JP2001330560A - Measuring method using total reflection attenuation and its device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To measure a large number of samples in a short time in a measuring device using total reflection attenuation such as a surface plasmon resonance measuring device or the like. SOLUTION: In this measuring device, a measuring unit 10 is equipped with a dielectric block, a metal layer formed on a surface of the dielectric block and a holding mechanism for the sample. A plurality of the measuring units 10 are supported by a supporting body 20, which is moved by a driving means 50 for the supporting body, and are sequentially sent to a measuring part. The measuring part comprises an optical system (such as a focusing lens 32) and a light detecting means 40 in which the optical system applies a light beam 30 emitted from a light source 31 to the measuring unit A condition of surface plasmon resonance is detected by measuring intensity of the light beam 30, which is subjected to total internal reflection on an interface between the dielectric block and the metal layer.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a measuring apparatus utilizing attenuated total reflection, such as a surface plasmon resonance measuring apparatus for quantitatively analyzing a substance in a sample by utilizing the generation of surface plasmons. The present invention relates to a measuring apparatus using attenuated total reflection which can measure a large number of samples in a short time.

[0002] The present invention also relates to a measuring method using a measuring apparatus utilizing such attenuated total reflection.

[0003]

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.

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

A surface plasmon resonance measuring apparatus using the above system basically includes, for example, a dielectric block formed in a prism shape, a metal film formed on one surface of the dielectric block and brought into contact with a sample, A light source for generating a beam, and the light beam is applied to a dielectric block so that various incident angles including a surface plasmon resonance condition can be obtained under a condition of total reflection at an interface between the dielectric block and the metal film. And an optical detection means for measuring the intensity of the light beam totally reflected at the interface to detect the state of surface plasmon resonance.

In order to obtain various angles of incidence as described above, a relatively narrow light beam may be deflected to be incident on the interface, or a component which is incident on the light beam at various angles may be included. As described above, a relatively thick light beam may be incident so as to be focused at the interface. In the former case, the light beam whose reflection angle changes with the deflection of the light beam is detected by a small photodetector that moves synchronously with the deflection of the light beam, or by an area sensor extending along the direction of the change in the reflection angle. Can be detected. 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.

[0007] In the surface plasmon resonance measurement device with the above structure, has when a light beam impinges the total reflection angle or more specific angle of incidence theta _SP the metal film, the electric field distribution in the sample in contact with the metal film An evanescent wave is generated, and surface plasmon is excited at the interface between the metal film and the sample by the evanescent wave. 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, both are in a resonance state, and the light energy is transferred to the surface plasmon, so that all of them are at the interface between the dielectric block and the metal film. 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.

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.

If the wave number of the surface plasmon is known from the incident angle θ _{SP at} which the attenuated total reflection (ATR) occurs, the dielectric constant of the sample can be obtained. That is, the wave number of the surface plasmon is K
_{When SP} and the angular frequency of the surface plasmon are ω, c is the speed of light in vacuum, ε _m and ε _s are the metal and the dielectric constant of the sample, respectively, there is the following relationship.

[0010]

(Equation 1) If the dielectric constant ε _s of the sample is known, the concentration of the specific substance in the sample can be determined based on a predetermined calibration curve or the like, so that the incident angle (attenuated total reflection angle) θ _{SP at which the} reflected light intensity decreases eventually
, A specific substance in the sample can be quantitatively analyzed.

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 leak mode sensor is basically formed of, for example, a dielectric block formed in a prism shape, a clad layer formed on one surface of the dielectric block, and formed on the clad layer and brought into contact with a sample. An optical waveguide layer, a light source for generating a light beam, and the light beam with respect to the dielectric block, a condition of total reflection is obtained at an interface between the dielectric block and the cladding layer, and the light is guided by the optical waveguide layer. An optical system that enters at various angles so that total reflection attenuation can be caused by excitation of the wave mode, and the intensity of the light beam totally reflected at the interface is measured to determine the excitation state of the waveguide mode, that is, the total reflection attenuation state. And a light detecting means for detecting.

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 on the optical waveguide layer, the refractive index of the sample and the characteristics of the sample related thereto are analyzed by knowing the specific incident angle at which the total reflection attenuation occurs. be able to.

The surface plasmon resonance measurement device and the leak mode sensor described above are used in the field of drug discovery research and the like.
It is sometimes used for random screening to find a specific substance that binds to a predetermined sensing substance. In this case, a sensing substance is fixed as a substance to be measured on the thin film layer (a metal film in the case of a surface plasmon resonance measuring apparatus, and a cladding layer and an optical waveguide layer in the case of a leakage mode measuring apparatus). The sample liquid containing the subject is dropped on the top, and the above-mentioned total reflection attenuation angle θ _SP 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 total reflection attenuation angle θ _SP is measured at every elapse of a predetermined time, the binding state between the subject and the sensing substance is measured based on the value, and based on the result, the specific substance is combined with the sensing substance based on the result. It can be determined whether or not there is. Examples of such a combination of the specific substance and the sensing substance include an antigen and an antibody. As a specific measurement relating to such a substance, for example, a rabbit anti-human IgG antibody is used as the sensing substance, And the determination of the presence or absence of binding to human IgG antibodies in the sample and the quantification thereof.

In order to measure the binding state between the subject and the sensing substance, it is not always necessary to detect the total reflection attenuation angle θ _SP itself. For example, it is also possible to add a sample liquid to a sensing substance, measure the angle change of the total reflection attenuation angle θ _SP thereafter, and measure the binding state based on the magnitude of the angle change.

[0016]

In a conventional measuring apparatus utilizing total reflection attenuation, such as the above-mentioned surface plasmon resonance measuring apparatus and leak mode sensor, it is difficult to measure a large number of samples. There is a problem that it takes time. In particular, when a measurement is performed several times at a time interval on one sample in order to detect a change in sample properties due to, for example, an antigen-antibody reaction or a chemical reaction, the measurement on the one sample is completed. If not, it will not be possible to measure a new sample, and it will take a very long time to measure the entire sample.

The present invention has been made in view of the above circumstances, and has as its object to provide a measuring apparatus utilizing attenuated total reflection which can measure a large number of samples in a short time.

The present invention also provides a method for measuring total reflection attenuation, which can efficiently perform measurement on a large number of samples in a short time, especially when one measurement is performed several times at intervals. The purpose is to provide a measurement method used.

[0019]

According to one aspect of the present invention, there is provided a measuring apparatus utilizing attenuated total reflection, comprising: a dielectric block as described above; a thin film formed on one surface of the dielectric block; A plurality of measurement units each including a sample holding mechanism for holding a sample on the surface of the sample, a support supporting the plurality of measurement units, a light source for generating a light beam, and the light beam to the dielectric block. On the other hand, an optical system to be incident at various angles of incidence so as to obtain a total reflection condition at the interface between the dielectric block and the thin film layer, and measuring the intensity of the light beam totally reflected at the interface,
A light detecting means for detecting a state of attenuated total reflection; and the support, the optical system, and the light detector so that the total reflection condition and various incident angles are sequentially obtained for each dielectric block of the plurality of measurement units. Relative to the means,
And a drive unit for sequentially arranging each measurement unit at a predetermined position with respect to the optical system and the light detection unit.

Further, another measuring apparatus utilizing attenuated total reflection according to the present invention comprises a dielectric block, a thin film layer formed on one surface of the dielectric block, A plurality of measurement units each including a sensing substance that interacts with a specific component and a sample holding mechanism for holding a sample on the surface of the sensing substance; a support supporting the plurality of measurement units; and a light beam. A light source to be generated, an optical system for causing the light beam to enter the dielectric block at various angles of incidence so as to obtain a total reflection condition at an interface between the dielectric block and the thin film layer, and A light detecting means for measuring the intensity of the light beam totally reflected in the step (a) to detect a state of attenuated total reflection; and Driving means for relatively moving the support, the optical system and the light detecting means so as to obtain various angles of incidence, and sequentially arranging the measuring units at predetermined positions with respect to the optical system and the light detecting means. And characterized in that:

[0021] Further, another measuring apparatus utilizing attenuated total reflection according to the present invention is configured so as to perform measurement by utilizing attenuated total reflection due to the above-mentioned surface plasmon resonance. A plurality of measurement units each including a thin film layer made of a metal film formed on one surface of the dielectric block, and a sample holding mechanism for holding a sample on the surface of the thin film layer; and supporting the plurality of measurement units. A support, a light source for generating a light beam, and the light beam with respect to the dielectric block at various angles of incidence so as to obtain total reflection conditions at an interface between the dielectric block and the metal film. An optical system to be incident, light detecting means for measuring the intensity of the light beam totally reflected at the interface to detect a state of attenuated total reflection due to surface plasmon resonance, and the plurality of measurement units. As sequentially the total reflection condition and various incident angles with respect to each dielectric block of the bets is obtained,
A drive unit for relatively moving the support, the optical system and the light detection unit, and sequentially arranging each measurement unit at a predetermined position with respect to the optical system and the light detection unit. Things.

Further, another measuring apparatus using total reflection attenuation according to the present invention is configured to perform measurement using total reflection attenuation due to surface plasmon resonance in the same manner as described above. A block, a thin film layer made of a metal film formed on one surface of the dielectric block, a sensing substance arranged on the surface of the thin film layer to interact with a specific component in a sample, and a thin film layer formed on the surface of the sensing substance. A plurality of measurement units including a sample holding mechanism for holding a sample, a support supporting the plurality of measurement units, a light source that generates a light beam, and the light beam with respect to the dielectric block. An optical system for incidence at various angles of incidence so that total reflection conditions are obtained at the interface between the dielectric block and the metal film, and the intensity of the light beam totally reflected at the interface is measured. A light detecting means for detecting a state of attenuated total reflection by surface plasmon resonance, and the support so that the total reflection condition and various incident angles are sequentially obtained for each dielectric block of the plurality of measurement units. The optical system and the light detecting means are relatively moved, and a driving means for sequentially arranging each measuring unit at a predetermined position with respect to the optical system and the light detecting means is provided.

[0023] Still another measuring apparatus using attenuated total reflection according to the present invention is configured so as to perform measurement using the attenuated total reflection caused by excitation of a waveguide mode in the optical waveguide layer. A dielectric block, a cladding layer formed on one surface of the dielectric block and a thin film layer formed of an optical waveguide layer formed thereon, and a sample holding mechanism for holding a sample on the surface of the thin film layer. A plurality of measurement units provided, a support supporting the plurality of measurement units, a light source that generates a light beam, and the light beam with respect to the dielectric block, the dielectric block and the cladding layer. An optical system that is incident at various angles of incidence so that the condition of total reflection is obtained at the interface, and the intensity of the light beam totally reflected at the interface is measured, and the waveguide mode of the optical waveguide layer is measured. Light detecting means for detecting a state of attenuated total reflection, and the support and the optical system, so that the total reflection condition and various incident angles are sequentially obtained for each dielectric block of the plurality of measurement units. And a drive unit for relatively moving the light detection unit and sequentially arranging each measurement unit at a predetermined position with respect to the optical system and the light detection unit.

Further, another measuring apparatus utilizing total reflection attenuation according to the present invention is configured to perform measurement using total reflection attenuation caused by excitation of a waveguide mode in the optical waveguide layer. , A dielectric block, a cladding layer formed on one surface of the dielectric block, and a thin film layer composed of an optical waveguide layer formed thereon, which is disposed on the surface of the thin film layer and interacts with a specific component in a sample. And a plurality of measurement units including a sample holding mechanism for holding a sample on the surface of the sensing substance, and a support that supports the plurality of measurement units,
A light source that generates a light beam, and an optical system that causes the light beam to enter the dielectric block at various angles of incidence so as to obtain a total reflection condition at an interface between the dielectric block and the cladding layer. A light detecting means for measuring the intensity of the light beam totally reflected at the interface to detect a state of attenuated total reflection due to excitation of a waveguide mode in the optical waveguide layer; and a dielectric member of each of the plurality of measurement units. The support and the optical system and the light detecting means are relatively moved so that the total reflection condition and various incident angles are sequentially obtained with respect to the blocks, and each measuring unit is sequentially moved with respect to the optical system and the light detecting means. And a driving means arranged at a predetermined position.

In the measuring apparatus using attenuated total reflection of the present invention, for example, the optical system and the light detecting means are kept stationary, and the driving means moves the support. It is said.

In this case, the support is a turntable that supports the plurality of measurement units on a circumference around a rotation axis, and the driving means rotates the turntable intermittently. It is desirable that this be done.
Further, in this case, as the support, one that linearly arranges and supports the plurality of measurement units is used, and as the driving unit, the support is intermittently linearly arranged in the direction in which the plurality of measurement units are arranged. What is moved may be applied.

On the other hand, contrary to the above, the support may be kept stationary, and the driving means may move the optical system and the light detecting means.

In this case, the support supports the plurality of measurement units on a circumference, and the driving means controls the optical system and the light detecting means by the plurality of support units supported by the support. It is desirable that the device be rotated intermittently along the measurement unit. Further, in this case, as the support, one that linearly arranges and supports the plurality of measurement units is used, and as the driving unit, the optical system and the light detection unit are supported by the plurality of units supported by the support. The one that intermittently moves linearly along the measurement unit of (1) may be applied.

On the other hand, when the driving means has a rolling bearing for supporting the rotating shaft, the driving means rotates the rotating shaft in one direction to perform a series of operations for the plurality of measuring units. When the measurement is completed, the rotation axis is returned in the other direction by the same amount as the rotation amount, and then the rotation axis is rotated in the one direction for the next series of measurements. It is desirable to be done.

Further, in the measuring apparatus utilizing attenuated total reflection of the present invention, the plurality of measuring units are connected in a row by a connecting member to form a unit connected body, and the supporter connects the unit connected body. It is desirable to be configured to support.

In the measuring apparatus utilizing attenuated total reflection of the present invention, means for automatically supplying a predetermined sample is provided to each sample holding mechanism of the plurality of measuring units supported by the support. It is desirable.

Further, in the measuring apparatus utilizing attenuated total reflection of the present invention, the dielectric block of the measuring unit is fixed to the support, and the thin film layer of the measuring unit and the sample holding mechanism are integrated to form the measuring chip. It is preferable that the measuring chip is formed so as to be exchangeable with respect to the dielectric block.

