JP4535975B2 - Data calibration method in surface plasmon resonance spectrum measuring apparatus - Google Patents

Description

  The present invention relates to a data calibration method in a surface plasmon resonance spectrum measuring apparatus, and more specifically, an optical measurement unit for detecting light absorption and emission and a surface plasmon for monitoring a change in refractive index of a sample using a near field as probe light. The present invention relates to a data calibration method in a surface plasmon resonance spectrum measuring apparatus for correcting a refractive index sensitivity generated by a temperature environment at the time of measurement in an optical sensor that measures a resonance (Surface plasmon resonance: SPR) spectrum.

The SPR sensor is a means for detecting a refractive index change in the vicinity of the metal surface from the shift of the SPR spectrum. The change in refractive index can be derived from the shift amount Δθ _SPR of the SPR angle θ _SPR that is the incident angle corresponding to the minimum point of the reflectance of the SPR spectrum. This Δθ _SPR is known to be sensitive to changes in temperature environment during measurement. Therefore, a change in the refractive index sensitivity of the SPR sensor included in the optical measurement unit may occur due to a temperature change caused by the external environment at the time of measurement and a heat source in the optical measurement unit. In particular, if a highly sensitive measurement with a refractive index change of 10 ⁻⁵ to 10 ⁻⁷ [RIU] (RIU: unit of relative refractive index sensitivity) is to be performed, it is necessary to suppress the change in refractive index sensitivity due to temperature. is there.

  Therefore, conventionally, a method has been used in which a thermostat using a liquid or a Peltier element is incorporated in the optical measurement unit so that the temperature of the optical measurement unit is always constant.

“Channel system”, [online], Biacore, Inc. [searched August 3, 2005], Internet <URL: http://www.biacore.co.jp/3_1_2.shtml>

  However, in an SPR measurement apparatus incorporating a thermostatic apparatus system that maintains such a temperature environment, the apparatus becomes large and expensive, and as a result, it is difficult to perform high-precision measurements in a portable size. There was a problem.

  An object of the present invention is to provide a data calibration method in an SPR spectrum measuring apparatus that can measure with high accuracy even if it is miniaturized and suppresses the cost.

The present invention, in order to achieve the above object, an invention according to claim 1, in the data calibration method in the surface plasmon resonance spectrometer, at least one or more predetermined temperature in the range of Jo Tokoro by measuring the spectrum of the substance with respect to the steps of acquiring first spectral data relating to the spectrum of the material at predetermined temperature, the first spectral data, correspondence to a predetermined temperature, stored as reference spectral data and creating a database, by measuring the spectrum of an unknown sample, and obtaining the measurement temperature when the spectral measurement of the second spectral data and unknown samples for spectrum the unknown sample, the surface plasmon resonance spectrometer multilayer Makumo a multilayer membrane model that models the measurement system configuration has Acquiring Rudeta performs optical calculation based on the multi-layer film model data, acquiring theoretical curve data in the form of the spectrum, with reference to the database, corresponding to the measured temperature of the reference spectral data, the reference spectrum acquiring third spectral data relating to the theoretical curve data, and linear transformation so that the maximum value and the minimum value matches the maximum value and the minimum value of the third spectral data respectively, the related theoretical spectra Obtaining the spectrum data of 4 and performing Fourier transform on the third spectrum data and the fourth spectrum data, and dividing the Fourier spectrum-transformed fourth spectrum data by the Fourier-transformed third spectrum data and, step of acquiring first data related to device functions If, acquiring a third data divided by the second data acquired by the first data and the second spectral data to Fourier transform, and the inverse Fourier transform of the third data, the Rukoto to have a acquiring fifth spectral data relating to the calibration spectra is characterized.

According to a second aspect of the present invention, in the data calibration method in the surface plasmon resonance spectrum measuring apparatus according to the first aspect, the calibration spectrum is applied to a polynomial based on the fifth spectral data, and the angle corresponding to the minimum point is used. The method further includes the step of obtaining a surface plasmon resonance angle.

The invention according to claim 3 is the data calibration method in the surface plasmon resonance spectrum measuring apparatus according to claim 2, wherein the step of obtaining the surface plasmon resonance angle creates a calibration curve using the surface plasmon resonance angle, The method has a step of correcting the refractive index sensitivity of the surface plasmon resonance spectrum.

