JP4898588B2 - Scanning microscope - Google Patents

Description

  The present invention relates to a scanning microscope.

  In general, a scanning microscope and a confocal microscope are known as techniques for scanning and imaging a sample at high speed (see Patent Document 1). In the confocal microscope according to Patent Document 1, light from a light source is linearly focused and incident on a light modulation member such as DMD or liquid crystal. Next, as the linear light is modulated by the light modulation member into light having a predetermined brightness and darkness along the direction of the light beam, the sample body is irradiated with light. Light (reflected light, fluorescence, etc.) generated from the sample body is returned to the light modulation member, and then separated from the illumination light path and detected by a photodetector.

JP 2004-199063 A

  However, according to the technique disclosed in Patent Document 1, it is difficult to change the light condensing range in accordance with the on / off switching of the small mirror on the light modulation member (for example, DMD). In addition, there is a problem that a loss occurs due to the amount of light irradiated to the off portion.

  The present invention has been made in view of such circumstances, and a scanning microscope that can easily reduce the loss of light quantity by changing the incident range of light condensed on the light modulation member. The purpose is to provide.

In order to solve the above problems, the present invention employs the following means.
The present invention includes a light source, a plurality of micro light modulation elements arranged in a two-dimensional array, and a micro light modulation element array that switches the micro light modulation element that is activated to an on state in one direction. An objective lens that collects the illumination light that has passed through the minute light modulation element array and irradiates the sample; a photodetector that detects light from the sample, the pixel having at least a one-dimensional array; It is disposed between the light source and the minute light modulation element array, has a different curvature in two axial directions perpendicular to the optical axis, and restricts an irradiation width in a direction perpendicular to the switching direction of the minute light modulation elements. An aspect ratio conversion optical system that includes an irregular curvature lens that irradiates illumination light onto the minute light modulation element array, and that is configured to be movable in the optical axis direction, and the minute light modulation element array includes: The objective lens Located at a position optically conjugate with the sample-side focal position, the curvature lens is provided on the minute light modulation element array side of the aspect ratio conversion optical system, and keeps the angle of the light beam with respect to the optical axis direction constant. A scanning microscope that irradiates the minute light modulation element array with the illumination light is provided.

  According to the present invention, the illumination light emitted from the light source passes through the inflection lens of the aspect ratio conversion optical system and enters the minute light modulation element array. Of the illumination light incident on the minute light modulation element array, the illumination light that has passed through the minute light modulation element activated in the ON state is condensed on the sample by the objective lens. Moreover, the light emitted from the sample is detected by a photodetector.

  By operating the micro light modulation element array, the micro light modulation element operated in the ON state is switched in one direction, so that the illumination light can be scanned on the sample at least one-dimensionally. By setting the micro light modulation element array to a position optically conjugate with the sample-side focal position of the objective lens, the micro light modulation element operated in the ON state can function as a confocal pinhole. A clear image of the sample along the sample-side focal plane of the lens can be acquired.

  Here, the different curvature lens of the aspect ratio conversion optical system refers to an anamorphic lens or a cylindrical lens. In this case, for example, the illumination light with the irradiation width limited in the direction orthogonal to the switching direction of the minute light modulation elements can be irradiated to the minute light modulation element array by a cylindrical lens or the like. As a result, even if the range of the micro light modulation element operated in the on state in the micro light modulation element array is limited to be small in the direction orthogonal to the switching direction of the micro light modulation element, the collection of illumination light is matched to the range. The light range can be set, and the loss of light quantity can be reduced.

  In addition, by operating the aspect ratio conversion optical system, the curvature of curvature lens is moved along the optical axis direction from the state in which the focal position coincides with the minute light modulation element array, so that the illumination to the minute light modulation element array is performed. The incident range of light can be changed easily. Further, the irregular curvature lens is provided on the minute light modulation element array side of the aspect ratio conversion optical system, and is configured to irradiate the illumination light to the minute light modulation element array while maintaining a constant angle with respect to the optical axis of the light beam. Yes. In other words, the different curvature lens is arranged so as not to interpose an optical system having a condensing function with the minute light modulation element array. Thereby, even if the irregular curvature lens moves in the optical axis direction, the incident range of the illumination light in the minute light modulation element array is not greatly changed with respect to the arrangement length in the switching direction of the minute light modulation elements. it can.

