JP2006189397A - Measuring apparatus using total reflection attenuation, and method for identifying sensor unit thereof - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To achieve compactness and low price for a measuring apparatus, capable of identifying sensor units and that utilizes total reflection attenuation. <P>SOLUTION: A metal film 13, which generates SPR (surface plasmon resonance) when such light as to satisfy conditions of total reflection is made incident, is formed in the upper surface of a prism 14. One end of the upper surface of the prism 14 is provided with a bar code 18, in which the production number is recorded as individual recognition information for identifying each sensor unit 12. A measuring machine of an SPR measuring apparatus reads the bar code 18 by a light source and a detector to be used for SPR measurement, when the sensor unit 12 is at a measuring position. It is thereby possible to eliminate the need for separately providing a bar code reader, etc. and achieve compactness and low price for the apparatus. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a measuring apparatus using total reflection attenuation such as surface plasmon resonance, and more particularly to the measuring apparatus capable of identifying a sensor unit used in the apparatus and its identifying method.

  2. Description of the Related Art There are known measuring apparatuses that measure the reaction of a sample by using total reflection attenuation when measuring an interaction between biochemical substances such as proteins and DNA, screening drugs, and the like.

  One of the measuring devices that use such total reflection attenuation is a measuring device that uses a surface plasmon resonance phenomenon (hereinafter referred to as an SPR measuring device). The surface plasmon is generated by the collective vibration of free electrons in the metal, and is a density wave of free electrons traveling along the surface of the metal.

  For example, in an SPR measurement apparatus that employs a Kretschmann arrangement known from Patent Document 1 or the like, a surface of a metal film formed on a transparent dielectric (hereinafter referred to as a prism) is used as a sensor surface, and a sample is formed on the sensor surface. After reacting, the metal film is irradiated from the back side of the sensor surface through the prism so as to satisfy the total reflection condition, and the reflected light is measured.

  Of the light irradiated to the metal film so as to satisfy the total reflection condition, a small amount of light called an evanescent wave passes through the metal film without being reflected and oozes out to the sensor surface side. At this time, if the frequency of the evanescent wave coincides with the frequency of the surface plasmon, SPR is generated and the intensity of the reflected light is greatly attenuated. The SPR measurement device measures the reaction state of the sample on the sensor surface by capturing the attenuation of the reflected light.

  By the way, when measuring the interaction of a biological sample such as protein or DNA using the SPR measurement device as described above, a ligand is attached to a sample fixing group called a linker film formed on a metal film. Then, the analyte is brought into contact with the ligand and the interaction between them is measured, and the ligand fixation requires a long time of at least about 1 hour.

  For this reason, in the SPR measuring apparatus described in Patent Document 1, a plurality of sensor units each including the prisms are arranged on the turntable, and the ligand is fixed and measured simultaneously while rotating the turntable. Thus, the measurement of a plurality of samples is performed efficiently.

Furthermore, in this SPR measurement device, a barcode recorded with a serial number is attached to each sensor unit, and this barcode is read by a barcode reader connected to the device body before fixing or measuring a ligand. Yes. If multiple sensor units are processed simultaneously in parallel, it will be difficult to accurately grasp the correspondence between each sensor unit.For example, the analyte to be contacted may be wrong, or the measurement result may be different from the results of other sensor units. Problems such as misunderstanding can occur. In the SPR measurement device described in Patent Document 1, the serial number recorded on the barcode is read as described above, and each sensor unit is identified by this serial number, thereby preventing the occurrence of these problems.
JP 2001-330560 A

  However, when a bar code reader is provided as in the SPR measuring apparatus described in Patent Document 1, there is a problem that the apparatus becomes large and expensive by the amount of the reader.

  The present invention has been made in view of the above problems, and is a measuring apparatus using total reflection attenuation capable of identifying a sensor unit, and an object thereof is to reduce the size and the price.

  In order to achieve the above object, the measuring apparatus using total reflection attenuation according to the present invention irradiates a sensor unit comprising a dielectric block having a thin film layer on one surface so as to satisfy the total reflection condition. A light source that receives the reflected light from the sensor unit and photoelectrically converts the reflected light into an electrical signal. The sensor unit is provided with an identification code representing individual recognition information, and the light source and the detection unit The identification code is optically read.

  It is preferable to provide information decoding means for decoding the optically read identification code into individual recognition information for each sensor unit.

  The identification code is preferably provided on the same plane as the thin film layer of the dielectric block, and the identification code is preferably a barcode.

  Further, it is preferable that the individual recognition information includes a manufacturing number of the sensor unit.

  In the sensor unit identification method according to the present invention, an identification code representing each individual recognition information provided in the sensor unit is optically read by the light source and the detection means, and decoded into the individual recognition information. Thus, the sensor unit is identified.

  According to the measuring apparatus using total reflection attenuation of the present invention, the identification code representing the individual recognition information provided in the sensor unit is optically read by the light source and the detection means used for measurement. There is no need to separately provide a means for reading the image, and the apparatus can be reduced in size and price.

