JP2008197178A - Laser scanning type microscope - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a laser scanning type microscope equipped with a spectroscopic device to perform high-speed wavelength scanning. <P>SOLUTION: The laser scanning type microscope spectrally splitting and detecting observation light is equipped with: a KTN crystal which spectrally splits the observation light according to dispersion characteristic and emits it; a detector which detects emitted light emitted from the KTN crystal; and a slit which is arranged between the KTN crystal and the detector, and made open in a direction orthogonal to the inclination of the refractive index of the KTN crystal. By controlling voltage to be applied to the KTN crystal, the detection wavelength of the emitted light passing through the slit is dynamically switched. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a laser scanning microscope.

  Confocal laser scanning microscopes are widely used for observation and measurement of biological samples and semiconductor samples, for example, because they have better resolution and contrast than normal microscopes and also have resolution in the sample height direction. . A confocal laser scanning fluorescence microscope in which this is combined with a fluorescence microscope is a microscope that observes fluorescence of a fluorescent substance in a sample with a laser. In recent years, a confocal laser scanning fluorescence microscope often uses a method of irradiating a sample with lasers of a plurality of colors (multiple wavelengths) and observing the sample in multiple colors using a plurality of fluorescent substances.

  At this time, a method using a spectroscopic device is known as a method for accurately measuring multi-color (multi-wavelength) observation light. For example, in Patent Document 1, a method is adopted in which the observation light is dispersed using a diffraction grating (grating) and the diffraction distribution is further tilted to obtain an intensity distribution in a desired wavelength range.

  However, there is a big problem with the method of driving the diffraction grating as described above. That is, it takes time to drive the diffraction grating and the like physically. That is, although multicolor observation light can be measured, the simultaneity of measurement is not ensured.

In particular, in recent biological specimen observation, it is becoming mainstream to observe biological reactions and biological activities in living specimens. In such observations, ensuring simultaneity is a very important factor. Under such circumstances, a spectroscopic device that is driven at high speed has been demanded.
JP 2004-212600 A

  In view of the above technical problem, an object of the present invention is to provide a laser scanning microscope including a spectroscopic device capable of performing high-speed wavelength scanning.

  In the laser scanning microscope for spectroscopically detecting the observation light, the above-described problem of the present invention is to detect the KTN crystal that emits the observation light by dispersing the observation light according to dispersion characteristics, and the detection that detects the emission light emitted from the KTN crystal. And a slit that is disposed between the KTN crystal and the detector and that opens in a direction perpendicular to the refractive index gradient of the KTN crystal, and controls the voltage applied to the KTN crystal, This is achieved by dynamically switching the detection wavelength of the emitted light passing through the slit. Since this configuration does not involve mechanical driving when switching the detection wavelength, the operation is very fast.

  Further, in a laser scanning microscope for spectroscopically detecting the observation light, a KTN crystal that divides and emits the observation light according to a dispersion characteristic, and a KTN crystal for detecting the emission light emitted from the KTN crystal. This can also be achieved by providing a one-dimensional array detector arranged in the gradient direction of the refractive index and dynamically switching the detection wavelength of the emitted light by controlling the voltage applied to the KTN crystal.

  Furthermore, it is desirable that the observation light incident on the KTN crystal is combined with a polarizing beam splitter and a λ / 2 plate, converted into linearly polarized light in the effective direction of the KTN crystal, and then incident on the KTN crystal. With this configuration, it is possible to reduce the loss caused by the polarization characteristics of the KTN crystal.

  Further, in the laser scanning microscope for spectroscopically detecting the observation light, the first KTN crystal that emits the observation light by dispersing the observation light according to a dispersion characteristic, and the emission light that is split by the first KTN crystal, A second KTN crystal that further diverges and emits light in a direction orthogonal to the spectral direction of the emitted light, and a two-dimensional array detector for detecting the emitted light emitted from the second KTN crystal. desirable. According to this configuration, it is possible to simultaneously detect the wavelength distribution of the observation light and the polarization ratio.