  Therefore, change the incident range of illumination light according to the pinhole diameter when changing the pinhole diameter of the confocal pinhole by changing the size of the micro light modulation element group operated in the ON state. Therefore, even if the inflection lens is moved in the optical axis direction, the incident light incident range can be limited to the minimum region including the micro light modulation element group in the on state, thereby reducing the loss of light amount. be able to.

In the said invention, it is good also as providing the control part which moves the position of the said curvature-of-curvature lens to an optical axis direction according to the pupil diameter of the said objective lens.
With this configuration, it is possible to prevent a decrease in light amount even when it is desired to observe while changing the size of the ON region of the minute light modulation element array. For example, when the pupil diameter of the objective lens is large, in order to obtain an image without impairing the resolution in the optical axis direction of the objective lens, the confocal pinhole is reduced by reducing the ON region of the micro light modulation element array. It is necessary to reduce the pinhole diameter. In this case, the operation of the control unit moves the inflection lens in a direction to narrow the incident range of the illumination light to the minute light modulation element array. Thereby, even if the area | region of the micro light modulation element operated to an ON state is made small, the fall of light quantity can be prevented, maintaining the resolution | decomposability of the optical axis direction which an objective lens has enough.

  On the other hand, when the pupil diameter of the objective lens is small, even if the pinhole diameter of the confocal pinhole is increased, the resolution of the objective lens in the optical axis direction is not significantly impaired. Therefore, by operating the control unit, the inflection lens is moved in a direction to widen the incident range of the illumination light to the minute light modulation element array. Thereby, the area | region of the micro light modulation element act | operated to an ON state can be enlarged, and the quantity of the light which returns from a sample can be increased. As a result, a bright image can be obtained while maintaining the resolution in the optical axis direction of the objective lens. In addition, by doing this, it is possible to reduce the number of scans and increase the speed of image acquisition.

In the above invention, the control unit may move the irregular curvature lens to a predetermined position on the optical axis associated with the pupil diameter of the objective lens.
By configuring in this way, when moving the inflection lens on the optical axis by the operation of the control unit, the inflection to the optimal position that can prevent a decrease in the amount of light while maintaining the resolution in the optical axis direction. The lens can be moved automatically.

The present invention includes a light source, a plurality of micro light modulation elements arranged in a two-dimensional array, and a micro light modulation element array that switches the micro light modulation element that is activated to an on state in one direction. An objective lens that collects the illumination light that has passed through the minute light modulation element array and irradiates the sample; a photodetector that detects light from the sample, the pixel having at least a one-dimensional array; It is disposed between the light source and the minute light modulation element array, has a different curvature in two axial directions perpendicular to the optical axis, and restricts an irradiation width in a direction perpendicular to the switching direction of the minute light modulation elements. An aspect ratio conversion optical system having a plurality of different curvature lenses with different irradiation widths for irradiating illumination light onto the minute light modulation element array, and configured to selectively switch the plurality of different curvature lenses. , The micro light change Element array, wherein disposed on the sample-side focal position optically conjugate with the position of the objective lens, the plurality of different curvatures lens is provided on the micro-optical modulation element array side of the front Symbol aspect ratio conversion optical system, Provided is a scanning microscope that irradiates the minute light modulation element array with the illumination light while keeping the angle of the light beam with respect to the optical axis constant.

  According to the present invention, the illumination light emitted from the light source passes through the inflection lens of the aspect ratio conversion optical system and enters the minute light modulation element array. Of the illumination light incident on the minute light modulation element array, the illumination light that has passed through the minute light modulation element activated in the ON state is condensed on the sample by the objective lens. Moreover, the light emitted from the sample is detected by a photodetector.