  According to the present invention, in an apparatus equipped with a thermometer for monitoring a refractive index and measuring an SPR spectrum, a theoretical curve obtained by optical calculation and a measured spectrum of a substance having a small refractive index change due to a temperature change near room temperature. And a database that records the theoretical spectrum of the output intensity of the detector derived from the above and the measured spectrum of a substance with a small refractive index change due to a temperature change near room temperature at each indicated value of the thermometer in the apparatus. Thus, it is possible to calibrate the deviation of the refractive index sensitivity depending on the temperature environment at the time of measurement without requiring expensive constant temperature equipment.

  The present invention is a method for correcting a deviation in refractive index sensitivity of an SPR sensor caused by a change in temperature environment during measurement by performing data processing with reference to a database without applying a thermostatic apparatus system in an SPR measurement apparatus. It is. Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings.

  FIG. 1 shows the configuration of an SPR measurement device according to an embodiment of the present invention. This SPR measurement device includes a Kretschmann type or Otto type optical measurement unit M and a calculation and result display unit P. The optical measuring unit M totally reflects the light emitted from the light source 1 through the prism on the metal thin film formed on the glass substrate 2 on which the sample is arranged, and detects the reflected light with the detector 4. Although not shown in FIG. 1, a mirror, a lens, and the like can be arranged in the optical path between the light source 1 and the detector 4 according to the design of the apparatus. Further, the temperature of the optical measurement unit M at the time of measurement is measured by the thermometer 3. Spectral data measured in the optical measurement unit M and data relating to the temperature at the time of measurement are processed and processed in the measurement data, and the calculation and result display unit P includes a personal computer 5 that displays the result on a display unit such as a display. Sent.

  Furthermore, the optical measurement unit M includes a first control unit (not shown) that controls the entire optical measurement unit M. The first control unit includes a CPU that executes control, a ROM that stores a control program for the CPU, and a first storage unit that includes a RAM that provides a work area for the CPU. The configuration is also integrated and controlled by this control unit. The personal computer 5 of the calculation and result display unit P also includes a second control unit that controls the calculation and result display unit P. The second control unit includes a CPU that executes control, and a second storage unit that includes a ROM that stores a control program for the CPU and a RAM that provides a work area for the CPU. These components are also integrated and controlled by this control unit. The personal computer 5 also includes an input operation unit (not shown) including a keyboard or various switches for inputting predetermined commands or data, etc., connected to the second control unit.

FIG. 2 shows a flowchart of calibration spectrum creation processing according to an embodiment of the present invention. The flowchart shown in FIG. 2 is a procedure performed by the first and second CPUs according to the programs stored in the first and second storage units, respectively. Before measuring an unknown sample, an actual spectrum for a substance (in this embodiment, water is used) that has a small refractive index change due to a temperature change near room temperature, such as water or air, disposed in the flow path 2 in advance. Measure for each indicated value of thermometer 3. The first control unit acquires actual spectrum data relating to the actual spectrum measured for the measured water, and transmits the acquired actual spectrum data to the personal computer 5. When the personal computer 5 receives the measured spectrum data, the second control unit uses these measured spectra as a reference at temperatures T _i (i = 1, 2,..., N) in the assumed measured temperature range. A database is prepared in advance as the spectrum B (T _i ). That is, the second control unit acquires reference spectrum data related to the reference spectrum B (T _i ) at the temperature T _i based on the actually measured spectrum data transmitted from the optical measurement unit M, creates a database, and stores the second spectrum. Memorize in means. (Step 201).

  On the other hand, a theoretical curve represented as a reflectance [%] with respect to the measurement angle [degree] (incident angle of the light emitted from the light source 1 with respect to the metal thin film) obtained by optical calculation is detected with respect to the measurement angle [degree]. Light intensity [A. U. ] To obtain theoretical spectrum A data obtained in accordance with the procedure of FIG. 3 (described later) (step 204). Next, the theoretical spectrum A data is Fourier transformed. That is, the second control unit acquires Fourier transform theoretical spectrum A data related to the actual measurement spectrum A transmitted from the optical measurement unit M, and stores it in the second storage unit (step 205).

When unknown sample measurement is performed here, the first control unit detects the instruction temperature T _S data related to the instruction temperature T _S of the thermometer 3 and the measured spectrum data of the measurement sample detected by the thermometer 3. Is transmitted to the personal computer 5. The second control unit stores the reference spectrum B (T _S ) corresponding to the indicated temperature T _S of the thermometer 3 at the time of measurement into a database in which the reference spectrum B (T _i ) stored in the second storage means is stored. Extract from data and Fourier transform. The second control unit acquires a Fourier transform reference spectrum B (T _S) data regarding the Fourier transformed reference spectrum B (T _S), is stored in the second storage means (step 203).