  Further, in the laser scanning microscope for spectroscopically detecting the observation light, the first KTN crystal that emits the observation light by dispersing the observation light according to a dispersion characteristic, and the emission light that is split by the first KTN crystal, A second KTN crystal that further diverges and emits light in a direction orthogonal to the spectral direction of the emitted light, a first one-dimensional array detector arranged in a refractive index gradient direction of the first KTN crystal, and the first It is also desirable to have a second one-dimensional array detector arranged in the direction of refractive index tilt of the second KTN crystal. Even with this configuration, it is possible to simultaneously detect the wavelength distribution of the observation light and the ratio of polarization.

  Further, in a laser scanning microscope for spectroscopically detecting observation light, two-dimensional array detection for detecting the KTN crystal that emits the observation light by dispersing the observation light according to dispersion characteristics and the emission light emitted from the KTN crystal It is also desirable to detect the wavelength distribution and its time change by rotating the spectral direction of the emitted light emitted from the KTN crystal by rotating the electric field applied to the KTN crystal. According to this configuration, while acquiring the wavelength distribution by spectroscopy, the time change can be acquired by rotating the spectral direction at a high speed.

  Further, in a laser scanning microscope for spectroscopically detecting the observation light, the KTN crystal that divides and emits the observation light according to dispersion characteristics, a polygon mirror that reflects the emission light emitted from the KTN crystal, and the polyhedral A lens for condensing the reflected light reflected from the mirror, and a one-dimensional array detector arranged on the light condensing surface of the lens and arranged in the spectral direction of the observation light, and a voltage applied to the KTN crystal. It is also preferable to dynamically switch the dispersion of the observation light by controlling.

  Further, in the laser scanning microscope for spectroscopically detecting the observation light, the KTN crystal that emits the observation light by dispersing the observation light according to a dispersion characteristic, and the first polarized light component that is emitted by deflecting the polarization component in the effective direction of the KTN crystal. The first polyhedral mirror that reflects the emitted light of the second and the polarization plane of the second emitted light that is emitted without deflecting the polarization component orthogonal to the effective direction of the KTN crystal is rotated by 90 degrees and again the KTN crystal A combination of a λ / 4 plate and a reflecting member for incidence on the second polygon mirror, a second polygon mirror for deflecting the linearly polarized light incident from the reflecting member and reflecting the second emitted light, and the first A lens that simultaneously collects the reflected light reflected from the polygon mirror and the reflected light reflected from the second polygon mirror, and is arranged on the condensing surface of the lens and arranged in the spectral direction of the observation light A one-dimensional array detector, By controlling the voltage applied to the N crystal, dynamically switches the variance of the observation light, it is also possible to detect the ratio of the wavelength distribution polarization simultaneously.

  Further, in the laser scanning microscope for spectroscopically detecting the observation light, the KTN crystal that emits the observation light by dispersing the observation light according to a dispersion characteristic, and the first polarized light component that is emitted by deflecting the polarization component in the effective direction of the KTN crystal. The first concave mirror that reflects the emitted light and the polarization plane of the second emitted light that is emitted without deflecting the polarization component orthogonal to the effective direction of the KTN crystal are rotated by 90 degrees to form the KTN crystal again. A combination of a λ / 4 plate for incidence and a reflecting member, a second concave mirror for deflecting the linearly polarized light incident from the reflecting member and reflecting the second emitted light, and the first concave mirror The slit is arranged on the common optical path of the reflected light reflected from the second concave mirror and the reflected light reflected from the second concave mirror and opens in a direction orthogonal to the spectral direction, and a desired wavelength is selected and detected by the slit. detection The provided, by controlling the voltage applied to the KTN crystal, it is conceivable to dynamically switch the detection wavelength of the emitted light passing through said slit.