  FIG. 1 is a block diagram showing a schematic configuration of an SPR measurement device 2 as a measurement device using total reflection attenuation. The SPR measuring device 2 includes a fixing device 4 for fixing the ligand (fixing step), a measuring device 6 for adding an analyte to the immobilized ligand and measuring the reaction state of both (measurement step), and the measuring device 6 And a data analyzer 8 that performs analysis (data analysis process) of data obtained by the above. Further, the fixing step and the measuring step are performed on the sensor unit 12 which is a separate body, so that a plurality of samples can be measured smoothly.

  FIG. 2 is an exploded perspective view showing a schematic configuration of the sensor unit 12. The sensor unit 12 includes a prism (dielectric block) 14 having a metal film (thin film layer) 13 formed on the upper surface, a channel member 41 having a channel 16 for feeding a liquid to the metal film 13, and a prism. 14 and the bottom surface of the flow path member 41 are held in pressure contact with each other, and a lid member 43 is disposed above the holding member 42.

  The channel member 41 is formed in a long shape, and three channels 16 are provided along the longitudinal direction thereof. The flow path 16 is a liquid feeding pipe bent into a substantially U shape, and has an inlet 16a for injecting liquid and an outlet 16b for discharging it. The diameter of the channel 16 is about 1 mm, and the interval between the inlet 16a and the outlet 16b is about 10 mm. Further, the bottom of the flow path 16 is open, and this open part is covered and sealed with the metal film 13 when being pressed against the prism 14. Hereinafter, a portion of the metal film 13 surrounded by the open portion (see FIG. 3) is referred to as a sensor cell 17.

  The flow path member 41 is molded from, for example, an elastic material such as rubber or PDMS (polydimethylsiloxane) in order to improve the adhesion with the metal film 13. Thus, when the flow path member 41 is pressed against the upper surface of the prism 14, the flow path member 41 is elastically deformed to fill the gap between the contact surfaces with the metal film 13, and the open bottom portion of each flow path 16 is watertight together with the prism 14. Cover. In this example, an example in which the number of the flow paths 16 is three has been described. However, the number of the flow paths 16 is not limited to three, and may be one or two, or may be four or more. .

  On the upper surface of the prism 14, a strip-shaped metal film 13 facing each flow path 16 is formed by, for example, vapor deposition. For example, gold or silver is used for the metal film 13, and the film thickness is, for example, 50 nm. This film thickness is appropriately selected according to the material of the metal film 13, the wavelength of the irradiated light, and the like.

  A linker film 22 for fixing the ligand is formed at a position corresponding to each flow path 16 on the metal film 13 (in the sensor cell 17). Hereinafter, of the metal film 13, the surface on which the linker film 22 is formed is referred to as a sensor surface 13a, and the back surface (the surface in contact with the prism 14) is referred to as a light incident surface 13b. The prism 14 condenses incident light toward the light incident surface 13b, and the metal film 13 is formed on the sensor surface 13a when light is incident on the light incident surface 13b so as to satisfy the total reflection condition. Generate SPR.

  Engaging claws 14 a that engage with the engaging portions 42 a of the holding member 42 are provided on both side surfaces of the long side of the prism 14. By these engagements, the flow path member 41 is sandwiched between the holding member 42 and the prism 14, and is held in a state where the bottom surface and the top surface of the prism 14 are in pressure contact with each other. Further, protrusions 14 b are provided at both ends of the prism 14. As will be described later, the sensor unit 12 is fixed in a state of being housed in a holder 52 (see FIG. 4). The protrusion 14 b engages with the holder 52 to position the sensor unit 12 at a predetermined storage position in the holder 52.

  The prism 14 is made of, for example, optical glass such as borosilicate crown (BK7) or barium crown (Bak4), polymethyl methacrylate (PMMA), polycarbonate (PC), amorphous polyolefin (APO), or the like. Representative optical plastics can be used.

  A bar code (identification code) 18 in which a production number is recorded as individual recognition information for identifying each sensor unit 12 is provided at one end of the upper surface of the prism 14. The bar code 18 is formed by, for example, forming a pattern by masking when forming the metal film 13 by vapor deposition, forming an uneven pattern by a mold when molding the prism 14, ink, or the like. Provided by marking the pattern with Alternatively, a concave surface that is recessed by one step may be formed on the upper surface of the prism 14, and a sticker on which the barcode 18 is recorded may be attached thereto.

  On the upper surface of the holding member 42, a receiving port 42b through which the tip of a liquid feeding means such as a pipette enters is formed at a position corresponding to the inlet 16a and the outlet 16b of each channel 16. These receiving ports 42b are formed in a funnel shape so that the liquid discharged from the pipette is guided to each inlet 16a. In addition, each receiving port 42 b comes into contact with each inlet 16 a and outlet 16 b and is connected to the channel 16 when the holding member 42 sandwiches the channel member 41 and engages with the prism 14.

  In addition, columnar bosses 42c are provided on both sides of each receiving port 42b. These bosses 42 c are fitted into holes 43 a formed in the lid member 43 to position the lid member 43. The lid member 43 is affixed to the upper surface of the holding member 42 by a double-sided tape 44 having holes at positions corresponding to the receiving ports 42b and the bosses 42c.

  The lid member 43 covers the receiving port 42 b that communicates with the flow path 16, thereby preventing the liquid injected into the flow path 16 from evaporating. The lid member 43 is made of, for example, an elastic material such as rubber or plastic, and a cross-shaped slit 43b is formed at a position corresponding to each receiving port 42b. As a result, the lid member 43 elastically deforms the slit 43b to allow the pipette to be inserted, and when the pipette is removed, the lid member 43 maintains the initial state and closes the receiving port 42b.