The correlation coefficient obtained by dividing the theoretical spectrum A subjected to Fourier transform by applying the Fourier self-deconvolution method and the reference spectrum B (T _S ) subjected to Fourier transform, that is, the device function H (T _S ). Ask for. In other words, the second control unit uses the Fourier transform theoretical spectrum A data and the Fourier transform reference spectrum B (T _S ) data stored in the second storage unit to generate a device function H related to the device function H (T _S ). (T _S ) data is acquired and stored in the second storage means (step 206). Specifically, since the data (data before Fourier transform) relating to the theoretical spectrum A and the reference spectrum B (T _i ) is obtained as a primary matrix of the light intensity from the detector, these are respectively obtained in steps 202 and 205. And a device function H (T _S ) is obtained from the following equation 1 in step 206.
H (T _S ) = F [A] / F [B (T _S )] (Formula 1)
The symbol F in the above formula represents the Fourier transform.

Note that steps 201 to 203 and steps 204 and 205 may be performed before step 206, respectively. The actual unknown sample measurement may be performed after step 201 and before steps 202 and 204. As described above, the actual unknown sample measurement results, indicated temperature T _S data of the measured spectral data and at that time for the measurement sample is sent to the personal computer 5 from the optical measuring system M. The second control unit performs Fourier transform on the measured spectrum S (T _S ) of the measurement sample transmitted from the optical measurement unit M at the indicated temperature T _S of the thermometer 3 at the time of the measurement, and the Fourier The Fourier transform actual measurement spectrum S (T _S ) data related to the actual measurement spectrum S (T _S ) for the measurement sample that has been converted is acquired and stored in the second storage means (steps 207 and 208). The device function H (T _S ) is divided by the Fourier-transformed measured spectrum F [S (T _S )]. That is, the second control unit obtains division data from the device function H (T _S ) data and the Fourier transform actually measured spectrum S (T _S ) data stored in the second storage unit, It is stored in the storage means (step 209). The Fourier-transformed actual spectrum F [S (T _S )] divided by the device function H (T _S ) is subjected to inverse Fourier transform (step 210), thereby obtaining a calibration spectrum S ′ at each indicated temperature ( Step 211). Here, measured spectrum S (T _S ) data obtained by measuring an unknown sample is also obtained as a linear matrix of light intensity. Therefore, calibration spectrum S ′ data is obtained as shown in Equation 2 from the device function H (T _S ) data and the Fourier transform actually measured spectrum S (T _S ) data. Note that InverseF in Equation 2 represents an inverse Fourier transform.
S ′ = InverseF {H (T _S ) / F [S (T _S )]} (Formula 2)
That is, the second control unit uses the device function H (T _S ) data and the Fourier transform actually measured spectrum S (T _S ) data stored in the second storage unit, so that the calibration spectrum S ′ represented by Expression 2 is used. Data is acquired and stored in the second storage means.

FIG. 3 shows a flowchart of a theoretical spectrum A creation process according to an embodiment of the present invention. First, in order to obtain a theoretical curve, a multilayer film model is created. In this specification, the multilayer model refers to a system composed of a plurality of media that includes incident light and reflected light at each interface between adjacent media, and analyzes the spectrum of the incident light and reflected light for the entire system. It is a calculation model for. Note that the multilayer film model in the present embodiment is modeled for the same configuration as the optical measurement system M. That is, it is a multilayer film model based on a system consisting of three layers of BK7 glass as a medium, a gold thin film, and air, water, or a modified film as a sample on the gold thin film. The second control unit acquires multilayer film model data related to the multilayer film model input from the input operation unit included in the second control unit, and stores the multilayer film model data in the second storage unit (step 301). 302). Next, in the personal computer 5, optical calculation based on the multilayer film model data is performed, and a theoretical curve is derived in the form of an SPR spectrum expressed as reflectance [%] with respect to the measurement angle [degree] (step 303). Extract the maximum and minimum reflectance. That is, the second control unit acquires theoretical curve data related to the theoretical curve from the multilayer film model data stored in the second storage unit, and stores the theoretical curve data in the second storage unit. Further, the second control unit acquires the maximum value data and the minimum value data of the theoretical curve related to the maximum value and the minimum value of the reflectance from the theoretical curve data, and stores them in the second storage means (step 304). On the other hand, the second control unit obtains reference spectrum B (T _S ) data corresponding to the indicated temperature T _S from the database-generated reference spectrum B (T _i ) data stored in the second storage unit. Extraction (step 305), the maximum value and the minimum value of the light intensity are extracted from the data. The first control unit acquires the actual measured spectrum data of the water related to the measured actual spectrum of water, and transmits the acquired actual measured spectrum data of water to the personal computer 5. The second control unit acquires the maximum value data and the minimum value data of the measured water spectrum regarding the maximum value and the minimum value of the light intensity from the measured spectrum data of the water transmitted from the optical measuring unit M, and stores the second data It is stored in the means (step 306). Next, a linear conversion formula for the theoretical curve is derived that linearly converts the theoretical curve so that the maximum and minimum values of the theoretical curve match the maximum and minimum values of the measured spectrum. The second control unit includes the maximum value data and minimum value data of the theoretical curve stored in the second storage means, and the maximum value data and minimum value of the actual measured spectrum of water stored in the second storage means. From the data, linear transformation formula data relating to the theoretical curve is acquired and stored in the second storage means (step 307). The theoretical spectrum A can be obtained by applying the above linear transformation formula to the theoretical curve. The second control unit obtains theoretical spectrum A data related to the theoretical spectrum A from the theoretical curve data and linear transformation equation data stored in the second storage means, and stores the theoretical spectrum A data in the second storage means (step 308). .

