Method and apparatus for measuring surface-enhanced raman scattering

Abstract

The present invention relates to a method and an apparatus for measuring surface-enhanced Raman scattering. The present invention provides an apparatus for measuring surface-enhanced Raman scattering, comprising: a Raman scattering measurement unit capable of detecting an optical signal resulting from incident light in a plurality of polarization states applied to a sample including nanoparticles enabling surface-enhanced Raman scattering measurement; a spectrometer for generating a spectral signal by spectroscopically analyzing the optical signal detected by a detector of the Raman scattering measurement unit; and a correction value calculation unit for calculating a correction value for the spectral signal, on the basis of the spectral signal resulting from irradiation with light in the plurality of polarization states.

Description

Surface-enhanced Raman scattering measurement method and device

[0001] The present invention relates to a method and device for measuring surface-enhanced Raman scattering. More specifically, the present invention relates to a method and device for measuring surface-enhanced Raman scattering that can be adjusted or compensated for according to the movement of nanoparticles.

[0002] Raman scattering is a phenomenon in which, when a material is illuminated with light of a certain frequency, light is scattered at a frequency that differs by the natural vibrational energy or rotational energy of a molecule or the lattice vibrational energy of a crystal. The analysis technique using surface-enhanced Raman scattering (SERS) is a highly sensitive analysis technique that obtains information about a material by enhancing the signal compared to conventional Raman scattering through surface treatment of nanoparticles.

[0003] SERS is related to surface plasmon resonance, a phenomenon in which electrons vibrate due to the interaction between light and electrons on a metal surface, and shows superior sensitivity compared to conventional Raman spectroscopy analysis.

[0004] These SERS can be used for highly sensitive and selective molecular identification in biosensing and chemical sensing. For example, Korean Patent No. 1361652 presents a high-speed screening device for multiple drugs using surface-enhanced Raman scattering. Furthermore, Korean Patent No. 1153748 presents a Au / Ag core-shell composite that can be used as a biosensor, demonstrating high selectivity and sensitivity by enhancing surface-enhanced Raman scattering.

[0005] However, when nanoparticles are dispersed in a liquid for molecular diagnosis using SERS, there is a problem in that the SERS intensity varies depending on whether the directions of the nanoparticles and the detector are aligned, which reduces the reproducibility of quantitative analysis.

[0006] Figure 1 is a drawing illustrating the movement of nanoparticles when applying surface-enhanced Raman scattering.

[0007] Referring to (a) of Fig. 1, when a laser for SERS measurement is irradiated on a nanoparticle (1), it shows that the nanoparticle (1) migrates from the laser focus (2) position and thus moves away from the laser focus (2). Referring to (b) of Fig. 1, when a laser for SERS measurement is irradiated on a nanoparticle (1), it shows that the nanoparticle (1) is positioned within the laser focus (2) range, but the nanoparticle (1) rotates.

[0008] When SERS detection of nanoparticles (1) is performed in a moving or rotated state as in Fig. 1, there is a problem that the reproducibility of SERS detection is reduced because the SERS intensity changes.

[0009] [Prior Art Literature]

[0010] (Patent Document 1) Patent Registration No. 1361652

[0011] (Patent Document 2) Patent Registration No. 1153748

[0012] (Non-patent Document 1) Dong-Kwon Lim et al., “Nanogap-engineerable Raman-active nanodumbbells for single-molecule detection”, Nature materials 9 (2010), 60-67

[0013] (Non-patent Document 2) Kim, Jongwoo, et al., “SERS-based particle tracking and molecular imaging in live cells: toward the monitoring of intracellular dynamics.” Nanoscale 11.45 (2019): 21724-21727

[0014] The purpose of the present invention is to provide a surface-enhanced Raman scattering measurement device and method capable of identifying the movement of nanoparticles and performing adjustment or correction accordingly in order to solve the problem of anisotropy of SERS signals due to the movement of nanoparticles.

[0015] The present invention provides a surface-enhanced Raman scattering measurement device comprising: a Raman scattering measurement unit capable of detecting an optical signal by incident light of multiple polarization states for a sample including nanoparticles capable of surface-enhanced Raman scattering measurement; a spectrometer for generating a spectral signal by spectroscopically analyzing the optical signal detected from a detector of the Raman scattering measurement unit; and a correction value calculation unit for calculating a correction value for the spectral signal based on the spectral signal according to the irradiation of light of the multiple polarization states.