  As shown in FIG. 3, the linker film 22 formed on the metal film 13 has a measurement region 22a (hereinafter referred to as an “act region”) in which a ligand is fixed and a reaction between the analyte and the ligand occurs, and a ligand. A reference region (hereinafter referred to as a “ref region”) 22b for obtaining a reference signal when measuring the signal in the act region 22a is provided without being fixed. The ref region 22b is formed when the linker film 22 is formed. As the formation method, for example, the linker film 22 is subjected to a surface treatment, and about a half of the linker film 22, the bonding group that binds to the ligand is deactivated, so that the half of the linker film 22 becomes the act region. 22a, and the other half becomes the ref region 22b.

  FIG. 4 is a perspective view schematically showing the configuration of the fixing machine 4. A mounting space 51 for mounting the plurality of sensor units 12 is secured on the housing base 50 that serves as a base of the fixed machine 4. The sensor unit 12 is subjected to a fixing process while being placed in the placement space 51.

  The sensor unit 12 is placed in the placement space 51 while being housed in the holder 52. The holder 52 can store a plurality of sensor units 12 (eight in this example). The holder 52 is provided with a slit for positioning the sensor unit 12 by fitting with the protrusion 14 b of the sensor unit 12. In addition, an opening is formed in the bottom of the holder 52 except for support portions that support both ends of the sensor unit 12. As will be described later, when the sensor unit 12 is taken out from the holder 52 in the measurement process, a push-up member 81a (see FIG. 5) is inserted from the opening, and the sensor unit 12 is pushed up.

  In the mounting space 51, for example, ten holders 52 can be arranged side by side, and pedestals 53 corresponding to the number of the holders 52 are provided. Each pedestal 53 is formed with a positioning boss for positioning the holder 52.

  Further, the fixing machine 4 is provided with a pipette head 54 in which three pairs of pipettes 19 are connected. The pipette head 54 accesses the sensor units 12 arranged in the placement space 51 via the holder 52 to inject and discharge liquid. Each pair of pipettes 19 includes a pair of pipettes 19 a and 19 b (see FIG. 7), and each pipette 19 a and 19 b is inserted into each of the inlet 16 a and the outlet 16 b of each sensor unit 12. Each pipette 19a, 19b has a function of injecting liquid into the flow path 16 and sucking out from the flow path 16, and when one is performing an injection operation, the other performs a suction operation. So that they work together. Since three pipette pairs 19 are provided in the pipette head 54, liquid can be simultaneously injected into or discharged from the three flow paths 16 included in one sensor unit 12. Further, the fixing device 4 is provided with a controller (not shown). The suction and discharge operations of the pipette head 54 are controlled by this controller, and the suction amount, discharge amount, timing, etc. are determined for each pipette pair 19.

  The housing base 50 is provided with a head moving mechanism 56 that moves the pipette head 54 in three directions of X, Y, and Z. The head moving mechanism 56 is a known moving mechanism including, for example, a conveyance belt, a pulley, and a motor. The head moving mechanism 56 is an elevating mechanism that moves the pipette head 54 up and down, and the pipette head 54 can be moved in the Y direction together with the elevating mechanism. A Y-direction moving mechanism including a guide rail 58 to be held and an X-direction moving mechanism that holds the guide rail 58 at both ends and moves the pipette head 54 in the X direction together with the guide rail 58. The head moving mechanism 56 is controlled by a controller, and the controller drives the head moving mechanism 56 to control the vertical and horizontal positions of the pipette head 54.

  In addition to the mounting space 51, a plurality of liquid storage units 61 for storing various liquids to be injected into the flow path 16 and a pipette tip storage unit 63 for storing the pipette tips 62 are provided on the housing base 50. And a well plate 64 in which a plurality of well-shaped ridges are arranged in a matrix.

  The number of the liquid storage units 61 is determined according to the type of liquid to be used, such as a ligand solution, a cleaning liquid, a fixing buffer liquid, a drying prevention liquid, an activation liquid, and a blocking liquid. Each liquid storage unit 61 is provided with six insertion ports in a straight line. The number of the insertion openings and the arrangement pitch are determined according to the number of pipettes of the pipette head 54 and the arrangement pitch. When the pipette head 54 injects the liquid into the flow path 16, the liquid storage unit 61 is accessed to suck in a desired liquid, and then moves to the placement space 51 to be injected into each flow path 16.

  The pipette tips 62 stored in the pipette tip storage section 63 are attached to the tip portions of the pipettes 19a and 19b in a replaceable manner. Since the pipette tip 62 is in direct contact with the liquid, it is exchanged for each liquid to be used so that different kinds of liquids are not mixed through the pipette tip 62. Each pipette 19a, 19b incorporates a mechanism for picking up and releasing the pipette tip 62 so that the pipette tip 62 is automatically replaced. When exchanging the pipette tip 62, the pipette head 54 first releases the used pipette tip 62 at a disposal unit (not shown), and then accesses the pipette tip storage unit 63 to access an unused pipette. The chip 62 is picked up.

  The well plate 64 is used for temporarily storing the liquid sucked up by the pipettes 19a and 19b, or for preparing a mixed liquid by mixing a plurality of types of liquids.

