JP2002372490A - Sensor utilizing total reflection attenuation and measurement chip assembly - Google Patents

Abstract

PROBLEM TO BE SOLVED: To prevent sample liquid being supplied to a measurement unit from being evaporated and to improve measurement accuracy in a sensor utilizing total reflection attenuation due to surface plasmon resonance or the like. SOLUTION: A cup-shaped measurement unit 10 is arranged on a turntable 20 by a loading/unlociding mechanism 65, and measurement liquid is dropped and supplied to the measurement unit 10 by a sample liquid supply mechanism 64. The turntable 20 is rotated, and a small amount of oil is dropped and supplied at a specific position by an oil supply mechanism 80. Further, the turntable 20 is rotated, light beams 30 are allowed to enter at various angles so that total reflection conditions can be obtained at the interface between a metal film formed on the inner bottom surface of a measurement unit 10 and a dielectric block under the metal film, and the light beams 30 being totally reflected on the interface are detected by a photodetector 40 and the characteristics of sample liquid are obtained by measuring the state of total reflection attenuation based on the light detection value. The surface of sample liquid in the measurement unit 10 is covered with oil, thus preventing the sample liquid from being evaporated and improving measurement accuracy.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a sensor and a measuring chip assembly utilizing attenuation of total reflection, such as a surface plasmon sensor for analyzing the characteristics of a substance in a sample solution by utilizing the generation of surface plasmons, and in particular, to a detailed description. The present invention relates to a sensor and a measurement chip assembly using attenuated total reflection provided with one or a plurality of sample liquid holding portions for holding a sample liquid on a measurement chip.

[0002]

2. Description of the Related Art In a metal, free electrons vibrate collectively to generate a compression wave called a plasma wave. And, the quantization of this compression wave generated on the metal surface is
It is called surface plasmon.

Conventionally, various surface plasmon sensors for quantitatively analyzing a substance in a sample liquid by utilizing the phenomenon that surface plasmons are excited by light waves have been proposed. Among them, a particularly well-known one uses a system called a Kretschmann configuration (see, for example, JP-A-6-167443).

A surface plasmon sensor using the above-mentioned system is basically composed of a dielectric block formed, for example, in a prism shape, a thin film layer made of a metal film formed on the upper surface of the dielectric block, and a sample formed on the thin film layer. A measurement chip having a sample liquid holding unit for holding a liquid, a light source for generating a light beam, and the light beam with respect to a dielectric block, the condition of total reflection being obtained at an interface between the dielectric block and the thin film layer. And an optical system for measuring the intensity of the light beam totally reflected at the interface to detect the state of surface plasmon resonance, that is, the state of attenuated total reflection. Things.

In order to obtain various angles of incidence as described above, a relatively narrow light beam may be incident on the interface by changing the angle of incidence, or may be incident on the light beam at various angles. A relatively thick light beam may be incident on the interface in a convergent light state or a divergent light state so that the component is included. In the former case, the light beam whose reflection angle changes according to the change in the incident angle of the incident light beam is detected by a small photodetector that moves in synchronization with the change in the reflection angle, or the direction in which the reflection angle changes. Can be detected by an area sensor extending along. On the other hand, the latter case can be detected by an area sensor extending in a direction in which all light beams reflected at various reflection angles can be received.

In the surface plasmon sensor having the above structure, when a light beam is incident on the thin film layer at a specific incident angle equal to or larger than the total reflection angle, an evanescent wave having an electric field distribution is formed in the sample liquid in contact with the thin film layer. This evanescent wave causes surface plasmons to be excited at the interface between the thin film layer and the sample solution. When the wave number vector of the evanescent light is equal to the wave number of the surface plasmon and the wave number matching is established, the two are in a resonance state, and the light energy is transferred to the surface plasmon. The intensity of the reflected light drops sharply. This decrease in light intensity is generally detected as a dark line by the light detection means.

[0007] The above resonance occurs only when the incident beam is p-polarized. Therefore, it is necessary to set in advance so that the light beam is incident as p-polarized light.

When the wave number of the surface plasmon is known from the incident angle at which the attenuated total reflection (ATR) occurs, that is, the total reflection elimination angle θ _SP , the dielectric constant of the sample liquid is obtained. That wave number K _SP of the surface _plasmon, the angular frequency of the surface plasmon omega, the speed of light in vacuum c, thin films epsilon _m and epsilon _s respectively, when the dielectric constant of the sample solution, the following relationship.

[0009]

(Equation 1) Knowing the dielectric constant epsilon _s of the sample liquid, the apparent concentration of a specific substance in a sample solution based on a predetermined calibration curve or the like, after all, by knowing the incident angle theta _SP that the reflected light intensity is lowered, the sample Properties related to the dielectric constant of the liquid, ie, the refractive index, can be determined.

As similar sensors utilizing attenuated total reflection (ATR), for example, “Spectroscopy”, Vol. 47, No. 1 (1998), pp. 21-23 and 26-27.
A leak mode sensor described on the page is also known. This leaky mode sensor is basically composed of a dielectric block formed in a prism shape, a clad layer formed on the upper surface of the dielectric block and an optical waveguide layer formed thereon, and a thin film layer formed on the thin film layer. A measuring chip provided with a sample liquid holding section for holding a sample liquid, a light source for generating a light beam, and the light beam being totally reflected on the dielectric block at an interface between the dielectric block and the cladding layer. An optical system for incidence at various angles so as to obtain conditions, and an optical detection means for measuring the intensity of the light beam totally reflected at the interface and detecting the excited state of the guided mode, that is, the state of attenuated total reflection. It is provided with.

In the leakage mode sensor having the above configuration,
When a light beam is incident on the cladding layer through the dielectric block at an incident angle equal to or greater than the total reflection angle, only light of a specific incident angle having a specific wave number is transmitted to the optical waveguide layer after passing through the cladding layer. The light propagates in a guided mode. When the waveguide mode is excited in this way, most of the incident light is taken into the optical waveguide layer, and thus the total reflection attenuation occurs in which the intensity of the light totally reflected at the interface sharply decreases.
Since the wave number of the guided light depends on the refractive index of the sample liquid on the optical waveguide layer, knowing the specific incident angle at which the total reflection attenuation occurs determines the refractive index of the sample liquid and the characteristics of the sample liquid related thereto. Can be analyzed.

The above-described surface plasmon sensor, leak mode sensor, and the like are sometimes used in the field of drug discovery research to study the interaction between a desired sensing substance and a sample solution. For example, it is used for measuring an interaction such as a binding action between a sensing substance and a specific substance contained in a sample liquid and a dissociation action of a specific substance from a binding substance into a sample liquid. Such interactions include protein-
It includes protein interaction, DNA-protein interaction, sugar-protein interaction, protein-peptide interaction, lipid-protein interaction and chemical substance binding.