  Further, in the laser scanning microscope for spectroscopically detecting the observation light, the KTN crystal that emits the observation light by dispersing the observation light according to the dispersion characteristic, and the first polarized light component that is emitted by deflecting the polarization component in the effective direction of the KTN crystal. The first concave mirror that reflects the emitted light and the polarization plane of the second emitted light that is emitted without deflecting the polarization component orthogonal to the effective direction of the KTN crystal are rotated by 90 degrees to form the KTN crystal again. A combination of a λ / 4 plate for incidence and a reflecting member, a second concave mirror for deflecting the linearly polarized light incident from the reflecting member and reflecting the second emitted light, and the first concave mirror A desired wavelength is selected and detected by a one-dimensional array detector arranged on the common optical path of the reflected light reflected from the light and the reflected light reflected from the second concave mirror and arranged in the spectral direction, and the slit. Detector Comprising, by controlling the voltage applied to the KTN crystal, it is conceivable to dynamically switch the detection wavelength of the emitted light passing through said slit.

According to the present invention, a laser scanning microscope provided with a spectroscopic device capable of performing high-speed wavelength scanning is provided.
Further, in one embodiment of the laser scanning microscope according to the present invention, polarization anisotropy and wavelength distribution can be acquired simultaneously.

  Furthermore, in one form of the laser scanning microscope according to the present invention, wavelength distribution and time-resolved measurement can be performed simultaneously.

Hereinafter, embodiments of the present invention will be described with reference to the drawings.
FIG. 1 is a schematic diagram of a laser scanning microscope having a general shape used in the practice of the present invention. The microscope main body 1 in the figure observes the sample 3 on the stage 2 using the objective lens 4. A scanning unit 5 is connected to the microscope main body 1, and a laser is introduced from the laser unit 6 through the fiber cable 7 to the scanning unit 5, and laser scanning is performed inside. The reason for the above configuration is that a general laser unit is a large apparatus and cannot be directly attached to the microscope body. Furthermore, a computer terminal 8 is provided, which is used to control the scan unit 5 and the laser unit 6 and to visualize the detection results obtained.

  FIG. 2 is a schematic diagram illustrating the internal configuration of a conventional laser scanning microscope. In an ordinary laser scanning microscope, a plurality of laser light sources 9 such as an argon ion laser (488 nm), a He—Ne green laser (543 nm), and a He—Ne laser (633 nm) are mounted as a laser light source, and a dichroic mirror 10 is combined. Thus, one laser unit 6 is configured by combining them in the same optical path. The laser combined in one optical path is introduced into the fiber cable 7 via the fiber coupling mechanism 13 after adjusting the light amount by an AOTF (Acousto-optic tunable filters) 12.

  The laser introduced from the fiber cable 7 into the scan unit 5 is reflected by the dichroic mirror 15 toward the objective lens 4. In the middle of the optical path, a galvanometer mirror 16, a pupil projection lens 17, and an imaging lens 18 are arranged and have a function of scanning a laser.

  Observation light from the sample passes through the reverse optical path, passes through the dichroic mirror 15 and enters the detection optical path. An imaging lens 20 is disposed in the detection optical path, and the observation light is imaged on a confocal pinhole 21. The observation light that has passed therethrough is dispersed by a diffraction grating (grading) 22, and then a detector (photomultiplier). ) 25 to detect observation light having a desired wavelength. Here, reference numerals 23 and 24 in the figure denote a concave mirror and a slit, respectively. At this time, the acquisition wavelength band can be changed by changing the angle between the diffraction grating 22 and the concave mirror 23, or by making the slit 24 variable.

Reference numerals 11, 14, and 19 in the figure are mirrors for bending the optical path.
The present invention greatly reduces the time loss associated with driving the diffraction grating 22, the concave mirror 23, or the slit 24 with KTN (potassium tantalate niobate) crystals. Therefore, the function of the KTN crystal will be described with reference to FIG.

  The KTN crystal 26 is an optical element that changes its refractive index (photoelectric effect) when a voltage is applied. The principle is as follows. When a voltage is applied by the two electrodes 27, an electric field gradient 28 is generated. As a result, a refractive index distribution is generated in the KTN crystal 26. The incident light is deflected by this refractive index distribution.