When such a measuring chip is applied, a cassette accommodating a plurality of the measuring chips and a chip supply for taking out the measuring chips one by one from the cassette and supplying them in a state of being combined with the dielectric block are provided. Preferably, means are provided.

Alternatively, the dielectric block, the thin film layer and the sample holding mechanism of the measuring unit may be integrated to form a measuring chip, and the measuring chip may be formed so as to be exchangeable with respect to the support.

When the measuring chip has such a structure, a cassette containing a plurality of the measuring chips, and a chip supplying means for taking out the measuring chips one by one from the cassette and supplying the chips in a state of being supported by a supporter Is desirably provided.

On the other hand, the optical system is configured to make the light beam enter the dielectric block in the form of convergent light or divergent light, and the light detecting means exists in the totally reflected light beam. It is desirable to be configured to detect the position of a dark line due to attenuated total reflection.

Preferably, the optical system is configured to cause the light beam to enter the interface in a defocused state. In such a case, it is desirable that the beam diameter in the moving direction of the support at the interface of the light beam be 10 times or more the mechanical positioning accuracy of the support.

Further, in the measuring apparatus utilizing attenuated total reflection of the present invention, the measuring unit is supported above the support, and the light source emits the light beam downward from a position above the support. It is preferable that the optical system is provided with a reflecting member that reflects the light beam emitted downward and that travels toward the interface.

In the measuring apparatus utilizing attenuated total reflection according to the present invention, the measuring unit is supported above the support, and the optical system makes the light beam incident on the interface from below the interface. And the light detection means is disposed with the light detection surface facing downward at a position above the support, and reflects upward the light beam totally reflected at the interface, It is desirable to provide a reflecting member that advances toward the light detecting means.

On the other hand, in the measuring apparatus utilizing the attenuated total reflection according to the present invention, the temperature for maintaining the measuring unit before and / or after being supported by the support at a predetermined set temperature. Preferably, an adjusting means is provided.

Further, in the measuring apparatus utilizing attenuated total reflection according to the present invention, the sample stored in the sample holding mechanism of the measuring unit supported by the support member is detected before the state of the attenuated total reflection is detected. It is desirable to provide a means for stirring.

Further, in the measuring apparatus utilizing the attenuated total reflection according to the present invention, at least one of the plurality of measuring units supported by the support is provided with a reference liquid having optical characteristics related to the optical characteristics of the sample. A reference liquid supply unit for supplying the reference liquid is provided, and the data indicating the state of the total reflection attenuation regarding the sample obtained by the light detection unit is based on the data indicating the state of the total reflection attenuation regarding the reference liquid. It is desirable to provide a correction means for correcting.

In such a case, if the sample is prepared by dissolving the sample in a solvent, it is preferable that the reference liquid supply means supplies the solvent as a reference liquid.

Further, the measuring apparatus using the attenuated total reflection according to the present invention comprises: a mark provided to each of the measuring units, the mark indicating individual identification information; and reading means for reading the mark from the measuring unit used for measurement. Input means for inputting sample information about the sample supplied to the measurement unit,
A display unit for displaying the measurement result, connected to the display unit, the input unit, and the reading unit, and storing the individual identification information and the sample information of each measurement unit in association with each other; Control means for displaying the measurement result obtained for the sample held in the display unit on the display means in association with the individual identification information and the sample information stored for the measurement unit.

A measuring method using attenuated total reflection according to the present invention uses the above-described measuring apparatus utilizing attenuated total reflection according to the present invention.
After detecting the state of attenuated total reflection with respect to the sample in one, the support and the optical system and the light detection means are relatively moved to detect the state of attenuated total reflection with respect to the sample in another measurement unit, and thereafter, It is characterized in that the support, the optical system and the light detecting means are relatively moved, and the state of total reflection attenuation is detected again for the sample in the one measurement unit.

[0046]

The measuring apparatus using the total reflection attenuation according to the present invention is a dielectric block and a thin film layer (a metal film in the case of using surface plasmon resonance) as described above. And a plurality of measurement units provided with a sample holding mechanism are supported on a support, and the support, the optical system, and the light detecting means are connected to each other. Since the measurement unit is moved relative to each other so that each measurement unit can be sequentially arranged at a predetermined position with respect to the measurement optical system and the light detection means, the sample held by each sample holding mechanism of the plurality of measurement units is supported by the above-described support. Measurements can be made one after another as the body moves. Thus, according to the measuring apparatus using the attenuated total reflection of the present invention, it is possible to perform measurement on a large number of samples in a short time.

More specifically, as described above, a turntable that supports a plurality of measurement units on a circumference around the rotation axis is used as a support, and the turntable is intermittently rotated by driving means. In the case where a plurality of measurement units are linearly arranged in a line and supported as a support, the support is intermittently linearly moved in a direction in which the plurality of measurement units are arranged by a driving unit. In the case of a configuration in which the sample is moved, measurement of a large number of samples can be performed very efficiently.

Contrary to the above, a support for supporting a plurality of measuring units on the circumference is used, and the optical system and the light detecting means are intermittently rotated by the driving means along the plurality of measuring units. In the case where the optical system and the light detection unit are intermittently driven along the plurality of measurement units by a driving unit, or by using a support for linearly arranging a plurality of measurement units and supporting the plurality of measurement units. Even in the case of a linear movement, it is possible to measure a large number of samples very efficiently.

In the measuring apparatus using the attenuated total reflection according to the present invention, a sensing substance which interacts with a specific component in the sample is arranged on the surface of the thin film layer, and the sample is placed on the surface of the sensing substance. In the configuration in which the sample is held, the above-described interaction changes the state of surface plasmon resonance or the excited state of the guided mode, that is, the state of attenuated total reflection. And a specific reaction between the substance and the sensing substance can be detected.

On the other hand, when the driving means has a rolling bearing for supporting the rotating shaft, the driving means rotates the rotating shaft in one direction to perform a series of operations for the plurality of measuring units. When the measurement is completed, the rotation axis is returned in the other direction by the same amount as the rotation amount, and then the rotation axis is rotated in the one direction for the next series of measurements. If a certain measuring unit is arranged at the measuring position, the revolving position of the rolling element of the rolling bearing is always constant. Thus, it is possible to prevent a decrease in measurement accuracy due to the fact that the revolving positions of the rolling elements are different at the time of measurement.

In the case where a plurality of measuring units are connected in a row by a connecting member to form a unit connected body, and the support is configured to support this unit connected body, Since the position accuracy of the above measurement unit is easily obtained, and since there is no need to hold a small measurement unit, it is excellent in handling properties and can contribute to improvement in the efficiency of measurement processing.

In the measuring apparatus utilizing attenuated total reflection according to the present invention, when means for automatically supplying a sample to each sample holding mechanism of a plurality of measurement units supported by a support is provided, the sample supply is performed. The time required is also shortened, so that measurements on a large number of samples can be performed in a shorter time.

In the measuring apparatus utilizing attenuated total reflection of the present invention, the dielectric block of the measuring unit is fixed to the support, and the thin film layer of the measuring unit and the sample holding mechanism are integrated to constitute a measuring chip. If this measurement chip is formed so as to be replaceable with the dielectric block, remove the measurement unit holding the sample whose measurement has been completed from the dielectric block, and combine a new measurement unit with the dielectric block. Accordingly, new samples can be successively provided for measurement, and measurement of a large number of samples can be performed in a shorter time.

When such a measuring chip is applied, a cassette accommodating a plurality of the measuring chips and chip supply means for taking out the measuring chips one by one from the cassette and supplying the chips in a state of being combined with the dielectric block. Is provided, the work of supplying the measuring chip is streamlined,
The time required for measuring a large number of samples can be further reduced.

Further, when the dielectric block, the thin film layer and the sample holding mechanism of the measuring unit are integrated to form a measuring chip, and the measuring chip is formed so as to be exchangeable with respect to the support, measurement can be performed. By removing the measurement unit holding the completed sample from the support and supporting a new measurement unit on the support, new samples can be used for measurement one after another, and the measurement of many samples can be further improved. This can be done in a short time.

When such a measuring chip is applied, a cassette accommodating a plurality of the measuring chips, and a chip supply means for taking out the measuring chips one by one from the cassette and supplying the chips in a state supported by a supporter Is provided, the work of supplying the measurement chip is streamlined also in this case, and the time required for measuring a large number of samples can be further reduced.

When the above-mentioned support is mechanically moved by the driving means, it is inevitable that a positioning error occurs. When this positioning error occurs, the relative stop position of the measuring unit supported thereon relative to the optical system fluctuates. If the position of the measurement unit with respect to the optical system fluctuates in this manner, an error occurs in detecting the state of attenuated total reflection. More specifically, an error occurs in the position measurement of a dark line due to attenuation of total reflection. As a cause of the error in the state of the total reflection attenuation due to the positioning error, the film thickness of the thin film layer, the film thickness of the sensing substance, and the reaction amount between the sensing substance and the subject are uneven in position. There is something.

However, if the optical system is configured to cause the light beam to be incident on the interface in a defocused state, the error in the state of attenuated total reflection (for example, the position measurement of the dark line) is averaged. And the measurement accuracy is improved.

In such a case, if the beam diameter in the moving direction of the support at the interface of the light beam is set to 10 times or more the mechanical positioning accuracy of the support, the measurement accuracy is higher. It will be. The reason is as follows. In this case, even if the positioning error occurs,
Since the amount is at most 1/10 of the beam diameter, and the remaining 9/10 is always included in the measurement range, the signal error caused by the positioning error can be reduced to 1/10. In a general quantitative analysis of a sample, if this type of measurement error can be suppressed to 1/10, there is no practical problem.

Further, in the measuring apparatus utilizing attenuated total reflection according to the present invention, the measuring unit is supported on the upper side of the support, and the light source is arranged so as to emit a light beam downward from a position above the support. Provided, the optical system includes a reflecting member that reflects the light beam emitted downward toward the interface upward and reflects the light beam emitted downward.
There is no need to consider the interference between the moving support and the light source and the optical system, and the degree of freedom in the layout of the light source and the optical system, and further, the layout of other elements that need to be disposed close to the support is reduced. Get higher.

Further, in the measuring apparatus utilizing attenuated total reflection of the present invention, the measuring unit is supported on the upper side of the support, and the optical system is configured to make the light beam enter the interface from below the interface. The light detection means is disposed with the light detection surface facing downward at a position above the support, and
When a reflecting member that reflects the light beam totally reflected at the interface upward and travels toward the light detecting means is provided, it is necessary to consider interference between the moving support and the light detecting means. And the layout of the light detection means,
Furthermore, the layout flexibility of other elements that need to be arranged near the support is increased.

On the other hand, in the measuring apparatus utilizing attenuated total reflection of the present invention, a temperature adjusting means for maintaining the measuring unit before and / or after being supported by the support at a predetermined set temperature is provided. When provided, it is possible to prevent a decrease in measurement accuracy due to a temperature change of the sample in the measurement unit.

In the measuring apparatus utilizing attenuated total reflection according to the present invention, means for stirring the sample stored in the sample holding mechanism of the measuring unit supported by the support before detecting the state of the attenuated total reflection. Is provided, it is possible to prevent a decrease in measurement accuracy due to non-uniform concentration of the analyte in the sample.

In the measuring apparatus utilizing attenuated total reflection according to the present invention, a reference liquid having optical characteristics related to the optical characteristics of the sample is supplied to at least one of the plurality of measuring units supported by the support. Reference liquid supply means is provided, and correction means for correcting data indicating the state of attenuated total reflection regarding the sample, obtained by the light detection means, based on data indicating the state of attenuated total reflection regarding the reference liquid is provided. In this case, changes in the refractive index of the sample solvent and changes in the characteristics of the measurement optical system due to environmental conditions such as temperature can be compensated, and the characteristics of the subject in the sample can be accurately measured.

Further, in the measuring apparatus utilizing attenuated total reflection according to the present invention, a mark indicating individual identification information provided to each of the measuring units, and a reading means for reading the mark from the measuring unit used for measurement are provided. Input means for inputting sample information relating to the sample supplied to the measurement unit, display means for displaying the measurement result, and this display means,
Connected to the input means and the reading means, and stores the individual identification information for each measurement unit and the sample information in association with each other, the measurement result obtained for a sample held in a certain measurement unit, When there is provided control means for displaying on the display means in association with the individual identification information and the sample information stored for the measurement unit, the individual identification information of the measurement unit, the sample information, and the measurement result are provided. By being managed in association with each other, it is possible to prevent the measurement from being performed by mistake in the combination of the measurement unit and the sample, or to prevent the measurement result of one sample from being mistakenly displayed as the measurement result of another sample. .

On the other hand, in the measuring method using attenuated total reflection according to the present invention, after detecting the state of attenuated total reflection with respect to the sample in one of the measuring units, the support and the optical system and the light detecting means are moved relative to each other. Then, the state of attenuated total reflection is similarly detected with respect to the sample in another measurement unit, and thereafter, the support and the optical system and the light detection means are relatively moved, so that the total of the sample in the one measurement unit is again measured. Since the state of return loss is detected, if several measurements are made at one time interval for one sample, use the time between those measurements to measure another sample. This makes it possible to efficiently measure a large number of samples in a short time.

[0067]

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

As shown in FIG. 1, the surface plasmon resonance measuring apparatus generates a plurality of measurement units 10, a turntable 20 supporting the plurality of measurement units 10, and a light beam (laser beam) 30 for measurement. A laser light source 31 such as a semiconductor laser and a condenser lens 32 forming an incident optical system
A photodetector 40; a support driving means 50 for intermittently rotating the turntable 20; and a drive signal for controlling the driving of the support driving means 50 and the output signal S of the photodetector 40.
The controller 60 includes a controller 60 that performs a process described below in response to the request, and an automatic sample supply mechanism 70.

As shown in FIG. 2, the measuring unit 10 includes, for example, a transparent dielectric block 11 formed in a rectangular parallelepiped shape, and gold, for example, formed on the upper surface of the dielectric block 11.
It comprises a metal film 12 made of silver, copper, aluminum or the like, and a sample holding frame 13 made of a cylindrical member defining a space closed on the side on the metal film 12. In the sample holding frame 13, for example, a liquid sample 15
Is stored.