In the case where the surface plasmon sensor or the leak mode sensor is used for random screening for finding a specific substance that binds to a sensing substance, the sensing substance is fixed on the thin film layer, and Various analytes are added with sample solutions dissolved in a solvent, and the state of total reflection attenuation is measured every predetermined time. If the analyte in the sample solution binds to the sensing substance, the binding changes the refractive index of the sensing substance with time. Therefore, the state of attenuated total reflection is measured every time a predetermined time elapses, and by measuring whether or not a change has occurred, whether or not the analyte and the sensing substance are bonded, that is, the It can be determined whether or not the specific substance binds. Examples of such a combination of the specific substance and the sensing substance include an antigen and an antibody, or an antibody and an antibody. Specifically, rabbit anti-human Ig
The G antibody can be immobilized on a measurement chip as a sensing substance, and a human IgG antibody can be used as a specific substance.

The applicant of the present invention has proposed, in Japanese Patent Application No. 2001-397411, a sensor utilizing attenuated total reflection which performs measurement using a measurement chip provided with a plurality of sample liquid holding portions. By using a sensor having such a configuration, measurement of a large number of samples can be performed in a short time.

[0015]

However, in a sensor using attenuated total reflection, which has been conventionally provided, a sample liquid holding portion having a cup-shaped or petri dish-like shape in which a thin film layer is previously formed on an inner bottom surface is provided. , And the state of the total reflection attenuation described above is measured.

For this reason, the sample liquid evaporates during the actual measurement after the supply of the sample liquid, the concentration of the sample liquid changes, and the measurement accuracy in the state of attenuated total reflection decreases. There was a fear.

In particular, when measuring the change over time of the interaction between the sensing substance disposed on the measurement chip and the sample liquid, the measurement time is often long, during which the sample liquid evaporates, There was a possibility that an accurate temporal change could not be measured.

The present invention has been made in view of the above circumstances, and has as its object to provide a sensor using attenuated total reflection that can prevent evaporation of a sample liquid and improve the measurement accuracy of the state of attenuated total reflection. .

[0019]

A sensor using attenuated total reflection according to the present invention includes a light source for generating a light beam,
A measurement chip including a dielectric block transparent to the light beam, a thin film layer formed on the upper surface of the dielectric block, and one or more sample liquid holding units for holding a sample liquid on the thin film layer An optical system for causing the light beam to enter the dielectric block at various angles of incidence such that total reflection conditions are obtained at the interface between the dielectric block and the thin film layer; and A sensor using total reflection attenuation, comprising: a light detection unit that detects the intensity of the light beam that has been subjected to the detection; and a measurement unit that measures the state of total reflection attenuation based on the detection result of the light detection unit. The sample liquid holding section has an opening at an upper portion thereof, and the opening is provided with a lid supply means for providing a lid for preventing evaporation of the sample liquid.

As such a sensor, the above-mentioned surface plasmon sensor using a metal film as the thin film layer, a clad layer formed on one surface of a dielectric block,
There is the above-mentioned leaky mode sensor or the like using a layer composed of the optical waveguide layer formed on the cladding layer as the thin film layer.

In the sensor according to the present invention, there are various methods for analyzing the substance in the sample liquid by measuring the intensity of the light beam totally reflected at the interface by the light detecting means. The light is incident at various angles of incidence at which the total reflection condition is obtained at the interface, the intensity of the light beam totally reflected at the interface is measured for each position corresponding to each angle of incidence, and the position of the dark line generated by the total reflection attenuation is measured. (Angle) may be detected for analysis, or DVNoort, K. johans
en, C.-F.Mandenius, Porous Gold in SurfacePlasmon R
esonance Measurement, EUROSENSORS XIII, 1999, pp.5
As described in No. 85-588, light beams of a plurality of wavelengths are incident on the interface at an incident angle at which the condition of total reflection is obtained, and the intensity of the light beam totally reflected at the interface is measured for each wavelength. The analysis may be performed by detecting the degree of total reflection attenuation for each wavelength.

Also, PINikitin, ANGrigorenko, AAB
eloglazov, MVValeiko, AISavchuk, OASavchuk, Sur
face Plasmon Resonance Interferometry for Micro-Ar
rayBiosensing, EUROSENSORS XIII, 1999, pp.235-238
A light beam is incident at an angle of incidence where total reflection conditions are obtained at the interface, and a portion of the light beam is split before the light beam is incident on the interface, as described in The sample analysis may be performed by causing the split light beam to interfere with the light beam totally reflected at the interface, and measuring the intensity of the light beam after the interference.

A sensing substance which interacts with the sample liquid is disposed on the thin film, and the measuring means detects attenuated total reflection based on a plurality of detection results detected at intervals by the light detecting means. It may measure the change over time of the state.

In the case where oil is used as the lid means, the lid means supply means may supply oil, for example, dropwise to the opening.

[0025] If the lid means uses a lid, the lid means supply means may cover the opening with the lid. As the lid, one having a small opening smaller than the opening,
It may be formed in the shape of a stretchable cord or in the form of a sheet.

The amount of water evaporation per hour in the sample liquid holding section provided with the lid means in the opening is preferably 2% or less. It is more preferable that the amount of water evaporated per hour is 0.5% or less.

Further, as a measurement chip assembly of the present invention, a dielectric block, a thin film layer formed on an upper surface of the dielectric block, and a sample liquid held on the thin film layer,
A measuring chip formed of one or more sample liquid holding portions having an opening and used for a sensor utilizing attenuated total reflection; and a lid provided at the opening of the measuring chip. It is a feature.

The lid may have a small opening smaller than the opening, may be formed in a stretchable cord shape, or may be formed in a sheet shape. .

The amount of water evaporation per hour in the sample liquid holding section provided with the lid at the opening is preferably 2% or less. It is more preferable that the amount of water evaporated per hour is 0.5% or less.

The sample liquid holding section raises the periphery of the portion where the thin film layer of the dielectric block made of, for example, transparent resin or the like is formed, and forms a truncated cone-shaped space having an opening at the top. It can be formed by providing it above. One sample liquid holding section may be provided in one measurement chip, or a plurality of sample liquid holding sections may be provided. When a plurality of sample liquid holding units are provided in one measurement chip, the sample liquid holding units may be arranged in a line,
Alternatively, they may be arranged in a plurality of rows. When a plurality of sample liquid holding units are provided in the measurement chip,
The light source, the optical system, the light detecting means, and the measuring means may be provided separately for each sample liquid holding unit, or may be a single unit. When a single device is used, the light source, the optical system, the light detection means, and the measurement means may be sequentially moved and used, or the measurement chip may be moved.

When a measurement chip provided with a plurality of sample liquid holding sections is used, the lid attached to the measurement chip is formed individually for each opening of each sample liquid holding section. Alternatively, they may be integrally formed so as to cover the openings of all the sample liquid holding sections.