In the data calibration method in the SPR spectrum measuring apparatus according to one embodiment of the present invention, the theoretical spectrum A is used to influence all the effects on various optical system components that are difficult to specify due to changes in the temperature environment. The calibration spectrum S ′ from which the deviation caused by the temperature change included in the actual measurement spectrum S (T _S ) is eliminated can be obtained by renormalization in the device function H (T _S ) using the measured value T _S as an index. .

  FIG. 4 shows a flowchart for SPR angle derivation according to an embodiment of the present invention. After obtaining the calibration spectrum S ′ in step 211, the calibration spectrum S ′ is applied to a polynomial to detect the minimum point of the light intensity (step 404). From the angle corresponding to this minimum point, the calibrated SPR angle of the unknown sample can be obtained (step 405). Here, a two-dimensional graph is drawn in which the calibrated SPR angle and refractive index of a sample with a known refractive index (for example, water, air, or oil whose refractive index is matched) are plotted on the vertical axis and the horizontal axis, respectively. To create a calibration curve. The calibrated SPR angle of the sample with the known refractive index is obtained by the same procedure as that for obtaining the calibrated SPR angle of the unknown sample. By using the calibration curve for the sample with known refractive index and the calibrated SPR angle of the unknown sample, the refractive index sensitivity of the SPR spectrum of the unknown sample can be corrected.

  In the embodiment of the present invention, the measurement system used for obtaining the theoretical spectrum has thermal stability within the measurement temperature range, and forms the metal thin film material and thickness, the refractive index of the prism of the measuring device, and the metal thin film. If there is no change in the substrate material to be processed, it is possible to calibrate spectra of samples in different materials or refractive index ranges in one theoretical spectrum.

  However, when changing the type of metal thin film used for measurement and the material and thickness of the substrate on which the metal thin film is formed, and the matching method between the prism and the substrate, the measured spectrum is measured in each case, and the maximum It is necessary to obtain the value and minimum value and update the theoretical spectrum.

  Further, if the temperature at which the theoretical spectrum A is measured is always kept constant, the refractive index can be determined by using the calibration spectrum S ′ derived from the unknown sample spectrum S (T) measured at any temperature, when the sample and the measurement system are the same. Not only the sensitivity but also the absolute value of the refractive index itself is always equal.

  The optical measurement unit M used for SPR measurement may change due to deterioration of the optical material. Therefore, at the time of actual measurement, it is desirable to prepare for obtaining the theoretical spectrum A by using a stable sample near the room temperature such as water or air each time the apparatus is started. Since the theoretical spectrum A in the present embodiment is obtained with reference to the actually measured spectrum, it includes an element of the apparatus temperature. Therefore, the absolute value of the refractive index obtained is a value including the influence of temperature at the time of actual measurement of the actual measurement spectrum used when deriving the theoretical spectrum.

When the optical measurement unit M and the sample used for _{obtaining the} reference spectrum B (T _i ) are thermally stable within the measurement temperature range, the measurement was performed at different temperatures within the assumed measurement temperature range. Even in this case, the refractive index sensitivity (unit: [degree / RIU]) is almost equal. Therefore, by performing the data processing according to the embodiment of the present invention, it is possible to remove the influence of the spectrum shift due to the temperature environment change from the actually measured spectrum. In a system in which the sample itself changes depending on the temperature, for example, a protein or polymer material, it is possible to extract a change due to the temperature of the measurement object as a change in refractive index, excluding a measurement temperature error.

