JP5488360B2 - Spectrofluorometer - Google Patents

Description

  The present invention relates to a spectrofluorometer that measures fluorescence generated when a sample is irradiated with excitation light.

When light is emitted to a molecule in the ground state, the molecule transitions to an excited state having a high energy level. The excited state molecules lose part of their energy due to vibration or heat, and then return to the ground state due to deactivation due to light emission. At this time, the light emitted from the molecule is fluorescence.
Therefore, in order to examine what the fluorescence emitted by the molecule is, the sample is irradiated with light of the set excitation wavelength λ _{EX1 and} the spectrum (light wavelength distribution) emitted from the sample at that time is measured. A spectrofluorometer has been developed (see, for example, Patent Document 1).

FIG. 5 is a schematic diagram showing an example of a conventional spectrofluorometer.
The spectrofluorometer 101 controls the light source unit 10 that emits light having a set excitation wavelength λ _EX1 , the sample chamber 20 in which the sample S is disposed, the detection unit 30 that measures the spectrum, and the entire spectrofluorometer 101. And a computer 150.

The light source unit 10 includes a light source 11 such as a high-brightness xenon lamp or a flash lamp that emits white light, and a concave diffraction grating 12a that wavelength-decomposes white light and irradiates the sample S with light having a set excitation wavelength λ _EX1. The excitation spectroscope 12 which has this, the excitation side slit 13, the mirror 15, and the monitor detector 14 (Si-PD or a photomultiplier tube) are provided. The concave diffraction grating 12a is rotated by the computer 150 so that the sample S can be irradiated with light having an arbitrary set excitation wavelength λ _EX1 .

The detection unit 30 wavelength-decomposes the light emitted from the sample S and emits light having a target wavelength λ _{n to} the photodetector 32, and a fluorescence spectrometer 31 having a concave diffraction grating 31a and a target wavelength λ _n . A photodetector (photomultiplier tube) 32 for detecting the light intensity and a fluorescence side slit 34 are provided. Concave diffraction grating 31a is adapted to be rotated by the computer 150, and is capable of emitting light of an arbitrary target wavelength lambda _n to light detector 32. As a result, the concave diffraction grating 31a can measure the spectrum by emitting light of the target wavelength λ _{n in} a predetermined region to the photodetector 32 while being rotated by the computer 150.

  The computer 150 includes a CPU (control unit) 151 and a memory 154, and an input device 52 having a keyboard, a mouse, and the like, and a display device 53 are connected to each other. Further, the functions processed by the CPU 151 will be described as a block. A light source control unit 151 a that controls the light source unit 10, a photodetector control unit 151 b that acquires a light intensity signal from the photodetector 32, and the display device 53. And a display control unit 151c for displaying the spectrum.

According to such a spectrofluorometer 101, when an analyst performs analysis of the sample S, the excitation is set for the sample S by controlling the light source unit controller 151a using the input device 52. Irradiation with light having a wavelength λ _EX1 (for example, 350 nm) is performed. The photodetector control unit 151b is emitted from the sample S by acquiring the light intensity signal from the photodetector 32 in a target wavelength range (for example, 300 nm to 800 nm) while rotating the concave diffraction grating 31a. Get the spectrum.
When the analysis of the sample S is completed, the display control unit 151c displays the spectrum on the display device 53. FIG. 6 is a diagram illustrating an example of the obtained spectrum. The vertical axis is the light intensity, and the horizontal axis is the wavelength.

JP-A-10-160673

However, in the spectrum obtained by the spectrofluorometer 101, a peak A of 350.1 nm, a peak B of 397.0 nm, a peak C of 450.1 nm, and a peak D of 700.5 nm appear as shown in FIG. ing. That is, in addition to the peak indicating fluorescence, the peak indicating the Rayleigh scattered light having the same wavelength λ _EX1 as the set excitation wavelength λ _EX1 used for exciting the sample S, the peak indicating the higher-order light of the Rayleigh scattered light, and the vibration of the molecule In some cases, a peak indicating Stokes Raman scattering light due to the above appears. Since a molecule loses a part of its energy due to vibration or heat and then returns to the ground state by deactivation due to light emission, the energy due to light emission is smaller than the excitation energy, so the peak that shows fluorescence is at the set excitation wavelength λ Appears on the longer wavelength side than _EX1 . However, a peak indicating Stokes Raman scattering light also appears on the long wavelength side in the same manner. In addition, a peak indicating higher-order light of Rayleigh scattered light also appears on the long wavelength side. That is, the spectrum obtained by the spectrofluorometer 101 may be complicated by overlapping a plurality of peaks A to D as shown in FIG. Therefore, the analyst may have a glance at the spectrum shown in FIG. 6 and may not know which of the peaks A to D indicates the fluorescence.