The measuring unit 10 is provided with a dielectric block 11
The sample holding frame 13 and the sample holding frame 13 are integrally formed from, for example, a transparent resin or the like. To be exchangeable,
For example, the measurement unit 10 may be fitted and held in a through hole formed in the turntable 20. In this example, the sensing substance 14 is fixed on the metal film 12, which will be described later in detail.

A plurality of turntables 20 (11 in this example)
) Of the measuring units 10 are supported at equal angular intervals on a circumference around the rotation shaft 20a. The support driving means 50 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 measurement unit 10.

The condenser lens 32 has a light beam as shown in FIG.
The light 30 is condensed and passed through the dielectric block 11 in a convergent light state, and is incident on the interface 11a 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 for total reflection of the light beam 30 at the interface 11a is obtained and surface plasmon resonance can occur.

The light beam 30 enters the interface 11a 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 composed of a line sensor in which a large number of light receiving elements are arranged in one row, and the light receiving elements are arranged such that the arrangement direction of the light receiving elements is 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 50 based on a predetermined sequence. The controller 60 includes a signal processing unit 61 that receives the output signal S of the photodetector 40, and a display unit 62 that receives an output from the signal processing unit 61.

The automatic sample supply mechanism 70 includes, for example, a pipette 71 for sucking and holding a predetermined amount of a liquid sample, and a pipette 71.
Means for moving the sample from the sample container 73 set at a predetermined position to the pipette 71 by suction and holding the sample in the sample holding frame 13 of the measurement unit 10 at a predetermined stop position. The sample is supplied dropwise.

Hereinafter, a sample analysis by the surface plasmon resonance measuring apparatus having the above configuration will be described. At the time of sample analysis, the turntable 20 is provided with the support driving means 50 as described above.
Is rotated intermittently. And turntable
The sample automatic supply mechanism 70 puts the sample on the sample holding frame 13 of the measurement unit
15 is supplied.

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

Since the light beam 30 is incident on the dielectric block 11 in a convergent light state as described above, it includes components incident on the interface 11a 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 11a, 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 11a to the metal film 12 side. When the light beam 30 enters the interface 11a 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.

Therefore, the light amount detection signal output from the photodetector 40 is
Check the amount of light detected for each light receiving element from signal S to detect dark lines.
The incident angle (total reflection attenuation)
Angle) θ _SPAnd enter the reflected light intensity I obtained in advance.
Based on the relationship curve with the angle of incidence θ, the specific substance in sample 15
Quantitative analysis can be performed. Controller 60 signal processing
The part 61 determines the specific substance in the sample 15 based on the above principle.
The quantity is analyzed, and the analysis result is displayed on the display unit 62.

When the measurement is performed only once for one sample 15, the measurement is completed by the above operation. Therefore, the measurement unit 10 having completed the measurement is manually operated from the turntable 20, or the automatic discharging means is provided. It may be used and discharged. On the other hand, when performing measurements on one sample 15 multiple times,
Turn table of measurement unit 10 as it is after measurement
If the sample 15 is held by the turntable 20, the sample 15 held by the measurement unit 10 can be measured again after one turn of the turntable 20.

As described above, this surface plasmon resonance measurement apparatus includes a plurality of measurement units 10
Since each turntable 20 is configured to move the turntable 20 to sequentially arrange each measurement unit 10 at the measurement position, the sample 15 held in each sample holding frame 13 of the plurality of measurement units 10 is With the movement of the turntable 20, the measurement can be provided one after another. Thus, according to the surface plasmon resonance measurement device, it is possible to perform measurement on a large number of samples 15 in a short time.

In the surface plasmon resonance measuring apparatus according to the present embodiment, the time required for supplying the sample is reduced by providing the automatic sample supply mechanism 70, and the measurement of a large number of samples 15 is performed in a shorter time. It becomes possible.

In the present embodiment, the dielectric blocks 11,
Measurement unit by integrating metal film 12 and sample holding frame 13
Since the measurement unit 10 is configured as a measurement chip and can be replaced with the turntable 20, the measurement unit 10 holding the sample 15 whose measurement has been completed is removed from the turntable 20 and a new measurement unit 10 is formed. Is supported by the turntable 20 so that a new sample
15 can be successively used for measurement, and the measurement of a large number of samples 15 can be performed in a shorter time.

The sensing substance 14 fixed on the surface of the metal film 12 binds to a specific substance in the sample 15. Examples of such a combination of the specific substance and the sensing substance 14 include an antigen and an antibody. In that case, the antigen-antibody reaction can be detected based on the total reflection attenuation angle θ _SP .

In other words, in this case, the refractive index of the sensing substance 14 changes in accordance with the binding state between the specific substance and the sensing substance 14, and the characteristic curve of FIG. An antigen-antibody reaction can be detected according to the total reflection attenuation angle θ _SP . In this case, the sample
Both 15 and the sensing substance 14 are samples to be analyzed.

Next, a second embodiment of the present invention will be described with reference to FIG. FIG. 4 shows a side surface shape of a main part of the surface plasmon resonance measuring apparatus according to the second embodiment. In FIG. 4, the same elements as those in FIG. 2 are denoted by the same reference numerals, and a description thereof will be omitted unless otherwise required (the same applies hereinafter).

This surface plasmon resonance measuring apparatus is shown in FIG.
The configuration of the measurement unit is different from that shown in FIG. That is, the measurement unit 10 'used here is composed of the dielectric block 11' fixed to the turntable 20, the sample holding frame 13 'and the metal film 12 integrated with each other. The sample holding frame 13 'is formed in a cylindrical shape with a bottom using a transparent dielectric, and the metal film 12 is fixed on the bottom surface thereof, and the both constitute a measurement chip.

The measuring chip is a dielectric block.
It can be removed as needed from 11 'and replaced. When combining the measurement chip with the dielectric block 11 ',
Preferably, a refractive index matching oil is interposed between the sample holding frame 13 'and the dielectric block 11'. Also, in this case, the sample holding frame 13 'is moved together with the dielectric block 11' to 1
One dielectric block is formed, and an interface 13a between the sample holding frame 13 'and the metal film 12 is irradiated with the light beam 30.

When using the measuring unit 10 'having the above structure, the measuring chip holding the sample 15 whose measurement has been completed is removed from the dielectric block 11', and a new measuring chip is combined with the dielectric block 11 '. By doing
New samples 15 can be successively used for measurement, and measurement of many samples 15 can be performed in a short time.

Next, a third embodiment of the present invention will be described with reference to FIG. FIG. 5 shows a side surface shape of a main part of the surface plasmon resonance measuring apparatus according to the third embodiment. This surface plasmon resonance measurement apparatus differs from the apparatus shown in FIG. 2 in the manner of irradiating the light beam 30 to the interface 11a between the dielectric block 11 and the metal film 12. That is, in the present apparatus, the light beam 30 condensed by the condenser lens 32 is applied to the interface 11a in a defocused state.

Since the turntable 20 is mechanically moved by the support driving means 50, it is inevitable that a positioning error occurs. When this positioning error occurs, the relative stop position of the measuring unit 10 supported thereon relative to the light beam 30 emitted from the condenser lens 32 fluctuates. If the position of the measurement unit 10 with respect to the light beam 30 fluctuates, an error occurs in detecting the state of surface plasmon resonance. More specifically, an error occurs in the position measurement of the dark line due to the total reflection attenuation described above.

However, if the light beam 30 is incident on the interface 11a in a defocused state as described above, the error of the state detection of the surface plasmon resonance (here, the position measurement of the dark line) is averaged. And the measurement accuracy is improved.

As in this embodiment, the light beam 30 is applied to the interface 11a.
When the light is incident on the interface 11a of the light beam 30 in the defocused state, the beam diameter in the moving direction of the turntable 20 at the interface 11a of the light beam 30 is one of the mechanical positioning accuracy of the turntable 20
It is desirable to set it to 0 times or more. The reason is as detailed above.

Next, a fourth embodiment of the present invention will be described with reference to FIGS. FIGS. 6 and 7 show a side surface shape and a plane shape of the surface plasmon resonance measuring apparatus of the fourth embodiment, respectively, and FIG. 8 shows a side surface shape of a main part thereof.

In this surface plasmon resonance measurement apparatus, as shown in FIGS.
The four measurement units 80 are supported at an angular interval of, and the turntable 20 is intermittently rotated in the direction of arrow R by 90 °. Therefore, when the turntable 20 stops, each measurement unit support of the turntable 20 stops at four positions sequentially. These positions are referred to as a measurement unit supply position P1, a sample supply position P2, a measurement position P3, and a measurement unit discharge position P4, respectively. The measurement unit 80 will be described later in detail.

The measurement unit 80 taken out by the chip supply means 76 is supplied to the measurement unit supporting portion which is stationary at the measurement unit supply position P1 from the cassette 75 accommodating a plurality of measurement units 80. The chip supply means 76 is constituted by a known air suction cup and a mechanism for moving the same, such as a cassette.
The measuring units 80 can be sucked and taken out one by one from a take-out port provided at the bottom of the base 75, and can be moved to be supplied to the measuring unit support.

The sample is supplied to the measurement unit 80 supported by the measurement unit support portion that is stationary at the sample supply position P2 by using the automatic sample supply mechanism 70.

Next, with respect to the measuring unit 80 supported by the measuring unit supporting portion stationary at the measuring position P3,
The sample held by the measurement unit 80 is analyzed using the surface plasmon resonance measurement means 77. This sample analysis will be described later in detail with reference to FIG.

Next, the measuring unit 80 supported by the measuring unit supporting portion which is stationary at the measuring unit discharge position P4.
Is discharged from the turntable 20 by the chip discharging means 78. When the turntable 20 is rotated 90 ° next time, the measurement unit support vacated in this way stops at the measurement unit supply position P1, and thereafter, the same processing as described above is repeated.

Hereinafter, a sample analysis by the surface plasmon resonance measuring means 77 will be described with reference to FIG. As shown here, the measurement unit 80 has a transparent dielectric block 81, a metal film 82, and a sample holding frame 83 integrally formed as in the measurement unit 10 described above.

The light beam 30 emitted from the laser light source 31 is condensed by the condenser lens 90, reflected by the mirror 91, and enters the interface 81a between the dielectric block 81 and the metal film 82. The light beam 30 totally reflected at the interface 81a is reflected by a mirror 92, collimated by a collimator lens 93, and received by a photodetector 40. The output signal S of the photodetector 40 is
The signal is input to a signal processing unit 61 (see FIG. 6) and provided for sample analysis. The sample analysis based on this output signal S
This is performed in the same manner as described in the first embodiment.

As described above, in this embodiment,
Chip supply means for taking out the measuring units 80 one by one from a cassette 75 containing a plurality of measuring units 80 serving as measuring chips and supplying the measuring units 80 in a state supported by the turntable 20
With the provision of the electrodes 76, the operation of supplying the measurement chips is streamlined, and the time required for measuring a large number of samples 15 can be sufficiently reduced.

As described above, the transparent dielectric block 81,
Not only the measurement unit 80 in which the metal film 82 and the sample holding frame 83 are integrally formed, but also the metal film 12 as shown in FIG.
Also, when applying a measurement unit 10 'in which the sample holding frame 13' is exchangeably formed with respect to the dielectric block 11 ', the metal film 12 and the sample holding frame 13' are integrated into a measurement chip, Is stored in a cassette and supplied automatically, as in the case described above, the work of supplying the measuring chip can be performed efficiently.

Further, in each of the embodiments described above, the rotating turntable 20 is used as a support for supporting the measuring unit, but the shape and the moving method of the support are not limited thereto. For example, the support supporting a plurality of measurement units is configured to reciprocate linearly,
With the movement, a plurality of measurement units may be sequentially set in the measurement unit.

In this case, when it is necessary to perform a plurality of measurements on one sample, a plurality of measuring units each including an optical system for irradiating the measuring unit with a light beam and light detecting means may be provided. For example, the measuring units are sequentially set in the measuring section with the movement of the support, and the measurement can be performed in each measuring section. Alternatively, only one such measurement unit is provided, the support is moved in one direction, the measurement unit is set on the measurement unit, and after the measurement is performed, the support is moved in the opposite direction to remove the measurement unit. Set it on the measuring section again,
The measurement may be performed.

The method of moving the support in the normal and reverse directions is applicable to the case where the turntable 20 described above is used. When this turntable 20 is used, a plurality of measuring units are provided, and one turntable 20 is provided.
It is also possible to configure so that a plurality of measurements are performed on one measurement unit during rotation.

Next, a fifth embodiment of the present invention will be described. FIG. 9 shows a side view of a main part of a measuring apparatus using attenuated total reflection according to a fifth embodiment of the present invention. In FIG. 9, the same elements as those in FIG. 2 are denoted by the same reference numerals as those in FIG.

The measuring apparatus of the present embodiment is the leak mode sensor described above, and is also configured to use the measuring unit 110 formed as a measuring chip in this example. A cladding layer 111 is formed on one surface (upper surface in the figure) of the dielectric block 11 constituting the measurement unit 110, and an optical waveguide layer 112 is 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 111 is formed in a thin film using a dielectric material having a lower refractive index than the dielectric block 11 or a metal such as gold. The optical waveguide layer 112 is also formed into a thin film using a dielectric material having a higher refractive index than the cladding layer 111, for example, PMMA. The thickness of the cladding layer 111 is, for example, 36.5 nm when formed from a gold thin film, and the thickness of the optical waveguide layer 112 is, for example, PM.
When formed from MA, the thickness is 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 111 at an incident angle equal to or greater than the total reflection angle through 11, the light beam 30 is totally reflected at the interface 11a between the dielectric block 11 and the cladding layer 111, but is transmitted through the cladding layer 111. Light having a specific wave number incident on the optical waveguide layer 112 at a specific incident angle propagates through the optical waveguide layer 112 in a guided mode. When the waveguide mode is excited in this manner, most of the incident light is taken into the optical waveguide layer 112, and thus the total reflection attenuation occurs in which the intensity of the light totally reflected at the interface 11a sharply decreases.