  FIG. 5 shows a theoretical curve obtained by the SPR measurement device according to one embodiment of the present invention. This theoretical curve is derived by performing optical calculations in a multilayer model assuming a system in which water is present on the gold thin film using BK7 glass having a 50 nm thick gold thin film formed on the substrate 2.

  On the other hand, while the optical measuring unit M is put in a thermostat and the temperature in the optical measuring unit M is changed, the first control unit measures the actual spectrum of water at each indicated value indicated by the thermometer 3 in the optical measuring unit M. The reference spectrum B (T) data related to the reference spectrum B (T) is acquired, and the reference spectrum B (T) data is transmitted to the personal computer 5. The second control unit records the transmitted reference spectrum B (T) data and creates a database.

Next, FIG. 6 shows a theoretical spectrum A obtained by the SPR measurement device according to one embodiment of the present invention. First, the first control unit, indicated temperature thermometer 3 in the optical measuring unit M is T _S, using the BK7 glass thickness 50nm gold thin film is formed on the substrate 2, the water present on the gold thin film The measured spectrum data of water related to the measured spectrum is acquired and transmitted to the second control unit. The second control unit, the measured spectrum data of water at the measurement temperature T _S, and acquires the maximum value data and the minimum value data of the measured spectrum of water on the maximum and minimum values of the light intensity. Then, the second control unit, the maximum value data of the theoretical curves of FIG. 5 (91.243%) and the minimum value data (0.603%), the maximum value data of the measured spectrum of water at the measurement temperature _{T S} The theoretical spectrum A is derived by linearly transforming the theoretical curve data so that the minimum value data matches the minimum value data.

Next, as an unknown sample, a KCl aqueous solution prepared at concentrations of 0.25, 0.5, 0.75, and 1.0 M is prepared. The first control unit obtained the measured spectra S (T _S ) of the KCl aqueous solutions having different concentrations when the indicated values of the thermometer 3 were 26.45 and 26.54 [° C.], respectively. Measured spectrum S (T _S ) data regarding each KCl aqueous solution is transmitted to the personal computer 5. The second control unit, with respect to the measured spectrum S (T _S ) data regarding each KCl aqueous solution, is the water reference spectrum B (T _S ) data (T _S = 26.45, 26.54) corresponding to the indicated temperature. ) And theoretical spectrum A, data processing according to the Fourier self-deconvolution method is performed to obtain eight calibration spectrum data. Note that the theoretical spectrum A, and the measured temperature is a T _S, the maximum value data and the minimum value data of the light intensity from the measured spectrum of the water obtained by performing a measurement just before measurement of the KCl aqueous solution, and the theoretical curve data The one obtained from the linear transformation formula obtained from is used.

FIG. 7 shows a calibration spectrum S ′ (a in FIG. 7), an actual measurement spectrum S (T _S ) in the KCl aqueous solution (b in FIG. 7), and a theoretical spectrum A (in FIG. 7) when using a 0.25M KCl aqueous solution as a sample. An example of c) is shown. The difference between the theoretical spectrum A and the calibration spectrum S ′ represents the difference in the physical refractive index between the water sample and the 0.25 M KCl aqueous solution sample from which the measurement temperature environment and the effects associated therewith have been removed.

The second control unit fits the vicinity of the light intensity minimum point of the calibration spectrum S ′ and the measured spectrum S (T _S ) of the KCl aqueous solution with a quadratic curve, and sets the SPR angle θ as an angle corresponding to the minimum value of the equation. _SPR is calculated.

FIG. 8 shows the refractive index dependence of the SPR angle θ _SPR obtained from the measured spectrum S (T _S ) data and the calibration spectrum S ′ data of the KCl aqueous solution obtained by the SPR measurement device according to one embodiment of the present invention. . The graph shown in FIG. 8 shows the KPR aqueous solution with the SPR angle θ _SPR obtained from the calibration spectrum S ′ data for each indicated value of the thermometer 3 at the time of measurement and the measured spectrum S (T _S ) data of the KCl aqueous solution as the vertical axis. The refractive index of the sample was plotted on the horizontal axis. As a result, in the SPR angle θ _SPR obtained from the measured spectrum S (T _S ) data of the KCl aqueous solution, the indicated value of the thermometer 3 is 26.45 ° C. (a in FIG. 8) and 26.54 ° C. The slope of the straight line is different from (b in FIG. 8), which indicates that the refractive index sensitivity changes with temperature. On the other hand, in the case of calibration angle data θ _SPR obtained from the calibration spectrum S ′, the slope of the straight line when the indicated temperature is 26.45 ° C. (c in FIG. 8) and 26.54 ° C. (d in FIG. 8). Are almost the same. From this, it can be confirmed that by applying the data processing method according to the Fourier self-deconvolution method of the present invention, the deviation of the refractive index sensitivity due to the measurement temperature environment is suppressed.

