and as a result, the time required for inspection can be greatly shortened. Even if the number of the optical systems 150 is smaller than the number of the liquid feeding systems 90, no problem occurs in the inspection.

  In general, as shown in FIG. 6, the primary reaction step c, the secondary reaction step d, and the signal measurement step e are performed in this order on the sample chip 20 in which the initial measurement step b has been completed. Go. If the n sample chips 20 are examined in this order, it is possible to prevent the optical system 150 from being performed in the same time zone as shown in FIG.

  On the other hand, the secondary reaction step d is performed before the primary reaction step c, that is, after labeling a non-detection substance (specific antigen) with a fluorescent substance, the formed complex is brought into contact with the capturing body 6 to obtain an antigen antibody It is also possible to cause the reaction to occur.

  An appropriate waiting time can be ensured between the steps as required. Such adjustment is controlled by a program of the control unit 58.

  Further, since it is not necessary to provide an optical system more than necessary in the fluorescence analyzer 10, it is possible to reduce the size and cost of the apparatus.

  As mentioned above, although preferable embodiment of this invention was described, this invention is not limited to the said embodiment.

  For example, in FIG. 6A, the initial measurement process b of the first sample chip, the initial measurement process b of the second sample chip, and the initial measurement process b of the third sample chip cannot be overlapped. Therefore, the initial measurement process b of the second sample chip and the initial measurement process b of the third sample chip are performed after a predetermined time interval from the initial measurement process b of the first sample chip. On the other hand, the primary reaction step c, the secondary reaction step d, and the signal measurement step e in each sample chip are continuously performed after the initial measurement step b is completed. While the n-1 th sample chip performs the primary reaction step c to the secondary reaction step d, the n th sample chip initial measurement step b is performed. In the meantime, the nth signal measurement step e can be performed. In FIG. 6A, the signal measurement step e of the n-1st sample chip is performed while performing the n-th primary reaction step c to the secondary reaction step d.

  By doing in this way, with respect to each sample chip, it is possible to eliminate or reduce the waiting time between each step of the primary reaction step c, the secondary reaction step d, and the signal measurement step e, and continuously execute it. Therefore, dissociation of the antigen-antibody reaction due to an extra waiting time can be prevented, higher sensitivity can be realized, and a shorter time can be achieved in the inspection of a plurality of specimen chips. In addition, since the time from the start of the primary reaction step c to the start of the signal measurement step e can be made the same for all the sample chips, the change after the antigen-antibody reaction does not vary in the measurement conditions among the plurality of sample chips. Can be made uniform and highly accurate inspection can be performed.

  In this invention, it can replace with such an aspect of FIG. 6 (A), and can also carry out in the aspect of FIG. 6 (B).

  That is, in FIG. 6B, the first sample chip initial measurement step b, the second sample chip initial measurement step b, and the third sample chip initial measurement step b are separated by a predetermined time interval. The process from the start to the end is the same as in FIG. 6A, but the primary reaction step c and the secondary reaction step d of each sample chip are performed simultaneously. In FIG. 6B, since the liquid feeding in the primary reaction step c and the secondary reaction step d is performed at a similar timing for a plurality of sample chips, it is possible to drive the liquid feeding system in common. Thus, further cost reduction and downsizing can be achieved.

  As described above, in the present embodiment, an example in which the initial measurement process (b) for the first sample chip 20 is started after all three sample chips 20, 20, 20 are incorporated in the housing 820 will be described. However, instead of this, first, the two sample chips 20 and 20 are inserted, and in this state, the initial measurement step b for the first sample chip 20 is started, and then the third sample chip 20 is inserted. The inspection can be continued.

  That is, a surface plasmon resonance fluorescence analysis system in which such a program is incorporated in the control unit 58 can be employed.

  With such a setting, the sample chips 20 that have been tested are sequentially removed from the housing 820, and the remaining sample chips 20 that have not been tested are newly inserted, and the newly inserted samples The chip 20 can be placed at the standby position. Thereby, for example, even an analyzer equipped with three sample chip insertion ports 830 can be operated continuously, and a large number of sample chips can be continuously inspected by one unit.