Here, the spectrum shown in FIG. 6 includes a peak A indicating Rayleigh scattered light having a set excitation wavelength λ _EX1 , a peak D indicating secondary light of Rayleigh scattered light having a wavelength λ _EX1 × 2, and a peak B indicating fluorescence. And a peak C indicating Stokes Raman scattering light appears. However, the peak judged by the analyst to indicate fluorescence may not be actually fluorescence, but may be peaks A and D indicating Rayleigh scattered light and peak C indicating Stokes Raman scattered light. It was.

In order to solve the above-mentioned problems, the present inventors have determined whether the peak in the obtained spectrum is a peak indicating fluorescence, a peak indicating Rayleigh scattered light, or a peak indicating Stokes Raman scattered light. We examined the identification method to identify whether there is.
First, it was found that a primary determination for determining whether or not a peak in the obtained spectrum is a peak indicating Rayleigh scattered light based on the set excitation wavelength λ _EX1 was performed. Specifically, by referring to the setting excitation wavelength lambda _EX1, if the same wavelength lambda _EX1 peak and setting the excitation wavelength lambda _EX1, to display suggesting that a peak indicating the Rayleigh scattered light, further setting If there is a peak with a wavelength λ _EX1 × n which is n times the excitation wavelength λ _EX1 , a display indicating that the peak indicates the nth-order light of Rayleigh scattered light is performed. In the primary determination, a peak other than the peak indicating Rayleigh scattered light is displayed to indicate the possibility of being a peak indicating fluorescence.

Next, a secondary determination for determining whether or not a peak other than the peak indicating Rayleigh scattered light is a peak indicating fluorescence based on a change in the wavelength position of the peak when the set excitation wavelength λ _EX1 is changed. Found to do. Here, when the set excitation wavelength λ _EX1 irradiated to the sample S is changed to the longer set excitation wavelength λ _EX2 , the peak wavelength position indicating fluorescence does not change, but the peak indicating Stokes Raman scattered light is not changed. It is known that the wavelength position shifts to the long wavelength side. FIG. 7 is a diagram illustrating an example of changes in peak wavelength positions between a plurality of spectra. As shown in FIG. 7, the set excitation wavelength λ _EX1 is changed in order so that (λ _EX1 +5) nm, (λ _EX1 +10) nm, (λ _EX1 +15) nm, and (λ _EX1 +20) nm. As it goes on, the wavelength position of the peak showing fluorescence does not change. On the other hand, the wavelength position of the peak indicating Stokes Raman scattering light is shifted to the long wavelength side.
Therefore, the sample S is irradiated with light having wavelengths λ _EX2 , λ _EX3 ,... Different from the set excitation wavelength λ _EX1 stepwise to the long wavelength side, and the spectrum emitted from the sample S at that time is measured. . And if the wavelength position of the peak between multiple spectra is shifted to the long wavelength side, a display suggesting that it is a peak indicating Stokes Raman scattering light is performed, and the wavelength position of the peak between multiple spectra If is not shifted, a display suggesting that it is a peak indicating fluorescence is performed.
Note that, when performing the secondary determination, if a spectrum in the target wavelength range of 300 nm to 800 nm emitted from the sample S is measured, it takes measurement time. Therefore, it has also been found that when performing a secondary determination, a peak is efficiently and clearly identified in a very short measurement time by measuring a spectrum in the minimum target wavelength range.