[0032]

In the sensor utilizing attenuated total reflection according to the present invention, a cover is provided at the opening of the sample liquid holding portion of the measuring chip to prevent evaporation of the sample liquid, thereby supplying the liquid to the sample liquid holding portion. It is possible to prevent the sample liquid from being evaporated and improve the measurement accuracy.

When measuring the time-dependent change in the interaction between the sensing substance disposed on the measurement chip and the sample liquid, evaporation of the sample liquid is prevented even when the measurement time is long. Therefore, an accurate temporal change can be measured.

If the oil dropped and supplied to the opening is used as the lid, the lid can be easily provided at the opening.

If the lid covered by the opening is used as lid means, evaporation of the sample liquid is prevented, and dust or the like may enter the sample liquid supplied to the sample liquid holding section. Can be prevented.

If the lid has a small opening, the sample liquid can be supplied through the small opening. Therefore, even before the sample liquid is supplied to the sample liquid holding section, the sample liquid can be supplied. A lid can be attached to the opening of the liquid supply mechanism.

If the lid is formed in a stretchable blind shape, the sample liquid can be supplied from the gap in the state of the screen. In this case, the lid can be attached to the opening of the sample liquid supply mechanism, and there is no need for precise positioning when supplying the sample liquid.

Further, if the lid is formed in a sheet shape, it is not necessary to perform precise positioning when attaching the lid to the opening, and the lid can be easily attached.

If the amount of water evaporation per hour in the sample liquid holding portion having the opening means provided with the lid means or the lid body is 2% or less, the refractive index of the sample liquid caused by the evaporation of the sample liquid. The change hardly occurs, and the measurement can be performed with high accuracy. If the amount of water evaporation per hour is 0.5% or less, the amount of evaporation of the sample liquid is smaller, and the measurement can be performed with higher accuracy.

When using a measurement chip provided with a plurality of sample liquid holding portions, an integrally formed lid attached to the opening of each sample liquid holding portion can be used. In a single operation, the lid can be attached to the opening of each sample liquid holding section, and the lid can be attached efficiently.

[0041]

Embodiments of the present invention will be described below in detail with reference to the drawings. FIG. 1 shows an overall shape of a surface plasmon sensor according to a first embodiment of the present invention, and FIG. 2 shows a side shape of a main part of the device.

As shown in FIG. 1, the surface plasmon sensor includes a plurality of measurement units 10, a turntable 20 supporting the plurality of measurement units 10, and a moving means for intermittently rotating the turntable 20. Support means 21, a laser light source 31 such as a semiconductor laser for generating a measurement light beam (laser beam) 30, a condenser lens 32 forming an incident optical system, a photodetector 40, A controller 60 that controls the driving of the light source 31 and the support driving means 21 and receives the output signal S of the photodetector 40 and performs processing described below; and an access mechanism 65 that moves the measurement unit 10 into and out of the turntable 20. , Sample liquid supply mechanism
70 and an oil supply mechanism 80 as lid means supply means. Note that the measurement unit 10 functions as a measurement chip of the present invention.

As shown in FIG. 2, the measuring unit 10 includes a dielectric block 11 having, for example, a substantially quadrangular pyramid shape, and gold, silver, or the like formed on the upper surface of the dielectric block 11.
And a metal film 12 made of copper, aluminum, or the like.

The dielectric block 11 is made of, for example, a transparent resin, and has a raised portion around the portion where the metal film 12 is formed. The raised portion is the sample liquid holding frame 13.
Is a frustum-shaped space in which the upper part expands, and functions as a sample liquid holding unit 13b for storing the sample liquid 15. An opening 13a is formed above the sample liquid holding section 13b.

On the turntable 20, a plurality (12 in this example) of through-holes 22 for fitting and holding the measuring unit 10 is provided.
Are provided at equal angular intervals on the circumference of the turntable 20 around the rotation axis 20a. The measurement unit 10 is held in the turntable 20 in a replaceable state.
The support driving means 21 is constituted by a stepping motor or the like, and intermittently rotates the turntable 20 by an angle equal to the arrangement angle of the through holes 22.

The condenser lens 32 has a light beam as shown in FIG.
30 is condensed and passed through the dielectric block 11 in a convergent light state, and is incident on the interface 12a between the dielectric block 11 and the metal film 12 so that various incident angles can be obtained. The range of the incident angle is a range including an angle range in which the condition of total reflection of the light beam 30 at the interface 12a is obtained and surface plasmon resonance can occur.

The light beam 30 enters the interface 12a as p-polarized light. In order to do so, the laser light source 31 may be disposed in advance so that the polarization direction thereof becomes a predetermined direction. Alternatively, the direction of polarization of the light beam 30 may be controlled by a wave plate or a polarizing plate.

The photodetector 40 is a line sensor in which a large number of light receiving elements are arranged in one line, and is arranged such that the light receiving elements are arranged in the direction of arrow X in FIG.

On the other hand, the controller 60 is
Receives an address signal A indicating the rotation stop position, and outputs a drive signal D for operating the support driving means 21 based on a predetermined sequence. Further, it comprises a measuring means 61 for receiving the output signal S of the photodetector 40, and a display section 62 for receiving an output from the measuring means 61. Further, it is connected to the access mechanism 65, the sample liquid supply mechanism 70, and the oil supply mechanism 80, and controls its operation as needed.

The access mechanism 65 includes a holding section 66 for holding the measuring unit 10 and means 67 for moving the holding section 66.

The sample liquid supply mechanism 70 is composed of a pipette 71 for sucking and holding a predetermined amount of the sample liquid and a means 72 for moving the pipette 71, and a sample liquid container 73 set at a predetermined position. Then, the sample liquid is sucked and held by the pipette 71, and the sample liquid is dropped and supplied to the sample liquid holding portion 13b of the measurement unit 10 at a predetermined stop position on the turntable 20.

The oil supply mechanism 80 is composed of a pipette 81 for sucking and holding a predetermined amount of the oil 50 and a means 82 for moving the pipette 81. The oil supply mechanism 80 moves the oil 50 from the oil container 83 set at a predetermined position. The sample liquid holding portion 13b of the measurement unit 10 to which the sample liquid 15 is supplied is disposed on the turntable 20, and the opening 13 is held by the pipette 81.
The oil 50 is supplied dropwise to a. As the oil 50, a mineral oil manufactured by Applied Biosystems is used.

Hereinafter, a sample analysis by the surface plasmon sensor having the above configuration will be described. First, the prepared measurement units 10 are arranged, for example, in a 96-well cassette. Each of the measurement units 10 is sequentially arranged in the through hole 22 on the turntable 20 by the access mechanism 65. At the time of sample analysis, the turntable 20 is, as described above, a support driving means.
It is intermittently rotated by 21. Then, when the turntable 20 stops, the measurement unit 10 that has stopped at a predetermined position
The sample liquid supply mechanism 70 supplies the sample liquid 15 to the sample liquid holding unit 13b.

