JP4720146B2 - Spectroscopic apparatus and spectral system - Google Patents

Description

  The present invention relates to a spectroscopic device and a spectroscopic system including a spectroscopic device and a confocal microscope.

  The confocal microscope not only has a high two-dimensional resolution of the specimen, but also a high resolution performance in the third dimension along the optical axis, compared to conventional microscopes. The use is spreading. In particular, in a confocal microscope equipped with a laser light source and performing observation by scanning a laser beam on a specimen that is an object to be examined, the light detection unit of the microscope is a pinhole and a photodetector (photomultiplier tube: PMT) and easy connection to the spectroscopic device. For this reason, a microscope capable of obtaining not only the three-dimensional shape observation of the test object but also the spectral information can be configured relatively easily.

  The wavelength dispersion element provided in the spectroscopic device emits incident light in different directions for each light wavelength. For this reason, there has been proposed an apparatus that includes a photodetector having a plurality of detection cells and acquires spectral information of a test object in a short time by simultaneously detecting light of different wavelengths (for example, Patent Documents). 1).

  In the spectrum by the confocal microscope, the fluorescence is captured and the spectrum is performed, but the excitation laser light reflected by the test object or reflected by the cover glass (hereinafter, this excitation laser light is referred to as stray light). Also enters the spectroscope. The intensity of fluorescence is extremely weak compared with the intensity of stray light. For example, the intensity of stray light may be several tens to 100 times the intensity of fluorescence.

  Therefore, if the detection sensitivity of the photodetector (PMT) is matched with the fluorescence intensity, the detector may be damaged because the intensity of the excitation laser light is extremely high. On the other hand, if the detection sensitivity of the photodetector is matched with the intensity of the excitation laser light, the intensity of light emission / fluorescence to be detected is too small to make detection almost impossible. Therefore, in the above-described conventional apparatus, a line filter (strip-shaped light shielding plate) is fixed in accordance with the bright line position of the excitation laser light, and the excitation laser light is blocked by the fixed light shielding plate so that the excitation laser light is detected by the photodetector. So as not to enter.

US Patent Application Publication No. 2002/0020819

  By the way, when the angle of the light dispersion element provided in the spectroscopic device is changed, when the light dispersion element is replaced, or when the wavelength of the excitation laser light is changed, the incident position of the excitation laser light on the photodetector Changes. Therefore, in the conventional spectroscopic device described above, it is necessary to prepare a fixed light shielding plate for each different condition. However, when a plurality of fixed light-shielding plates are provided in this way, there is a possibility that the light to be detected may be blocked, leading to a decrease in detection accuracy.

The spectroscopic device according to the first aspect of the present invention is included in an irradiation unit that irradiates the test object with irradiation light having a predetermined wavelength selected from a plurality of wavelengths, and light from the test object that is irradiated by the irradiation unit. A wavelength dispersion element having a plurality of diffraction gratings having different dispersion characteristics for dispersing the detection target light and the irradiation light reflected by the test object for each wavelength , and any one of the plurality of diffraction gratings, An element switching means selectively disposed in an optical path of light from the test object; and a photodetector comprising a plurality of light detection cells for detecting light dispersed by the selectively disposed diffraction grating ; Shielding light from a predetermined light detection cell of the photodetector so that irradiation light reflected by the test object dispersed by the selectively disposed diffraction grating is not detected by the photodetector. means, with a predetermined wavelength information said selected Based on the information of the dispersion characteristics of the selectively disposed diffraction grating, characterized by comprising a position changing means for changing the light blocking position of the light shielding means.

  According to the present invention, since the light shielding means is changed by the changing means according to the wavelength characteristics of unnecessary light, it is possible to suppress the light to be detected from being shielded as much as possible, and to prevent a reduction in detection accuracy. Can do.