  Further, when the fixing device 4 performs the fixing process, the housing base 50 is covered with the cover, and the inside of the fixing device 4 including the placement space 51 is shielded. The sensor unit 12 is stored in the mounting space 51 for a predetermined time until the ligand immobilization is completed. At this time, the degree of progress of immobilization depends on the temperature. Therefore, the fixing machine 4 keeps the internal temperature at a predetermined temperature by a temperature controller (not shown) while fixing. The set temperature, standing time, and the like are appropriately determined according to the type of ligand to be immobilized.

  FIG. 5 is a perspective view schematically showing the configuration of the measuring device 6. The measuring machine 6 includes a holder transport mechanism 71, a pickup mechanism 72, a head moving mechanism 73, and a measurement stage 74, and these parts are accommodated in a housing 75. The holder conveyance mechanism 71 includes a conveyance belt 76, a carriage 77 attached to the conveyance belt 76, and a plate 78 that is attached to the carriage 77 and on which the holder 52 is placed. The holder transport mechanism 71 moves each sensor unit 12 accommodated in the holder 52 to a pickup position where the pickup mechanism 72 picks up by moving the plate 78 on which the holder 52 is placed in the X direction.

  The pickup mechanism 72 is a mechanism for picking up the sensor unit 12 from the holder 52, a push-up mechanism 81 that pushes up the sensor unit 12 accommodated in the holder 52 from below to above, and the push-up mechanism 81 above the holder 52. A handling head 82 sandwiches and holds the pushed sensor unit 12 from both sides. As described above, an opening is provided at the bottom of the holder 52, and the plate 78 is formed on the frame so as to expose the opening. The push-up mechanism 81 enters the opening of the holder 52 via the plate 78, contacts the bottom surface of the sensor unit 12, pushes up the push-up member 81a, and push-up member that drives the push-up member 81a up and down And a drive mechanism 81b.

  The handling head 82 is provided with a pair of claws that sandwich the sensor unit 12, and the sensor unit 12 is gripped by the claws. The handling head 82 has a head body 82 a attached to a ball screw 86 via a nut 84, and is provided so as to be movable between a pickup position above the holder 52 and the measurement stage 74. The handling head 82 grips the sensor unit 12 at the pickup position, moves in the Y direction in that state, and carries the sensor unit 12 to the measurement stage 74. Further, after the measurement is completed, the sensor unit 12 is released by returning to the pickup position, and the measured sensor unit 12 is returned to the holder 52.

  The measurement stage 74 is provided with a measurement unit 31 including an illumination unit 32 and a detector (detection means) 33 below the position where the sensor unit 12 is disposed. The sensor unit 12 has a plurality of sensor cells 17, and measurement is performed for each sensor cell 17. The handling head 82 sequentially moves the sensor cells 17 to the measurement positions on the optical path of the illumination unit 32 by moving the sensor units 12 in the Y direction at the arrangement pitch of the sensor cells 17. In addition, as shown in this figure and FIG. 3, the direction of the incident light ray irradiated to the sensor unit 12 and the reflected light ray reflected by the sensor surface 13a, and the arrangement direction of each sensor cell 17, that is, each flow channel 16 horizontal portion The illuminating unit 32 and the detector 33 are arranged so that the flow direction is perpendicular to the other.

  The reaction state between the ligand and the analyte appears as a change in the resonance angle (the incident angle of light irradiated on the sensor surface 13a). The illumination unit 32 includes, for example, a light source 34 and an optical system 36 including a condensing lens, a diffusion plate, a polarizing plate, and the like (see FIG. 7). The arrangement position and the installation angle are all incident angles of illumination light. It is adjusted to satisfy the reflection conditions.

  As the light source 34, for example, a light emitting element such as an LED (Light Emitting Diode), an LD (Laser Diode), or an SLD (Super Luminescent Diode) is used. The light source 34 uses one such light emitting element, and irradiates light from the single light source toward each light incident surface 13b. The diffusion plate diffuses light from the light source 34 and suppresses unevenness in the amount of light in the light emitting surface. The polarizing plate passes only p-polarized light (linearly polarized light having an oscillating electric field parallel to the incident surface) that causes SPR in the irradiated light. In addition, when using LD, when the direction of polarization of the irradiation light itself emitted from the light source 34 is uniform, the polarizing plate is unnecessary. Further, even when a light source with uniform polarization is used, if the direction of polarization becomes uneven due to passing through the diffusion plate, the direction of polarization is aligned using a polarizing plate. The light thus diffused and polarized is condensed by the condenser lens and applied to the prism 14. Thereby, the light which does not vary in light intensity and can have various incident angles can be incident on the light incident surface 13b.

  The detector 33 receives the light reflected by the light incident surface 13b and outputs an electrical signal having a level corresponding to the light intensity. Since light of various angles is incident on the light incident surface 13b, the light is reflected on the light incident surface 13b at a reflection angle corresponding to each incident angle. The detector 33 receives light of these various reflection angles. For example, a CCD area sensor or a photodiode array is used as the detector 33, and the reflected light reflected at various reflection angles on the light incident surface 13b is received and photoelectrically converted, and this is converted into an SPR signal as a data analyzer. 8 is output.