Since the wave number of the guided light in the optical waveguide layer 112 depends on the refractive index of the sample 15 on the optical waveguide layer 112, by knowing the specific incident angle at which the total reflection attenuation occurs,
The refractive index of the sample 15 and the characteristics of the sample 15 related thereto can be analyzed. The signal processing unit 61 quantitatively analyzes the specific substance in the sample 15 based on the above principle, and the analysis result is displayed on a display unit (not shown).

The leak mode sensor according to the present embodiment is also configured such that a plurality of measurement units 110 are supported on a turntable 20, and the turntable 20 is moved so that each measurement unit 110 is sequentially arranged at a measurement position. Thus, the samples 15 held in the sample holding frames 13 of the plurality of measurement units 110 can be used for the measurement one after another as the turntable 20 moves. Thus, according to the leak mode sensor, it is possible to perform measurement on a large number of samples 15 in a short time.

Also in the present embodiment, the dielectric block 11,
The measuring unit 110 is configured by integrating the cladding layer 111 and the optical waveguide layer 112, and the measuring unit 110 is exchangeable with the turntable 20 as a measuring chip, so that the sample 15 for which measurement has been completed is held. Measuring unit
By removing the 110 from the turntable 20 and supporting the new measurement unit 110 on the turntable 20, new samples 15 can be successively provided for measurement, and the measurement of many samples 15 can be performed in a shorter time. It becomes possible.

Next, a sixth embodiment of the present invention will be described. FIG. 10 shows a side surface shape of a main part of a measuring apparatus using attenuated total reflection according to a sixth embodiment of the present invention. In FIG. 10, the same elements as those in FIG. 9 are denoted by the same reference numerals as those in FIG.

The measuring device of the present embodiment is also a leak mode sensor, and is configured to use the measuring unit 120 formed as a measuring chip. A cladding layer 111 is formed on one surface (the upper surface in the figure) of the dielectric block 11 constituting the measurement unit 120, an optical waveguide layer 112 is formed thereon, and the sensing substance 14 is further fixed thereon. ing. The measuring apparatus of the present embodiment basically includes a measuring unit 12
The only difference from FIG. 9 is that the sensing substance 14 is fixed at 0.

The sensing substance 14 binds to a specific substance in the sample 15 similarly to the sensing substance 14 in the apparatus shown in FIG. Such specific substances and sensing substances
As a combination with 14, also in this example, for example, an antigen and an antibody can be mentioned. In that case, the total reflection attenuation angle θ _SP
The antigen-antibody reaction can be detected based on the

That is, also in this case, the interface 11a of the light beam 30
The relationship between the incident angle θ and the reflected light intensity I is basically the same as that shown in FIG. 3, but the refractive index of the sensing substance 14 varies depending on the bonding state between the specific substance and the sensing substance 14. Since the effective refractive index of the optical waveguide layer 112 changes, and this changes the relationship, the antigen-antibody reaction can be detected according to the attenuated total reflection angle θ _SP .

Next, a seventh embodiment of the present invention will be described. FIG. 11 shows a side surface shape of a main part of a measuring apparatus utilizing attenuated total reflection according to a seventh embodiment of the present invention. In FIG. 10, the same elements as those in FIGS. 1 and 2 are denoted by the same reference numerals.

The measuring apparatus according to the present embodiment is a surface plasmon resonance measuring apparatus, which is used by holding the measuring unit 10 storing the sample 15 on the measuring unit holding section 128 formed on the upper surface of the turntable 20. Is configured. The measuring unit 10 has basically the same configuration as the measuring unit 10 used in the apparatus shown in FIGS. 1 and 2 except that the shape of the dielectric block 11 is slightly different.

In the present apparatus, the point that the light beam 30 is incident on the interface 11a (see FIG. 2) between the dielectric block 11 and the metal film 12 from below is the same as the configuration in FIG. A mirror that reflects the light beam 30 totally reflected at the interface 11a
The light beam 30 that is totally reflected there and travels upward is detected by the light detector 40 disposed above the mirror 121 with the light detection surface facing downward. .

As described above, if the totally reflected light beam 30 is reflected by the mirror 121 and the photodetector 40 is disposed at a position higher than the turntable 20, the rotating turntable 20 and the light detection It is not necessary to consider the interference with the detector 40, and the layout of the photodetector 40 and the layout of other elements that need to be arranged near the turntable 20 are increased in flexibility.

Next, an eighth embodiment of the present invention will be described. FIG. 12 shows a side surface shape of a main part of a measuring apparatus using attenuated total reflection according to an eighth embodiment of the present invention. The measurement device of the present embodiment is also a surface plasmon resonance measurement device, and is different from the device of FIG. 11 in that a mirror 122 that reflects the light beam 30 is provided,
The difference is that the laser light source 31 is disposed above the mirror 122.

Thus, together with the photodetector 40, the laser light source
If the position 31 is also provided above the turntable 20, the moving turntable 20, the photodetector 40 and the laser light source 31
There is no need to consider interference with the
And the layout of both the laser light source 31, and further,
The layout flexibility of other elements that need to be arranged near the turntable 20 is increased.

The layout of the photodetector 40 and the laser light source 31 in the seventh and eighth embodiments described above is not limited to the surface plasmon resonance measuring device,
Of course, the present invention can be applied to a leak mode sensor as shown in FIG. 0, and in such a case, the same effect as described above can be obtained.

In addition, such a photodetector 40 and a laser light source
The layout of the 31 is not limited to a surface plasmon resonance measurement device or a leak mode sensor using a turntable, and as described above, a surface plasmon resonance measurement device or a leak mode Of course, the present invention can be applied to a sensor, and in such a case, the same effect as described above can be obtained.

By the way, the turntable 20 shown in FIG.
More specifically, as shown in FIG.
The motor 50 is connected to a drive source 50a such as a stepping motor via a rotation shaft 20a, and is rotated by the drive source 50a. Generally, at least one rotating shaft 20a
Usually, it is supported via a plurality of rolling bearings 130 on a shaft holding portion 50b fixed to the main body of the support driving means 50.

The above-mentioned rolling bearing 130 is provided with the rotating shaft 20a.
, An outer ring 130b fitted and fixed to the shaft holder 50b, and an inner ring 130a and an outer ring 130b
Rolling element 130c consisting of a ball or roller rolling between
(The figure shows a sphere). In the rolling bearing 130 having such a configuration, the rotating shaft 20a
Is rotated by the drive source 50a, the inner ring 130a rotates integrally therewith, and the rolling element 130c revolves while rotating along with the inner ring 130a.

Therefore, if the roundness of the rolling element 130c is inferior or the surface roughness of the inner ring 130a or the outer ring 130b is large, the inner ring 130a, Means that the radial position and inclination of the rotating shaft 20a fluctuate. In such a case, the rotation center of the turntable 20 is shifted, and its posture is inclined from a horizontal state. Turntable 20
When such a behavior occurs, the posture of the measurement unit 10 (see FIG. 2) supported thereon changes, and
The incident angle of the light beam 30 with respect to 11a fluctuates,
This can cause errors in the measurement results for 15.

Hereinafter, the rotation control of the turntable 20 for preventing such a measurement error from occurring will be described. Here, it is assumed that, for example, 16 measurement units 10 are supported on the turntable 20 at equal angular intervals, that is, at intervals of 22.5 °. Also, their measurement units 10
The position on the turntable 20 where
(Channel), 2ch, 3ch... 16ch.

Measurement unit 10 supported on each channel
As described above, when the turntable 20 is stopped, irradiation of the light beam 30 and detection of the intensity of the total reflection light are performed by the configuration shown in FIG. Hereinafter, the stop position of the measurement unit 10 that receives this operation is referred to as a “detection position”.

In this example, every 5 ch, that is, 112.5
It is assumed that the measurement unit 10 of each channel is set to the above detection position at the intermittent rotation angle of
In the first rotation of 20, 1ch → 6ch → 11ch → 16c
In the order of h, in the second rotation, 5ch → 10ch → 15ch
In the order of →, in the third rotation, 4ch → 9ch → 14ch, in the fourth rotation, 3ch → 8ch → 13ch, in the fifth rotation, 2ch → 7ch → 12ch,
The measurement unit 10 of each channel is sequentially set to the detection position. The directions of these five rotations are all the same.

Thus, when the measurement unit 10 of each channel is set at the detection position, and the irradiation of the light beam 30 and the detection of the total reflection light intensity are performed on the measurement unit 10, the sample 15 is moved to the measurement unit 10 of another channel. Another operation such as supply is performed as described above.

As described above, when the turntable 20 is intermittently rotated five times and a predetermined operation is performed on all of the 16-channel measurement units 10, the turntable 20 is then rotated in the five rotation directions. Is continuously rotated 5 times in the opposite direction. Turntable 20 intermittently 5 for measurement
While being rotated, the rolling element 130c of the rolling bearing 130 revolves on the inner ring 130a as described above, and then when the turntable 20 is continuously rotated five times in the opposite direction, the rolling element 130c becomes It is returned to the initial revolution position at the start of this series of measurements.

Therefore, when performing a plurality of measurements on one sample 15 in order to check the change over time of the characteristic shown in FIG.
If five intermittent rotations for measurement are performed after five rotations in the opposite direction of 20, when the same measurement unit 10 is set to the detection position a plurality of times, the rolling bearing 130 rotates in each rotation. The revolving positions of the moving body 130c are all the same. In such a case, the same measurement unit 10 is kept in a common posture in each measurement,
It is possible to prevent a measurement error from occurring due to a change in the incident angle of the light beam 30 with respect to the interface 11a for each measurement.

In order to prevent the occurrence of measurement errors as described above, the number of channels set on the turntable 20, the angle of intermittent rotation of the turntable 20,
The number of rotations and the like in a series of measurements are not limited to those in the example described above, and can be set as appropriate.

Further, the control of the rotation of the turntable 20 described above is not limited to the surface plasmon resonance measuring device,
Also, the present invention can be applied to the leak mode sensor as shown in FIG. 10 and FIG. 10, and in such a case, the effect of preventing the occurrence of the measurement error can be obtained as described above.

By the way, the values measured by the surface plasmon resonance measuring device and the leak mode sensor described above tend to fluctuate according to changes in conditions such as the temperature around the device.
That is, the refractive index of the sample liquid changes depending on the temperature, etc.,
This is because a minute change in characteristics occurs in the measuring optical system of the apparatus. Hereinafter, a ninth embodiment of the present invention will be described in which a variation in measured value due to such a cause can be prevented.

FIG. 14 shows a side view of a main part of a measuring apparatus utilizing attenuated total reflection according to a ninth embodiment of the present invention. The measuring device of the present embodiment is a surface plasmon resonance measuring device. For example, a dielectric block 210 having a shape in which a part of a substantially quadrangular pyramid is cut off, and one surface (the upper surface in FIG. And a metal film 212 formed of, for example, gold, silver, copper, aluminum, or the like.

The dielectric block 210 is made of, for example, a transparent resin or the like, and has a raised portion around a portion where the metal film 212 is formed. The raised portion 210 a serves as a sample holding portion for storing a liquid sample 211. Function. In this example, the sensing substance 230 is fixed on the metal film 212. The sensing substance 230 will be described later.

The dielectric block 210 and the metal film 212
A disposable measurement unit (measurement chip) 222 is formed, and is fitted and supported one by one in a plurality of chip holding holes 231a provided on a turntable 231 similar to the turntable 20 of the apparatus of FIG. After the dielectric block 210 is supported by the turntable 231 in this manner, the turntable 231 is intermittently rotated by a predetermined angle, and the liquid sample 211 is dropped onto the dielectric block 210 stopped at a predetermined position. The liquid sample 211 is held in the sample holding unit 210a. Thereafter, when the turntable 231 is further rotated by a predetermined angle, the dielectric block 210 is sent to the measurement position shown in FIG. 14 and stops there.

The surface plasmon resonance measuring apparatus according to the present embodiment includes, in addition to the dielectric block 210, a laser light source 214 composed of a semiconductor laser or the like for generating one light beam 213, An optical system 215 that passes through the block 210 and enters the interface 210b between the dielectric block 210 and the metal film 212 so that various incident angles can be obtained, and a light beam 213 totally reflected at the interface 210b is parallelized. A collimator lens 16 for lightening, a light detecting means 17 for detecting the parallel light beam 213, a differential amplifier array 18 connected to the light detecting means 17, and a driver 19
And a signal processing unit 220 composed of a computer system or the like.
And a display unit 21 connected to the signal processing unit 220.

FIG. 15 is a block diagram showing an electrical configuration of the surface plasmon resonance measuring apparatus. As shown in the figure, the driver 19 is connected to each differential amplifier 18 of the differential amplifier array 18.
a and 18b, 18c,... sample and hold the sample and hold circuits 22a, 22b, 22c,. An A / D converter 24, a multiplexer 23, and a sample-and-hold circuit 22a for digitizing the output and inputting it to the signal processing unit 220;
, And a controller 26 that controls the operation of the drive circuit 25 based on an instruction from the signal processing unit 220.

As shown in FIG. 14, a light beam 213 emitted in a divergent light state from a laser light source 214 is focused on an interface 210 b between a dielectric block 210 and a metal film 212 by the action of an optical system 215. Therefore, the light beam 213 includes components incident on the interface 210b 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 213 is totally reflected at the interface 210b, and the reflected light beam 213 contains components reflected at various reflection angles.

The light beam 213 is applied to the interface 210b by p
Inject with polarized light. In order to do so, the laser light source 214 may be disposed in advance so that the polarization direction is a predetermined direction. In addition, the direction of polarization of the light beam 213 may be controlled by a wavelength plate or a polarizing plate.

After being totally reflected at the interface 210 b, the light beam 213 parallelized by the collimator lens 16 is detected by the light detecting means 17. Light detection means 17 in this example
Is a plurality of photodiodes 17a, 17b, 17c...
This is a photodiode array that is arranged in a row, and in a plane shown in FIG.
Are arranged so that the direction in which the photodiodes are arranged is substantially perpendicular to the traveling direction. Therefore, the above interface
The components of the light beam 213 totally reflected at various reflection angles at 210b are respectively converted into different photodiodes 17a and 17a.
b, 17c... receive light.