  Hereinafter, the best mode for carrying out the present invention will be described with reference to the drawings. FIG. 1 is a diagram showing an embodiment of a spectroscopic system according to the present invention, and is a block diagram showing a schematic configuration of a spectroscopic system using a scanning confocal microscope. In the spectroscopic system of FIG. 1, the microscope is an epi-illumination microscope, and can capture reflected light, fluorescence, scattered light, etc. in the optical reaction. Hereinafter, fluorescence observation will be described as an example. Further, when configured with a transmission microscope, it is possible to grasp the degree of absorption due to fluorescence, scattered light, and transmitted light amount.

  The spectroscopic system shown in FIG. 1 includes a microscope unit 1, a light source unit 2, a scanning optical system 3, and a spectroscopic unit 4. The light source unit 2 and the scanning optical system 3 are connected by an optical fiber 5. Similarly, the scanning optical system 3 and the spectroscopic unit 4 are connected by an optical fiber 6. The light source unit 2 is provided with laser light sources 21a and 21b having different output wavelengths. In the example shown in FIG. 1, two laser light sources are provided, but three or more laser light sources may be provided. The laser beams emitted from the laser light sources 21 a and 21 b are aligned on the same optical axis using the total reflection mirror 22 and the dichroic mirror 23, and then collected by the condenser lens 24 and incident on the optical fiber 5.

  The laser beam transmitted to the scanning optical system 3 by the optical fiber 5 is emitted from the end face of the fiber with a predetermined NA, and is converted into parallel light by the collimator lens 31 in the scanning optical system 3. The parallel light from the collimator lens 31 is reflected by the dichroic mirror 32 and is incident on the scanning unit 33 constituted by a galvanometer or the like. The scanning unit 33 includes a pair of movable mirrors and can scan the incident laser light two-dimensionally in two directions orthogonal to the optical axis of the objective lens 12 by tilting them together.

  The laser light emitted from the scanning unit 33 is imaged on the primary image plane by the scanning lens 34 and then imaged on the sample 7 which is the test object by the second objective lens 11 and the objective lens 12. The image of the laser beam formed on the sample 7 is a point image and is focused to a size determined by the NA of the objective lens 12. When the laser light is scanned by the scanning unit 33, the laser light L (point image) imaged on the sample 7 is scanned two-dimensionally.

  When the sample 7 is irradiated with laser light, reflection, absorption, fluorescence, scattering, and the like occur in the irradiated region due to the optical characteristics of the sample 7. When fluorescent observation is performed on a biological tissue, the tissue of the sample 7 may be observed by staining with a plurality of fluorescent reagents. Then, by switching between the laser light sources 21a and 21b, the respective fluorescence is observed by irradiating the laser light corresponding to the fluorescent reagent.

  Reflected light, fluorescence, scattered light and the like generated in the irradiation region of the sample 7 are imaged on the primary image plane by the objective lens 12 and the second objective lens 11, and then converted into parallel light by the scanning lens 34. Is incident on. The light from the sample 7 is descanned by being scanned again by the scanning unit 33, and reverses the same optical path as the laser light incident on the scanning unit 33 from the dichroic mirror 32 (which may be a half mirror having a reflectance of 20%). become.

  The light from the sample 7 that has entered the dichroic mirror 32 from the scanning unit 33 passes through the dichroic mirror 32 and is collected by the lens 35 onto the pinhole 36 a of the pinhole light shielding plate 36. The pinhole light-shielding plate 36 is in a positional relationship conjugate with the spot-like laser light (point image) on the sample 7, and the condensing point on the pinhole 36a is an image of the spotlight. Therefore, even if light is generated from other regions on the sample 7, since the light does not form an image on the pinhole 36a, most of the light is blocked by the pinhole light shielding plate 36 and passes through the pinhole 36a. Absent.

  The light that has passed through the pinhole 36 a is collected by the lens 37, enters the optical fiber 6, and is transmitted to the spectroscopic unit 4 through the optical fiber 6. One end of the optical fiber 6 is connected to face the entrance slit 41 of the spectroscopic unit 4, and the light from the sample 7 is emitted at an emission angle determined by the NA of the optical fiber 6, and is split from the entrance slit 41. The light enters the portion 4.