  By operating the micro light modulation element array, the micro light modulation element operated in the ON state is switched in one direction, so that the illumination light can be scanned on the sample at least one-dimensionally. By setting the micro light modulation element array to a position optically conjugate with the sample-side focal position of the objective lens, the micro light modulation element operated in the ON state can function as a confocal pinhole. A clear image of the sample along the sample-side focal plane of the lens can be acquired.

  In this case, the micro light modulation element array is irradiated with illumination light whose irradiation width is limited in a direction orthogonal to the switching direction of the micro light modulation elements by an inflection lens of the aspect ratio conversion optical system, for example, a cylindrical lens. Can do. As a result, even if the range of the micro light modulation element operated in the on state in the micro light modulation element array is limited to be small in the direction orthogonal to the switching direction of the micro light modulation element, the collection of illumination light is matched to the range. The light range can be set, and the loss of light quantity can be reduced.

  In addition, by operating the aspect ratio conversion optical system and selectively switching the different curvature lens to another different curvature lens having a different irradiation width, the incident range of the illumination light with respect to the minute light modulation element array can be easily changed. be able to. Further, the irregular curvature lens is provided on the minute light modulation element array side of the aspect ratio conversion optical system, and is configured to irradiate the illumination light to the minute light modulation element array while maintaining a constant angle with respect to the optical axis of the light beam. Yes. In other words, the different curvature lens is arranged so as not to interpose an optical system having a condensing function with the minute light modulation element array. This prevents the illumination light incident range in the micro light modulation element array from changing greatly with respect to the array length in the switching direction of the micro light modulation elements even when switching to another lens having a different irradiation width. Can do.

  Therefore, even when switching to a different curvature lens with a different irradiation width, such as when changing the pinhole diameter of the confocal pinhole by changing the size of the micro light modulation element group that is activated in the on state, Since the incident range of the illumination light can be limited to the minimum region including the minute light modulation element group in the state, the loss of the light amount can be reduced.

In the above invention, the aspect ratio conversion optical system includes a rotation unit having a rotation axis parallel to the optical axis of the light, and the plurality of different curvature lenses having different irradiation widths are separated from the rotation axis in the rotation unit. It is good also as arrange | positioning at equal distance.
With this configuration, by rotating the rotating unit, any one of the plurality of different curvature lenses can be arranged on the optical axis of the illumination light. Therefore, switching of the different curvature lens can be performed by a simple operation without accompanying a lens replacement operation or the like.

In the above-mentioned invention, it is good also as providing the rotation control part which adjusts the rotation angle of the rotation part according to the pupil diameter of the objective lens while detecting the information on the pupil diameter of the objective lens.
With this configuration, the information on the pupil diameter of the objective lens is detected by the operation of the rotation control unit, and the rotation angle of the rotation unit is adjusted according to the pupil diameter of the objective lens based on the information. Therefore, it is possible to automatically select a different curvature lens suitable for obtaining a confocal effect with respect to the pupil diameter of the objective lens from a plurality of different curvature lenses having different irradiation widths.

Further, in the above-mentioned invention, it may be provided with a scanning unit that is arranged between the minute light modulation element array and the objective lens and scans the illumination light in a direction orthogonal to a switching direction of the minute light modulation element. Good.
With this configuration, the illumination light that has passed through the minute light modulation element that has been activated in the illumination light incident on the minute light modulation element array is scanned by the scanning means, and is applied to the sample by the objective lens. Focused. Here, since the illumination light is scanned in the direction orthogonal to the switching direction of the minute light modulation elements by the operation of the scanning means, the illumination light can be scanned two-dimensionally on the sample.