The photodiodes 17a, 17b, 17c,...
… Each output of each differential amplifier 18 of the differential amplifier array 18
a, 18b, 18c... At this time, outputs of two photodiodes adjacent to each other are input to a common differential amplifier. Therefore, each differential amplifier 18a, 18b,
The output of 18c is a plurality of photodiodes 17a, 17
It can be considered that the light detection signals output from b, 17c... are differentiated with respect to their juxtaposition directions.

The outputs of the differential amplifiers 18a, 18b, 18c... Are respectively sampled and held by circuits 22a, 22b, 22c.
Are sampled and held at predetermined timings and input to the multiplexer 23. The multiplexer 23 is provided for each of the sampled and held differential amplifiers 18a, 18b, 18c,.
Are input to the A / D converter 24 in a predetermined order. The A / D converter 24 digitizes these outputs and inputs them to the signal processing unit 220.

FIG. 16 shows the light intensity at each incident angle θ of the light beam 213 totally reflected at the interface 210b and the differential amplifiers 18a and 18a.
b, 18c... are explained. Here, it is assumed that the relationship between the incident angle θ of the light beam 213 to the interface 210b and the light intensity I is as shown in the graph of FIG.

The light incident on the interface 210b at a specific incident angle θ _SP excites the surface plasmon at the interface between the metal film 212 and the sample 211.
Drops sharply. That is, θ _SP is the total reflection attenuation angle,
At this angle θ _SP , the reflected light intensity I takes a minimum value.
This decrease in the reflected light intensity I is represented by D in FIG.
Observed as dark lines in the reflected light.

FIG. 16B shows a photodiode.
17a, 17b, 17c,... Are shown in parallel, and as described above, these photodiodes 17a, 17b, 17c
.. correspond to the above incident angle θ uniquely.

Then, the photodiodes 17a, 17b, 17c
…… in the juxtaposition direction, that is, the incident angle θ and the differential amplifier
The relationship between 18a, 18b, 18c... And the output I ′ (differential value of the reflected light intensity I) is as shown in FIG.

[0154] The signal processing unit 220, based on the value of the differential value I 'input from the A / D converter 24, outputs the differential amplifier 18a,
From among 18b, 18c..., The one that provides the output closest to the differential value I ′ = 0 corresponding to the total reflection attenuation angle θ _SP (in the example of FIG. 16, the differential amplifier 18d) is selected. Then, the differential value I ′ output from the output unit is subjected to a correction process described later, and the value is displayed on the display unit 21. In some cases, there is a differential amplifier that outputs the differential value I ′ = 0, and in that case, the differential amplifier is naturally selected.

Thereafter, the differential value I 'output from the selected differential amplifier 18d every time a predetermined time elapses is displayed on the display means 21 after receiving the correction processing. This differential value I ′ changes the dielectric constant, that is, the refractive index of the substance in contact with the metal film 212 (see FIG. 14) of the measurement unit 222, and the curve shown in FIG. When it changes in shape, it rises and falls accordingly. Therefore, by continuously measuring the differential value I ′ with the passage of time, it is possible to examine the change in the refractive index of the substance in contact with the metal film 212, that is, the change in the characteristic.

In particular, in the present embodiment, the sensing substance 230 that binds to a specific substance (analyte) in the liquid sample 211 is fixed to the metal film 212, and the refractive index of the sensing substance 230 depends on the bonding state. Changes, the state of the change of the coupling state can be examined by continuously measuring the differential value I ′. That is, in this case, both the liquid sample 211 and the sensing substance 230 are samples to be analyzed. Examples of such a combination of the specific substance and the sensing substance 230 include an antigen and an antibody.

As is clear from the above description, in the present embodiment, a plurality of photodiodes 17a,
Since a photodiode array in which 17b, 17c... Are arranged in a line is used, FIG.
Even if the curve shown in Fig. 6 (1) changes to a large extent to some extent moving in the left-right direction, dark line detection is possible. In other words, by using such an array of light detecting means 17,
A large measurement dynamic range can be ensured.

Note that a plurality of differential amplifiers 18a, 18b, 18c
Instead of using the differential amplifier array 18 composed of..., One differential amplifier is provided, and photodiodes 17a, 17b, and 17 are provided.
.. may be switched by a multiplexer, and two adjacent outputs may be sequentially input to the one differential amplifier.

Next, the above-described correction processing performed on the differential value I 'will be described in detail. As shown in FIG. 14, the measurement unit 222 fixed to the turntable 231
Above one stop position, an automatic reference liquid supply mechanism 221 is provided. This automatic reference liquid supply mechanism 221
Has a basic structure similar to that of the automatic sample supply mechanism 70 shown in FIG. 1, and supplies the reference liquid dropwise to one of the plurality of measurement units 222 supported by the turntable 231. The liquid sample 211 is not supplied to the measurement unit 222 to which the reference liquid has been supplied.

Here, the liquid sample 211 of the present example is obtained by dissolving the test sample in a solvent, and this solvent is used as the reference liquid. For example, when the subject is biotinylated insulin and PBS (phosphate buffer solution) containing 0.1% BSA solution is used as a solvent, PBS containing this 0.1% BSA solution is used as a reference solution.

As described above, the turntable 23
The reference liquid is supplied to one of the plurality of measurement units 222 supported by 1, and the liquid sample 211 is supplied to the remaining measurement units 222. The liquid sample 211 stored in each measurement unit 222 is supplied to The measurement of the differential value I ′ is performed as follows. At the same time, the differential value I ′ is similarly measured for the reference liquid stored in one measurement unit 222.

Here, (1) and (2) in FIG. 17 show that the differential value I ′ of the liquid sample 211 stored in a certain measuring unit 222 and the differential value I ′ of the reference liquid with the passage of time. An example of the change will be described. In the figure, the former is indicated by black circles as measured data, and the latter is indicated by white circles as correction data.

By knowing the actual measurement data, that is, the change with time of the differential value I ′ of the liquid sample 211, as described above, the specific substance (subject) and the sensing substance 230 in the liquid sample 211 are compared with each other. However, the actual measurement data is affected by conditions such as the temperature around the device. It depends on the temperature of the liquid sample
This is because the refractive index of the solvent 211 changes and a minute position change occurs in the measuring optical system of the apparatus.

The correction process performed by the signal processing section 220 on the differential value I ′ as the actual measurement data is a process of subtracting the differential value I ′ as the correction data from the differential value I ′. That is, the data after correction is as shown by continuous curves in (1) and (2) of FIG. Note that the above subtraction is performed between the differential values I ′ of the liquid sample 211 and the reference liquid, which have the same reaction time with the sensing substance 230.

The differential value I ′ relating to the reference liquid is represented by
Regardless of the specific substance (analyte) in 1, a value corresponding to the refractive index of the solvent of the reference liquid, that is, the liquid sample 211, is taken, and if there is a small positional change in the optical system of the apparatus, it is also reflected. Is what it is. Therefore, if a correction is performed by subtracting such a differential value I ′ relating to the reference liquid from the differential value I ′ as the actually measured data, the corrected data can be used as a result of a change in the refractive index of the solvent due to environmental conditions such as temperature or optical measurement. By compensating for the change in the characteristic of the system, the characteristic of the analyte in the liquid sample 211 is purely indicated.

Next, a tenth embodiment of the present invention will be described. FIG. 18 shows a plan shape of a main part of a measuring apparatus using attenuated total reflection according to a tenth embodiment of the present invention. The measuring device of this embodiment is a surface plasmon resonance measuring device. In FIG. 18, the same elements as those in FIG. 7 are denoted by the same reference numerals as those in FIG. That is, 20 is a turntable that rotates intermittently in the direction of arrow R, 70 is an automatic sample supply mechanism, 76 is chip supply means, 77 is surface plasmon resonance measurement means, 78
Is a chip discharging means.

In the present example, the turntable 20 supports 96 measuring units 80 and supports 3.75 ° (= 360 °).
96) so as to rotate intermittently at angular intervals. Then, the chip supply means 76 receives the well plate 240 containing the 96 measurement units 80 in the setting unit 241 and supplies the measurement units 80 one by one from there onto the turntable 20 sequentially. In this example, the measurement unit
A sensing substance similar to the sensing substance 14 shown in FIG. 2 is fixed on the metal film 80 in advance, and the measurement unit 80 is supplied on the turntable 20 in that state.

The liquid sample 15 is also a 96-well plate.
Pre-stored in each well of 242, this well plate 242
Is set in the set section 243 of the automatic sample supply mechanism 70, from which each sample 15 is transferred to the measuring unit on the turntable 20.
Supplied to 80. The measurement by the surface plasmon resonance measuring means 77 is performed in the same manner as described above. And
After being subjected to the measurement, the measuring unit 80
By 78, it is discharged from the turntable 20 into the collection well plate 244.

The apparatus having the above configuration can be suitably used for the above-described random screening for finding a specific substance that binds to a predetermined sensing substance.

As described above, when the refractive index of the liquid sample 15 changes due to a change in the temperature around the apparatus, the value measured by the surface plasmon resonance measurement apparatus changes. Hereinafter, a configuration for preventing the temperature change of the liquid sample 15 will be described.

FIG. 19 shows a side view of the setting section 243 of the automatic sample supply mechanism 70 described above. As shown, the set portion 243 has 96 concave portions 245 for accommodating each well (portion for storing the liquid sample 15) of the well plate 242.
a metal block 245 having a.a. The metal block 245 has a temperature control means 246 including a Peltier element or a heater for changing the temperature, and a metal block 245.
And a temperature sensor 247 for detecting the temperature. The temperature detection signal S1 of the temperature sensor 247 is input to the controller 248, and the controller 248 controls the driving of the temperature adjusting means 246 based on the temperature detection signal S1, so that the temperature of the metal block 245 is set to a predetermined value. Kept at temperature. The metal block 245 is desirably formed of, for example, copper having a high thermal conductivity.

As described above, if the temperature of the metal block 245 is maintained at the set temperature, the liquid sample 15 stored in each well of the well plate 242 and supplied to the measurement unit 80 is also maintained at the set temperature. In addition, it is possible to prevent the refractive index of the liquid sample 15 from changing in accordance with the temperature and changing the value measured by the surface plasmon resonance measurement device.

In order to maintain the temperature of the metal block 245 at the set temperature, as described above, the temperature adjusting means 246 including a Peltier element, a heater, and the like attached to the metal block 245.
Instead of using, a mechanism for circulating a temperature-controlled liquid may be used as in the surface plasmon resonance measurement apparatus according to the eleventh embodiment of the present invention shown in FIG.

That is, in the surface plasmon resonance measurement apparatus shown in FIG. 20, a liquid flow path 245b passing through the entire area of the metal block 245 is formed, and the liquid flow path 245b has an upstream end and a downstream end. Is the circulation passage 250
Is connected. And in the middle of the circulation passage 250, for example, a pump 251 for circulating and pumping a liquid such as water,
A heater 252 for heating the liquid is provided.

The driving of the heater 252 is controlled by a temperature sensor 247 and a controller 248 similar to those shown in FIG. 19, whereby the liquid flowing through the circulation passage 250 and sent to the liquid flow passage 245b of the metal block 245 is controlled. Is maintained at a predetermined set temperature, and thus the metal block 24
The temperature of 5 is kept at this set temperature. By maintaining the temperature of the metal block 245 at the set temperature in this manner, also in this case, it is possible to prevent the refractive index of the liquid sample 15 from changing according to the temperature and the measured value from changing.

In order to prevent the measured value from being changed by the temperature change of the liquid sample 15 as described above, the measuring unit after being supplied onto the turntable 20 is used.
It is desirable to also suppress the temperature change of the liquid sample 15 in 80. Hereinafter, a surface plasmon resonance measuring apparatus according to a twelfth embodiment of the present invention, which is formed as described above, will be described.

FIG. 21 is a perspective view showing a part of a surface plasmon resonance measuring apparatus according to a twelfth embodiment of the present invention. In this figure, the same elements as those shown in FIGS. 18 and 19 are denoted by the same reference numerals as those in those figures.

In the apparatus according to the present embodiment, a cover 260 whose upper opening of a cylindrical member having a low height is closed by a disk is disposed on the turntable 20. Air is trapped inside the cover 260 and is difficult to circulate outside the cover 260.
However, this cover 260 has a minimum
A small opening 260a for receiving the sample dropping pipette 71, and an opening 26 for receiving the surface plasmon resonance measuring means 77 in which the optical system is housed in the case 262.
0b, a small opening 260c for receiving a chip supply means (not shown), and a small opening 260d for receiving a chip discharge means also not shown.

The cover 260 has a temperature control means 246 comprising a Peltier element or a heater for changing the internal temperature, and a temperature sensor 24 for detecting the temperature inside the cover 260.
7 and are attached. The temperature detection signal S1 of the temperature sensor 247 is input to the controller 248, and the controller 248 generates the temperature control signal 2 based on the temperature detection signal S1.
By controlling the driving of 46, the temperature in cover 260 is maintained at a predetermined set temperature.

By keeping the temperature inside the cover 260 at the set temperature as described above, the measuring unit 80 on the turntable 20
Since the liquid sample 15 in is also maintained at the set temperature, the refractive index of the liquid sample 15 changes according to the temperature, and it is possible to prevent the measurement value by the surface plasmon resonance measurement device from changing. .

The turntable 20 as described above is used.
A mechanism for maintaining the liquid sample 15 in the upper measurement unit 80 at the set temperature, and a mechanism for maintaining the liquid sample 15 before being supplied to the measurement unit 80 at the set temperature (same as the above set temperature as shown in FIGS. ) May be applied together, and in such a case, the effect of preventing the measured value from changing due to the temperature becomes even more remarkable.

Further, as described above, the mechanism for keeping the sample 15 at the set temperature is not limited to the surface plasmon resonance measuring apparatus.
The present invention is also applicable to the leak mode sensor as shown in FIGS. 9 and 10, and in such a case, the effect of preventing the measured value from being changed by the temperature can be obtained in the same manner as described above.