  FIG. 2 is an enlarged detailed view of the spectroscopic unit 4. The light incident from the incident slit 41 is incident on the reflection mirror 42 constituting the incident optical system. The reflection mirror 42 is a concave mirror having the entrance slit 41 as a focal point, and the light incident on the reflection mirror 42 is reflected by the reflection mirror 42 to become parallel light and enters the diffraction grating 43A that is a wavelength dispersion element. The rotary stage 46 to which the diffraction grating 43A is fixed is provided with four diffraction gratings 43A to 43D having different dispersion characteristics.

The light incident on the diffraction grating 43A is emitted from the diffraction grating 43A at a diffraction angle corresponding to the light wavelength. When the wavelength of incident light is λ, the incident angle is α, and the output angle is β, the relationship shown in the following equation (1) holds. Here, m is the diffraction order, and d is the grating interval of the diffraction grating 43A.
sinα + sinβ = mλ / d (1)

  FIG. 2 shows two types of diffracted lights L1 and L2 having different wavelengths. When performing fluorescence observation, most of the light L that passes through the pinhole 26a and enters the spectroscopic unit 4 is fluorescent, but some excitation laser light reflected by the sample 7 or reflected by the cover glass is also included. Yes.

  The fluorescence is diffracted by the diffraction grating 43A, and is emitted in a predetermined angle range according to the wavelength included in the fluorescence. In FIG. 2, the diffracted light L1 shows what is emitted in a predetermined direction among the diffracted fluorescence. On the other hand, the diffracted light L2 represents the diffracted light of the excitation laser light. A reflection mirror 44 that is an emission optical system is provided in the emission direction of the diffracted lights L1 and L2. The reflection mirror 44 is a concave mirror that focuses on the light detection surface of the photodetector 45 that detects the dispersed light. Of the light diffracted by the diffraction grating 43 </ b> A, light in a predetermined wavelength range enters the reflection mirror 44 and is collected on the light detection surface of the photodetector 45 by the reflection mirror 44.

  Diffracted beams L1 and L2 having different diffraction angles are respectively reflected by the reflection mirror 44 and condensed at different positions on the light detection surface of the photodetector 45. As will be described later, in this embodiment, a movable light shielding plate 404 is provided in front of the incident surface of the photodetector 45 (upward in the drawing), and this light shielding plate 404 is disposed at the incident position of the diffracted light L2. The Therefore, the configuration is such that the diffracted light L2 of the excitation laser light is blocked by the light shielding plate 404 and does not enter the photodetector 45.

  On the other hand, when the angle of the rotary stage 46 is changed to change the tilt of the diffraction grating 43A, the wavelength range of the light diffracted in the direction of the reflection mirror 44 changes. That is, the wavelength band of the fluorescence detected by the photodetector 45 can be changed by changing the inclination of the diffraction grating 43A. Further, as described above, since the plurality of diffraction gratings 43A to 43D having different dispersion characteristics can be fixed to the rotary stage 46, for example, when the rotary stage 46 is rotated 180 degrees in FIG. 2, the diffraction grating 43A is replaced. The diffraction grating 43C is ready for use.

  Thus, by changing the dispersion characteristics by switching the diffraction gratings 43 </ b> A to 43 </ b> D, for example, the resolution can be changed by changing the width of the detection wavelength band of the photodetector 45. Here, up to four diffraction gratings can be attached to the rotary stage 46, but the number of attachments is not limited to four.

  FIG. 3 is a perspective view showing the appearance of the photodetector 45. The photodetector 45 includes a photodetector array 45a composed of a plurality of photodetector cells (32-channel PMT) 400 and a shielding mechanism 45b. On the light receiving surface of the photodetector array 45a, the light receiving surfaces of the respective light detection cells 400 are arranged one-dimensionally in the left-right direction in the figure, and light having different diffraction angles (that is, having different wavelengths) by the respective light detection cells 400. Are detected respectively. And by measuring the detection signal output from each photodetection cell 400, the light intensity distribution of the incident light can be obtained. The photodetector 45 is not limited to the PMT, and may be a sensor such as a CCD.