[0016] In one embodiment, the Raman scattering measurement unit may include a stage on which the sample is placed, a light source that outputs light, a polarization adjustment unit that adjusts the polarization state of the light output from the light source, and a detector that detects the light signal emitted from the sample by the light irradiated to the sample.

[0017] In addition, the correction value calculation unit can calculate the relative directionality of the nanoparticles with respect to the polarization direction of the incident light based on the first spectral signal by the incident light in the first polarization state and the second spectral signal by the incident light in the second polarization state, and calculate a correction coefficient according to the relative directionality to calculate a correction value for the first spectral signal or the second spectral signal.

[0018] In one embodiment, the relative directionality may be an angle formed by the polarization direction of the incident light and the binding direction of the Raman reporter to the nanoparticle.

[0019] Additionally, the angle can be calculated by the polarization direction of the first polarization state, the polarization direction of the second polarization state, the intensity of the first spectral signal, and the intensity of the second spectral signal.

[0020] In one embodiment, the intensity may be a time-averaged pointing vector in the near field or the far field.

[0021] In addition, the correction value calculation unit can calculate the correction coefficient by comparing the intensity of the spectral signal with the reference intensity for the nanoparticle.

[0022] In one embodiment, the surface-enhanced Raman scattering measurement device may further include a device control unit including a polarization control unit that controls the correction value calculation unit and the polarization adjustment unit to adjust the polarization state of the incident light, and a result value output unit that outputs the correction value calculated by the correction value calculation unit.

[0023] In addition, the present invention provides a surface-enhanced Raman scattering measurement method for processing an optical signal emitted from a sample including nanoparticles capable of surface-enhanced Raman scattering measurement by incident light of multiple polarization states, the method comprising: receiving a first spectral signal according to a first polarization state and a second spectral signal according to a second polarization state; calculating a relative directionality of the nanoparticles with respect to a polarization direction of the incident light based on the first spectral signal and the second spectral signal; and calculating a correction value for the first spectral signal or the second spectral signal based on the relative directionality.

[0024] According to the present invention, there is an effect of providing a uniform SERS detection result by irradiating a sample with light of different polarization states to measure a SERS signal, calculating the relative directionality of nanoparticles according to the polarization direction, and correcting and outputting the SERS signal of the detected nanoparticles.

[0025] Figure 1 is a drawing illustrating the movement of nanoparticles when applying surface-enhanced Raman scattering.

[0026] FIG. 2 is a diagram schematically illustrating a configuration of a surface-enhanced Raman scattering measurement device according to one embodiment of the present invention.

[0027] Figure 3 is a drawing showing an example of a nanoparticle with a Raman reporter attached and polarized light, in which the binding direction of the Raman reporter to the nanoparticle is known.

[0028] Figure 4 is a drawing showing an example of a nanoparticle with a Raman reporter attached and polarized light, in which the binding direction of the Raman reporter to the nanoparticle is unknown.

[0029] FIG. 5 is a flowchart illustrating a surface-enhanced Raman scattering measurement method according to one embodiment of the present invention.

[0030] Figure 6 is a drawing exemplifying the application of the present invention.

[0031] Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the attached drawings. First, when assigning reference numerals to components in each drawing, it should be noted that identical components are assigned the same numerals as much as possible even if they are shown in different drawings. Furthermore, in describing the present invention, if a detailed description of a related known structure or function is judged to obscure the gist of the present invention, the detailed description thereof will be omitted. In addition, although preferred embodiments of the present invention will be described below, it should be understood that the technical idea of ​​the present invention is not limited thereto and can be modified and implemented in various ways by those skilled in the art.

[0032] FIG. 2 is a diagram schematically illustrating a configuration of a surface-enhanced Raman scattering measurement device according to one embodiment of the present invention.