In addition, the peak indicating the nth-order light of the Rayleigh scattered light may overlap with the peak indicating fluorescence. FIG. 8 is a diagram illustrating an example of the obtained spectrum. In the spectrum shown in FIG. 8A, a peak indicating the Rayleigh scattered light having the set excitation wavelength λ _EX1 and a peak indicating the secondary light of the Rayleigh scattered light having the wavelength λ _EX1 × 2 appear. And the peak which shows the secondary light of the Rayleigh scattered light of wavelength (lambda) _EX1 * 2 and the peak which shows fluorescence have overlapped. Therefore, even when the analyst looks at the spectrum shown in FIG.
Therefore, it has been found that the peak indicating the Rayleigh scattered light is removed by using an optimal Rayleigh cut filter. Here, when a Rayleigh cut filter that blocks light having a cutoff wavelength (for example, (λ _EX1 +10) nm) or less is arranged in front of the fluorescence spectrometer 31, Rayleigh scattered light having a set excitation wavelength λ _{EX1 in} the obtained spectrum. And a peak indicating the secondary light of the Rayleigh scattered light having the wavelength λ _EX1 × 2 can be removed. Therefore, as shown in FIG. 8B, with reference to the set excitation wavelength λ _EX1 , a Rayleigh cut filter that blocks light having a cutoff wavelength or less is arranged. Thereby, a spectrum as shown in FIG. 8C is obtained.
When executing the secondary determination, as shown in FIG. 9, a plurality of Rayleigh cut filters having different cutoff wavelengths are provided, and each specified by λ _EX1 , λ _EX2 , λ _EX3,. Of the cut-off wavelengths, a Rayleigh cut filter that is longer than the set excitation wavelength and has the smallest difference in the number of wavelengths from the excitation wavelength can be selected more accurately by selecting from a plurality of Rayleigh cut filters. .

That is, the spectrofluorometer of the present invention includes a light source that emits white light and an excitation spectrometer having a diffraction grating that wavelength-decomposes the white light and irradiates the sample with light having a set excitation wavelength. A fluorescence spectrometer having a diffraction grating that wavelength-decomposes light emitted from the sample and emits light of a target wavelength to a photodetector; and A detection unit comprising a photodetector for detecting light intensity; and a control unit for acquiring a spectrum by detecting the light intensity in a target wavelength region by the photodetector by controlling the excitation spectrometer; A spectrofluorometer comprising a display device and an input device, wherein the control unit, based on the set excitation wavelength, each peak in the obtained spectrum is a peak indicating Rayleigh scattered light, or fluorescent also Display and suggesting either could be a peak indicating the Raman light, for said fluorescent or peak was displayed suggesting the possibility that a peak indicating the Raman light, the secondary judgment together with the display In addition, the control unit performs stepwise light with a wavelength different from the set excitation wavelength by designating a peak for executing the secondary determination with the input device. In addition to irradiating the sample on the longer wavelength side, the light detector detects the light intensity in the wavelength region that is narrower than the target wavelength region and includes the wavelength of the specified peak, thereby acquiring a plurality of spectra. Based on the change in the wavelength position of the peak among the plurality of spectra, the designated peak is a peak indicating Stokes Raman scattering light or a peak indicating fluorescence. The display suggesting either a click so that performed on the display device.
Here, the “set excitation wavelength” is an arbitrary wavelength determined in advance by an analyst or the like before analysis, and is, for example, 350 nm or the like.
The “target wavelength region” is an arbitrary wavelength region determined in advance by an analyst or the like before analysis, and is, for example, 300 nm to 800 nm.

  As described above, according to the spectrofluorometer of the present invention, it is possible to identify whether the peak in the obtained spectrum is a peak indicating fluorescence or a peak indicating Rayleigh scattered light.

(Means and effects for solving other problems)
In the above invention, the control unit controls the fluorescence spectrometer to irradiate the sample with light having a wavelength different from the set excitation wavelength in a stepwise manner to the long wavelength side, and to emit a plurality of spectra. Based on the change in the wavelength position of the peak between multiple spectra, a display indicating either a peak indicating Stokes Raman scattering light or a peak indicating fluorescence is performed. May be.
As described above, according to the spectrofluorometer of the present invention, it is possible to identify whether the peak in the obtained spectrum is a peak indicating fluorescence or a peak indicating Stokes Raman scattering light.

Further, in the above invention, the control unit irradiates the sample with a light having a wavelength different from the set excitation wavelength stepwise to the long wavelength side when a peak is designated by the input device. In doing so, the spectrum may be acquired by detecting the light intensity in a wavelength region that is narrower than the target wavelength region and includes the wavelength of the designated peak by the photodetector.
As described above, according to the spectrofluorometer of the present invention, it is possible to quickly identify whether the peak in the obtained spectrum is a peak indicating fluorescence or a peak indicating Stokes Raman scattering light. it can.