  The shielding mechanism 45b includes a feed screw 401 extending in the left-right direction in front of the photodetector array 45a, a stepping motor 402 that rotationally drives the feed screw 401, and a photodetector array 45a by rotation of the feed screw 401. A slider 403 that moves in the left-right direction, and a light-shielding plate 404 standing on the slider. The light shielding plate 404 is a metal plate that has been subjected to a black matte treatment, but a mirror may be used. When a mirror is used, the light is trapped at the reflected point. The width dimension of the light shielding plate 404 is made slightly larger than the spot projection image by the emission optical system (reflection mirror 44).

  In general, a stepping motor (a) has a small rotation angle error and is less likely to accumulate errors. (B) A high torque can be generated during positioning by flowing an excitation current. (C) Feedback control This has the advantage that speed control and position control are possible without performing this, and these advantages can be utilized when driving the light shielding plate 404. In particular, even in open loop control, the light shielding plate 404 can be moved to the excitation laser light incident position with high accuracy. Of course, a DC servo motor may be used in place of the stepping motor 402. In this case, however, a sensor and a feedback circuit for positioning the DC servo motor are required, which increases the size and cost of the apparatus. End up.

  As shown in FIG. 2, the light shielding plate 404 prevents the diffracted light L <b> 2 of the excitation laser light that is unnecessary light from entering the light detection cell 400. The arrangement position of the light shielding plate 404, which is an incident position of unnecessary light, is determined by the wavelength of the laser light, the type of the diffraction grating, and the inclination of the diffraction grating. Since the wavelength band incident on the photodetector array 45a is changed by changing the inclination of the diffraction grating 43A, the inclination of the diffraction grating 43A is determined by determining the wavelength band to be detected.

  Returning to FIG. 1, the microscope unit 1, the light source unit 2, the scanning optical system 3, and the spectroscopic unit 4 provided in the spectroscopic system are respectively controlled by the control unit 8. An input unit 9 for inputting setting conditions and the like is connected to the control unit 8. As described above, the user input information necessary for determining the position of the light shielding plate 404 is the laser wavelength, the type of diffraction grating, and the wavelength band to be detected. These information are input from the input unit 9 to the CPU 81 of the control unit 8. .

  The CPU 81 selects one of the laser light sources 21a and 21b as a light source based on the input information, drives the rotary stage 46 via the stage controller 83, and moves the light shielding plate 404 to a predetermined position via the light shielding plate controller 84. Move to. The rotation stage 46 is provided with a rotation sensor such as a rotary encoder, and the sensor signal is input to the stage controller 83. Reference numeral 85 denotes a detector controller that controls and drives the photodetector 45, and an output signal of the photodetector 45 is output to the CPU 81 via the detector controller 85. Data necessary for control is stored in advance in the memory 82 of the control unit 8.

《Explanation of movement of shading plate》
FIG. 4 is a flowchart showing an example of the detection operation by the spectroscopic system. First, in step S1, user input information necessary for the operation of the shielding mechanism 45b is input by the input unit 9 and stored in the memory 82. User input information includes laser wavelength (laser light source) selection information, diffraction grating 43A-43D selection information, and wavelength band selection information. The user inputs such information from the input unit 9.

  In step S2, the rotary stage 46 is rotationally driven based on the user input information in step S1, and the designated diffraction grating is moved to the use position as in the diffraction grating 3A in FIG. 2, and then according to the wavelength band. Position at a predetermined angle. In step S3, the position where the light shielding plate 404 should be moved, that is, the light shielding plate position, is calculated based on the input user input information. In the present embodiment, a light shielding plate position reference table (look-up table) in which light shielding plate positions corresponding to laser wavelengths, diffraction grating types, and wavelength bands are converted into data is stored in the memory 82 in advance, and input by a user. The light shielding plate position (that is, the excitation laser light incident position) is obtained from the information and the light shielding plate position reference table.

  In step S4, the number of pulses necessary for driving to the light shielding plate position calculated in step S3 is given to the stepping motor 402 to drive the stepping motor 402, and the light shielding plate 404 is moved to the light shielding plate position obtained in step S3. To move. For example, when a stepping motor 204 with 360 pulses per rotation and a feed screw 401 with two fine threads per mm are used, if the moving distance is 10 mm, 10 × 2 × 360 = 7200 pulses are sent to the stepping motor 402. By sending out, the light shielding plate 404 is moved to a predetermined position.