Further, the turntable 20 is rotated,
The measuring unit 10 to which the liquid 15 has been supplied is the oil supply mechanism 80
Is stopped at the position where the oil supply mechanism 80 is provided.
As a result, the opening 13 of the sample liquid holding portion 13b of the measurement unit 10
The oil 50 is supplied dropwise to a. In the sample liquid holder 13b
The amount of oil supplied dropwise is determined by the amount of
It is set in advance according to the surface area of the liquid material 15. Usually a trial
Surface area of material 15 20mm ²More than 20 micro oils per
When the oil is supplied, the entire surface of the sample liquid 15 is
Covered. For this reason, the sample liquid holding portion 13b under the oil 50
Means that water evaporation per hour is less than 2%
The sample liquid 15 hardly evaporates during the time.

Thereafter, when the turntable 20 is rotated several times and then stopped, the measuring unit 10 holding the sample liquid 15 in the sample liquid holding section 13b receives the light beam 30 incident on the dielectric block 11. At a measurement position (the position of the measurement unit 10 on the right side in FIG. 2). In this state, the laser light source 31 is driven by a command from the controller 60, and the light beam 30 emitted therefrom is converged as described above, and the interface 12 between the dielectric block 11 and the metal film 12 is conveyed.
a. The light beam 30 totally reflected at the interface 12a
Is detected by the photodetector 40.

Since the light beam 30 enters the dielectric block 11 in a convergent light state as described above, the light beam 30 includes components incident on the interface 12a at various incident angles θ. The incident angle θ is an angle equal to or larger than the total reflection angle. Therefore, the light beam 30 is totally reflected at the interface 12a, and the reflected light beam 30 contains components reflected at various reflection angles.

As described above, when the light beam 30 is totally reflected,
An evanescent wave exudes from the interface 12a to the metal film 12 side. When the light beam 30 enters the interface 12a at a specific incident angle θ _SP , the evanescent wave resonates with the surface plasmon excited on the surface of the metal film 12, so that the reflected light intensity I drops sharply.
FIG. 3 schematically shows the relationship between the incident angle θ and the reflected light intensity I when this total reflection attenuation phenomenon occurs.

Then, in the measuring means 61, as the state of total reflection attenuation, the amount of light detected for each light receiving element is checked from the output signal S output from the photodetector 40, and based on the position of the light receiving element where the dark line is detected, An incident angle (attenuation angle of total reflection) θ _SP is obtained. In addition, the specific substance in the sample liquid 15 can be quantitatively analyzed based on a relationship curve between the reflected light intensity I and the incident angle θ which is obtained in advance. Based on the above principle, the quantitative analysis of the specific substance in the sample liquid 15 is performed, and the analysis result is displayed on the display unit 62. After the measurement is completed, the measurement unit 10 that has completed the measurement is removed from the turntable 20 by the access mechanism 65.

In this embodiment, the sample liquid 15 is stored in the sample liquid holding section.
Since the oil 50 is supplied to the sample liquid holding portion 13b after being supplied to the sample liquid 13b, the surface of the sample liquid 15 is covered with the oil 50. Under the oil 50, the water evaporation amount per hour is 2 % Or less. Therefore, during the measurement time,
15 hardly evaporates, the concentration of the sample liquid is kept constant, and the generation of heat of vaporization due to evaporation can be prevented, so that the measurement accuracy in the state of attenuated total reflection is improved. Further, since oil is used as the lid means, the lid means can be easily provided in the opening 13a of the sample liquid holding section 13b. In addition,
More preferably, the amount of water evaporation is 0.5% or less.

In each of the embodiments described above, the rotating turntable 20 is used as a support for supporting the measuring unit 10. However, the moving method and the shape of the support are not limited to this. . For example, turntable
Instead of rotating 20 in one direction, move the measurement unit in the opposite direction and set the measurement unit again on the light measurement mechanism consisting of the light source 31, the condenser lens 32 and the photodetector 40, and perform measurement. Is also good. Also, by providing a plurality of light measurement mechanisms,
It is also possible to configure so that a plurality of measurements are performed for one measurement unit while the turntable 20 makes one rotation.

Alternatively, a support supporting a plurality of measurement units is configured to reciprocate linearly or move in a plane in the XY directions, and the plurality of measurement units are moved to one or a plurality of optical measurement mechanisms in accordance with the movement. It may be set sequentially.

As a modification of the first embodiment, the support is fixed and the optical measurement mechanism is moved to measure each of the measurement units 10 sequentially, or the support and the optical measurement mechanism are used. It is also conceivable to move both of them to sequentially measure the measurement units 10. Also in this case, it is possible to perform measurement on a large number of measurement units in a short time.

As another modified example of the present embodiment,
There is a type in which a sensing substance is fixed on the metal film 12, and whether or not the analyte contained in the sample liquid is a specific substance that binds to the sensing substance is determined. In this case,
The temporal change of the total reflection attenuation angle θ _SP due to surface plasmon resonance is measured, and when the total reflection attenuation angle θ _SP greatly changes, it is determined that the subject is a specific substance. In order to make such a determination, it is necessary to perform measurement for one measurement unit 10 a plurality of times. In this case, if the measurement unit 10 is supported on the turntable 20 as it is even after the first measurement, the measurement unit 10 can be subjected to the measurement again by rotating the turntable 20. As described above, when the measurement of one measurement unit 10 is performed for a long time, there is a possibility that the concentration of the sample liquid 15 may change, particularly due to the evaporation of the sample liquid 15,
Since the oil covers the surface of the sample liquid, the evaporation of the sample liquid is prevented, and the measurement accuracy of the change over time in the total reflection state is improved.

Next, a second embodiment of the present invention will be described with reference to FIGS. FIG. 4 shows the overall shape of the surface plasmon sensor according to the second embodiment,
FIG. 5 shows the shape of the lid 51 used in the present embodiment. In FIG. 4, the same elements as those in FIG. 1 are denoted by the same reference numerals, and description thereof will be omitted unless otherwise required.

In this surface plasmon sensor, the lid 51 is used as a lid supplying means, and the lid 51 is connected to the sample liquid holding section of the measuring unit 10.
A lid supply mechanism 84 is used to cover the opening 13a of 13b. As shown in FIG. 5A, a top view, and FIG. 5B, a side view thereof, the lid 51 has an opening 13a of the sample liquid holding portion 13b.
It is a rectangular lid having a slightly smaller, truncated cone-shaped convex portion 52 at the center, and one side of the rectangular shape is a sample liquid holding frame 13.
Is the same as the length of one side of the upper part of. The lid supply mechanism 84 includes a holder 85 for holding the lid 51 and a means 86 for moving the holder 85, and its operation is controlled by the controller 60.