Here, when the liquid sample 15 is, for example, the one obtained by dissolving a test object in a solvent as described above, the liquid sample 15 is treated with a surface plasmon in order to accurately determine the characteristics of the test object. It is desired that the concentration of the analyte be uniformed by sufficiently stirring before subjecting to resonance measurement. This is true not only for surface plasmon resonance measurement but also for measurement using a leaky mode sensor as shown in FIGS.

In order to sufficiently stir the liquid sample 15 as described above, for example, taking the apparatus of FIG. 21 as an example,
If a predetermined number of measurement units 80 are supported on the turntable 20, before turning on the surface plasmon resonance measurement, the turntable 20 can be continuously rotated in one direction at high speed,
Alternatively, it is effective to rotate the turntable 20 alternately several times in the forward direction and the reverse direction.

Further, taking the apparatus of FIG. 21 as an example, the sample dropping pipette of the automatic sample supply mechanism 70 will be described.
After the liquid sample 15 is dropped and supplied into the measurement unit 80 by 71, the tip of the pipette 71 is advanced into the liquid sample 15 stored in the measurement unit 80, and the pipette is
Even if the operation of sucking the liquid sample 15 into the 71 and then discharging the liquid sample 15 into the measuring unit 80 is performed several times, the liquid sample 15 can be satisfactorily stirred.

All of the devices of the embodiments described above use a turntable as a support for supporting a plurality of measurement units. Next, an embodiment using a reciprocating linear type support or the like will be described. I do.

FIG. 22 shows a perspective view of a surface plasmon resonance measuring apparatus according to a thirteenth embodiment of the present invention. 2, the same elements as those shown in FIG. 2 are denoted by the same reference numerals as those in FIG.

The surface plasmon resonance measuring apparatus according to the present embodiment is slidably engaged with two guide rods 400, 400 arranged in parallel with each other as a support for supporting the measuring unit 10, and can be viewed along the guide rods. A slide block 401 is used that can move linearly in the direction of the arrow Y inside. The slide block 401 has the guide rod 4
A precision screw 402 arranged in parallel with 00,400 is screwed together, and this precision screw 402 is rotated forward and reverse by a pulse motor 403 constituting a support driving means together therewith.

The driving of the pulse motor 403 is controlled by a motor controller 404. That is, the output signal S40 of a linear encoder (not shown) which is incorporated in the slide block 401 and detects the position of the slide block 401 in the longitudinal direction of the guide rods 400, 400 is input to the motor controller 404. The driving of the pulse motor 403 is controlled based on this signal S40.

Also, below the guide rods 400, 400,
The laser light source 31 and the condenser lens 32 sandwich the slide block 401 that moves along
And a photodetector 40. The laser light source 31, the condenser lens 32, and the photodetector 40 are basically the same as those in the apparatus of FIG.

Here, in the present embodiment, a stick-shaped unit connecting body 410 in which eight measuring units 10 are connected and fixed is used as an example.
The slide blocks 401 are set in a state of being arranged in a line. FIG. 23 shows this unit connected body.
14 shows a detailed structure of 410. As shown here, the unit connected body 410 is formed by connecting eight measurement units 10 similar to those shown in FIG.

As shown in FIG. 24, for example, as shown in FIG.
It is set to 420 and transported and handled in this state. That is, the measurement unit 10 is transported and handled in 96 units by this one plate 420.

When a plurality of measuring units 10 are collectively handled as a unit connected body 410, a dispenser 430 as shown in FIG. 25 is used as a means for supplying a liquid sample to the measuring unit 10. Is desirable. The dispenser 430 is connected to the measurement unit 10
As many dispensing nozzles 431 as the number of
Since it is supported by the support member 432 at the same pitch as the arrangement pitch of 0, and the liquid sample can be simultaneously dispensed to the plurality of measurement units 10 of one unit connected body 410,
The efficiency of the sample supply operation can be improved.

When using the surface plasmon resonance measuring apparatus shown in FIG. 22, one unit connected body 410 is set in the unit setting section 401a of the slide block 401 from the plate 420 shown in FIG. You. As shown, the slide block 401 is formed of a frame-shaped member having an opening 401b penetrating in the left-right direction (up-down direction in the figure), so that the laser light source 31 The emitted measuring light beam 30 may be irradiated. Further, the light beam 30 totally reflected at the interface between the dielectric block 11 and the metal film 12 of the measurement unit 10 can be detected by the photodetector 40.

When the surface plasmon resonance measurement is performed in the present apparatus, the slide block 401 is moved by driving the pulse motor 403, and the slide block 401 is first moved.
The first, that is, the rightmost measuring unit in FIG.
10 is stopped at a position where the measurement light beam 30 is irradiated. Then, the surface plasmon resonance measurement on the sample of the measurement unit 10 is performed in the same manner as described above.

When the measurement is completed, the slide block 401 is moved rightward in FIG. 22 by the same distance as the arrangement pitch of the measurement units 10 by the rotation of the pulse motor 403, and stopped at that position. As a result, the second measurement unit 10 can be irradiated with the measurement light beam 30, and the surface plasmon resonance measurement is similarly performed on the sample of this measurement unit 10. Similarly,
The intermittent movement of the slide block 401 and the surface plasmon resonance measurement are repeated, and the surface plasmon resonance measurement is performed on all the samples of the eight measurement units 10.

When the above operation is completed, the pulse motor 40
3 is driven to rotate in the opposite direction to the above case, and the slide block 401 is returned to the predetermined left end position. If surface plasmon resonance measurement is performed only once for one sample, slide block is used at that position using a means not shown.
The unit connected body 410 is taken out from 401. On the other hand, when the surface plasmon resonance measurement is performed a plurality of times at a predetermined time interval on one sample, the slide block 401 is used.
Is intermittently moved rightward again from the left end position, and surface plasmon resonance measurement is performed on each sample of the eight measurement units 10 in the same manner as described above. Thereafter, by repeating the operation of returning the slide block 401 to the left end position and the operation of the surface plasmon resonance measurement, the measurement of the sample of one measurement unit 10 can be performed any number of times.

The slide block 401 described above
Of course is not limited to the surface plasmon resonance measurement device, but can be applied to a leak mode sensor as shown in FIGS. 9 and 10 as well.

Also, instead of the unit connected body 410, a unit connected body as shown in FIG. 26 or FIG. 27 can be used. Unit connection body 44 shown in FIG.
In the case of 0, for example, one prismatic dielectric bar 441 is formed by using the same material as the dielectric block 11 shown in FIG. 23, and a plurality of bottomed holes 442 are formed in the dielectric bar 441. The bar 441 serves as a sample holding unit, and the metal film 12 and the sensing substance 14 are formed on the bottom surface of the hole 442. That is, in this unit connection body 440, a measurement unit is configured for each hole 442.

The unit connected body 440 having the above-described structure is used.
Can be manufactured more easily and can achieve cost reduction as compared with a configuration in which the measuring units 10 are formed one by one as in the unit connected body 410 shown in FIG. .

On the other hand, unit linked body 450 shown in FIG.
The measurement unit 460 is fitted and supported in a plurality of unit support holes 452 formed in the unit support plate 451. The measurement unit 460 is basically
3 is the same as the measurement unit 10 shown in FIG. 3, but is formed by cutting off a part of a square pyramid so as to fit into the tapered unit support hole 452 so as not to pass through the unit support hole 452. Has become.

When such a unit connected body 450 is applied, a plurality of measurement units 460 are respectively supported on a single unit support plate 451 in advance, and a plurality of such unit support plates 451, for example, as shown in FIG. It may be set on the plate 420 and handled. Alternatively, one unit support plate 451 is fixed to, for example, the slide block 401 of the apparatus in FIG. 22, and a plurality of measurement units 460 are supplied to the unit support plate 451 one by one. May be removed from the unit support plate 451, and a plurality of new measurement units 460 may be supplied to the unit support plate 451.

Compared with the above-described unit connected body 450, the unit connected body 410 in FIG. 23 and the unit connected body 440 in FIG. 26 have a surface plasmon Position accuracy on the resonance measurement device is easily obtained, and since there is no need to hold a small measurement unit, it is excellent in handling properties and can contribute to improvement in efficiency of measurement processing.

Next, a surface plasmon resonance measuring apparatus according to a fourteenth embodiment of the present invention will be described. FIG.
Shows a side surface shape of the surface plasmon resonance measurement device according to the fourteenth embodiment of the present invention. 2, the same elements as those shown in FIG. 2 are denoted by the same reference numerals as those in FIG.

In the surface plasmon resonance measuring apparatus of the present embodiment, a support bar 500 linearly extending in the direction of arrow Y in the figure is used as a support for supporting the measurement unit 10. A plurality of unit support holes 501 are formed in the support bar 500 at predetermined intervals along the length thereof, and the measurement units 502 are fitted and supported one by one in the unit support holes 501. It has become. The measurement unit 502 is basically the measurement unit 1 shown in FIG.
It is the same as 0, but has a shape obtained by cutting off a part of a quadrangular pyramid and fits into the tapered unit support hole 501 so as not to pass through the unit support hole 501.

A guide rail 520 extending parallel to the support bar 500 is disposed below the support bar 500.
20 is combined with a slide block 521 that is movable along the guide rail 520, that is, in the direction of the arrow Y. The slide block 521 receives driving force from driving means 531 mounted thereon, and
It is possible to reciprocate along 20.

On the slide block 521,
A guide rail 522 extending in the X direction perpendicular to the arrow Y direction is fixed, and a slide block 523 movable along the guide rail 522 is combined with the guide rail 522. The slide block 523 is capable of reciprocating along a guide rail 522 by receiving a driving force from a driving unit 533 mounted thereon.

An optical unit 524 is fixed on the slide block 523. This optical unit 524
A laser light source 31 that emits a light beam 30 for measurement, a collimator lens 525 that parallelizes the light beam 30 emitted in a divergent light state from the laser light source 31, and reflects the light beam 30 that has become parallel light. Then, the mirror 52 is moved toward the interface between the dielectric block 11 and the metal film 12 of the measurement unit 502.
6, a condenser lens 527 for condensing the light beam 30 reflected by the mirror 526, a mirror 528 for reflecting the light beam 30 totally reflected at the interface, and a light detection for detecting the light beam 30 reflected by the mirror 528. The container 40 is mounted.

When performing the surface plasmon resonance measurement by the apparatus of this embodiment, the optical unit 524 is first set to the standby position indicated by the two-dot chain line in the figure. This position is Y
The direction is aligned with the first, that is, the rightmost measurement unit 502 in the figure. In the X direction, the first measurement unit 502 is positioned laterally (to the rear or the front in the figure). It is out of position.

In this state, the plurality of measurement units 502
The liquid sample 15 is supplied to each of the plurality of unit support holes 501 of the support bar 500 using a supply unit (not shown). Next, the driving unit 533 is driven in the forward direction, the slide block 523 is moved by a predetermined distance in the X direction, and the optical unit 524 moves the light beam to the interface between the dielectric block 11 and the metal film 12 of the first measurement unit 502. It is set to the X direction position where 30 can be irradiated.

Next, the laser light source 31 is driven to irradiate the interface with the light beam 30, and the light beam totally reflected at the interface is
30 is detected by the photodetector 40, and a surface plasmon resonance measurement is performed. This measurement itself is performed in the same manner as in each of the embodiments described above.

When the measurement of the liquid sample 15 by the first measurement unit 502 is completed, the driving means 533 is driven in the reverse direction, and the slide block 523 is moved by a predetermined distance in the X direction (the direction opposite to the above). The optical unit 524 is returned to the standby position. Next, the driving means 531 is driven, and the slide block 521 is moved leftward in the figure by the same distance as the arrangement pitch of the unit support holes 501 of the support bar 500. Next, the driving unit 533 is driven in the forward direction, the slide block 523 is moved by a predetermined distance in the X direction, and the optical unit 524 moves the light beam to the interface between the dielectric block 11 and the metal film 12 of the second measurement unit 502. X that can irradiate 30
Set to the direction position.

Next, the laser light source 31 is driven to irradiate the interface with the light beam 30, and the light beam totally reflected at the interface is
30 is detected by the photodetector 40, and a surface plasmon resonance measurement is performed.

Thereafter, by repeating the same operation, the surface plasmon resonance measurement is performed on the liquid sample 15 of each of the third, fourth, fifth,... Measurement units 502. In this way, when the measurement of the liquid sample 15 by all the measurement units 502 is completed, the driving unit 531 is driven in the opposite direction to the above-described measurement, and the slide block 521 is moved rightward in the drawing. The optical unit 524 is returned to the standby position.

Thereafter, by repeating the same operation as described above, a plurality of predetermined measurements can be performed on the liquid sample 15 of one measurement unit 502. After the predetermined number of measurements have been completed, the optical unit 524 is returned to the standby position, and then all of the measurement units 502 are ejected from the support bar 500 by ejection means (not shown).

Thereafter, the measuring units 502 are supplied to the unit support holes 501 of the support bar 500 in the same manner as described above, and when the unmeasured liquid samples 15 are supplied thereto, the same applies to the liquid samples 15. The surface plasmon resonance measurement is performed.

As described above, the mechanism for moving the optical unit 524 while keeping the plurality of measurement units 502 stationary is not limited to the surface plasmon resonance measurement apparatus, and is shown in FIGS. 9 and 10. The same is applicable to such a leak mode sensor. In addition, a plurality of measurement units may be supported along a circular arc on a circular support, and the measurement optical system may be circularly moved along the measurement units to sequentially perform measurement on a sample of each measurement unit. It is possible.

Next, a description will be given of a surface plasmon resonance measuring apparatus according to a fifteenth embodiment of the present invention, which can prevent a measurement result of each measuring unit from being erroneously evaluated.

FIG. 29 shows a structure of a main part of a surface plasmon resonance measuring apparatus according to a fifteenth embodiment of the present invention. This apparatus basically performs surface plasmon resonance measurement using a turntable 20 and a measurement unit 80 similar to those shown in FIG. The liquid sample to be measured is stored and supplied in each well of the well plate 242 similar to that shown in FIG.
Is dispensed to the measurement unit 80 using the automatic sample supply mechanism 70 provided with

The apparatus according to the present embodiment is used as an example in the above-described random screening, in which the sensing substance is fixed on a metal film formed on one surface of the dielectric block of the measurement unit 80. A sample that may contain a specific substance (analyte) that binds to the sensing substance is subjected to measurement. Each measurement unit 80 is provided with a bar code 80b indicating a production number (serial number) for identifying each of them. In the following, the sensing substance is referred to as a receptor, and the sample is referred to as a ligand.