  When the light shielding plate 404 is moved to a predetermined light shielding plate position, the exciting current of the stepping motor 402 is stopped. In subsequent step S5, the corresponding laser light source is caused to emit light based on the laser wavelength selection information input from the input unit 9, and the fluorescence from the sample 7 is detected by the photodetector 45 to perform fluorescence spectroscopic measurement.

  The reason why the excitation current of the stepping motor 402 is stopped in step S4 is as follows. In the spectroscopic unit 4, the distance between the main surface of the reflection mirror 44 that is an output optical system and the photodetector 45 is an important factor that determines the resolution and spectral range in spectroscopic measurement. Assuming that the dispersion of light in the diffraction grating 43A is dβ / dλ, it spreads on the detection surface of the photodetector 45 as (distance) × (dβ / dλ). Therefore, in order to improve the reproducibility of spectroscopic measurement, it is necessary to improve the reproducibility of “distance”. Therefore, in a spectroscopic device, the base of the device is generally constructed using a low thermal expansion material, and in principle, the spectroscopic device is used under a constant temperature condition.

  By the way, the excitation current for stopping the rotation of the stepping motor, which is generally performed, causes the temperature of the spectroscopic unit 4 to rise due to the increase of the motor temperature, and may cause the spectroscopic unit 4 to be out of order. is there. Therefore, in the present embodiment, the excitation current is stopped during the stop after the light shielding plate 404 is driven. Therefore, hard grease with high viscosity is used for the feed screw 401 so that the stop position is maintained, the number of threads per unit length of the feed screw 401 is increased, or a positioning mechanism such as a click mechanism is used. Anyway.

  As described above, in the present embodiment, since the light shielding plate 404 can be moved, even when the setting condition is changed by switching the laser light sources 21a and 21b and the incident position of the excitation laser light that is unnecessary light is changed, By moving the light shielding plate 404 according to the setting conditions, it is possible to prevent the excitation laser light from entering the photodetector 45. As a result, it is possible to perform fluorescence measurement with higher accuracy by increasing the sensitivity of the photodetector 45.

  On the other hand, in the conventional spectroscopic device described above, since the light shielding plate is fixed, when a plurality of laser light sources are used, the light shielding plate is arranged at each excitation laser light incident position determined by the wavelength of each laser light source. There is a need. When a plurality of fixed light-shielding plates are arranged in this way, a light-shielding plate for a laser light source that is not used shields part of the fluorescent diffracted light to be detected, resulting in a measurement error. End up.

  FIGS. 5A to 5C are diagrams for explaining the shielding of such fluorescence diffracted light. When the laser light source 21a is used, excitation laser light and fluorescence on the detection surface of the photodetector 45 are illustrated. The intensity distribution is schematically shown. 5A to 5C, the horizontal axis represents the position (and wavelength) in the arrangement direction of the light detection cells 400 (see FIG. 3), and the vertical axis represents the diffracted light intensity.

  The distribution curve C1 shown in FIG. 5A represents the diffracted light of the excitation laser light when the sample 7 is irradiated with the laser light source 21a, and the distribution curve C2 represents the fluorescence diffracted light at that time. A distribution curve C3 shows the diffracted light of the excitation laser light when the laser light 21b is used for comparison. Actually, the height of the distribution curves C1 and C3 with respect to the distribution curve C2 is about several tens to 100 times.

  In the case of the conventional apparatus, it is necessary to arrange fixed light-shielding plates at both positions x1 and x2 of the distribution curve C1 and the distribution curve C3 so as to correspond to both the laser light sources 21a and 21b. In this case, the photodetector 45 measures the fluorescent diffracted light represented by the distribution curve C21 as shown in FIG. In other words, since not only the excitation laser beam but also the fluorescence diffracted light is shielded by the fixed light shielding plate arranged at the positions x1 and x2, the fluorescence diffracted light in the areas indicated by reference numerals A1 and A2 is not measured. Therefore, the spectroscopic measurement is affected, and the influence increases as the number of fixed light-shielding plates increases.