However, this action of the KTN crystal 26 has polarization characteristics. That is, the above-described action occurs only in the linearly polarized light in the effective incident direction.
FIG. 4 is a diagram for explaining the operation when random polarized light is incident on the KTN crystal 26. In the figure, a solid line indicates effective linearly polarized light, which is deflected in the electric field direction as described above. On the other hand, the component orthogonal to the effective incident direction behaves differently. Most of this orthogonal component is orthogonal, and the rest is deflected opposite to the effective linear polarization (approximately 1/8 swing angle). That is, when random polarized light enters the KTN crystal 26, the light beam is separated in the direction of the electric field.

  Next, a basic configuration for performing wavelength selection using the KTN crystal 26 will be described with reference to FIG. Like a general optical element, the KTN crystal 26 also has dispersion. That is, when multi-wavelength light is incident, it is decomposed and emitted for each wavelength. Therefore, by extracting a part of the emitted light through the slit 29 or the opening, it is possible to select only a light beam having a desired wavelength. In addition, since the refractive index distribution of the KTN crystal 26 can be changed by controlling the voltage applied to the electrode 27, the selected wavelength can be changed without moving the slit 29. Since this operation does not involve material driving, the operation is fast.

  Hereinafter, an embodiment of the present invention in which the KTN crystal 26 is used for spectroscopy in a laser scanning microscope will be described. Here, only the internal configuration of the scan unit 5 will be described, but other portions may have the same configuration as the conventional one.

  FIG. 6 is a schematic diagram of an internal configuration of the scan unit 5 according to the embodiment of the present invention. In the figure, the multi-wavelength laser light introduced from the laser unit through the fiber cable is reflected by the dichroic mirror 15 toward the objective lens 4. On the contrary, the observation light from the sample passes through the dichroic mirror 15 and is focused on the confocal pinhole 21 by the imaging lens 20.

  In this embodiment of the present invention, the KTN crystal 26 is arranged at the subsequent stage of the confocal pinhole 21. As described above, the KTN crystal 26 has a function of separating and emitting incident light. The split observation light is collimated by the concave mirror 23 and applied to the slit 24. At this time, a lens or the like may be used instead of the concave mirror 23. However, a mirror that does not generate chromatic aberration is preferable to handle the dispersed light.

  Since the observation light irradiated to the opening of the slit 24 is limited to a certain wavelength band, the observation light limited to a certain wavelength band is provided to the detector 25 (preferably photomultiplier) arranged at the subsequent stage of the slit 24. Is detected. At this time, the wavelength band applied to the opening of the slit 24 can be adjusted by controlling the voltage applied to the KTN crystal 26.

  FIG. 7 is a schematic diagram of the internal configuration of the scan unit 5 according to an embodiment of the present invention. This configuration is a configuration that prevents loss of light amount due to the polarization characteristics of the KTN crystal in the configuration of the first embodiment. Therefore, the description of the same components as those in the first embodiment is omitted.

  This embodiment of the present invention has a configuration in which a mechanism for converting observation light into linearly polarized light is disposed between the confocal pinhole 21 and the KTN crystal 26. Specifically, a configuration in which the polarization beam splitter 30 and the λ / 2 plate 31 are combined can be considered. First, the polarizing beam splitter 30 separates the linearly polarized light in the effective direction of the KTN crystal 26 and the linearly polarized light in the orthogonal direction thereof. Thereafter, the polarization plane of the linearly polarized light orthogonal to the effective direction is rotated by 90 degrees using the λ / 2 plate 31. Thereafter, the two linearly polarized lights are synthesized by the synthesis optical member 32. The optical path from the polarization beam splitter 30 to the combining optical member 32 must be appropriately guided by a mirror or the like, but is omitted in the drawing.

  According to this embodiment, it is possible to suppress the loss of light amount due to the polarization characteristics of the KTN crystal.

  FIG. 8 is a schematic diagram of the internal configuration of the scan unit 5 according to an embodiment of the present invention. In this configuration, in the configuration of the first embodiment, a one-dimensional detector 33 such as a line sensor is disposed instead of the detector 25 such as a photomultiplier. With this configuration, the wavelength distribution of the dispersed observation light can be acquired at once.