In the above invention, the objective lens further collects light returning from the sample, and returns between the minute light modulation element array and the photodetector via the minute light modulation element array. May be provided with other scanning means for scanning in the same direction in synchronization with the scanning means.
With this configuration, the same objective lens illuminates the sample and receives light from the sample. In this case, the light emitted from the sample and returning through the objective lens and the micro light modulation element array can be guided to a two-dimensional region on the photodetector by the operation of other scanning means. Therefore, since a two-dimensional imaging unit such as a CCD can be used as a photodetector and an image of the entire sample can be formed on the two-dimensional imaging unit, a line scan image is acquired. Image acquisition can be speeded up compared to the case of constructing a two-dimensional image.

In the invention described above, the light source may be a laser light source. Since the amount of light can be increased by using laser light, a decrease in the amount of light detected in each pixel of the photodetector can be prevented even when scanning is performed at high speed.
In the invention described above, the minute light modulation element may be a micromirror.

  According to the present invention, it is possible to change the incident range of light incident on the minute light modulation element array in accordance with the range of the minute light modulation element operated in the ON state, thereby reducing the loss of light amount. There is an effect that can be.

[First Embodiment]
Hereinafter, a scanning microscope 10 according to a first embodiment of the present invention will be described with reference to the drawings.
As shown in FIG. 1, a scanning microscope 10 according to the present embodiment includes a laser light source (light source) 1 and a cylindrical lens (different optical lens) that limits the irradiation width of laser light (illumination light) emitted from the laser light source 1. A curvature lens) 23, an aspect ratio conversion optical system 3 having the cylindrical lens 23, and a DMD (micro light modulation element array, digital micro) that reflects laser light that has passed through the cylindrical lens 23 of the aspect ratio conversion optical system 3. A mirror array) 7, a galvano mirror (scanning means) 9 that scans the laser light reflected by the DMD 7 in one direction, and the laser light reflected by the galvano mirror 9 is condensed and irradiated on the sample 19. The objective lens 11 that condenses the fluorescence emitted from the sample 19 and the fluorescence condensed by the objective lens 11 are imaged. It includes a first image forming lens 13 and the second imaging lens 15, a photodetector 17 for detecting the fluorescence focused by the second imaging lens 15.
In the figure, reference numeral 5 denotes a dichroic mirror.

For example, the laser light source 1 emits substantially parallel laser light having a substantially circular cross section as illumination light.
The aspect ratio conversion optical system 3 is disposed between the laser light source 1 and the DMD 7 and expands the light beam diameter of the laser light emitted from the laser light source 1 toward the laser light source 1 so as to obtain a large parallel light beam. The collimator 21 is provided, the DMD 7 side has different curvatures in two axial directions orthogonal to the optical axis direction, and orthogonal to the irradiation width of the laser light, specifically, the switching direction of the micromirror (micro light modulation element) 29. A cylindrical lens 23 is provided for limiting the irradiation width in the direction in which the light is emitted.

The cylindrical lens 23 is provided so as to be movable in a direction along the optical axis by a moving mechanism (not shown).
Further, as shown in FIG. 1, the cylindrical lens 23 is disposed so as not to interpose an optical system having a condensing function with the DMD 7, and is arranged with respect to the optical axis of the off-axis light beam included in the laser beam. The DMD 7 is irradiated with laser light while keeping the angle constant.

  The laser light incident on the aspect ratio conversion optical system 3 is enlarged to a light beam diameter of a predetermined size by the collimator 21, and then an axis perpendicular to the light beam diameter in one axial direction is maintained by the cylindrical lens 23. It is focused only in the direction. As a result, the aspect ratio of the cross-sectional shape of the laser beam is converted, and the irradiation width in the direction orthogonal to the switching direction of the micromirror 29 is limited. As a result, the incident range of the laser beam on the DMD 7 is changed by moving the cylindrical lens 23 along the optical axis direction.

  That is, when the focal position of the cylindrical lens 23 coincides with the DMD 7, the incident range of the laser light on the DMD 7 is linear, and when it is arranged at a position shifted in any optical axis direction from that position, The incident light is incident on the DMD 7 in a wider incident range.