[0033] A surface-enhanced Raman scattering (SERS) measurement device (10) according to one embodiment of the present invention includes a stage (12) on which a sample (14) to be detected by SERS is placed, a light source (20) that outputs light, a polarization adjustment unit (22) that adjusts the polarization state of the light output from the light source (20), a detector (28) that detects an optical signal generated in the sample by light irradiated on the sample, a spectrometer (30) that analyzes the optical signal obtained from the detector (28) to generate a spectral signal, and a device control unit (40) that controls the operation of the device. Here, the stage (12), the light source (20), the polarization adjustment unit (22) and the detector (28), as well as the beam splitter (24) and the objective lens (26) described below, can be understood to constitute a Raman scattering measurement unit.

[0034] The stage (12) is used to load a sample (14), and in one embodiment, the stage (12) can be moved in a horizontal direction (left-right direction in FIG. 2) or a vertical direction (front-back direction of the ground in FIG. 2). The sample (14) includes nanoparticles for applying surface-enhanced Raman scattering.

[0035] A light source (20) outputs light to be irradiated onto a sample (14) placed on a stage (12). The light source may be a laser light source in the near-infrared or visible light band.

[0036] The polarization adjustment unit (22) adjusts the polarization of the light output from the light source (20). It is preferable that the polarization adjustment unit (22) be capable of adjusting the light output from the light source (20) to at least two polarization states of a first polarization and a second polarization. The polarization adjustment unit (22) may be configured to rotate a linear polarizer using a motor, or may be configured to control a liquid crystal variable phase retarder (LCVPR) with an electric signal by providing a liquid crystal variable phase retarder (LCVPR) between a fixed linear polarizer and a λ / 4 wavelength plate. In addition, in one embodiment, the polarization adjustment unit (22) may use a Fiber Polarization Controller (FPC) product of Thorlabs, inc.

[0037] Light whose polarization has been adjusted in the polarization adjustment unit (22) is reflected from the beam splitter (24) and passes through the objective lens (26) to be irradiated onto the sample (14). An optical signal generated in the sample (14) by the irradiated light passes through the beam splitter (24) and enters the detector (28).

[0038] The detector (28) can detect an optical signal generated by light irradiated onto the sample (14) (in the description of the present invention, the optical signal is sometimes referred to as a 'SERS signal'). In one embodiment, the detector (28) can be configured to include a Raman filter and an image sensor such as a Charge Coupled Device (CCD) or Complementary Metal Oxide Semiconductor (CMOS) that acquires the optical signal.

[0039] A spectrometer (30) can analyze an optical signal acquired from a detector (28) to generate a spectral signal. The spectral signal may be a Raman spectrum or an intensity of a signal acquired from the detector (28).

[0040] A device control unit (40) may be provided to control the overall operation of the surface-enhanced Raman scattering measurement device (10). In one embodiment, the device control unit (40) includes a main control unit (42), a polarization control unit (44), a correction value calculation unit (46), and a result value output unit (48). The device control unit (40) may be configured to include one or more microprocessors that operate according to control logic.

[0041] The main control unit (42) can control the sequential operation of the polarization control unit (44), the correction value calculation unit (46), and the result value output unit (48).

[0042] The polarization control unit (44) controls the polarization adjustment unit (22) so that the light output from the light source (20) can be adjusted to a first polarization state or a second polarization state. In the following description, the first spectral signal and the second spectral signal detected by irradiating light of the first polarization state and light of the second polarization state from the light source (20) are described as examples, but in the practice of the present invention, it is also possible to irradiate light of three or more different polarization states to the sample (14), obtain the detection signals, and then calculate the correction value in the correction value calculation unit (46).

[0043] A correction value calculation unit (46) calculates a correction value of a spectral signal based on a first spectral signal of a spectrometer (30) based on an optical signal acquired from a detector (28) in a first polarization state and a second spectral signal of a spectrometer (30) based on an optical signal acquired from a detector (28) in a second polarization state. The correction value can be calculated by multiplying a correction coefficient for either the first spectral signal or the second spectral signal.

[0044] The result value output unit (48) outputs the spectral signal corrected by the correction value calculation unit (46).

[0045] Meanwhile, in FIG. 2, the polarization state of the light output from the light source (20) is adjusted using the polarization adjustment unit (22) to irradiate the polarized light to the sample (14), but in the practice of the present invention, the method of adjusting the polarization of the light irradiated to the sample (14) can be configured in various ways. For example, the polarization direction of the light irradiated to the sample (14) can be adjusted by configuring the light irradiation configuration including the objective lens (26) to be rotatable with respect to the sample (14). Alternatively, by providing a plurality of light sources (20) and making the angles at which the light output from each light source (20) is irradiated to the sample (14) different, it may be possible for light with different polarization states to be irradiated to the nanoparticles included in the sample (14).