In the present embodiment, a computer system 600 for managing information related to surface plasmon resonance measurement, and a computer system for managing this information and outputting a measurement result based on the output signal S of the photodetector 40 610 are provided. Computer system 600
Is composed of, for example, a general-purpose personal computer including a main body unit 601, a display 602, a keyboard 603, and the like. A first bar code reader 604 for reading the bar code 80b of the measurement unit 80 is connected to the main body 601 of the computer system 600. The first bar code reader 604 is connected to the measuring unit 8
It is installed at the place where the above-mentioned receptor is fixed at 0.

On the other hand, the computer system 610 is also constituted by a general-purpose personal computer having, for example, a main body 611, a display 612, a keyboard 613 and the like. A second bar code reader 614 for reading the bar code 80b of the measurement unit 80 is connected to the main body 611 of the computer system 610. This second
The barcode reader 614 is installed at a place where the ligand is dispensed to the measurement unit 80.

The well plate 242 has a bar code 242b indicating the type of ligand stored in each well.
The bar code 242b is read by a third bar code reader 620. The third barcode reader 620 is connected to the main unit 611 of the computer system 610.

Hereinafter, the operation of this apparatus will be described with reference to FIG. 30 showing the flow of the measurement operation. In FIG. 30, the flow of information processing related to measurement performed in the computer systems 600 and 610 is shown in the right column.
The flow of operations related to the processing of this information is shown in the left column.

First, as a flow of operation, first, a measuring unit 80 is manufactured by a manufacturer, and at this time, a bar code indicating the aforementioned serial number is given to the measuring unit 80.
80b is attached (step P1). At this time, the serial number indicated by each bar code 80b is registered in a predetermined storage unit of the computer system 600 by using the keyboard 603 or the like for production management of the measurement unit (step Q1). Then, after the bar code 80b is attached, the above-described metal film is formed on the dielectric block constituting the measuring unit 80 (Step P2).

Next, the receptor is fixed to the measuring unit 80 (step P3). At this time, the bar code 8 of the measurement unit 80 is read by the first bar code reader 604.
The serial number indicated by 0b is read, and information F indicating the type of the fixed receptor is input using the keyboard 603 or the like. Stored and registered in the storage means (step Q
2). The computer system 600 transfers the information F to the computer system 610 using, for example, the Internet or a dedicated communication line. Computer system 610
Stores this information F in a predetermined storage means of the main body 611.

The measuring unit 80 to which the receptor was fixed,
The apparatus is supported by the turntable 20 of the apparatus shown in FIG. 29 (Step P4). At this time, the measuring unit 80 is
Before or after supporting the second bar code reader
The serial number indicated by the barcode 80b of the measurement unit 80 is read by 614 and input to the computer system 610. Based on the stored information F, the computer system 610 reads out the type of receptor corresponding to the input serial number and causes the display 612 to display it. Thereby, the user of the apparatus can confirm whether or not the correct receptor is stuck to the measuring unit 80 to be used from now on (step Q3).

Next, the ligand is dispensed to the measurement unit 80 supported by the turntable 20 (step P).
5). At this time, the bar code 242b indicating the type of ligand stored in each well of the well plate 242 is read by the third bar code reader 620.

The ligand stored in each well of the well plate 242 is dispensed to the measurement unit 80 in a predetermined order, and the measurement unit 80 also passes through the second bar code reader 614 after passing through the second bar code reader 614. This is sent to the ligand dispensing position according to a predetermined sequence. Therefore, the computer system 610 may use the second barcode reader 614.
And the type of ligand dispensed to each measurement unit 80 based on the information G and H from the third barcode reader 620, and associates it with the serial number of the measurement unit 80 in the storage means. It is stored and registered (step Q4).

Next, after a lapse of a predetermined time for allowing the receptor to react with the ligand, surface plasmon resonance measurement is performed on the measurement unit 80 supported by the turntable 20 (step P6). In the case of the present embodiment, this measurement is performed to find a ligand that binds to the receptor as described above.
As in the ninth embodiment shown in FIGS. 4 and 15, the determination can be made based on the output signal S of the photodetector 40.

The computer system 610 obtains the result of the reaction between each receptor and the ligand based on the output signal S, further determines the binding state between the receptor and the ligand based on the result of the reaction, and transmits the result of the measurement to the measuring unit. Stored in the storage means in association with the serial number of 80,
It is registered (step Q5).

The measurement result thus obtained is displayed on display 612 of computer system 610 (step Q6). A display example of the measurement result is shown in (Table 1) below.

[0233]

[Table 1] As described above, in the present embodiment, all of the receptor, the ligand, and the measurement result are stored in the storage unit in association with the serial number of the measurement unit 80.
This prevents the measurement of an unintended combination of a receptor and a ligand, or prevents the measurement result of one of the combinations from being mistakenly displayed as the measurement result of another combination.

The means for identifying the measurement unit is not limited to the barcode 80b described above, but may be any other character such as a magnetic recording layer, a semiconductor memory, or a printed character read by an OCR.

In the above example, the measurement result is displayed on the display 612 of the computer system 610. However, the measurement result may be output from a printer and recorded on paper.

Next, an embodiment of the surface plasmon resonance measuring method according to the present invention will be described. FIG. 31 is a diagram illustrating a flow of an operation when a sample is analyzed by one embodiment of the surface plasmon resonance measurement method of the present invention.

In FIG. 31, reference numerals 100 and 200 indicate a measuring unit and a turntable, respectively. The measuring unit 100 is basically the measuring unit 1 in FIG.
As in the case of 0, the dielectric block, the metal film, and the sample holding frame are integrated into a chip to form a chip. Hand turntable 200
Has 96 chip supporting portions 300 as an example arranged at equal angular intervals (the number does not match because they are schematically shown in the figure), and each angle is equal to the angular interval of the chip supporting portions 300. It rotates intermittently in the counterclockwise direction in the figure. The time interval of this intermittent rotation is 2 seconds in this example.

Also, positions F1, F2, F indicated by arrows in the figure
Reference numerals 3, F4 and F5 indicate positions where the processing of supply of the measurement chip, supply of the sensing substance, measurement, injection of the sample, and discharge of the measurement chip are performed. The above-mentioned sensing substance binds to the specific substance in the sample similarly to the sensing substance 14 shown in FIG. 2, and the specific examples are also as described above.

First, as shown in FIG. 1A, a measuring chip 100 having a sensing substance fixed film attached in advance on a metal film is placed on a chip supporting portion 300 of a turntable 200 one by one from a measuring chip supply position F1. Supply and support there.

Next, when the turntable 200 is rotated once, the measuring chip 1 supported by the chip supporting portion 300 is rotated.
Reference numeral 00 denotes a sensing substance supply position F2 shown in FIG.

The supply of the sensing substance is performed in parallel with the supply of the measurement chip 100. Then, when the turntable 200 is rotated 96 times, that is, once, the measuring chip 100 containing the sensing substance is moved to all the chip supporting portions 300.
The turntable 200 returns to the initial position while being supported by the. Thus, the time required for the turntable 200 to return to the initial position is 2 × 96 = 192 seconds. FIG. 3C shows this state. After that, it stands by for a predetermined time if necessary.

Next, as shown in FIG.
In the above, the same operation as the surface plasmon resonance measurement operation described above is performed. However, this measurement operation is performed on the measurement chip 100 to which the sample has not been supplied yet, and the output signal S (for example, see FIG. 2) of the photodetector 40 under that state is detected as the measurement baseline.

Next, when the turntable 200 rotates once, the measuring chip 10 which has been subjected to the above-described operation of detecting the baseline is measured.
0 is arranged at a sample injection position F4 shown in FIG. 5 (5), where a sample is injected into the measurement chip 100.

This sample injection is performed in parallel with the above-described baseline detection operation. And turntable 200 is 9
After six rotations, that is, one rotation, the detection operation of the baseline and the sample injection are performed on all the measurement chips 100, and the turntable 200 returns to the initial position.

Next, as shown in FIG.
, A surface plasmon resonance measurement operation is performed. This measurement operation is made to the measurement chip 100 the sample is supplied, it is required ATR angle theta _SP described above.
When the turntable 200 is rotated 96 times, that is, once, the measurement is completed once for all the measurement chips 100, and the turntable 200 returns to the initial position.

Then, by continuing the rotation of the turntable 200 and performing the surface plasmon resonance measurement at the measurement position F3, the measurement is sequentially performed a plurality of times for each measurement chip 100, such as the second time, the third time, and so on. obtain.

As described above, in this measurement method, a certain 1
After detecting the state of surface plasmon resonance with respect to the sample on one measurement chip 100, the turntable 200 is rotated once and the next measurement is performed on the sample, and then the samples on another measurement chip 100 are successively repeated in the same manner. Since the state of surface plasmon resonance can be detected, measurement of a large number of samples can be performed efficiently in a short time.

When the turntable 200 enters the predetermined n-th rotation, the n-th measurement is performed on the sample of each measurement chip 100. The measurement chip 100 that has completed the n-th measurement is the same as FIG. It is discharged from the turntable 200 at the measurement chip discharge position F5 shown in 7).

When the sample injection has been completed for all the measurement chips 100 on the turntable 200, the turntable 200 is stopped if necessary before starting the measurement, and the sample and the sample The reaction with the sensing substance may be waited.

FIG. 32 shows a sample (specimen) obtained by the above method.
2 shows two examples of multi-point measurement results. Here, the sampling time on the horizontal axis indicates the time until each measurement is performed after sample injection. The amount of binding on the vertical axis is the amount of the specific substance in the sample that specifically binds to the sensing substance, and corresponds to the change in the surface plasmon resonance detection signal, that is, the amount of movement of the dark line described above.

As shown here, by performing multi-point measurement, unlike the case where only the final amount of binding is obtained, the time until the amount of binding reaches saturation can be known.

The surface plasmon resonance measuring method applied to the case where the turntable 200 is used has been described above. However, the surface plasmon resonance measuring method according to the present invention is not limited to this, and the above-described support for reciprocating linear movement can be used. The present invention is applicable to the case where it is used, and in such a case, a similar effect can be obtained.

[Brief description of the drawings]

FIG. 1 is an overall view of a surface plasmon resonance measuring apparatus 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 resonance measuring apparatus of FIG. 1;

FIG. 3 is a graph showing a schematic relationship between an incident angle of a light beam and a light intensity detected by a photodetector in a surface plasmon resonance measuring apparatus.

FIG. 4 is a partially cutaway side view showing a main part of a surface plasmon resonance measuring apparatus according to a second embodiment of the present invention.

FIG. 5 is a partially broken side view showing a main part of a surface plasmon resonance measuring apparatus according to a third embodiment of the present invention.

FIG. 6 is a side view of a surface plasmon resonance measurement apparatus according to a fourth embodiment of the present invention.

FIG. 7 is a plan view of a surface plasmon resonance measurement apparatus according to the fourth embodiment.

FIG. 8 is a partially cutaway side view showing a main part of the surface plasmon resonance measuring apparatus according to the fourth embodiment.

FIG. 9 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. 10 is a partially broken side view showing a main part of a leakage mode sensor according to a sixth embodiment of the present invention.

FIG. 11 is a side view of a surface plasmon resonance measuring apparatus according to a seventh embodiment of the present invention.

FIG. 12 is a side view of a surface plasmon resonance measuring apparatus according to an eighth embodiment of the present invention.

FIG. 13 is a partially broken side view showing a structural example of a drive shaft portion of a turntable in the surface plasmon resonance measurement apparatus of the present invention.

FIG. 14 is a partially broken side view showing a main part of a surface plasmon resonance measuring apparatus according to a ninth embodiment of the present invention.

FIG. 15 is a block diagram showing an electrical configuration of the surface plasmon resonance measuring device of FIG. 14;

16 is a schematic diagram showing the relationship between the light beam incident angle and the detected light intensity and the relationship between the light beam incident angle and the differential value of the light intensity detection signal in the surface plasmon resonance measurement device of FIG.

FIG. 17 is a graph illustrating a change in measured values in the surface plasmon resonance measurement apparatus in FIG.

FIG. 18 is a plan view showing a surface plasmon resonance measurement apparatus according to a tenth embodiment of the present invention.

FIG. 19 is a partially cutaway side view showing a main part of the surface plasmon resonance measuring apparatus of FIG. 18;

FIG. 20 is a partially broken side view showing a main part of a surface plasmon resonance measurement apparatus according to an eleventh embodiment of the present invention.

FIG. 21 is a partially cutaway perspective view showing a main part of a surface plasmon resonance measurement apparatus according to a twelfth embodiment of the present invention.

FIG. 22 is a perspective view showing a surface plasmon resonance measuring apparatus according to a thirteenth embodiment of the present invention.

FIG. 23 is a perspective view showing a part of the surface plasmon resonance measuring apparatus of FIG. 22;

FIG. 24 is a plan view showing a part of the surface plasmon resonance measuring apparatus of FIG. 22;

FIG. 25 is a front view showing a part of the surface plasmon resonance measuring apparatus of FIG. 22;

FIG. 26 is a perspective view showing an example of a connected measurement unit used in the surface plasmon resonance measurement apparatus of the present invention.

FIG. 27 is a perspective view showing another example of a connected measurement unit used in the surface plasmon resonance measurement apparatus of the present invention.

FIG. 28 is a partial side view showing a surface plasmon resonance measurement apparatus according to a fourteenth embodiment of the present invention.

FIG. 29 is a schematic configuration diagram showing a surface plasmon resonance measurement device according to a fifteenth embodiment of the present invention.

30 is a diagram showing a flow of operations and information processing in the surface plasmon resonance measurement apparatus of FIG. 29.

FIG. 31 is an explanatory diagram illustrating an operation flow in one embodiment of the surface plasmon resonance measurement method of the present invention.