  FIG. 9 is a schematic diagram of an internal configuration of the scan unit 5 according to the embodiment of the present invention. In this configuration, in the configuration of the second embodiment, a one-dimensional detector 33 such as a line sensor is arranged instead of the detector 25 such as a photomultiplier. With this configuration, it is possible to acquire the wavelength distribution of the dispersed observation light at a time, and to suppress the loss of light amount due to the polarization characteristics of the KTN crystal.

  FIG. 10 is a schematic diagram of the internal configuration of the scan unit 5 according to an embodiment of the present invention. In this configuration, by using the polarization characteristics of the KTN crystal, the ratio between the wavelength distribution and the polarization can be obtained at one time.

  The KTN crystal can bend incident light in a direction in which a voltage is applied and splits the light in that direction. Moreover, the KTN crystal acts only on the linearly polarized light in the effective direction. Therefore, the observation light is dispersed for each orthogonal component by the two KTN crystals 26 and 26 'in which the voltage application directions are orthogonal to each other. As a result, the emitted observation light is separated in two directions and separated. That is, if the detection is performed by a two-dimensional detector 34 such as a CCD, an observation result dispersed in an L shape can be obtained. At this time, the light intensity in the vertical and horizontal directions represents the degree of polarization.

  According to this configuration, since the ratio between the wavelength distribution and the polarization can be acquired at one time, it is suitable for biological observation applications such as fluorescence polarization and Homo-FRET.

  FIG. 11A is a schematic diagram of the internal configuration of the scan unit 5 according to the embodiment of the present invention. In this configuration, by controlling the electric field applied to the KTN crystal, the wavelength distribution and its time change can be acquired at once.

  The structure of the KTN crystal is cubic and has no optical anisotropy. That is, the refraction direction of incident light is determined only by the direction of the applied electric field. In this configuration, as shown in FIG. 11B, electrodes 26 are arranged on the upper, lower, left and right sides when viewed from the incident surface, and the electric field in all directions in the KTN crystal 26 is controlled by controlling the upper and lower potential differences and the left and right potential differences. The configuration is such that can be generated. For example, at this time, if the upper and lower potential difference and the left and right potential difference are oscillated while being shifted by a quarter phase, the incident random polarized light is emitted while rotating. At this time, the emitted light is split in the radial direction of rotation. That is, if a two-dimensional array detector 34 such as a CCD is arranged on the exit side of the KTN crystal 26 as shown in FIG. 11A, the wavelength distribution and its time change can be acquired at once.

  FIG. 12 illustrates one mode of a spectroscopic method using a KTN crystal. This figure shows an embodiment in which the dispersion on the detection surface is different when the deflection angle of the emitted light is changed by controlling the voltage applied to the KTN crystal 26.

  In the configuration example shown in the figure, the observation light that has passed through the imaging lens 20 and the confocal pinhole 21 is incident on the KTN crystal 26. The spectral light beam emitted from the KTN crystal 26 is reflected by the polygon mirror 35 and detected by the one-dimensional array detector 31 through the lens 36. Spectral rays emitted from the KTN crystal 26 are representatively shown in red (R), green (G), and blue (B). When the deflection angles are different for two types of solid arrows and broken arrows, respectively. The optical path is shown.

  The exit light having the deflection angle indicated by the solid line arrow and the exit light having the deflection angle indicated by the broken line arrow are reflected by different surfaces of the polygon mirror 35. At this time, the angle of the polygon mirror 35 is set so that the reflected light of the center wavelength (G light beam in this example) is parallel. Then, the light beam having the central wavelength is always condensed at the same point of the detector by the lens 36 in the subsequent stage. On the other hand, since the angle when the light beams other than the central wavelength are reflected by the polygon mirror 35 is not kept constant, the position where the light is condensed by the lens 36 is not kept constant. That is, the dispersion on the detection surface differs depending on the difference in deflection angle in the KTN crystal 26.

  That is, according to the above configuration, the acquired wavelength region per pixel of the one-dimensional array detector 31 can be controlled by controlling the voltage applied to the KTN crystal 26.