[0046] The configuration in which the correction value calculation unit (46) calculates a correction value for the spectral signal based on the first spectral signal of the spectrometer (30) based on the optical signal acquired from the detector (28) in the first polarization state and the second spectral signal of the spectrometer (30) based on the optical signal acquired from the detector (28) in the second polarization state is described in more detail below.

[0047] Figure 3 is a drawing showing an example of a nanoparticle with a Raman reporter attached and polarized light, in which the binding direction of the Raman reporter to the nanoparticle is known.

[0048] In (a) of Fig. 3, the nanoparticle (1) is exemplarily shown as a dipole, and a Raman reporter (3) is attached to the nanoparticle (1). In (a) of Fig. 3, "a" represents the radius of the nanoparticle (1), is a unit vector indicating the binding direction of the Raman reporter (3) to the silver nanoparticle (1). = <n x , n y , n z > can be displayed, and based on the coordinate system shown in Fig. 3, =<0, 1, 0>. That is, the example in Fig. 3 assumes that the binding direction of the Raman reporter (3) to the nanoparticle (1) is known.

[0049] Figure 3 (b) shows incident light (21) irradiated on nanoparticles (1). is a unit vector indicating the direction of propagation of incident light (21), is a unit vector representing the vibration direction of the electric field of incident light (21). = <k x , k y , k z > can be displayed as, = <E x , E y , E z > can be displayed. Based on the coordinate system shown in Fig. 3, =<1, 0, 0>, =<0, E y , E z > In the device configuration of Fig. 2, the polarization direction of the incident light (21) by the polarization adjustment unit (22) is controllable, can be viewed as a known value.

[0050] In the example of Figure 3, =<0, 1, 0>, =<0, E y , E z>Therefore, class If the angle formed is θ1, a relationship similar to mathematical expression 1 is established, and θ1 is derived as in mathematical expression 2.

[0051]

[0052]

[0053] For the light emitted from the Raman reporter (3) of the nanoparticle (1) according to the incident light (21), the time-averaged Poynting vector (P(r,θ)) in the near field can be expressed as in mathematical equation 3.

[0054]

[0055] In mathematical expression 3, r is the distance between the center of the nanoparticle and the center of the Raman reporter, θ is the angle formed by the electric field direction of the irradiating photon and the vector formed by the center of the nanoparticle and the center of the Raman reporter, ω is the angular frequency of the photon irradiating the sample, ε0 is the permittivity in vacuum, ε m represents the relative permittivity.

[0056] In mathematical expression 3, when θ = 0, P(r,θ) is the maximum value (P Max ) and when the Time-Averaged Pointing Vector at θ1 in Equation 2 is P1, P Max The relationship between P and P1 is as shown in mathematical formula 4, and P in mathematical formula 4 Max The value of / P1 becomes the calibration factor.

[0057]

[0058] By irradiating the nanoparticle (1) with incident light (21) and multiplying the correction coefficient of Equation 4 by P1, which is the Time-Averaged Pointing Vector of the SERS signal detected by the detector (28), a uniform spectral signal can be output even if the binding direction of the Raman reporter (3) to the nanoparticle (1) and the polarization state of the incident light (21) are different.

[0059] The example in Fig. 3 is explained assuming that the binding direction of the Raman reporter (3) to the nanoparticle (1) is known, but in general situations, the binding direction of the Raman reporter (3) to the nanoparticle (1) is unknown. Next, the calculation of the correction factor in a situation where the binding direction of the Raman reporter (3) to the nanoparticle (1) is unknown is explained.

[0060] Figure 4 is a drawing showing an example of a nanoparticle with a Raman reporter attached and polarized light, in which the binding direction of the Raman reporter to the nanoparticle is unknown.