FIG. 32 is a graph showing an example of measurement results obtained by the surface plasmon resonance measurement method of the present invention.

[Explanation of symbols]

 10, 10 'Measuring unit 11, 11' Dielectric block 11a Interface between dielectric block and metal film 12 Metal film 13, 13 'Sample holding frame 13a Interface between dielectric block and metal film 14 Sensing substance 16 Collimator lens 17 Detection means 18 Differential amplifier array 19 Driver 20 Turntable 21 Display means 30 Light beam 31 Laser light source 32 Condensing lens 40 Photodetector 50 Support driving means 60 Controller 61 Signal processing unit 62 Display unit 70 Automatic sample supply mechanism 75 Cassette 76 Chip supply means 77 Surface plasmon resonance measurement means 78 Chip ejection means 80 Measurement unit 80b Barcode 81 Transparent dielectric block 81a Interface between dielectric block and metal film 82 Metal film 83 Sample holding frame 90 Condensing lenses 91, 92 Mirror 93 Collimator lens 100 Measurement chip 110 Measurement unit 111 Cladding layer 112 Optical waveguide layer 120 Measurement unit 121, 122 Mirror 128 Measuring unit holder 130 Rolling bearing 200 Turntable 210 Dielectric block 210a Sample holder 210b Interface between dielectric block and metal film 211 Liquid sample 212 Metal film 213 Light beam 214 Laser light source 215 Optical system 220 Signal processing unit 221 Reference Automatic liquid supply mechanism 222 Measurement unit 230 Sensing substance 231 Turntable 240, 242 Well plate 241, 243 Well plate set 244 Recovery well plate 245 Metal block 245b Liquid flow path 246 Temperature control means 247 Temperature sensor 248 Controller 250 Circulation passage 251 Pump 252 Heater 260 Cover 300 Chip support 400 Guide rod 401 Slide block 402 Precision screw 403 Pulse motor 404 Motor controller 410 Unit connection 411 Connection member 420 Plate 430 Dispenser 431 Dispensing nozzle 432 Support member 440, 450 Unit connection 441 Dielectric bar 451 unit Support plate 452 Unit support hole 460 Measurement unit 500 Support bar 501 Unit support hole 502 Measurement unit 520 Guide rail 521 Slide block 522 Guide rail 523 Slide block 531, 533 Driving means 524 Optical unit 525 Collimator lens 526, 528 Mirror 527 Condenser Lens 600, 610 Computer system 601, 611 Body of computer system 602, 612 Display 603, 613 Keyboard 604 First barcode reader 614 Second barcode reader 620 Third barcode reader

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Nobuhiko Ogura 798 Address, Miyadai, Kaisei-cho, Ashigara-gun, Kanagawa Photo Film Co., Ltd. F-term (reference) in Film Co., Ltd. NN06 PP01

Claims (30)

[Claims] A plurality of measurement units each including a dielectric block, a thin film formed on one surface of the dielectric block, and a sample holding mechanism for holding a sample on the surface of the thin film layer; A support that supports the measurement unit, a light source that generates a light beam, and a light source that emits the light beam with respect to 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 incident angle of; an optical detection means for measuring the intensity of a light beam totally reflected at the interface to detect a state of attenuated total reflection; and sequentially for each dielectric block of the plurality of measurement units. The support and the optical system and the light detecting means are relatively moved so that the total reflection condition and various incident angles can be obtained, and each measuring unit is sequentially moved to the optical system and the light detecting means. Measuring apparatus utilizing attenuated total reflection and a drive means for placing in position against. 2. A dielectric block, a thin film layer formed on one surface of the dielectric block, a sensing substance arranged on the surface of the thin film layer to interact with a specific component in a sample, and a sensing substance of the sensing substance. A plurality of measurement units each including a sample holding mechanism for holding a sample on the surface; a support supporting the plurality of measurement units; a light source for generating a light beam; and the light beam with respect to the dielectric block. An optical system for incidence 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 measuring the intensity of the light beam totally reflected at the interface, Light detection means for detecting the state of attenuation; and the support and the light so that the total reflection condition and various incident angles are sequentially obtained for each dielectric block of the plurality of measurement units. System and a light-detecting means are moved relative to one another, the measuring apparatus that utilizes attenuated total reflection comprising a drive means for placing in position the measuring units for sequential optical system and light-detecting means. 3. A plurality of measurement units each including a dielectric block, a thin film layer formed of a metal film formed on one surface of the dielectric block, and a sample holding mechanism for holding a sample on the surface of the thin film layer. A support for supporting the plurality of measurement units, a light source for generating a light beam, and a condition for total reflection at the interface between the dielectric block and the metal film with respect to the dielectric block. An optical system that makes incident light at various angles of incidence so as to be able to detect the light, and a photodetector that measures the intensity of the light beam totally reflected at the interface to detect a state of attenuated total reflection due to surface plasmon resonance; The support and the optical system and the light detecting means are moved relative to each other so that the total reflection condition and various incident angles are sequentially obtained for each dielectric block of the unit. Tsu preparative successively the optical system and light-detecting means measuring apparatus utilizing attenuated total reflection and a drive means for placing in position relative. 4. A dielectric block, a thin film layer formed of a metal film formed on one surface of the dielectric block, a sensing substance disposed on the surface of the thin film layer and interacting with a specific component in a sample, and A plurality of measurement units each including a sample holding mechanism for holding a sample on the surface of the sensing substance; a support supporting the plurality of measurement units; a light source for generating a light beam; An optical system that enters the body block at various angles of incidence so as to obtain a total reflection condition at the interface between the dielectric block and the metal film, and the intensity of the light beam totally reflected at the interface is measured. Light detecting means for detecting a state of attenuated total reflection due to surface plasmon resonance; and a total reflection condition and various incident conditions for each dielectric block of the plurality of measurement units. Driving means for relatively moving the support, the optical system and the light detecting means, and sequentially arranging each measuring unit at a predetermined position with respect to the optical system and the light detecting means. A measuring device using total reflection attenuation. 5. A thin film layer comprising a dielectric block, a cladding layer formed on one surface of the dielectric block, and an optical waveguide layer formed thereon, and a sample holder for holding a sample on the surface of the thin film layer. A plurality of measurement units including a mechanism, a support supporting the plurality of measurement units, a light source for generating a light beam, and the light beam with respect to the dielectric block, the dielectric block and the clad. An optical system for incidence at various angles of incidence so as to obtain the condition of total reflection at the interface with the layer; and measuring the intensity of the light beam totally reflected at the interface to excite the guided mode in the optical waveguide layer. A light detecting means for detecting a state of attenuated total reflection by the support, so that the total reflection condition and various incident angles are sequentially obtained for each dielectric block of the plurality of measurement units. Serial and an optical system and light-detecting means are moved relative, measurement device using attenuated total reflection and a drive means for placing in position the measuring units for sequential optical system and light-detecting means. 6. A thin film layer comprising a dielectric block, a cladding layer formed on one surface of the dielectric block, and an optical waveguide layer formed thereon, and a thin film layer disposed on the surface of the thin film layer and specified in a sample. A plurality of measurement units each including a sensing substance that interacts with components and a sample holding mechanism for holding a sample on the surface of the sensing substance; a support supporting the plurality of measurement units; and a light beam generation. An optical system for causing the light beam to enter the dielectric block at various angles of incidence so as to obtain a total reflection condition at an interface between the dielectric block and the cladding layer; Light detecting means for measuring the intensity of the totally reflected light beam to detect a state of attenuated total reflection due to excitation of a waveguide mode in the optical waveguide layer; The support and the optical system and the light detecting means are relatively moved so that the total reflection condition and various incident angles are sequentially obtained with respect to the body block, and each measurement unit is sequentially moved to the optical system and the light detecting means. A measuring device using attenuated total reflection, comprising: a driving unit disposed at a predetermined position with respect to the total reflection attenuation. 7. The optical system according to claim 1, wherein the optical system and the light detecting unit are kept stationary, and the driving unit is configured to move the support. A measuring device using the total reflection attenuation described in the item. 8. The apparatus according to claim 8, wherein the support is a turntable that supports the plurality of measurement units on a circumference about a rotation axis, and the driving unit rotates the turntable intermittently. The measuring apparatus according to claim 7, wherein: 9. The supporter supports the plurality of measurement units in a line in a straight line, and the driving means intermittently supports the supporters in the direction in which the plurality of measurement units are arranged. 8. The measuring device using attenuated total reflection according to claim 7, wherein the measuring device is configured to move linearly. 10. The apparatus according to claim 1, wherein said support is kept stationary, and said driving means moves said optical system and light detecting means. A measuring device using the total reflection attenuation described in the item. 11. The supporter supports the plurality of measurement units on a circumference, and the driving unit controls the optical system and the light detection unit on a plurality of measurement units supported by the supporter. 11. The measuring device using attenuated total reflection according to claim 10, wherein the measuring device is rotated intermittently along the unit. 12. The supporter supports the plurality of measurement units in a line in a straight line, and the driving unit supports the optical system and the light detection unit on the supporter. 2. The method according to claim 1, wherein the linear movement is performed intermittently along a plurality of measurement units.
A measuring apparatus using the total reflection attenuation described in item 0. 13. When the driving means has a rolling bearing that supports the rotating shaft, and when the rotating shaft is rotated in one direction to complete a series of measurements on the plurality of measurement units, The rotation axis is returned in the other direction by the same amount as the rotation amount, and then the rotation axis is rotated in the one direction for the next series of measurements. A measuring device using the total reflection attenuation according to claim 8. 14. The plurality of measurement units are connected in a row by a connection member to form a unit connection body, and the support is configured to support the unit connection body. 13. A measuring apparatus using attenuated total reflection according to claim 9 or 12. 15. The apparatus according to claim 1, further comprising means for automatically supplying a predetermined sample to each sample holding mechanism of the plurality of measurement units supported by the support.
4. A measuring device using the attenuated total reflection according to any one of 4. 16. A measurement chip, wherein a dielectric block of the measurement unit is fixed to the support, and a thin film layer of the measurement unit and a sample holding mechanism are integrated to constitute a measurement chip. 16. The measuring device using attenuated total reflection according to claim 1, wherein the measuring device is formed so as to be exchangeable with respect to the block. 17. A cassette containing a plurality of the measurement chips, and chip supply means for taking out the measurement chips one by one from the cassette and supplying the chips in a state of being combined with the dielectric block. A measuring apparatus using the total reflection attenuation according to claim 16. 18. The dielectric block of the measurement unit,
The measuring chip is formed by integrating the thin film layer and the sample holding mechanism, and the measuring chip is formed so as to be exchangeable with respect to the support.
A measuring device using the total reflection attenuation described in the item. 19. A cassette comprising a plurality of the measurement chips, and a chip supply means for taking out the measurement chips one by one from the cassette and supplying the chips in a state supported by the support. 19. A measuring apparatus using attenuated total reflection according to claim 18. 20. The optical system according to claim 1, wherein the optical system causes the light beam to enter the dielectric block in a state of convergent light or divergent light, and the light detecting means is present in the totally reflected light beam. 20. The measuring apparatus using attenuated total reflection according to any one of claims 1 to 19, wherein the position of a dark line is detected by attenuated total reflection. 21. The measuring apparatus utilizing attenuated total reflection according to claim 20, wherein said optical system causes said light beam to enter said interface in a defocused state. 22. The apparatus according to claim 21, wherein a beam diameter in the moving direction of the support at the interface of the light beam is set to 10 times or more of a mechanical positioning accuracy of the support. A measuring device that uses total reflection attenuation. 23. The measurement unit is supported above the support, and the light source is arranged to emit the light beam downward from a position above the support, and the optical system includes: The total reflection attenuation according to any one of claims 1 to 22, further comprising a reflection member that reflects the light beam emitted downward to travel upward toward the interface. Measuring device using. 24. The measurement unit is supported above the support, the optical system is configured to make the light beam incident on the interface from below the interface, and the light detecting means is provided on the support. A light-reflecting member is provided at a position above the body with the light detection surface facing downward, and a reflecting member that reflects the light beam totally reflected at the interface upward and proceeds toward the light detection means is provided. 24. The measuring device using attenuated total reflection according to claim 1, wherein 25. A temperature adjusting means for maintaining the measuring unit before and / or after being supported by the support at a predetermined set temperature. 25. A measuring apparatus using attenuated total reflection according to any one of claims 24 to 24. 26. A device for stirring a sample stored in a sample holding mechanism of a measurement unit supported by the support before detecting the state of attenuated total reflection. 26. A measuring device utilizing attenuated total reflection according to any one of 1 to 25. 27. At least one of the plurality of measurement units supported by the support is provided with reference liquid supply means for supplying a reference liquid having optical characteristics related to the optical characteristics of the sample, A correction means for correcting data indicating the state of the total reflection attenuation for the sample obtained by the detection means based on the data indicating the state of the total reflection attenuation for the reference liquid is provided. Items 1 to 26
A measuring device using the attenuated total reflection according to any one of the preceding claims. 28. The method according to claim 27, wherein when the sample is obtained by dissolving an analyte in a solvent, the reference liquid supply means supplies the solvent as the reference liquid. Measurement device using total reflection attenuation. 29. A mark attached to each of the measurement units, the mark indicating individual identification information, reading means for reading the mark from the measurement unit used for measurement, and sample information on a sample supplied to the measurement unit. Input means for inputting, display means for displaying a measurement result, connected to the display means, the input means and the reading means, and storing the individual identification information and the sample information of each measurement unit in association with each other. And the measurement results obtained for the sample held in a certain measurement unit are
2. A control means for displaying on said display means in association with said individual identification information and said sample information stored for said measuring unit.
28. A measuring apparatus using the attenuated total reflection according to any one of to 28. 30. A measuring method using a measuring apparatus utilizing attenuated total reflection according to any one of claims 1 to 29, wherein a state of attenuated total reflection is detected for a sample in one of the measuring units. Thereafter, the support and the optical system and the light detection means are relatively moved to detect a state of attenuated total reflection with respect to the sample in another measurement unit, and thereafter, the support, the optical system, and the light detection means A measurement method using attenuated total reflection, wherein the state of the attenuated total reflection is detected again with respect to the sample in the one measurement unit by relative movement.