The spectrofluorometer of the present invention also includes a light source that emits white light and an excitation spectrometer having a diffraction grating that wavelength-decomposes the white light and irradiates the sample with light having a set excitation wavelength. A fluorescence spectrometer having a diffraction grating that wavelength-decomposes light emitted from the sample and emits light of a target wavelength to a photodetector; and A detection unit comprising a photodetector for detecting light intensity; and a control unit for acquiring a spectrum by detecting the light intensity in a target wavelength region by the photodetector by controlling the excitation spectrometer; A spectrofluorophotometer comprising a display device and an input device, wherein the detection unit includes a Rayleigh cut filter that blocks light below the cutoff wavelength in front of the fluorescence spectrometer, and blocks the Rayleigh cut filter. The wavelength is From serial setting excitation wavelength so that a long wavelength side.
As described above, according to the spectrofluorometer of the present invention, the peak showing Rayleigh scattered light can be removed, and the peak in the obtained spectrum can be identified as the peak showing fluorescence. .

In the above invention, the control unit controls the fluorescence spectrometer to irradiate the sample with light having a wavelength different from the set excitation wavelength in a stepwise manner to the long wavelength side, and to emit a plurality of spectra. Based on the change in the wavelength position of the peak between multiple spectra, a display indicating either a peak indicating Stokes Raman scattering light or a peak indicating fluorescence is performed. May be.
As described above, according to the spectrofluorometer of the present invention, it is possible to identify whether the peak in the obtained spectrum is a peak indicating fluorescence or a peak indicating Stokes Raman scattering light.

And in the said invention, when the said control part irradiates a sample by changing the light of the wavelength different from the said setting excitation wavelength to the long wavelength side stepwise by designating a peak with the said input device, A spectrum may be acquired by detecting the light intensity in a wavelength region that is narrower than the target wavelength region and includes a specified peak wavelength by a photodetector.
As described above, according to the spectrofluorometer of the present invention, it is possible to quickly identify whether the peak in the obtained spectrum is a peak indicating fluorescence or a peak indicating Stokes Raman scattering light. it can.
Furthermore, in the above invention, the detection unit includes a plurality of Rayleigh cut filters each having a different cutoff wavelength, and the control unit controls the detection unit based on the set excitation wavelength, thereby providing a plurality of Rayleigh filters. One Rayleigh cut filter may be selected from the cut filters, and the Rayleigh cut filter may be disposed in front of the fluorescence spectrometer.

1 is a schematic configuration diagram illustrating an example of a spectrofluorometer according to Embodiment 1. FIG. It is a figure which shows an example of the acquired spectrum and an identification table. It is a figure which shows an example of the acquired spectrum and an identification table. It is a schematic block diagram which shows an example of the spectrofluorometer which concerns on Embodiment 2. FIG. It is a schematic block diagram which shows an example of the conventional spectrofluorimeter. It is a figure which shows an example of the acquired spectrum. It is a figure which shows an example of the change of the wavelength position of the peak between several spectra. It is a figure which shows an example of the acquired spectrum. It is a figure for demonstrating several Rayleigh cut filters.

  Hereinafter, embodiments of the present invention will be described with reference to the drawings. The present invention is not limited to the embodiments described below, and includes various modes without departing from the spirit of the present invention.

<Embodiment 1>
FIG. 1 is a schematic configuration diagram illustrating an example of a spectrofluorometer according to the first embodiment. In addition, the same code | symbol is attached | subjected about the thing similar to the spectrofluorometer 101. FIG.
The spectrofluorometer 1 controls the light source unit 10 that emits light having a set excitation wavelength λ _EX1 , the sample chamber 20 in which the sample S is disposed, the detection unit 30 that measures the spectrum, and the entire spectrofluorometer 1. And a computer 50.

  The computer 50 includes a CPU (control unit) 51 and a memory 54, and an input device 52 having a keyboard, a mouse, and the like, and a display device 53 are connected to each other. Further, the functions processed by the CPU 51 will be described in a block form. A light source control unit 51a that controls the light source unit 10, a photodetector control unit 51b that acquires a light intensity signal from the photodetector 32, and a display device 53. A display control unit 51c for displaying the spectrum and an identification table display control unit 51d for displaying the identification table on the display device 53.