  As shown in FIG. 2, the DMD 7 includes a plurality of two-dimensionally arranged micromirrors 29, and one or more adjacent (3 × 3 = 9 in the example shown in FIG. 2) micromirrors 29 ′ are simultaneously turned on. The ON region that has been actuated on is switched so as to move in one direction. As a result, of the laser light incident on the DMD 7, the laser light irradiated on the micromirror 29 ′ in the on region is reflected.

  Since the laser light reflected in the on region of the DMD 7 is incident on the galvano mirror 9 via the first relay lens 25, the galvano mirror 9 is moved as the on region of the DMD 7 moves in one direction. The incident position of the laser beam also moves in one direction.

  Further, the DMD 7 is disposed at a position optically conjugate with the sample-side focal position of the objective lens 11. As a result, the ON region of the DMD 7 can function as a confocal pinhole.

  The galvanometer mirror 9 scans the laser beam reflected in the ON region of the DMD 7 in one direction orthogonal to the moving direction of the ON region. The laser beam reflected by the galvanometer mirror 9 is focused on the sample 19 via the second relay lens 27 and the objective lens 11. Then, by synchronizing the on / off switching operation of the micromirror 29 in the DMD 7 and the swinging operation of the galvanometer mirror 9, the two-dimensionally scanned laser light can be irradiated onto the sample 19. Yes.

  The fluorescence generated by irradiating the sample 19 with the laser light and exciting the fluorescent substance contained in the sample 19 is collected by the objective lens 11, then the first imaging lens 13, Through the second relay lens 27, the galvanometer mirror 9, the first relay lens 25 and the DMD 7, the laser beam returns in the opposite direction. The fluorescent light passes through the dichroic mirror 5 and is separated from the laser light, and then collected by the second imaging lens 15 and incident on the photodetector 17.

  The photodetector 17 is a line sensor having a plurality of pixels arranged one-dimensionally, and is a one-dimensional fluorescence image generated by moving the ON region of the DMD 7 at each swing position of the galvanometer mirror 9, that is, Line scan images are acquired sequentially. The acquired line scan image is stored in association with the swing position of the galvano mirror 9 so that a two-dimensional fluorescence image can be constructed later.

The operation of the scanning microscope 10 according to the present embodiment configured as described above will be described.
When laser light is emitted from the laser light source 1, the laser light first passes through the collimator 21 in the aspect ratio conversion optical system 3, so that its light beam diameter is expanded to become a large parallel light beam. Then, when the laser light passes through the cylindrical lens 23, the irradiation width of the laser light in the direction orthogonal to the switching direction of the micromirror 29 is limited. Thereby, for example, even when the ON region of the DMD 7 is limited to be small in the scanning direction by the galvanometer mirror 9, the condensing range of the laser light can be set in accordance with the range.

  In addition, the aspect ratio conversion optical system 3 changes the incident range of the laser beam on the DMD 7 by moving the cylindrical lens 23 along the optical axis direction from the state in which the focal position coincides with the DMD 7. In this case, the cylindrical lens 23 irradiates the DMD 7 with the laser light while keeping the angle of the off-axis light beam included in the light beam of the laser light constant with respect to the optical axis, as described above. Even if it moves to, the incident range of the laser beam in DMD7 does not change with respect to the arrangement length of the switching direction (for example, see FIG. 2) of the micromirror 29.

Thereby, for example, when the size of the ON region of the DMD 7 is changed and the pinhole diameter of the pinhole is changed, the cylindrical lens 23 is used to change the laser beam incident range in accordance with the pinhole diameter. Even if the laser beam is moved in the optical axis direction, the incident range of the laser beam is limited to the minimum region including the ON region.
In this way, the laser light that has passed through the aspect ratio conversion optical system 3 is reflected by the dichroic mirror 5 and enters the DMD 7.

Since the DMD 7 uses one or more adjacent micromirrors 29 ′ as the ON region (see FIG. 2), the laser beam reflected by the ON region is incident on the galvanomirror 9 via the first relay lens 25. The
In this case, since the DMD 7 is switched so as to move the ON region in one direction, the laser light is incident on the galvanometer mirror 9 so as to move in the ON region switching direction.