DESCRIPTION OF SYMBOLS 10 Fluorescence analyzer 12 Holding | maintenance part 14 Calculation part 16 Display part 20 Sample chip 21 Prism 22 Incidence surface 23 Deposition surface 24 Ejection surface 25 Conductor film 30 Channel member 31 Channel 32 Detection unit 33 Guide unit 35 Seal member 40 Light source 41 Light-Emitting Element 42 Angle Changing Unit 50 Light Measuring Unit 51 Light-Receiving Unit 52 Light-Receiving Element 53 Optical System 54 Optical Filter 55 Optical Filter 56 Position Switching Device 58 Control Unit 60 Reagent Chip 85 Memory Card Slot 86 Print Output Port 87 Operation Panel 88 Input / Output Terminal 90 Liquid feeding system 150 Optical system 820 Housing 830 Sample chip insertion port b Initial measurement process c Primary reaction process d Secondary reaction process e Signal measurement process θ Incident angle θ1 Resonance angle θ3 Measurement angle θ5 Incident angle

Claims (9)

The sample chip for holding a substance to be detected with a plurality, wherein the target substance is held respectively by the plurality of sample chips, continuously detecting surface plasmon resonance fluorescence analysis by surface plasmon resonance fluorescence analysis A method,
The sample chip has a prism and a conductor film formed on one surface of the prism,
When the plurality of sample chips are respectively inserted into fluorescence analyzers having a plurality of (n) sample chip insertion ports,
(B) an initial measurement step in which initial measurement is performed by irradiating the conductive film from the light source via the prism with light from the light source to the sample chip inserted into the sample chip insertion port of the fluorescence analyzer;
(C) injecting a sample solution containing the substance to be detected to the sample chip, and the sample solution is brought into contact with the catching捉体on the conductive film, the capturing member with the substance to be detected contained in the sample solution A primary reaction step of binding
(D) a secondary reaction step of labeling the substance to be detected with a fluorescent substance;
(E) irradiating the conductive film from the prism side with excitation light at a measurement angle, and measuring the amount of fluorescence emitted from the fluorescent substance that labels the substance to be detected, and a signal measurement step ,
The step (c) or the step (d) is performed after the step (b), and the step (e) is performed after the step (c) and the step (d) are completed.
Wherein among the plurality of sample chips, the 1 th of said step (c) or the step for the analyte chip (d) is completed Then simultaneously, the step (e) and two eyes for said one th sample chips of the started process (d) or the step (c) for the analyte chip, the (n-1) -th said step for the analyte chip (c) or said step (d) completing then simultaneously, the (n -1) A surface plasmon resonance fluorescence analysis method starting the step (e) for the first sample chip and the step (d) or the step (c) for the nth sample chip.   The surface plasmon resonance fluorescence analysis method according to claim 1, wherein the initial measurement step includes at least one of resonance angle adjustment and / or blank measurement.   The step (c) is performed after the step (b), the step (d) is performed after the step (c), and the step (e) is performed after the step (d). The surface plasmon resonance fluorescence analysis method described in 1.   The surface plasmon resonance fluorescence analysis method according to claim 1 or 2, wherein the step (d) is performed before the step (c). In a surface plasmon resonance fluorescence analyzer for injecting a fluid into a guide part of a sample chip and reacting a reagent and a substance to be detected and measuring a reaction result from the detection part of the sample chip using an optical system,
A plurality (n) of sample chip insertion ports for inserting the sample chips into a housing of the surface plasmon resonance fluorescence analyzer;
A liquid feeding system for injecting fluid into the specimen chip;
The inserted the specimen tip from the specimen tip insertion port, and a drive member for liquid-based feed said a flow body guide portion is fixed at a predetermined position for communicating,
An optical system for optically detecting the substance to be detected;
The number of the optical system is set to a number smaller than the number of the sample chip insertion ports ,
When a plurality of sample chips are inserted into each of the plurality of sample chip insertion ports,
(B) an initial measurement step of performing initial measurement by irradiating the conductor film with light from the light source through the prism to the sample chip inserted into the sample chip insertion port of the surface plasmon resonance fluorescence analyzer;