  FIG. 13 illustrates a development of the spectroscopic method using the KTN crystal of Example 7. In this embodiment, in addition to the effects of the seventh embodiment, the effects of the fifth embodiment can be obtained. That is, the ratio of polarization can be obtained at the same time as the amount of dispersion on the detection surface is changed. In the same figure as in FIG. 12, the rays separated as in FIG. 12 are representatively indicated by red (R), green (G), and blue (B) arrows.

  As described above, the KTN crystal 26 can deflect linearly polarized light in an effective direction, but transmits a polarized component that is not so. This embodiment is characterized in that the linearly polarized light that has been transmitted is incident on the KTN crystal 26 again.

  The observation light that has passed through the imaging lens 20 and the confocal pinhole 21 is incident on the KTN crystal 26. At this time, the optical path of linearly polarized light in the effective direction is the same as that in the seventh embodiment. The spectroscopic light beam emitted from the KTN crystal 26 is reflected by the polygonal mirror 35 and condensed on the one-dimensional array detector 31 by the lens 36. Here, the optical path of linearly polarized light in the effective direction is indicated by broken-line arrows in the drawing. It is also conceivable to use a two-dimensional array detector instead of the one-dimensional array detector 31.

  On the other hand, linearly polarized light having a component orthogonal to the effective direction is transmitted through the KTN crystal 26, then reflected by the λ / 4 + mirror 37 having a reflection film coating on the λ / 4 plate, and again incident on the KTN crystal 26 from the opposite direction. The At this time, the linearly polarized light having a component orthogonal to the effective direction passes through λ / 4 twice, thereby rotating the polarized light by 90 degrees. That is, this time, the linearly polarized light in the effective direction is incident on the KTN crystal 26 and is split by the KTN crystal 26. The optical path at this time is indicated by a solid line arrow in the figure, and becomes an optical path having a relation between a broken line arrow and a mirror image. That is, the spectroscopic light beam emitted from the KTN crystal 26 is reflected by the polygonal mirror 35 and condensed on the one-dimensional array detector 31 by the lens 36.

  According to the above configuration, it is possible to change the amount of dispersion on the one-dimensional array detector 31 and simultaneously obtain the polarization ratio. Instead of the one-dimensional array detector 31, two combinations of slits and detectors may be arranged.

  FIG. 14 illustrates a different form of spectroscopic method using a KTN crystal. In this embodiment, the idea of the seventh embodiment is added to the first embodiment. This embodiment is a different embodiment having the same effect as the second embodiment. That is, also in this embodiment, it is possible to prevent a light amount loss due to the polarization characteristics of the KTN crystal.

  The observation light that has passed through the confocal pinhole 21 is incident on the KTN crystal 26. At this time, the optical path of linearly polarized light in the effective direction is the same as that in the first embodiment. The spectroscopic light beam emitted from the KTN crystal 26 is reflected by the concave mirror 23, a desired wavelength is selected by the slit 24, and detected by the detector 25. Here, the optical path of linearly polarized light in the effective direction is indicated by a solid line arrow in the figure.

  On the other hand, linearly polarized light having a component orthogonal to the effective direction is transmitted through the KTN crystal 26, then reflected by the λ / 4 + mirror 37 having a reflection film coating on the λ / 4 plate, and again incident on the KTN crystal 26 from the opposite direction. The At this time, the linearly polarized light having a component orthogonal to the effective direction passes through λ / 4 twice, thereby rotating the polarized light by 90 degrees. That is, this time, the linearly polarized light in the effective direction is incident on the KTN crystal 26 and is split by the KTN crystal 26. The optical path at this time is indicated by a solid line arrow in the figure, and becomes an optical path having a relation between a broken line arrow and a mirror image. That is, the spectral light beam emitted from the KTN crystal 26 is reflected by the concave mirror 23, a desired wavelength is selected by the slit 24, and detected by the detector 25. At this time, the slit 24 and the detector 25 are shared by both optical paths, and the detector 25 is designed so that the dispersed wavelengths are overlapped, so that the detector 25 can receive a light amount with little loss.

  According to said structure, the loss of the light quantity by the polarization characteristic which a KTN crystal has can be prevented. In place of the detector 25 and the slit 24, a one-dimensional array detector may be arranged.