According to such a spectrofluorometer 1, when an analyst performs analysis of the sample S, the excitation is set for the sample S by controlling the light source unit control unit 51a using the input device 52. Irradiation with light having a wavelength λ _EX1 (for example, 350 nm) is performed. The photodetector control unit 51b acquires the spectrum emitted from the sample S by acquiring the light intensity signal from the photodetector 32 in the target wavelength range of 300 nm to 800 nm while rotating the concave diffraction grating 31a. To do.
Then, when the analysis of the sample S is completed, the display control unit 51 c displays the spectrum on the display device 53. Further, the identification table display control unit 51d refers to the set excitation wavelength λ _EX1 by executing the primary determination, and indicates the Rayleigh scattered light if there is a peak of the same wavelength λ _EX1 as the set excitation wavelength λ _EX1. Display indicating that it is a peak, and if there is a peak with a wavelength λ _EX1 × n that is n times the set excitation wavelength λ _EX1 , a display indicating that the peak indicates the n-order light of Rayleigh scattered light The identification table for performing is displayed.

FIG. 2 is a diagram illustrating an example of the obtained spectrum and the identification table. The vertical axis is the light intensity, and the horizontal axis is the wavelength. In the spectrum shown in FIG. 2, a peak A of 350.1 nm, a peak B of 397.0 nm, a peak C of 450.1 nm, and a peak D of 700.5 nm appear. Further, the identification table shown in FIG. 2 displays that the peak A of the set excitation wavelength λ _EX1 indicates that it is a peak indicating Rayleigh scattered light, and the peak D of the wavelength λ _EX1 × 2 indicates Rayleigh scattered light. Display indicating that it is a peak indicating secondary light, peak B is indicating that it may be a peak indicating fluorescence or Raman light, and peak C is a peak indicating fluorescence or Raman light. The display suggests the possibility of being. In addition, the indication “Δ mark” for performing the secondary determination is also performed for those that suggest the possibility of a peak indicating fluorescence or Raman light.

Further, when the analyst determines that the analyst performs the secondary determination for the predetermined peak using the input device 52, the identification table display control unit 51d has wavelengths λ _EX2 , λ _EX3 ,... Different from the set excitation wavelength λ _EX1. Is irradiated to the sample S in a stepwise manner to the long wavelength side, and a spectrum in a narrow wavelength range (λ _peak −30 nm) to (λ _peak +50 nm) emitted from the sample S at that time is measured. If the peak wavelength position is shifted to the longer wavelength side, a display indicating that the peak is indicative of Stokes Raman scattering light is performed. If the peak wavelength position is not shifted, the peak indicating fluorescence is displayed. Display to suggest that there is.
For example, first, when the analyst determines that the secondary determination is performed on the peak B of 397.0 nm using the input device 52, the wavelengths λ _EX2 , λ _EX3 ,..., Λ _EX5 different from the set excitation wavelength λ _EX1. The sample S is irradiated to the long wavelength side in 5 steps every 10 nm. The photodetector control unit 51b acquires the spectrum emitted from the sample S by acquiring the light intensity signal from the photodetector 32 in a narrow wavelength range of 370 nm to 450 nm while rotating the concave diffraction grating 31a. To do. And since the wavelength position of a peak does not shift, the display which suggests that it is a peak which shows fluorescence is performed.

Next, when the analyst determines that the secondary determination is performed on the peak C of 450.1 nm using the input device 52, the wavelength λ _EX2 , λ _EX3 ,..., Λ _EX5 is different from the set excitation wavelength λ _EX1 . The sample S is irradiated with light at 10-nm increments in five steps on the long wavelength side. The photodetector control unit 51b acquires the spectrum emitted from the sample S by acquiring the light intensity signal from the photodetector 32 in a narrow wavelength range of 420 nm to 500 nm while rotating the concave diffraction grating 31a. To do. And since the wavelength position of the peak shifted, the display which suggests that it is a peak which shows Stokes Raman scattering light is performed.
FIG. 3 is a diagram illustrating an example of the obtained spectrum and the identification table. The vertical axis is the light intensity, and the horizontal axis is the wavelength. In the spectrum shown in FIG. 3, a peak A of 350.1 nm, a peak B of 397.0 nm, a peak C of 450.1 nm, and a peak D of 700.5 nm appear. In addition, the identification table shown in FIG. 3 displays that the peak A of the set excitation wavelength λ _EX1 indicates that it is a peak indicating Rayleigh scattered light, and that peak B is a peak indicating fluorescence. Display indicating that peak C is a peak indicating Stokes Raman scattering light, and peak D of wavelength λ _EX1 × 2 is a peak indicating secondary light of Rayleigh scattered light. A suggestion is displayed. In addition, about the thing which suggested the possibility of being the peak which shows the fluorescence or Raman light shown in FIG. 2, the display "completed" for showing that secondary determination was performed is also performed.