When a sample is analyzed by the surface plasmon sensor having the above-described structure, the measurement units 10 arranged in a 96-well cassette (not shown) are moved by the access mechanism 65 as in the first embodiment shown in FIG. The sample liquid supply mechanism 70 sequentially arranges the sample liquid 15
Is supplied.

After the sample liquid is supplied, the turntable 20 is rotated, and when the measuring unit 10 to which the sample liquid 15 is supplied is stopped at the position where the lid supply mechanism 84 is provided, the lid supply mechanism 84 The lid 51 is put on the measurement unit 10. On this occasion,
The lid supply mechanism 84 holds one of the lids 51 stacked in advance so that the convex portions 52 face down by the holding unit 85, and
Is the opening of the sample liquid holding section 13b of the measurement unit 10.
The lid body 51 is put on the measuring unit 10 so as to be fitted in 13a.

By using the lid supply mechanism 84 having the above structure to cover the opening 13a of the sample liquid holding portion 13b with the lid 51, the evaporation of the sample liquid 15 is prevented, the measurement accuracy is improved, and the It is possible to prevent dust and the like from entering the sample liquid 15 supplied to the liquid holding unit 13b.

FIG. 6 shows a modified example of the lid 51 described above.
(A) is a top view, and (b) is a side view thereof. A square lid 102 mainly having a bowl-shaped convex portion 101 slightly larger than the opening 13a of the sample liquid holding portion 13b. Can be considered. If such a lid 102 is used, it is not necessary to perform precise positioning when the lid 102 is mounted on the measuring unit 10. 7A shows a top view and FIG. 7B shows a side view thereof as a modified example.
103 can be used, and positioning when the lid 103 is mounted on the measuring unit 10 is further facilitated. When such a lid 103 is used, if a measuring unit 105 having a flange 104 as shown in FIG.
A sufficient space for holding the sample liquid 15 can be ensured in the measurement unit 105.

If a soft resin or rubber is used as the material of each lid, the adhesion between the opening 13a and the lid is improved, and the evaporation of the sample liquid 15 can be further prevented. Further, if a sealing material such as oil is added between the upper part of the measuring unit and the lid, the adhesion is further improved.

Next, a third embodiment of the present invention will be described with reference to FIGS. FIG. 9 shows this third
10 shows the overall shape of the surface plasmon sensor according to the embodiment, and FIG. 10 shows the shape of a lid 53 used in the present embodiment. In FIG. 9, elements that are the same as the elements in FIG. 1 are given the same reference numerals, and descriptions thereof will be omitted unless otherwise required.

In this surface plasmon sensor, a lid for covering the opening 13a of the sample liquid holding section 13b of the measurement unit 10 before the measurement unit 10 is placed on the turntable 20 as a lid means supply means. A body supply mechanism 87 is used. The lid 53 has a top view in FIG.
As shown in the side view of (b), it is a square lid having a truncated cone-shaped convex portion 54 slightly smaller than the opening 13a of the sample liquid holding portion 13b at the center. , The length of one side of the upper portion of the sample liquid holding frame 13. A small opening 55 is provided at the center of the convex portion 54 such that the tip of the pipette 71 of the sample liquid supply mechanism 70 can be inserted.

The cover supply mechanism 87 includes a holding section 88 for holding the cover 53 and means 89 for moving the holding section 88, and its operation is controlled by the controller 60.

When a sample is analyzed by the surface plasmon sensor having the above-described structure, first, the lid supply mechanism 87 is attached to the measuring unit 10 arranged in a 96-well cassette (not shown).
Put 53 on. At this time, the lid supply mechanism 87 holds one of the lids 53 stacked in advance so that the convex portions 54 face down by the holding portion 88, and the lid 53 holds the convex portions 54 of the measurement unit 10 The lid is inserted into the opening 13a of the sample liquid holder 13b.
Put 53 on the measurement unit 10.

The access mechanism 65 sequentially arranges the measuring units 10 covered with the lid 53 in the through holes 22 on the turntable 20. The turntable 20 is rotated, and the measuring unit 10
Is stopped at a predetermined position, the sample liquid supply mechanism 70 supplies the sample liquid 15 to the sample liquid holding unit 13b of the measurement unit 10. When supplying the sample liquid, the pipette 71 of the sample liquid supply mechanism 70 is inserted into the small opening 55 of the lid 53, and the sample liquid 15 is supplied dropwise.

In the present embodiment, the lid 53 is placed over the opening 13a of the sample liquid holding portion 13b of the measuring unit 10 before the sample liquid 15 is supplied by using the lid supply mechanism 87 having the above configuration. Thus, the evaporation of the sample liquid 15 can be reliably prevented, the measurement accuracy is further improved, and dust and the like can be prevented from entering the sample liquid 15 supplied to the sample liquid holding unit 13b.

As a modified example of the lid 53, FIG.
As shown in FIG. 1A, a top view is shown, and a side view thereof is shown in FIG. In this case, since the sample liquid can be supplied by inserting the pipette 71 through the blind-shaped gap, it is not necessary to precisely determine the position when supplying the sample liquid.

Further, as another modified example of the lid, FIG.
(A) shows a top view, and (b) shows a side view of the lid, which is attached to the measurement unit 10 by a pivot 57.
58 can also be used. The lid 58 has a small opening 59 in the center.
The opening 13a of the measuring unit 10 is opened and closed by rotating the measuring unit 10 on the pivot 57 as a rotation axis. When the lid 58 is attached to the measurement unit 10 as described above, the lid 58 does not come off from the measurement unit 10 when the measurement unit 10 is moved. FIG. 13A shows a top view of a measurement unit using the lid 58.
It is also possible to use a measuring unit 96 having a fitting part 95 at the upper part as shown in the side view of FIG. The measuring unit 96 has a fitting portion 95 formed only on a pair of opposing wall surfaces on the upper part thereof. For this reason, the lid 58 is
The opening can be covered with the lid 58 by fitting and sliding on the 95. Once the lid 58 is attached to the measurement unit 96, the lid 58 does not easily come off the opening.

In each of the above embodiments, as described above, not only the measurement unit 10 in which the dielectric block 11, the metal film 12, and the sample liquid holding frame 13 are integrally formed, but also the metal film 12 and the sample It is also possible to apply a measurement unit in which the liquid holding frame 13 is integrated and is exchangeably formed for the dielectric block 11.

Next, a fourth embodiment of the present invention will be described with reference to FIG. 14, FIG. 15, and FIG. FIG. 14 is a plan view showing a schematic configuration of a surface plasmon sensor according to the fourth embodiment. FIG. 15 shows a side surface shape of a main part, and FIG. 16 shows a sheet supply mechanism as cover means supply means. It is a schematic block diagram. In FIGS. 14 and 15, the same elements as those in FIGS. 1 and 2 are denoted by the same reference numerals, and a description thereof will be omitted unless otherwise necessary.