It is a schematic diagram of a laser scanning microscope of a general shape. It is the schematic explaining the internal structure of the conventional laser scanning microscope. It is a figure explaining the function of a KTN crystal. It is a figure explaining the effect | action when random polarized light injects into a KTN crystal. It is a figure of the basic composition for performing wavelength selection using a KTN crystal. It is the schematic of the internal structure of the scan unit by Example 1 of this invention. It is the schematic of the internal structure of the scan unit by Example 2 of this invention. It is the schematic of the internal structure of the scan unit by Example 3 of this invention. It is the schematic of the internal structure of the scan unit by Example 4 of this invention. It is the schematic of the internal structure of the scan unit by Example 5 of this invention. It is the schematic of the internal structure of the scan unit by Example 6 of this invention. It is the schematic of the spectroscopy by Example 7 of this invention. It is the schematic of spectroscopy by Example 8 of this invention. It is the schematic of the spectroscopy by Example 9 of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 ... Microscope main body 2 ... Stage 3 ... Sample 4 ... Objective lens 5 ... Scan unit 6 ... Laser unit 7 ... Fiber cable 8 ... Computer terminal 9 ... Laser light source 10 ... Dichroic mirror 11 ... Mirror 12 ... AOTF
DESCRIPTION OF SYMBOLS 13 ... Fiber coupling mechanism 14 ... Mirror 15 ... Dichroic mirror 16 ... Galvano mirror 17 ... Pupil projection lens 18 ... Imaging lens 19 ... Mirror 20 ... Imaging Lens 21 ... Confocal pinhole 22 ... Dichroic mirror 23 ... Concave mirror 24 ... Slit 25 ... Detector 26 ... KTN crystal 27 ... Electrode 28 ... Electric field gradient 29 ... Slit 30 ... Polarizing beam splitter 31 ... λ / 2 plate 32 ... Synthetic optical member 33 ... One-dimensional array detector 34 ... Two-dimensional array detector 35 ... Polyhedral mirror 36 ... Lens 37 ... λ / 4 plate + mirror

Claims (10)