  Then, the galvano mirror 9 is swung so that the incident laser light is scanned in one direction orthogonal to the switching direction of the ON region of the DMD 7. Accordingly, the on-region switching operation of the DMD 7 and the oscillating operation of the galvano mirror 9 are synchronized, so that they are reflected by the galvano mirror 9 and collected on the sample 19 via the second relay lens 27 and the objective lens 11. The laser beam to be emitted is scanned two-dimensionally on the sample 19.

  When fluorescence is generated by irradiating the sample 19 with the laser beam, the fluorescence is collected by the objective lens 11, and then the first imaging lens 13, the second relay lens 27, the galvanometer mirror 9, It returns to the opposite direction to the laser beam via the relay lens 25 of 1 and the DMD 7. Then, after passing through the dichroic mirror 5 and being separated from the laser light, the fluorescence is condensed by the second imaging lens 15 and is incident on the photodetector 17.

  In this case, since the DMD 7 is disposed at a position optically conjugate with the sample-side focal position of the objective lens 11, the fluorescence is reflected by the same micromirror 29 'that reflects the laser light. , And enters the photodetector 17. Then, by setting the ON region of the DMD 7 to be sufficiently small, the ON region functions as a confocal pinhole, and a clear fluorescent image of the sample 19 along the sample-side focal plane of the objective lens 11, that is, the DMD 7 A line scan image generated by moving the ON region in one direction is acquired by the photodetector 17.

  In this way, the line scan image acquired by the photodetector 17 for each swing position of the galvanometer mirror 9 is stored in association with the swing position of the galvanometer mirror 9. Thereby, a two-dimensional fluorescence image can be constructed based on the stored line scan image.

  As described above, according to the scanning microscope 10 according to the present embodiment, the condensing range of the laser light is set in accordance with the range of the ON region of the DMD 7 by the cylindrical lens 23 in the aspect ratio conversion optical system 3. Can do. Further, when the cylindrical lens 23 is moved in the optical axis direction by the operation of the aspect ratio conversion optical system 3, the incident range of the laser light can be limited to a minimum region including the ON region of the DMD 7. As a result, it is possible to reduce the light loss.

The present embodiment can be modified as follows.
For example, the scanning microscope 10 shown in FIG. 1 may include a control unit (not shown) that moves the position of the cylindrical lens 23 in the optical axis direction according to the pupil diameter of the objective lens 11. This makes it possible to prevent a decrease in the amount of light even when the on-region size of the DMD 7 is changed for observation. For example, when the pupil diameter of the objective lens 11 is large, in order to obtain an image without impairing the resolution of the objective lens 11 in the optical axis direction, the pin area of the confocal pinhole is reduced by reducing the ON region of the DMD 7. It is necessary to reduce the hole diameter. In this case, the cylindrical lens 23 is moved in the direction of narrowing the incident range of the laser light to the DMD 7 by the operation of the control unit. As a result, even if the ON region of the DMD 7 is made small, it is possible to prevent a decrease in light amount while maintaining sufficient resolution in the optical axis direction of the objective lens 11.

  On the other hand, when the pupil diameter of the objective lens 11 is small, even if the pinhole diameter of the confocal pinhole is increased, the resolution of the objective lens 11 in the optical axis direction is not significantly impaired. Therefore, the cylindrical lens 23 is moved in a direction to widen the incident range of the laser light to the DMD 7 by the operation of the control unit. Thereby, the ON region of the DMD 7 can be enlarged, and the amount of light returning from the sample 19 can be increased. As a result, a bright image can be obtained while maintaining the resolution of the objective lens 11 in the optical axis direction. In addition, by doing this, it is possible to reduce the number of scans and increase the speed of image acquisition.