[0061] In (a) of Fig. 4, θ is a unit vector representing the vibration direction of the electric field of the incident light (21). is a unit vector indicating the binding direction of the Raman reporter (3) to the nanoparticle (1). This is the angle formed. I don't know = <n x , n y , n z > can be expressed as

[0062] A unit vector indicating the direction of propagation of incident light (21) based on the coordinate system shown in Fig. 4 =<1, 0, 0>, =<0, E y , E z >, and the polarization state of the incident light (21) is in the first polarization state. =<0, E 1y , E 1z > and in the second polarization state =<0, E 2y , E 2z > It is assumed that it is controlled by .

[0063] , and If the angles formed by these are θ1 and θ2, respectively, the relationship between θ1 and θ2 is as shown in mathematical expression 5.

[0064]

[0065] Referring to Equation 3 for the Time-Averaged Pointing Vector in the Near Field, the relationship between P1, which is the Time-Averaged Pointing Vector of the SERS signal measured in the first polarization state, and P2, which is the Time-Averaged Pointing Vector of the SERS signal measured in the second polarization state, can be expressed as Equation 6.

[0066]

[0067] , is a value that can be controlled by setting the first polarization state and the second polarization state, and since P1 and P2 are values ​​that can be known through measurement, θ1 and θ2 can be calculated from mathematical expressions 5 and 6.

[0068] Once θ1 and θ2 are calculated, a correction coefficient for each SERS signal can be calculated by applying θ1 or θ2 to Equation 4 for P1, which is the Time-Averaged Pointing Vector of the SERS signal in the first polarization state, or P2, which is the Time-Averaged Pointing Vector of the SERS signal in the second polarization state.

[0069] A simple example of obtaining the correction coefficient in the example of Fig. 4 is described. In Fig. 4 =<0, 1, 0>, If it is controlled as =<0, 0, 1>, then in mathematical expression 5, cos|θ2-θ1|=0 and |θ2-θ1|=π / 2. In addition, if the value of mathematical expression 6 is, for example, P1 / P2=4 as a result of the measurement of P1 and P2, the value of θ1 is solved as 0 or π from the two equations. In mathematical expression 3, since the relationship P(r, θ+π)=P(r, θ) is established, θ1 can be determined as 0, and this value can be substituted into mathematical expression 4 to obtain the correction coefficient.

[0070] Meanwhile, in mathematical expression 4, the correction coefficient is P Max It was set based on P Max can be understood as a reference value of a spectral signal that can be set as needed. That is, in the practice of the invention, the reference value of the spectral signal for calculating the correction coefficient is P, which is the maximum value of the spectral signal. Max It may also be possible to set it to a value smaller than or equal to .

[0071] In the above description, the time-averaged Poynting vector in the near field was used to calculate the magnitude of the first spectral signal for the optical signal in the first polarization state detected by the detector (28) and the second spectral signal for the optical signal in the second polarization state detected by the detector (28). However, in the practice of the present invention, it may also be possible to use the time-averaged Poynting vector in the far field when comparing the magnitudes of the first spectral signal and the second spectral signal. In addition, the time-averaged Poynting vector in the near field or the far field described above can be understood as one of the methods for calculating the intensity of the spectral signal. That is, the main feature of the present invention is that it confirms the direction of bonding between the nanoparticle and the Raman reporter by using the intensity of light emitted from the Raman reporter attached to the nanoparticle by light of a different polarization state irradiated on the nanoparticle, and corrects the spectral signal from the spectrometer (30) based on this, thereby outputting consistent results.

[0072] FIG. 5 is a flowchart illustrating a surface-enhanced Raman scattering measurement method according to one embodiment of the present invention.

[0073] The light source (20) is controlled to irradiate light onto a sample containing nanoparticles (S10).

[0074] The light output from the light source (20) is irradiated to the sample in a first polarization state and in a second polarization state different from the first polarization state, and a first spectral signal in the first polarization state and a second spectral signal in the second polarization state are measured (S20). The first spectral signal and the second spectral signal measured by the spectrometer (30) are transmitted to the correction value calculation unit (46).

[0075] The correction value calculation unit (46) calculates the relative directionality of the nanoparticles with respect to the polarization direction of light using the first spectral signal in the first polarization state and the second spectral signal in the second polarization state (S30). The relative directionality of the nanoparticles with respect to the polarization direction of light may be the angle formed by the polarization direction of light and the binding direction of the Raman reporter to the nanoparticles.