  On the other hand, in the present embodiment, since the light shielding plates 404 can be moved according to the setting conditions, one light shielding plate 404 is sufficient. When the laser light source 21a is used, the light shielding plate 404 is moved to the position x1, and when the laser light source 21b is used, the light shielding plate 404 is moved to the position x2. Therefore, the distribution curve of the fluorescent diffracted light measured when the laser light source 21a is used is C22. In this case, only the area A1 at the position x1 where the light shielding plate 404 is disposed is affected, and the number of affected areas is determined by the number of laser light sources, the type of diffraction grating, and the tilt angle of the diffraction grating. One regardless of the number of conditions. As described above, in this embodiment, since one light shielding plate 404 is moved and arranged at the excitation laser light incident position, the influence of the light shielding plate 404 on the spectroscopic measurement can be reduced as compared with the conventional case.

  The conventional apparatus is configured to serve as both the pinhole 36a of the scanning optical system 3 and the entrance slit 41 of the spectroscopic unit 4. Therefore, when the pinhole 36a is changed, the size of the spot light imaged on the photodetector 45 also changes. Therefore, it is necessary to set the width of the light shielding plate to be large so as to correspond to all of the plurality of used pinholes.

  On the other hand, in the spectroscopic system of the present embodiment, the scanning optical system 3 and the spectroscopic unit 4 are connected by an optical fiber 6 so that the light passing through the pinhole 36a is transmitted to the spectroscopic unit 4. Therefore, even if the size of the pinhole 36 a is changed to change the resolution of the scanning optical system 3, the light enters the spectroscopic unit 4 from the incident slit 41 with a diameter determined by the optical fiber 6. As a result, even if the pinhole 36a is changed, the size of the spot light on the photodetector 45 does not change, and it is not necessary to set the width of the light shielding plate 404 unnecessarily large.

  In the embodiment described above, only one set of the shielding mechanism 45b is provided, but a plurality of sets may be provided to provide a plurality of light shielding plates 404. For example, two sets of shielding mechanisms 45b are provided, and one light shielding plate 404 is responsible for shielding the left half region of the light receiving surface shown in FIG. 3, and the other light shielding plate 404 is responsible for shielding the right half region shown in the figure. To be configured.

  In addition, when the light passing through the pinhole 36a is directly incident on the spectroscopic unit 4, the number of shielding mechanisms 45b corresponding to the type of the diameter of the pinhole 36a is provided, and the width of each light shielding plate 404 is provided. May be set to a width corresponding to each pinhole diameter. When the diameter of the pinhole 36a is changed, the diffracted light of the excitation laser light is shielded using the light shielding plate 404 corresponding to the diameter. The light shielding plate 404 that is not used for light shielding is retracted out of the light shielding range by avoiding the front of the detection surface so as not to disturb measurement.

  Furthermore, if the light detector 45 can independently stop the output of each light detection cell 400 and reduce the sensitivity, the light shielding plate 404 can be used to block the excitation laser light incident on the light detection cell, instead of blocking the excitation laser light incident position. The output of the light detection cell 400 may be stopped or the sensitivity may be reduced. In addition, the present invention is not limited to the above embodiment as long as the characteristics of the present invention are not impaired.

  In the correspondence between the embodiment described above and the elements of the claims, the diffraction gratings 43A to 43D are wavelength dispersion elements, the sample 7 is a test object, the light shielding plate 404 is a light shielding means, a feed screw 401, and a stepping step. The motor 402, the slider 403, and the CPU 81 are changing means, the rotary stage 46 is an element switching means, the detector cell controller 85 is a cell control means, and the microscope unit 1, the light source unit 2 and the scanning optical system 2 are confocal microscopes. Constitute.