In a laser scanning microscope that spectroscopically detects observation light,
A KTN crystal that diverges and emits the observation light according to dispersion characteristics;
A detector for detecting the emitted light emitted from the KTN crystal;
A slit that is disposed between the KTN crystal and the detector and has an opening in a direction perpendicular to the refractive index gradient of the KTN crystal;
A laser scanning microscope characterized by dynamically switching the detection wavelength of the emitted light passing through the slit by controlling a voltage applied to the KTN crystal. In a laser scanning microscope that spectroscopically detects observation light,
A KTN crystal that diverges and emits the observation light according to dispersion characteristics;
A one-dimensional array detector arranged in the refractive index gradient direction of the KTN crystal for detecting the emitted light emitted from the KTN crystal;
A laser scanning microscope, wherein a detection wavelength of the emitted light is dynamically switched by controlling a voltage applied to the KTN crystal. In the laser scanning microscope according to claim 1 or 2,
The laser beam incident on the KTN crystal is converted into linearly polarized light in the effective direction of the KTN crystal by combining a polarizing beam splitter and a λ / 2 plate, and then incident on the KTN crystal. Type microscope. In a laser scanning microscope that spectroscopically detects observation light,
A first KTN crystal that splits and emits the observation light according to dispersion characteristics;
A second KTN crystal that splits and emits the emitted light separated by the first KTN crystal in a direction perpendicular to the spectral direction of the emitted light;
A two-dimensional array detector for detecting the emitted light emitted from the second KTN crystal;
A laser scanning microscope, wherein the wavelength distribution of the observation light and the ratio of polarization are detected simultaneously. In a laser scanning microscope that spectroscopically detects observation light,
A first KTN crystal that splits and emits the observation light according to dispersion characteristics;
A second KTN crystal that splits and emits the emitted light separated by the first KTN crystal in a direction perpendicular to the spectral direction of the emitted light;
A first one-dimensional array detector arranged in the refractive index gradient direction of the first KTN crystal; and a second one-dimensional array detector arranged in the refractive index gradient direction of the second KTN crystal;
A laser scanning microscope, wherein the wavelength distribution of the observation light and the ratio of polarization are detected simultaneously. In a laser scanning microscope that spectroscopically detects observation light,
A KTN crystal that diverges and emits the observation light according to dispersion characteristics;
A two-dimensional array detector for detecting light emitted from the KTN crystal;
A laser scanning microscope characterized by detecting a wavelength distribution and its time change by rotating a spectral direction of the emitted light emitted from the KTN crystal by rotating an electric field applied to the KTN crystal. In a laser scanning microscope that spectroscopically detects observation light,
A KTN crystal that diverges and emits the observation light according to dispersion characteristics;
A polygon mirror that reflects the emitted light emitted from the KTN crystal;
A lens for collecting the reflected light reflected from the polygon mirror;
A one-dimensional array detector arranged on the condensing surface of the lens and arranged in the spectral direction of the observation light;
A laser scanning microscope, wherein dispersion of the observation light is dynamically switched by controlling a voltage applied to the KTN crystal. In a laser scanning microscope that spectroscopically detects observation light,
A KTN crystal that diverges and emits the observation light according to dispersion characteristics;
A first polygon mirror that reflects the first emitted light that is emitted by deflecting the polarization component in the effective direction of the KTN crystal;
A λ / 4 plate and a reflecting member for rotating the polarization plane of the second outgoing light emitted without deflecting the polarization component orthogonal to the effective direction of the KTN crystal by 90 degrees and entering the KTN crystal again. Combination with
A second polygonal mirror that reflects the second emitted light that is emitted by deflecting the linearly polarized light incident from the reflecting member;
A lens that simultaneously collects the reflected light reflected from the first polygon mirror and the reflected light reflected from the second polygon mirror;
A one-dimensional array detector arranged on the condensing surface of the lens and arranged in the spectral direction of the observation light;
A laser scanning microscope characterized in that, by controlling a voltage applied to the KTN crystal, the dispersion of the observation light is dynamically switched to simultaneously detect the wavelength distribution and the ratio of polarization. In a laser scanning microscope that spectroscopically detects observation light,
A KTN crystal that diverges and emits the observation light according to dispersion characteristics;
A first concave mirror that reflects the first emitted light that is emitted by deflecting the polarization component in the effective direction of the KTN crystal;
A λ / 4 plate and a reflecting member for rotating the polarization plane of the second outgoing light emitted without deflecting the polarization component orthogonal to the effective direction of the KTN crystal by 90 degrees and entering the KTN crystal again. Combination with
A second concave mirror that reflects the second emitted light that is emitted by deflecting the linearly polarized light incident from the reflecting member;
A slit disposed on a common optical path of the reflected light reflected from the first concave mirror and the reflected light reflected from the second concave mirror, and opened in a direction perpendicular to the spectral direction;
A detector for selecting and detecting a desired wavelength by the slit;
A laser scanning microscope characterized by dynamically switching the detection wavelength of the emitted light passing through the slit by controlling a voltage applied to the KTN crystal. In a laser scanning microscope that spectroscopically detects observation light,
A KTN crystal that diverges and emits the observation light according to dispersion characteristics;
A first concave mirror that reflects the first emitted light that is emitted by deflecting the polarization component in the effective direction of the KTN crystal;
A λ / 4 plate and a reflecting member for rotating the polarization plane of the second outgoing light emitted without deflecting the polarization component orthogonal to the effective direction of the KTN crystal by 90 degrees and entering the KTN crystal again. Combination with
A second concave mirror that reflects the second emitted light that is emitted by deflecting the linearly polarized light incident from the reflecting member;
A one-dimensional array detector arranged on the common optical path of the reflected light reflected from the first concave mirror and the reflected light reflected from the second concave mirror, arranged in the spectral direction;
A detector for selecting and detecting a desired wavelength by the slit;
A laser scanning microscope characterized by dynamically switching the detection wavelength of the emitted light passing through the slit by controlling a voltage applied to the KTN crystal.