As shown in FIG. 14, this surface plasmon sensor is composed of six measurement units 10A, 10B, 10C,.
A surface plasmon sensor using surface plasmon resonance for detecting the state of attenuated total reflection by making light beams 30A, 30B, 30C... Incident in parallel on A, 10B, 10C.

The surface plasmon sensor has a measurement unit array 68 composed of six measurement units 10A, 10B, 10C,... And laser light sources 31A, 31B, 31C, which are light sources for generating light beams 30A, 30B, 30C,. And the light beam above
30A, 30B, 30C ... each measuring unit 10A, 10
, B, 10C, etc., and condenser lenses 32A, 32B, 32C.
A plurality of photodetectors 40A, 40B, 40C... Which receive the respective reflected light beams reflected by A, 10B, 10C... Corresponding to the measurement units 10A, 10B, 10C. , 40B, 40C... Output signals SA, S
B, SC ... and a controller 69 for performing signal processing;
A sample liquid supply mechanism 74 for supplying different types of sample liquid to the respective measurement units 10A, 10B, 10C,..., And a sheet supply mechanism 75 as lid means supply means. In addition,
The measurement unit array 68 functions as the measurement chip of the invention.

Each of the measurement units 10A, 10B, 10C,... Has the same structure as that of the other measurement units except that the dielectric block 11 and the sample liquid holding frame 13 are integrally formed with the dielectric block and the sample liquid holding frame. For example, the measurement unit shown in FIG.
It has the same configuration as 10. FIG. 15 shows a schematic configuration diagram of the measurement unit 10A as an example.

On the other hand, the controller 69
.. Which receive output signals SA, SB, SC from A, 40B, 40C,.
A, 61B, 61C,... The sample liquid supply mechanism 74 and the sheet supply mechanism
It is connected to 75 and controls its operation as needed.

As shown in FIG. 16, the sheet feeding mechanism 75 rotates the sheet holding portion 76, a flexible cover sheet 77 wound around the sheet holding portion 76, and the sheet holding portion 76. And a sheet supply unit 78 for attaching a cover sheet 77 to the opening 13a of each sample liquid holding unit 13b of the measurement unit array 68 as necessary. The operation of the sheet supply unit 78 is controlled by the controller 69.

The measurement of the state of attenuated total reflection with respect to the sample liquid supplied to the one measuring unit, for example, the measuring unit 10A, is performed by measuring the laser light source 31A, the condenser lens 32A,
Measuring unit 10A, photodetector 40A, and measuring means 61A
Is performed by a surface plasmon sensor unit 98A.

First, prior to the measurement, the measurement unit array 68 is set at the measurement position by a measurement unit array access mechanism (not shown). At this time, the cover sheet 77 is wound around the sheet holding section 76. Under the control of the controller 69, the sample liquid supply mechanism 74 supplies various sample liquids 15 to the sample liquid holding units 13b of the measurement units 10A, 10B, 10C,. Thereafter, under the control of the controller 69, the sheet supply unit 78 rotates the sheet holding unit 76 in the direction in which the bar sheet 77 is pushed out by a predetermined amount. For this reason, the cover sheet
The opening 13a of each of the measuring units 10A, 10B, 10C... Of the measuring unit array 68 is covered by the tip of the 77. By covering the opening 13a with the cover sheet 77, the sample liquid holding units of the measuring units 10A, 10B, 10C...
13b is sealed and the amount of water evaporation per hour is less than 2%. Therefore, the sample liquid 15 hardly evaporates within the measurement time.

As described above, each measurement unit 10A, 10A
With the openings 13a of B, 10C,... Covered by the front end of the cover sheet 77, the measurement of the state of attenuated total reflection is performed. Each surface plasmon sensor unit 98A, 98B, 98
Since the measurement operation in C ... is almost the same as the measurement operation in the first embodiment, the description of the measurement operation is omitted. When the measurement is completed, under the control of the controller 69,
The sheet supply unit 78 reversely rotates the sheet holding unit 76 by a predetermined amount. The cover sheet 77 is wound around the sheet holding unit 76.

In the present embodiment, the sample solution 15 is
After being supplied to 13b, the opening 13a of each measurement unit 10A, 10B, 10C ... is covered by the cover sheet 77,
The sample liquid holding unit 13b of each of the measurement units 10A, 10B, 10C,...
And the amount of evaporation per hour is 2% or less, so that the sample solution 15 hardly evaporates within the measurement time, the concentration of the sample solution is kept constant, and the heat of vaporization due to evaporation is reduced. Since it can be prevented, the measurement accuracy of the state of attenuated total reflection is improved. More preferably, the amount of water evaporation is 0.5
% Or less. Further, since the cover sheet 77 is used as the lid, it is not necessary to perform precise positioning when attaching the lid to the opening 13a, and the lid 13 can be easily attached.

Since one cover sheet 77 is used as a cover provided in a measurement unit array having a plurality of sample liquid holding units, each measurement unit 10 can be operated in one operation.
Covers (cover sheet 7)
7) can be attached, and the lid can be attached efficiently.

As a modified example of the fourth embodiment,
A surface plasmon sensor provided with a lid supply mechanism 45 as shown in FIG. 17 instead of the sheet supply mechanism 75 is also conceivable. The cover supply mechanism 45 includes a cover 46 and a cover supply unit 47 that covers the cover 46 over the opening 13a of the sample liquid holding unit 13b of the measurement unit array 68. The lid supply unit 47 includes a lid
A holding unit 48 for holding the holding unit 46, and a unit 49 for moving the holding unit 48, the operation of which is controlled by a controller 69.
Cover the opening 13a of the sample liquid holding section 13b. The lid 46 may be formed in a flat plate shape or a sample liquid holding unit.
A truncated cone-shaped protrusion slightly smaller than the opening 13a of 13b is
A plate-shaped member having a position corresponding to the opening 13a of the sample liquid holding portion 13b, or a bowl-shaped protrusion slightly larger than the opening 13a of the sample liquid holding portion 13b,
A plate formed at a position corresponding to the opening 13a of the opening 13b can be used. Further, if a soft resin or rubber is used as a material of each of these lids, the adhesion between the opening 13a and the lid is improved, and the sample liquid is further reduced.
15 evaporation can be prevented. If a sealing material such as oil is added between the upper portion of the measurement unit array 68 and the lid, the adhesion is further improved.

As another modified example of the fourth embodiment, a surface plasmon sensor provided with a resin sheet supply mechanism 111 as shown in FIG. The resin sheet supply mechanism 111 includes a resin sheet
112 and a sheet crimping portion 113 for thermocompression bonding the resin sheet 112 to the upper portion of the measurement unit array 68 by thermocompression. If such a resin sheet 112 is used as a lid, there is no risk that the lid will come off when the measurement unit array 68 is moved. Since the resin sheet 112 is attached to the upper part of the measurement unit array 68 by thermocompression bonding, when the resin sheet 112 becomes unnecessary, it can be easily peeled off.