[0076] The correction value calculation unit (46) calculates a correction coefficient using the angle formed by the polarization direction of light and the binding direction of the Raman reporter to the nanoparticles, and applies the correction coefficient to the measured spectral signal to correct the spectrum of the spectral signal (S40).

[0077] The result value output section (48) outputs the corrected spectrum information (S50).

[0078] Figure 6 is a drawing exemplifying the application of the present invention.

[0079] Pathological diagnosis can be classified into in-vivo diagnosis and in-vitro diagnosis. Among the in-vitro diagnosis methods, molecular diagnosis refers to pathological diagnosis through DNA base sequence detection. The present invention can be utilized in such molecular diagnosis. However, since FIG. 6 is an example showing the application target of the surface-enhanced Raman scattering measurement device and method according to the present invention, the present invention is not limited to the structure or diagnostic purpose of the nanoparticles illustrated in FIG. 6. The present invention can also be applied when a Raman reporter is attached to a single nanoparticle, and can be used for various molecular diagnosis and pathological examinations.

[0080] Figure 6 illustrates two individual nanoparticles (NP1, NP2), two single stranded DNAs (ssDNA1, ssDNA2), a Raman reporter for detecting Raman signals, and a target DNA to be detected. The upper part of Figure 6 shows a negative state where the target DNA is not detected, and the lower part shows a positive state where the target DNA is detected.

[0081] Initial State

[0082] At room temperature, ssDNA1 and ssDNA2 are combined to form double-stranded DNA (hereinafter, dsDNA), and a Raman reporter is positioned between the two nanoparticles (NP1, NP2). In this case, the SERS phenomenon can occur due to coupled plasmons from the two nanoparticles (NP1, NP2).

[0083] Denaturation

[0084] T to prepare for diagnosis MWhen heated to this temperature, dsDNA denatures and separates, unwinding into ssDNA1 and ssDNA2, increasing the distance between the two nanoparticles (NP1 and NP2). Furthermore, ssDNA1 and ssDNA2 can bind to other single-stranded DNA pairs. In this case, the SERS phenomenon can be triggered by plasmons in the nanoparticle (NP2) to which the Raman reporter is attached.

[0085] Hybridization

[0086] For diagnosis, insert target DNA and T A When the temperature is lowered to a certain degree, the paired ssDNAs bind. In the example of Fig. 6, the length of ssDNA2 is made shorter than the length of the target DNA, so that the target DNA has a stronger binding affinity to ssDNA1 than to ssDNA2.

[0087] In the upper case of Fig. 7, where no target DNA exists in the sample, two nanoparticles (NP1, NP2) recombine to generate a SERS phenomenon due to coupled plasmons.

[0088] In this regard, in the lower part of Fig. 7, where target DNA exists in the sample, the target DNA binds to ssDNA1, and the two nanoparticles (NP1, NP2) do not bind again. Accordingly, unlike the initial state, a SERS phenomenon occurs due to the plasmon of NP2.

[0089] Theoretically, the SERS signal from coupled plasmons is known to be several times stronger than the SERS signal from single nanoparticle plasmons. The presence of target DNA can be confirmed by examining the magnitude of this SERS signal.

[0090] The present invention, in detecting SERS signals of nanoparticles, changes the polarization state of light irradiated on the nanoparticles and irradiates the nanoparticles, applies a correction coefficient to the spectral signal that can be measured differently depending on the direction of the nanoparticles, and outputs it uniformly, thereby ensuring reproducibility and providing more accurate information to the tester.

[0091] The above description is merely an illustrative description of the technical idea of ​​the present invention, and those skilled in the art will appreciate that various modifications, changes, and substitutions may be made without departing from the essential characteristics of the present invention. Therefore, the embodiments disclosed in the present invention and the accompanying drawings are not intended to limit the technical idea of ​​the present invention, but rather to explain it, and the scope of the technical idea of ​​the present invention is not limited by these embodiments and the accompanying drawings. The scope of protection of the present invention should be interpreted by the following claims, and all technical ideas within a scope equivalent thereto should be interpreted as being included in the scope of the rights of the present invention.