  Further, as shown in FIG. 3, an act region 22 a and a ref region 22 b are formed on the linker film 22. The detector 33 outputs an SPR signal corresponding to the act region 22a as an act signal, and outputs an SPR signal corresponding to the ref region 22b as a ref signal.

  A well plate 88 for storing the analyte solution is placed beside the measurement stage. For example, each well of the well plate 88 contains different types of analyte solutions.

  The measuring device 6 is provided with a pipette pair 26 having the same function as the pipette pair 19 of the fixing device 4, and injects various liquids from the inlets 16 a of the sensor unit 12 into the flow path 16. The head moving mechanism 73 carries the pipette head 87 to the sensor unit 12 and the well plate 88 at the measurement position while moving the pipette head 87 having the pipette pair 26 in three directions of X, Y, and Z. . The head moving mechanism 73 has substantially the same configuration as the head moving mechanism 56 of the fixed machine 4. The pipette head 87 accesses the sensor cell 17 to be measured, and injects and discharges the liquid. Since the pipette head 87 is for injecting and discharging liquid with respect to a specific sensor cell 17 to be measured, unlike the pipette head 54 of the fixing machine 4, only one pair of pipettes 26 is provided. ing. Although not shown, the measuring device 6 has a well plate for storing a measurement buffer solution and a cleaning solution at a position accessible by the pipette pair 26, a pipette tip storage unit for storing a replacement pipette tip, and the like. Is provided.

  Further, when the measuring device 6 drives the handling head 82 to cause each sensor cell 17 to enter the measurement position on the optical path, the measuring device 6 measures the barcode 18 provided at one end of the sensor unit 12 prior to each sensor cell 17. Enter the position. As a positioning method of the bar code 18 and each sensor cell 17, for example, an optical sensor, a magnetic sensor, or a detection means such as a mechanical switch may be used to detect the position, or a motor that drives the handling head 82 may be rotated. Positioning by number may be used.

  The measuring instrument 6 that has caused the barcode 18 to enter the measurement position optically reads the pattern of the barcode 18 by the illumination unit 32 and the detector 33. The detector 33 outputs a signal representing the bar code 18 received and photoelectrically converted to the data analyzer 8 as an individual recognition signal.

  FIG. 6 is a block diagram schematically showing the configuration of the data analyzer 8. The data analyzer 8 includes, for example, an input device 91, a CPU (Central Processing Unit) 92, a ROM (Read Only Memory) 93, a RAM (Random Access Memory) 94, a monitor 95, an HDD (Hard Disk). Drive) 96, communication I / F 97, signal processing unit 98, decoder 99, and the like. The input device 91 is, for example, a keyboard or a mouse, and inputs an instruction from the user to the data analyzer 8. The CPU 92 comprehensively controls each unit of the data analyzer 8. The ROM 93 stores a control program, various setting data, and the like. The RAM 94 is used as a working memory when the CPU 92 or the signal processing unit 98 performs calculations. The HDD 96 is a known data storage device, and various programs such as a control program and a data analysis program are stored in the HDD 96. In addition, various measurement data measured by the measuring device 6 are recorded in the HDD 96. The communication I / F 97 receives the SPR signal and the individual recognition signal output from the detector 33. The signal processing unit 98 performs data analysis based on the acquired SPR signal. The monitor 95 displays the detected SPR signal and the analysis result based on the detected SPR signal.

  The ROM 93 or the HDD 96 has codes corresponding to the barcode 18 such as EAN (European Article Number), JAN (Japanese Article Number), NW7 (Narrow Wide 7), ITF (Iinterleaved 2 of 5), and CODE39. , Stored in advance. The decoder 99 as information decoding means identifies each sensor unit 12 by decoding the acquired individual recognition signal into a serial number for each sensor unit 12 with a predetermined algorithm based on the stored code. The decoder 99 displays the decoded serial number on the monitor 95.

  The HDD 96 stores a sensor management file as shown in Table 1, for example. This sensor management file is for collectively managing each sensor cell 17 based on the manufacturing number for each sensor unit 12 recorded in the barcode 18. Since three sensor cells 17 are provided in the sensor unit 12 of this example, the cell No. Is composed of a combination of a manufacturing number of the sensor unit 12 and a dash number representing each sensor cell 17. The sensor management file contains the cell No. For example, the sensor cells 17 are collectively managed by associating and storing the types of fixed ligands, the types of analytes, the names of electronic files storing measurement results and analysis results, and the like. The sensor management file may be automatically recorded when the process of each item to be stored is completed, or may be manually recorded via the input device 91.

  Next, the SPR measurement method of the SPR measurement device 2 having the above configuration will be described using the explanatory diagram shown in FIG.

  The fixing step of fixing the ligand to the linker film 22 is performed in a state where the sensor unit 12 is placed in the placement space 51 of the fixing machine 4 via the holder 52. The fixing device 4 drives the head moving mechanism 56 in response to a fixing start instruction from the user, and uses the pipette pair 19 to inject the ligand solution 21 in which the ligand 21a is dissolved in the solvent from the injection port 16a.

  As a pretreatment before performing the ligand immobilization process for injecting the ligand solution 21, the fixing machine 4 first feeds a fixing buffer solution to the linker film 22 to moisten the linker film 22, and then An activation process of the linker film 22 is performed to facilitate the binding of the ligand to the film 22. For example, in the amine coupling method, carboxymethyl dextran is used as the linker film 22, and the amino group in the ligand is directly covalently bonded to the dextran. As the activation liquid in this case, a mixed liquid of N ′-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) is used. The fixing device 4 cleans the flow path 16 with the fixing buffer solution after the activation process.