  As described above, according to the spectrofluorometer 1 of the present embodiment, the peak in the obtained spectrum is a peak indicating fluorescence, a peak indicating Rayleigh scattered light, or Stokes Raman scattered light. Whether it is a peak can be identified in a short time.

<Embodiment 2>
FIG. 4 is a schematic configuration diagram illustrating an example of a spectrofluorometer according to the second embodiment. In addition, the same code | symbol is attached | subjected about the thing similar to the spectrofluorometer 101. FIG.
The spectrofluorometer 201 controls the light source unit 10 that emits light of the set excitation wavelength λ _EX1 , the sample chamber 20 in which the sample S is disposed, the detection unit 230 that measures the spectrum, and the entire spectrofluorometer 201. And a computer 250.

  The computer 250 includes a CPU (control unit) 251 and a memory 54, and an input device 52 having a keyboard, a mouse, and the like, and a display device 53 are connected to each other. Further, the functions processed by the CPU 251 will be described as a block. A light source control unit 251 a that controls the light source unit 10, a photodetector control unit 251 b that acquires a light intensity signal from the photodetector 32, and the display device 53. And a display control unit 251c for displaying the spectrum, and an identification table display control unit 251d for displaying the identification table on the display device 53.

The detection unit 230 wavelength-decomposes the light emitted from the sample S and emits the light having the target wavelength λ _{n to} the photodetector 32, and the fluorescence spectrometer 31 having the concave diffraction grating 31a and the light having the target wavelength λ _n . It includes a photodetector (photomultiplier tube) 32 that detects light intensity, a fluorescence side slit 34, and a plurality of (for example, five) Rayleigh cut filters 233 that block light having a cutoff wavelength or less. The plurality of Rayleigh cut filters 233 have different cutoff wavelengths and are selected by the computer 250, so that an arbitrary Rayleigh cut filter 233 can be disposed in front of the fluorescence spectrometer 31. Yes. At this time, for example, based on the set excitation wavelength λ _EX1 , the computer 250 arranges a Rayleigh cut filter 233 that blocks light having a cutoff wavelength (for example, (λ _EX1 +10) nm) or less in front of the fluorescence spectrometer 31. (See FIG. 9).

According to such a spectrofluorometer 201, when an analyst performs analysis of the sample S, the excitation is set for the sample S by controlling the light source control unit 251a using the input device 52. Irradiation with light having a wavelength λ _EX1 (for example, 350 nm) is performed. Based on the set excitation wavelength λ _EX1 , the photodetector control unit 251b arranges an arbitrary Rayleigh cut filter 233 in front of the fluorescence spectrometer 31 and rotates the concave diffraction grating 31a while rotating the concave diffraction grating 31a. The spectrum emitted from the sample S is acquired by acquiring the light intensity signal in the target wavelength range of 300 nm to 800 nm.

Then, when the analysis of the sample S is completed, the display control unit 251c displays the spectrum on the display device 53. When the identification table display control unit 251d determines to execute the secondary determination for the predetermined peak, the light of the wavelengths λ _EX2 , λ _EX3 ,... Different from the set excitation wavelength λ _EX1 is gradually changed to the long wavelength side. The sample S is irradiated, and an arbitrary Rayleigh cut filter 233 is placed in front of the fluorescence spectrometer 31, and the narrow wavelength range (λ _peak −30 nm) to (λ _peak +50 nm) emitted from the sample S at that time is Measure the spectrum. If the peak wavelength position is shifted to the longer wavelength side, a display indicating that the peak is indicative of Stokes Raman scattering light is performed. If the peak wavelength position is not shifted, the peak indicating fluorescence is displayed. Display to suggest that there is.
For example, first, when it is determined that the secondary determination is performed on the peak B of 397.0 nm, light of wavelengths λ _EX2 , λ _EX3 ,..., Λ _EX5 different from the set excitation wavelength λ _EX1 is increased in five steps every 10 nm. The sample S is irradiated with the wavelength side. Light detector control unit 251b is set excitation wavelength lambda _EX2, lambda _EX3, · · ·, based on lambda _EX5, and place any Rayleigh cut filter 233 in front of the fluorescent spectroscope 31, a concave diffraction grating 31a The spectrum emitted from the sample S is acquired by acquiring the light intensity signal from the photodetector 32 in the narrow wavelength range of 370 nm to 450 nm while rotating. And since the wavelength position of a peak does not shift, the display which suggests that it is a peak which shows fluorescence is performed.