  Moreover, the data calibration method in the SPR spectrum measuring apparatus of the present invention includes a light emitter 1 serving as the light source 1 and a detector 4 serving as a two-dimensional imager, and absorption or fluorescence of light from the light source 1 by the sample. It can be applied to all systems in which the temperature change in the environment to which the optical measurement unit M belongs is superimposed on the signal of the detector 4.

It is a figure which shows the structure of the SPR measuring apparatus which concerns on one Embodiment of this invention. It is a figure which shows the flowchart of the calibration spectrum preparation process which concerns on one Embodiment of this invention. It is a figure which shows the flowchart of the theoretical spectrum A preparation process which concerns on one Embodiment of this invention. It is a figure which shows the flowchart about SPR angle derivation based on one Embodiment of this invention. It is a figure which shows the theoretical curve obtained by the SPR measuring apparatus which concerns on one Embodiment of this invention. It is a figure which shows the theoretical spectrum A obtained by the SPR measuring apparatus which concerns on one Embodiment of this invention. The calibration spectrum S ′ (FIG. 7a), the measured spectrum S (T _S ) (FIG. 7b) and the theoretical spectrum A (FIG. 7c) of the 0.25M KCl aqueous solution as a sample. It is a figure which shows an example. Illustrates a refractive index dependency of the measured spectrum S (T _S) data and calibration spectra S 'SPR angle theta _SPR derived from the data of KCl solution obtained by SPR measuring apparatus according to an embodiment of the present invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Light source 2 Glass substrate 3 Thermometer 4 Detector 5 Personal computer M Optical measurement part P Calculation and result display part

Claims (3)

In the data calibration method in the surface plasmon resonance spectrum measuring apparatus,
A step of measuring the spectrum of a substance to at least one or more predetermined temperature in the range of Jo Tokoro, to obtain the first spectral data relating to the spectrum of the material for each of the predetermined temperature,
And creating a pre-Symbol first spectral data, correspondence to the predetermined temperature, the database storing the reference spectrum data,
Measuring a spectrum of an unknown sample to obtain second spectrum data relating to the spectrum of the unknown sample and a measurement temperature at the time of spectrum measurement of the unknown sample;
Acquiring multilayer film model data relating to a multilayer film model modeling the configuration of a measurement system included in the surface plasmon resonance spectrum apparatus;
Performing optical calculations based on the multilayer film model data to obtain theoretical curve data in the form of spectra ;
Referring to the database, obtaining third spectrum data relating to a reference spectrum corresponding to the measured temperature among the reference spectrum data ;
Linearly transforming the theoretical curve data such that the maximum value and the minimum value thereof coincide with the maximum value and the minimum value of the third spectrum data , respectively, to obtain fourth spectrum data related to the theoretical spectrum;
A Fourier transform is performed on the third spectrum data and the fourth spectrum data, and the fourth spectrum data subjected to the Fourier transform is divided by the third spectrum data subjected to the Fourier transform. Obtaining first data; and
Obtaining third data obtained by dividing the first data by second data obtained by Fourier transforming the second spectral data;
It said third data by inverse Fourier transform, data calibration method in the surface plasmon resonance spectrum measuring apparatus according to claim Rukoto which have a acquiring fifth spectral data relating to the calibration spectra. Te data calibration method odor in the surface plasmon resonance spectrometer according to claim 1,
A method for calibrating data in a surface plasmon resonance spectrum measuring apparatus, further comprising the step of applying the calibration spectrum to a polynomial based on the fifth spectrum data and obtaining a surface plasmon resonance angle from an angle corresponding to a minimum point . Te data calibration method odor in the surface plasmon resonance spectrometer according to claim 2, obtaining the surface plasmon resonance angle step,
A method for calibrating data in a surface plasmon resonance spectrum measuring apparatus, comprising the step of creating a calibration curve using the surface plasmon resonance angle and correcting the refractive index sensitivity of the surface plasmon resonance spectrum.

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