  Examples of the buffer solution for fixation and the solvent (diluent) of the ligand solution 21 include, for example, various buffer solutions (buffer solutions), physiological salt solutions represented by physiological saline, pure water, and the like. Is used. The type of each of these liquids, the pH value, the type of mixture, the concentration thereof, and the like are appropriately determined according to the type of ligand 21a. For example, when a biological sample is used as the ligand 21a, a physiological saline whose pH is adjusted to near neutral is often used. However, in the amine coupling method, since the linker film 22 is negatively (negatively) charged by carboxymethyldextran, the protein is positively (positively) charged so as to easily bind to the linker film 22. In some cases, a phosphate buffer solution (PBS: phosphatic-buffered, saline) having a strong buffering action containing a high concentration of phosphate may be used.

  After performing the activation process and the washing, the fixing machine 4 injects the ligand solution 21 into the flow path 16 to perform the ligand immobilization process. When the ligand solution 21 is injected into the flow path 16, the ligand 21 a diffusing into the solution is precipitated and gradually deposited on the linker film 22. Thereby, the contacted ligand 21 a is bonded to the linker film 22, and the ligand 21 a is fixed on the linker film 22. The immobilization usually takes about one hour. During this time, the sensor unit 12 is stored in a state where environmental conditions such as temperature are set to predetermined conditions. Further, while the immobilization is proceeding, the ligand solution 21 in the flow channel 16 may be allowed to stand, but the ligand solution 21 in the flow channel 16 may be stirred and flowed. By doing so, the binding between the ligand 21a and the linker film 22 is promoted, and the amount of ligand 21a immobilized can be increased.

  When the fixation of the ligand 21a to the linker film 22 is completed, the fixing device 4 sucks out the ligand solution 21 with the pipette 19b and discharges it from the flow path 16, and then the cleaning liquid is injected into the flow path 16 to complete the fixation. The linker film 22 is washed. Moreover, the fixing machine 4 injects a blocking liquid as necessary, and performs a blocking process to deactivate the reactive group of the linker film 22 that has not been bonded to the ligand 21a. As the blocking liquid, for example, ethanolamine-hydrochloride is used. When this blocking process is performed, the flow path 16 is washed again. After the final cleaning, the fixing device 4 injects the drying prevention liquid into the flow path 16. The sensor unit 12 is stored in the mounting space 51 or another storage place until the measurement in a state where the linker film 22 is immersed in the drying prevention liquid.

  The measurement process starts from a state in which the sensor unit 12 is placed on the plate 78 of the measuring device 6 together with the holder 52. The measuring device 6 moves the plate 78 to the pickup position in accordance with a measurement start instruction from the user and drives the pickup mechanism 72 to carry the predetermined sensor unit 12 to the measurement stage 74.

  The measuring device 6 carrying the sensor unit 12 to the measurement stage 74 drives the head moving mechanism 73 to first inject the measurement buffer solution into the flow path 16. Thereafter, an analyte solution 27 in which the analyte is dissolved in a solvent is injected, and then the measurement buffer solution is injected again. Note that the flow path 16 may be washed once before the measurement buffer solution is injected for the first time. Data reading by the detector 33 is started immediately after the measurement buffer solution is first injected in order to detect a reference signal level. After the analyte solution 27 is injected, the measurement buffer solution is injected again. Until it is done. Thereby, it is possible to measure the SPR signal until the detection of the reference level, the reaction state between the analyte and the ligand, and the desorption by the injection of the buffer solution for measurement of the bound analyte and the ligand.

  Examples of the buffer solution for measurement and the solvent (diluent) of the analyte solution 27 include various buffer solutions (buffer solutions), physiological salt solutions typified by physiological saline, and pure water. used. The type of each of these liquids, the pH value, the type of mixture and its concentration, etc. are appropriately determined according to the type of ligand or analyte. For example, DMSO (dimethyl-sulfo-oxide) may be included in physiological saline in order to facilitate the dissolution of the analyte. This DMSO greatly affects the signal level. As described above, the measurement buffer solution is used for detecting the reference level. Therefore, when the analyte solution 27 contains DMSO, the measurement buffer solution having a DMSO concentration comparable to the DMSO concentration should be used. Is preferred.

  The analyte solution 27 is often stored for a long time (for example, one year). In such a case, a difference in concentration occurs between the initial DMSO concentration and the DMSO concentration at the time of measurement due to a change with time. May end up. When it is necessary to perform strict measurement, it is preferable to estimate such a concentration difference from the level of the ref signal when the analyte solution 27 is injected, and to correct the measurement data (DMSO concentration correction).

  The correction data for correcting the DMSO concentration is obtained by injecting a plurality of types of measurement buffer solutions having different DMSO concentrations into the flow path 16 before injecting the analyte solution 27, and ref according to the DMSO concentration change at this time. It is obtained by examining the respective changes in the signal level and the act signal level.