Then, when it is determined to execute the secondary determination for the peak C of 450.1nm is set excitation wavelength lambda _EX1 different wavelengths lambda _EX2, lambda _EX3, · · ·, long wavelength 10nm every five levels the light of lambda _EX5 The sample S is irradiated on the side. Light detector control unit 251b is set excitation wavelength lambda _EX2, lambda _EX3, · · ·, based on lambda _EX5, and place any Rayleigh cut filter 233 in front of the fluorescent spectroscope 31, a concave diffraction grating 31a The spectrum emitted from the sample S is acquired by acquiring the light intensity signal from the photodetector 32 in the narrow wavelength range of 420 nm to 500 nm while rotating. And since the wavelength position of the peak shifted, the display which suggests that it is a peak which shows Stokes Raman scattering light is performed.
As a result, in the identification table, a display indicating that the peak B is a peak indicating fluorescence is performed, and a display indicating that the peak C is a peak indicating Stokes Raman scattering light is performed.

  As described above, according to the spectrofluorometer 1 of the embodiment, it is possible to identify in a short time whether the peak in the obtained spectrum is a peak indicating fluorescence or a peak indicating Stokes Raman scattering light. Can do.

<Other embodiments>
(1) In the spectrofluorometer 1 described above, the primary determination is first performed, and the analyst performs the secondary determination using the input device 52. However, when the primary determination is performed, the first determination is automatically performed. It is good also as a structure which performs a next determination.
(2) In the spectrofluorometer 201 described above, the configuration in which the arbitrary Rayleigh cut filter 233 is automatically arranged in front of the fluorescence spectrometer 31 is shown, but the optimum type of the Rayleigh cut filter is displayed on the display device. The analyzer may arrange a Rayleigh cut filter.

  The present invention can be suitably used for a spectrofluorometer and the like.

DESCRIPTION OF SYMBOLS 10 Light source part 11 Light source 12 Excitation spectrometer 12a Concave diffraction grating 20 Sample chamber 30 Detection part 31 Fluorescence spectroscope 31a Concave diffraction grating 32 Photodetector 51 Control part

Claims (1)

A light source unit comprising: a light source that emits white light; and an excitation spectrometer having a diffraction grating that wavelength-decomposes the white light and irradiates the sample with light having a set excitation wavelength;
A sample chamber in which the sample is disposed;
A detection unit comprising a fluorescence spectrometer having a diffraction grating that wavelength-decomposes light emitted from the sample and emits light of a target wavelength to a photodetector, and a photodetector that detects light intensity of the target wavelength When,
By controlling the excitation spectroscope, the light detector detects the light intensity in the target wavelength region, thereby obtaining a spectrum;
A spectrofluorometer comprising a display device and an input device,
The control unit may determine whether each peak in the obtained spectrum based on the set excitation wavelength is a peak indicating Rayleigh scattered light, or a peak indicating fluorescence or Raman light. For the display that suggests the display and the peak that performed the display suggesting the possibility of being a peak indicating the fluorescence or Raman light, the display device performs a display for performing secondary determination together with the display,
Furthermore, the control unit is configured to irradiate the sample by changing the light having a wavelength different from the set excitation wavelength stepwise to the long wavelength side by designating a peak for performing the secondary determination with the input device, By detecting the light intensity of the wavelength region that is narrower than the target wavelength region and includes the wavelength of the specified peak with the photodetector, a plurality of spectra are acquired,
An indication indicating whether the designated peak is a peak indicating Stokes Raman scattering light or a peak indicating fluorescence based on a change in wavelength position of peaks between the plurality of spectra. The spectrofluorometer characterized by performing the above-mentioned on the display device .

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