It is a figure which shows one Embodiment of the spectroscopy system by this invention. 3 is an enlarged detail view of a spectroscopic unit 4. FIG. 2 is a perspective view showing an external appearance of a photodetector 45. FIG. It is a flowchart which shows an example of the detection operation by a spectroscopy system. It is a figure explaining light shielding of fluorescence diffracted light, (a) is distribution curve C1-C3, (b) is distribution curve C2 in the case of a fixed type light shielding plate, (c) is a distribution curve in the case of the present invention. C2 is shown respectively.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Microscope part 2 Light source part 3 Scanning optical system 4 Spectroscopic part 5,6 Optical fiber 7 Sample 8 Control part 9 Input part 21a, 21b Laser light source 36 Pinhole light-shielding plate 36a Pinhole 42,44 Reflection mirror 43A-43C Diffraction grating 45 Photodetector 45a Photodetection array 45b Shielding mechanism 81 CPU
85 Detector cell controller 400 Photodetection cell 401 Feed screw 402 Stepping motor 403 Slider 404 Light shielding plate

Claims (6)

An irradiation means for irradiating the test object with irradiation light of a predetermined wavelength selected from a plurality of wavelengths;
Wavelength dispersion having a plurality of diffraction gratings having different dispersion characteristics for dispersing, for each wavelength, light to be detected included in light from the object irradiated by the irradiation means and light reflected by the object. Elements,
Element switching means for selectively disposing any one of the plurality of diffraction gratings in an optical path of light from the test object;
A photodetector comprising a plurality of photodetector cells for detecting light dispersed by the selectively arranged diffraction grating ;
Light shielding means for shielding a predetermined light detection cell of the photodetector so that irradiation light reflected by the test object dispersed by the selectively disposed diffraction grating is not detected by the photodetector. When,
Position changing means for changing a light shielding position of the light shielding means based on information on the selected predetermined wavelength and information on dispersion characteristics of the selectively disposed diffraction grating. A spectroscopic device. The spectroscopic device according to claim 1,
The irradiation means irradiates the test object with excitation light as irradiation light of the predetermined wavelength,
The spectroscopic apparatus characterized in that the detection target light is fluorescence generated by irradiating the test object containing a fluorescent dye with the excitation light. The spectroscopic device according to claim 1 or 2 ,
The light shielding means is a light shielding plate for preventing the irradiation light reflected by the test object from entering the light detection cell of the photodetector,
The spectroscopic device , wherein the position changing means changes the light shielding position by moving and driving the light shielding plate by an actuator . The spectroscopic device according to any one of claims 1 to 3,
In place of the light shielding means, cell control means for reducing the sensitivity or stopping the operation of each light detection cell provided in the light detector for each light detection cell,
The cell control means is a position where irradiation light reflected by the test object is incident based on information on the selected predetermined wavelength and information on dispersion characteristics of the selectively disposed diffraction grating. spectroscopic apparatus characterized that you causing desensitization or deactivation of the light detecting cells in the. The confocal microscope which scans and irradiates the excitation light condensed on the test object by the objective lens and detects the fluorescence from the test object, and the spectroscopic device according to claim 1 used in the confocal microscope. In a spectroscopic system consisting of
  The plurality of light detection cells are arranged in an array,
  The light-shielding means is composed of a plate-like member that is wider than the width of the luminous flux of the irradiation light reflected by the test object,
  The position changing means is configured to cause the irradiation light reflected by the test object to be the light according to information on the selected predetermined wavelength and information on dispersion characteristics of the selectively disposed diffraction grating. A spectroscopic system, wherein the plate member is moved to a position incident on a detection cell. The confocal microscope which scans and irradiates the excitation light condensed on the test object by the objective lens and detects the fluorescence from the test object, and the spectroscopic device according to claim 1 used in the confocal microscope. In a spectroscopic system consisting of
  The plurality of light detection cells are arranged in an array,
  The light shielding means comprises a plurality of plate-like members wider than the width of the luminous flux of the irradiation light reflected by the test object, depending on the type of the luminous flux width,
  The position changing means is configured to cause the irradiation light reflected by the test object to be the light according to information on the selected predetermined wavelength and information on dispersion characteristics of the selectively disposed diffraction grating. A spectroscopic system, wherein the plate-like member is moved to a position incident on a detection cell according to the type of width of the light beam.

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