  The reaction state between the ligand and the analyte appears as a transition of the attenuation position of the reflected light within the light receiving surface of the detector 33. For example, before and after the analyte contacts the ligand, the refractive index on the sensor surface 13a is different, and the resonance angle at which SPR occurs is different. When the analyte comes into contact with the ligand and starts to react, the resonance angle of the reflected light starts to change accordingly, and the attenuation position of the reflected light in the light receiving surface starts to move. The measuring device 6 outputs an SPR signal indicating the reaction state of the sample thus obtained to the data analyzer 8.

  In FIG. 7, for the sake of convenience, the direction of the incident light beam on the light incident surface 13 b and the reflected light beam reflected there are parallel to the direction of the liquid flowing in the flow channel 16 so that the configuration of the measuring instrument 6 is clear. The illumination unit 32 and the detector 33 are arranged so that, as shown in FIGS. 3 and 5, the directions of the incident light beam and the reflected light beam are actually orthogonal to the liquid flowing direction. The illumination part 32 and the detection part 33 are arrange | positioned so that it may irradiate in a direction (direction orthogonal to a paper surface). However, there is no problem even if the measurement unit 31 is arranged as shown in FIG.

  In the data analysis step, the SPR signal obtained by the measuring device 6 is analyzed by the data analyzer 8 to quantitatively analyze the characteristics of the analyte. The data analyzer 8 receives the SPR signal from the detector 33 via the communication I / F 97 and sends this SPR signal to the signal processing unit 98.

  Based on a data analysis program stored in the ROM 93 or the HDD 96, the signal processing unit 98 performs data analysis by obtaining a difference or ratio between the act signal and the ref signal obtained by the measuring device 6. For example, difference data between the act signal and the ref signal is obtained, and the difference data is used as measurement data, and analysis is performed based on the difference data. By doing so, it becomes possible to cancel noise caused by disturbances such as individual differences of the sensor unit 12 and the linker film 22, mechanical fluctuations of the apparatus, changes in the temperature of the liquid, and the like. Good high-accuracy measurement can be performed. The signal processing unit 98 displays the analysis result on the monitor 95 and records it in the HDD 96.

  Next, the operation of the SPR measurement device 2 of the present invention will be described using the flowchart shown in FIG.

  The sensor unit 12 is placed in the placement space 51 of the fixing machine 4 while being fitted in the holder 52. At this time, for example, the serial number of the sensor unit 12 can be obtained by visually observing the serial number written on the box for packing the sensor unit 12 or by reading the barcode 18 with a handy type barcode reader (not shown). get. A dedicated bar code reader may be provided in the fixing device 4 so that the bar code 18 is automatically read when the holder 52 is placed. Further, the position in the mounting space 51 where the sensor unit 12 that has acquired the serial number is set is recorded in association with the above-described sensor management file, or registered in the controller of the fixed machine 4, for example. It is preferable to keep it.

  The fixing device 4 in which the sensor unit 12 is set performs a fixing process on each sensor cell 17 in accordance with a fixing start instruction from the user. Since the manufacturing number of the sensor unit 12 is known, the user records the type of the ligand fixed to each linker film 22 in this fixing step in the sensor management file.

  The sensor unit 12 in which the ligand is fixed is moved to the measuring machine 6 together with the holder 52 and placed on the plate 78. The measuring machine 6 having the sensor unit 12 mounted on the plate 78 moves the plate 78 to the pickup position in accordance with a measurement start instruction from the user and drives the pickup mechanism 72 to move the sensor unit 12 to the measurement stage 74. Carry to.

  The handling head 82 of the pickup mechanism 72 first causes the barcode 18 to enter the measurement position on the optical path. After confirming that the barcode 18 is at the measurement position, the measuring device 6 optically reads the barcode 18 with the illumination unit 32 and the detector 33 and outputs it to the data analyzer 8 as an individual recognition signal. The data analyzer 8 that has received the individual recognition signal via the communication I / F 97 sends this individual recognition signal to the decoder 99 and decodes it into the serial number to identify the sensor unit 12 at the measurement position. The data analyzer 8 that has decoded the individual recognition signal causes the monitor 95 to display the decoded serial number and sensor management file.

  When the identification of the sensor unit 12 is completed, the measuring device 6 drives the handling head 82 to sequentially enter each sensor cell 17 to the measurement position, and measures each sensor cell 17. At this time, the user uses the cell number recorded in the sensor management file based on the decoded serial number displayed on the monitor 95. Can be used to confirm the type of ligand immobilized on each linker film 22. This prevents, for example, injecting an incorrect analyte into each sensor cell 17.

  The measuring device 6 that has measured each sensor cell 17 outputs each SPR signal to the data analyzer 8. The data analyzer 8 that has received the SPR signal via the communication I / F 97 sends this SPR signal to the signal processing unit 98 to quantitatively analyze the characteristics of the analyte. In addition, it is preferable to record the type of the injected analyte, the name of the electronic file storing the measurement result and the analysis result, and the like in the sensor management file. Thereby, for example, it is possible to prevent a measurement result or an analysis result from being mistaken for a result of another sensor unit 12. These records may be manually input by the user via the input device 91, or may be automatically recorded by the data analyzer 8.

  Thus, according to the SPR measurement device 2 of the present embodiment, when the sensor unit 12 is identified by the measuring device 6, the barcode 18 provided at one end of the sensor unit 12 is replaced with the illumination unit 32 and the detector. Therefore, it is not necessary to separately provide a bar code reader or the like, and the apparatus can be reduced in size and cost.