(C) Injecting a sample solution containing the substance to be detected into the sample chip, bringing the sample solution into contact with a capturing body on the conductor film, and detecting the substance and the capturing body contained in the sample solution A primary reaction step of binding
(D) a secondary reaction step of labeling the substance to be detected with a fluorescent substance;
(E) performing a signal measurement step of irradiating the conductive film from the prism side with excitation light at a measurement angle and measuring the amount of fluorescence emitted from the fluorescent substance that labels the detection target substance. Composed,
The step (c) or the step (d) is performed after the step (b), and the step (e) is performed after the step (c) and the step (d) are completed.
At the same time when the step (c) or the step (d) for the first sample chip is completed among the plurality of sample chips, the step (e) and the second step for the first sample chip are completed. The step (d) or the step (c) for the sample chip is started, and at the same time the step (c) or the step (d) for the (n-1) -th sample chip is completed, the (n-1) ) A surface plasmon resonance fluorescence analyzer configured to start the step (e) for the first sample chip and the step (d) or the step (c) for the nth sample chip . A surface composed of a surface plasmon resonance fluorescence analyzer for injecting a fluid into the guide part of the sample chip, reacting the reagent and the substance to be detected, and measuring the reaction result from the detection part of the sample chip using an optical system, and a control unit A plasmon resonance fluorescence analysis system comprising:
  The sample chip has a prism and a conductor film formed on one surface of the prism,
  The surface plasmon resonance fluorescence analyzer is
  A plurality (n) of sample chip insertion ports for inserting the sample chips into a housing of the surface plasmon resonance fluorescence analyzer;
  A liquid feeding system for injecting fluid into the specimen chip;
  A driving member for fixing the sample chip inserted from the sample chip insertion port at a predetermined position where the fluid guide unit and the liquid feeding system communicate;
  An optical system for optically detecting the substance to be detected;
  The number of the optical system is set to a number smaller than the number of the sample chip insertion ports,
  When a plurality of the sample chips are inserted into each of the plurality of sample chip insertion ports, the control unit
  (B) an initial measurement step in which initial measurement is performed by irradiating the conductor film with light from the light source through the prism to the sample chip inserted into the sample chip insertion port of the surface plasmon resonance fluorescence analyzer;
  (C) Injecting a sample solution containing the substance to be detected into the sample chip, bringing the sample solution into contact with a capturing body on the conductor film, and detecting the substance and the capturing body contained in the sample solution A primary reaction step of binding
  (D) a secondary reaction step of labeling the substance to be detected with a fluorescent substance;
  (E) performing a signal measurement step of irradiating the conductive film from the prism side with excitation light at a measurement angle and measuring the amount of fluorescence emitted from the fluorescent substance that labels the detection target substance. Composed and
  The step (c) or the step (d) is performed after the step (b), and the step (e) is performed after the step (c) and the step (d) are completed.
  At the same time when the step (c) or the step (d) for the first sample chip is completed among the plurality of sample chips, the step (e) and the second step for the first sample chip are completed. The step (d) or the step (c) for the sample chip is started, and at the same time the step (c) or the step (d) for the (n-1) -th sample chip is completed, the (n-1) And) a surface plasmon resonance fluorescence analysis system configured to start the step (e) for the first sample chip and the step (d) for the nth sample chip.   The surface plasmon resonance fluorescence analysis system according to claim 6, wherein the initial measurement step includes at least one of resonance angle adjustment and / or blank measurement. The step (c) is performed after the step (b), the step (d) is performed after the step (c), and the step (e) is performed after the step (d). The surface plasmon resonance fluorescence analysis system described in 1. The surface plasmon resonance fluorescence analysis system according to claim 6 or 7, wherein the step (d) is performed before the step (c).

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