Claims

1. A Raman scattering measurement unit capable of detecting an optical signal by incident light of multiple polarization states for a sample containing nanoparticles capable of surface-enhanced Raman scattering measurement; A spectrometer that generates a spectral signal by spectroscopically analyzing the optical signal detected from the detector of the Raman scattering measurement unit; and A correction value calculation unit that calculates a correction value for the spectral signal based on the spectral signal according to the light irradiation of the plurality of polarization states; A surface-enhanced Raman scattering measurement device comprising:

2. In paragraph 1, The above Raman scattering measurement unit, The stage on which the above sample is placed, A light source that outputs light, A polarization adjustment unit for adjusting the polarization state of light output from the above light source, and A surface-enhanced Raman scattering measurement device characterized by including a detector that detects the optical signal emitted from the sample by light irradiated on the sample.

3. In paragraph 1 or 2, A surface-enhanced Raman scattering measuring device characterized in that the correction value calculating unit calculates the relative directionality of the nanoparticles with respect to the polarization direction of the incident light based on the first spectral signal by the incident light in the first polarization state and the second spectral signal by the incident light in the second polarization state, and calculates a correction coefficient according to the relative directionality to calculate a correction value for the first spectral signal or the second spectral signal.

4. In paragraph 3, A surface-enhanced Raman scattering measurement device, characterized in that the relative directionality is an angle formed by the polarization direction of the incident light and the binding direction of the Raman reporter to the nanoparticle.

5. In paragraph 4, A surface-enhanced Raman scattering measurement device, characterized in that the angle is calculated by the polarization direction of the first polarization state, the polarization direction of the second polarization state, the intensity of the first spectral signal, and the intensity of the second spectral signal.

6. In paragraph 5, A surface-enhanced Raman scattering measurement device, characterized in that the above intensity is a time-averaged pointing vector in a near field or a far field.

7. In paragraph 4, A surface-enhanced Raman scattering measurement device characterized in that the correction value calculation unit calculates the correction coefficient by comparing the intensity of the spectral signal with the reference intensity for the nanoparticle.

8. In paragraph 1 or 2, A surface-enhanced Raman scattering measurement device characterized in that it further includes a device control unit including a polarization control unit that controls the correction value calculation unit and the polarization adjustment unit to adjust the polarization state of the incident light, and a result value output unit that outputs the correction value calculated by the correction value calculation unit.

9. A surface-enhanced Raman scattering measurement method for irradiating a sample containing nanoparticles capable of surface-enhanced Raman scattering measurement with incident light of multiple polarization states and processing an optical signal emitted from the sample, A step of receiving a first spectral signal according to a first polarization state and a second spectral signal according to a second polarization state by a correction value calculation unit; A step in which the correction value calculation unit calculates the relative directionality of the nanoparticles with respect to the polarization direction of the incident light based on the first spectral signal and the second spectral signal; and A step in which the correction value calculation unit calculates a correction value for the first spectral signal or the second spectral signal according to the relative directionality; A surface-enhanced Raman scattering measurement method comprising:

10. In paragraph 9, In the step of calculating the above correction value, A surface-enhanced Raman scattering measurement method, characterized in that the correction value calculation unit calculates the relative directionality of the nanoparticles with respect to the polarization direction of the incident light based on the first spectral signal by the incident light in the first polarization state and the second spectral signal by the incident light in the second polarization state, and calculates a correction coefficient according to the relative directionality to calculate a correction value for the first spectral signal or the second spectral signal.

11. In paragraph 10, A surface-enhanced Raman scattering measurement method, wherein the relative directionality is an angle formed by the polarization direction of the incident light and the binding direction of the Raman reporter to the nanoparticle.

12. In paragraph 11, A surface-enhanced Raman scattering measurement method, characterized in that the angle is calculated by the polarization direction of the first polarization state, the polarization direction of the second polarization state, the intensity of the first spectral signal, and the intensity of the second spectral signal.

13. In paragraph 12, A surface-enhanced Raman scattering measurement method, characterized in that the above intensity is a time-averaged pointing vector in a near field or a far field.

14. In paragraph 11, In the step of calculating the above correction value, A surface-enhanced Raman scattering measurement method characterized in that the correction value calculation unit calculates the correction coefficient by comparing the intensity of the spectral signal with the reference intensity for the nanoparticle.

Images