  In the above embodiment, one bar code 18 is provided in the sensor unit 12 having the three sensor cells 17, and each sensor cell 17 is managed by the serial number of the sensor unit 12. However, the present invention is not limited to this. Instead, for example, a barcode 18 in which the ID number of the sensor cell 17 is recorded as individual recognition information may be provided corresponding to each sensor cell 17. Further, the barcode 18 is not limited to the upper surface of the prism 14, and may be any position that can be optically read by the illumination unit 32 and the detector 33, such as a side surface on the long side of the prism 14.

  In the above embodiment, the barcode 18 is used as the identification code. However, the present invention is not limited to this, and any character or symbol can be used as long as it can be optically read by the illumination unit 32 and the detector 33. It may be.

  Furthermore, in the above embodiment, the sensor unit 12 is moved by the handling head 82, and each sensor cell 17 and the barcode 18 are individually measured by irradiating light from a single light source such as an LED. For example, as shown in FIG. 9, each sensor cell 17 or barcode 18 is measured at a time using a so-called fan beam in which a single light source is fan-shaped with an incident-side convex lens 110 and an incident-side concave lens 111. May be. The reflected light is condensed on the detector 33 by the reflection side convex lens 112 and the reflection side concave lens 113.

  In addition, the sensor cell 17 and the barcode 18 may be measured at a time by using so-called line illumination having approximately the same length as the prism 14 for the illumination unit 32, or a single light source may be a polygon mirror or the like. You may make it measure by scanning.

  In the above embodiment, only one pair of pipettes 26 is provided on the pipette head 87 of the measuring device 6, but when measuring each sensor cell 17 and the barcode 18 at a time as described above, a fixing device is used. It is preferable that a plurality of pairs of pipettes 26 are provided in the pipette head 87 as in the case of the four pipette heads 54, and liquid is simultaneously injected into or discharged from the plurality of flow paths 16 included in one sensor unit 12. .

  In the above embodiment, an SPR measurement device is shown as an example of a measurement device using total reflection attenuation. However, for example, a leakage mode sensor is known as a measurement device using total reflection attenuation. ing. The leakage mode sensor is composed of a dielectric, and a thin film composed of a clad layer and an optical waveguide layer that are sequentially layered thereon. One surface of the thin film serves as a sensor surface, and the other surface receives light. It becomes a surface. When light is incident on the light incident surface so as to satisfy the total reflection condition, a part of the light is transmitted through the cladding layer and taken into the optical waveguide layer. In this optical waveguide layer, when the waveguide mode is excited, the reflected light at the light incident surface is greatly attenuated. The incident angle at which the waveguide mode is excited changes according to the refractive index of the medium on the sensor surface, similar to the resonance angle of SPR. By detecting the attenuation of the reflection angle, the chemical reaction on the sensor surface is measured.

It is a block diagram which shows schematic structure of a SPR measuring apparatus. It is a disassembled perspective view which shows schematic structure of a sensor unit. It is explanatory drawing which extracts and demonstrates one sensor cell. It is a perspective view which shows the structure of a fixing machine roughly. It is a perspective view which shows the structure of a measuring machine roughly. It is a block diagram which shows the structure of a data analyzer roughly. It is explanatory drawing of a SPR measuring method. It is a flowchart which shows the identification method of a sensor unit. It is explanatory drawing which illustrates a fan beam light source schematically.

Explanation of symbols

2 SPR measurement device (measurement device using total reflection attenuation)
4 Fixing machine 6 Measuring machine 8 Data analyzer 12 Sensor unit 13 Metal film (thin film layer)
14 Prism (dielectric block)
18 Bar code (identification code)
32 Illumination part 33 Detector (detection means)
34 Light source 99 Decoder (information decoding means)

Claims (6)

A sensor unit consisting of a dielectric block with a thin film layer formed on one side, and a light source that irradiates light so as to satisfy the total reflection condition, and receives the reflected light from the sensor unit and photoelectrically converts it into an electrical signal In a measuring device using total reflection attenuation provided with a detecting means for
An identification code representing individual recognition information is provided in the sensor unit,
A measuring apparatus using total reflection attenuation, wherein the identification code is optically read by the light source and the detection means.   2. The measuring apparatus using total reflection attenuation according to claim 1, further comprising information decoding means for decoding the optically read identification code into individual recognition information for each sensor unit.   3. The measuring apparatus using total reflection attenuation according to claim 1, wherein the identification code is provided on the same plane as the thin film layer of the dielectric block.   The measuring device using total reflection attenuation according to claim 1, wherein the identification code is a bar code.   The measuring apparatus using total reflection attenuation according to any one of claims 1 to 4, wherein the individual recognition information includes a manufacturing number of the sensor unit. A sensor unit consisting of a dielectric block with a thin film layer formed on one side, and a light source that irradiates light so as to satisfy the total reflection condition, and receives the reflected light from the sensor unit and photoelectrically converts it into an electrical signal A measuring device using total reflection attenuation provided with a detecting means for
An identification code representing each individual recognition information provided in the sensor unit is optically read by the light source and the detection means,
A sensor unit identification method, wherein the sensor unit is identified by decoding into the individual recognition information.

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