Further, as another modified example of the fourth embodiment, a surface plasmon sensor provided with a viscous fluid supply mechanism 115 as shown in FIG. The viscous fluid supply mechanism 115 includes a viscous fluid 116 such as grease, and a viscous fluid supply unit 117 that supplies the viscous fluid 116 to an upper portion of the measurement unit array 68. If such a viscous fluid 116 is used as a lid,
Insert the pipette of the sample liquid supply mechanism into the
Since the sample liquid 15 can be supplied to the sample liquid holding section 13b of the measurement unit array 68, the lid can be attached to the measurement unit array 68 before the sample liquid 15 is supplied.

Next, a fifth embodiment of the present invention will be described with reference to FIGS.

Since the overall configuration of the fifth embodiment is substantially the same as that of the first embodiment, only the numbers of the different components in FIG. 1 are appended in the figure. In FIG. 20, the same elements as those in FIG. 2 are denoted by the same reference numerals, and description thereof will be omitted unless otherwise required.

The sensor using the attenuated total reflection according to the fifth embodiment is the leak mode sensor described above.
It is configured to use a measurement unit 90 that functions as a measurement chip. A cladding layer 91 is formed on the upper surface of the dielectric block 11 of the measurement unit 90, and an optical waveguide layer 92 is further formed thereon.

The dielectric block 11 is made of, for example, synthetic resin or B
It is formed using an optical glass such as K7. On the other hand, the cladding layer 91 is formed in a thin film shape using a dielectric material having a lower refractive index than the dielectric block 11 or a metal such as gold. Further, the optical waveguide layer 92 is a dielectric having a higher refractive index than the cladding layer 91,
This is also formed in a thin film shape using, for example, PMMA. The thickness of the cladding layer 91 is, for example, 36.5 nm when it is formed from a gold thin film, and the thickness of the optical waveguide layer 92 is, for example, PMMA.
From about 700 nm.

In the leakage mode sensor having the above configuration,
Light beam 30 emitted from laser light source 31 is a dielectric block
When the light beam 30 is incident on the cladding layer 91 at an incident angle equal to or larger than the total reflection angle through the interface 11, the light beam 30 is totally reflected at the interface 91 a between the dielectric block 11 and the cladding layer 91.
The light of a specific wave number that has passed through the optical waveguide layer 92 and entered the optical waveguide layer 92 at a specific incident angle propagates through the optical waveguide layer 92 in a guided mode. When the waveguide mode is excited in this way, most of the incident light is taken into the optical waveguide layer 92, and thus the total reflection attenuation occurs in which the intensity of the light totally reflected at the interface 91a sharply decreases.

Since the wave number of the guided light in the optical waveguide layer 92 depends on the refractive index of the sample liquid 15 on the optical waveguide layer 92, knowing the specific incident angle at which the total reflection attenuation occurs gives the sample liquid 15 Can be measured.

Also in the present embodiment, the sample liquid 15 is stored in the sample liquid holding section.
Since the oil 50 is supplied to the sample liquid holding unit 13b after being supplied to the sample liquid 13b, the surface of the sample liquid 15 is covered with the oil 50. Therefore, evaporation of the sample liquid 15 is prevented,
Measurement accuracy is improved. In particular, when measuring the temporal change of the attenuated total reflection angle θ _SP , even if the measurement time is long, the evaporation of the sample liquid 15 is prevented, and the measurement accuracy of the temporal change of the state of the attenuated total reflection is reduced. improves. Note that, in the present embodiment, a modified example similar to that of the first embodiment is applicable, and a similar effect is obtained. Furthermore, a form similar to the second embodiment and the third embodiment is also possible, and similar effects can be obtained. Further, similarly to the fourth embodiment, a sensor using a measurement unit array having a plurality of sample liquid holding units can be configured.

Next, a sixth embodiment of the present invention will be described with reference to FIG. The overall shape of the surface plasmon sensor according to the present embodiment is the same as that of the surface plasmon sensor according to the first embodiment shown in FIG. The surface plasmon sensor of the present embodiment is different from the surface plasmon sensor of the first embodiment in the measurement method. In the present embodiment, the sensing substance 14 is fixed on the metal film 12, and it is determined whether or not the analyte contained in the sample liquid 15 is a specific substance that binds to the sensing substance 14. .

As shown in FIG. 21, a laser light source 120 and a CCD 121 are provided at the measurement positions of the surface plasmon sensor according to the present embodiment.
0 and the CCD 121, a collimator lens 122, an interference optical system 123, a condenser lens 124 and an aperture 125
Are arranged.

The interference optical system 123 includes a polarizing filter 131
, Half mirror 132, half mirror 133 and mirror 1
34. Further, the CCD 121 is connected to the measuring means 135, and the measuring means 135 is connected to the display unit 62.

Hereinafter, the measurement operation in the surface plasmon sensor according to the present embodiment will be described. Laser light source 12
0 is driven, and the light beam 140 is emitted in the state of divergent light. The light beam 140 is collimated by the collimator lens 122 and enters the polarization filter 131. The light beam 140 transmitted through the polarization filter 131 and incident on the interface 12a with p-polarized light is partially split by the half mirror 132 as a reference light beam 140R, and the remaining light transmitted through the half mirror 132 Beam 140S is interface 12a
Incident on. The light beam 140S totally reflected by the interface 12a and the reference light beam 140R reflected by the mirror 134 enter the half mirror 133 and are combined. The combined light beam 140 'is condensed by the condenser lens 124, and the aperture
After passing through 125, it is detected by the CCD 121. At this time, the light beam 140 ′ detected by the CCD 121 is
An interference fringe is generated according to the state of interference between 140S and reference light beam 140R.

Here, it is determined whether or not the sensing substance 14 fixed on the surface of the metal film 12 binds to the analyte in the sample solution 15, that is, whether or not the analyte is a specific substance that binds to the sensing substance. Whether or not there is a sample can be determined by continuously measuring after dropping the sample liquid 15 and detecting a change in interference fringes detected by the CCD 121.

That is, since the refractive index of the sensing substance 14 changes according to the binding state between the analyte in the sample solution 15 and the sensing substance 14, the light beam totally reflected at the interface 12a
140S and reference light beam 140R are half mirror 13
When the signal is synthesized by 3, the state of interference changes.
The presence or absence of a binding reaction can be detected according to the change in the interference fringes. The measuring means 135 detects the presence or absence of the above-described reaction based on the above principle, and the result is displayed on the display unit 62.

In order to make such a determination, it is necessary to perform measurement for one measurement unit 10 a plurality of times.
In this case, if the measurement unit 10 is supported on the turntable 20 as it is even after the first measurement, the measurement unit 10 can be measured again by the rotational movement of the turntable 20. As described above, when the measurement of one measurement unit 10 is performed for a long time, the concentration of the sample liquid 15 may change due to evaporation of the sample liquid 15 in particular. Since the surface of the liquid is covered, evaporation of the sample liquid is prevented, and the measurement accuracy of the change over time in the total reflection state is improved.

In the present embodiment, modifications similar to those of the first embodiment can be applied, and the same effects can be obtained. Furthermore, a form similar to the second embodiment and the third embodiment is also possible, and similar effects can be obtained. Further, similarly to the fourth embodiment, a sensor using a measurement unit array having a plurality of sample liquid holding units can be configured.

[Brief description of the drawings]

FIG. 1 is an overall view of a surface plasmon sensor according to a first embodiment of the present invention.

FIG. 2 is a partially cutaway side view showing a main part of the surface plasmon sensor of FIG. 1;

FIG. 3 is a graph showing a schematic relationship between an incident angle θ of a light beam and a reflected light intensity I in the surface plasmon sensor.

FIG. 4 is an overall view of a surface plasmon sensor according to a second embodiment of the present invention.

FIG. 5 is a top view and a side view showing the shape of the lid of FIG. 4;

FIG. 6 is a top view and a side view showing the shape of a modified example of the lid.

FIG. 7 is a top view and a side view showing the shape of a modified example of a lid.

FIG. 8 is a side view showing a modified example of the measuring unit.

FIG. 9 is an overall view of a surface plasmon sensor according to a third embodiment of the present invention.

10 is a top view and a side view showing the shape of the lid of FIG. 9;

FIG. 11 is a top view and a side view showing a shape of a modified example of a lid.

FIG. 12 is a top view and a side view showing a shape of a modified example of a lid.

FIG. 13 is a top view and a side view showing shapes of modified examples of the measurement unit and the lid.

FIG. 14 is an overall view of a surface plasmon sensor according to a fourth embodiment of the present invention.

15 is a partially cutaway side view showing a main part of FIG. 14;

FIG. 16 is a schematic configuration diagram of a sheet feeding mechanism.

FIG. 17 is a schematic configuration diagram of a lid supply mechanism.

FIG. 18 is a schematic configuration diagram of a resin sheet supply mechanism.

FIG. 19 is a schematic configuration diagram of a viscous fluid supply mechanism.

FIG. 20 is a partially cutaway side view showing a main part of a leakage mode sensor according to a fifth embodiment of the present invention.

FIG. 21 is a partially broken side view showing a main part of a surface plasmon sensor according to a sixth embodiment of the present invention.

[Explanation of symbols]

 10, 90, 97 Measurement unit 10A, 10B, 10C ... Measurement unit 11 Dielectric block 12 Metal film 12a Interface between dielectric block and metal film 13 Sample liquid holding frame 13a Opening 13b Sample liquid holding unit 14 Sensing substance 15 Sample Liquid 20 Turntable 21 Support driving means 30, 30A, 30B, 30C Light beam 31, 31A, 31B, 31C Laser light source 32, 32A, 32B, 32C Light collecting lens 40, 40A, 40B, 40C Light detection Container 50 Oil 45, 84, 87 Lid supply mechanism 46, 51, 53, 57, 58, 102, 103 Lid 47 Lid supply 61, 61A, 61B, 61C ... Measuring means 62 Display 64, 74 Sample liquid Supply mechanism 65 Access mechanism 68 Measurement unit array 75 Sheet supply mechanism 77 Cover sheet 80 Oil supply mechanism 91 Cladding layer 91a Interface between dielectric block and cladding layer 92 Optical waveguide layer

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) G01N 35/04 G01N 35/04 A 35/10 35/06 A (72) Inventor Masayuki Naya Ashigara-gun, Kanagawa 798, Miyagi, Kaisei-cho Fuji Photo Film Co., Ltd. 798 No. Fuji Photo Film Co., Ltd. F-term (reference) 2G057 AA02 AB04 AB07 AC01 BA01 BB01 BB06 BC07 HA04 2G058 AA09 CC17 CD04 CF12 EA02 ED03 GA02 2G059 AA02 BB04 BB12 CC16 DD12 DD13 EE01 EJ02 EJ01 GG03 PP04

Claims (13)

[Claims] 1. A light source for generating a light beam, a dielectric block transparent to the light beam, a thin film layer formed on an upper surface of the dielectric block, and one holding a sample liquid on the thin film layer Alternatively, a measurement chip including a plurality of sample liquid holding units, and various light incident on the dielectric block so that total reflection conditions can be obtained at an interface between the dielectric block and the thin film layer. An optical system for incidence at an angle, light detecting means for detecting the intensity of the light beam totally reflected at the interface, and measuring means for measuring a state of attenuated total reflection based on a detection result of the light detecting means. In the sensor using the attenuated total reflection, the sample liquid holding part of the measurement chip has an opening at an upper part, and the opening means has a lid means supply means for providing a lid means for preventing evaporation of the sample liquid. Having prepared Sensor utilizing attenuated total reflection, characterized. 2. A sensing substance that interacts with the sample liquid is disposed on the thin film, and the measuring unit performs total reflection based on a plurality of detection results detected by the light detecting unit at intervals. 2. A sensor using attenuated total reflection according to claim 1, wherein the sensor measures a change with time in the state of attenuation. 3. The apparatus according to claim 1, wherein said cover means is oil, and said cover means supply means supplies the oil dropwise to said opening. sensor. 4. The total reflection according to claim 1, wherein said lid means is a lid, and said lid supply means covers said opening with said lid. Sensor using attenuation. 5. The sensor using attenuated total reflection according to claim 4, wherein said lid has a small opening smaller than said opening. 6. The sensor using attenuated total reflection according to claim 4, wherein said lid is formed in a stretchable blind shape. 7. The sensor using attenuated total reflection according to claim 4, wherein said lid is formed in a sheet shape. 8. The amount of water evaporation per hour in the sample liquid holding unit provided with the lid means in the opening is 2 hours.
%. The sensor using attenuated total reflection according to any one of claims 1 to 7, wherein 9. A dielectric block, a thin film layer formed on an upper surface of the dielectric block, and one or a plurality of sample liquid holding portions that hold a sample liquid on the thin film layer and have an opening. A measurement chip assembly comprising: a measurement chip formed and used for a sensor utilizing the attenuated total reflection; and a lid provided at an opening of the measurement chip. 10. The apparatus according to claim 9, wherein the lid has a small opening smaller than the opening.
The measurement chip assembly as described. 11. The measuring chip assembly according to claim 9, wherein said lid is formed in a stretchable cord shape. 12. The measuring chip assembly according to claim 9, wherein said lid is formed in a sheet shape. 13. The amount of water evaporation per hour in the sample liquid holding section in which the lid is provided in the opening is 2
%. The measurement chip assembly according to claim 9, wherein