  Further, for example, as shown in FIG. 3, the scanning microscope 10A may be configured not to include the galvanometer mirror 9, that is, the scanning unit. In this case, it is desirable that the sample 19 is provided so as to be movable in a direction orthogonal to the switching direction of the micromirror 29. Specifically, the sample 19 is placed on the stage 35, and the sample 19 is reciprocated in a direction orthogonal to the switching direction of the micromirror 29 (see the X direction in FIG. 3) by the operation of the stage moving unit 37. Then, the laser beam reflected by the DMD 7 is irradiated onto the sample 19 via the first imaging lens 13 and the objective lens 11, and the fluorescence emitted from the sample 19 is reflected on the objective lens 11 and the first imaging lens. After passing through the lens 13, it may be arranged so as to enter the photodetector 17 through the DMD 7, the dichroic mirror 5, and the second imaging lens 15. In this case, the operation of the stage moving unit 37 synchronizes the on / off switching operation of the micromirror 29 in the DMD 7 and the above-described moving operation of the sample 19, whereby the two-dimensionally scanned laser light is applied to the sample 19. Can be irradiated.

  As shown in FIG. 4, the scanning microscope 10B may employ a transmissive element, for example, a liquid crystal element 39, instead of the DMD 7, as the minute light modulation element array. In this case, it is desirable that the liquid crystal element 39 is disposed at a position optically conjugate with the sample-side focal position of the objective lens 11. Specifically, after the laser light reflected by the dichroic mirror 5 passes through the liquid crystal element 39, the first relay lens 25, the galvano mirror 9, the second relay lens 27, the first imaging lens 13, and The sample 19 is irradiated through the objective lens 11. Then, the fluorescence emitted from the sample 19 is transmitted through the liquid crystal element 39 through the objective lens 11, the first imaging lens 13, the second relay lens 27, the galvanometer mirror 9, and the first relay lens 25. After that, it may be arranged so as to pass through the dichroic mirror 5 and the second imaging lens 15 and to enter the photodetector 17. By doing so, the liquid crystal element 39 can function as a confocal pinhole.

  Further, for example, as shown in FIG. 5, a halogen lamp may be adopted as the light source instead of the laser light source 1. In this case, for example, a halogen lamp 43 may be provided between the aspect ratio conversion optical system 3 ′ and the diffuser 41. Further, instead of the halogen lamp 43, a pulse light source (not shown) may be employed.

  For example, as shown in FIG. 6, the scanning microscope 10 </ b> C may be configured such that the fluorescence generated from the sample 19 does not pass through the DMD 7. In this case, it is desirable to employ a pulse laser as the illumination light. Specifically, the pulse laser emitted from the laser light source 1 ′ and passing through the aspect ratio conversion optical system 3 is the DMD 7, the first relay lens 25, the galvanometer mirror 9, the second relay lens 27, and the first connection. After being reflected by the dichroic mirror 5 ′ through the image lens 13, the light is condensed by the objective lens 11 and irradiated onto the sample 19. Then, the light emitted from the sample 19 is condensed by the objective lens 11 and transmitted through the dichroic mirror 5 ′, and then condensed by the second imaging lens 15 and incident on the CCD 45. That's fine. By doing in this way, the clear fluorescence image of the sample 19 in the sample side focal plane of the objective lens 11 is acquirable using the multiphoton excitation effect by a pulse laser.

[Second Embodiment]
Next, a scanning microscope 20 according to a second embodiment of the present invention will be described with reference to FIG.
In the description of the present embodiment, portions having the same configuration as those of the scanning microscope 1 according to the first embodiment described above are denoted by the same reference numerals and description thereof is omitted.

  In the scanning microscope 20 according to the present embodiment, the DMD 7 emits laser light emitted from the laser light source 1 on the DMD 7 side of the aspect ratio conversion optical system 35 while limiting the irradiation width in the direction orthogonal to the switching direction of the micromirror 29. A cylindrical lens 23 for irradiating the lens, and cylindrical lenses (anomalous curvature lenses) 33 and 34 having different irradiation widths from the cylindrical lens 23.

  Each cylindrical lens 23, 33, 34 is provided so as to be alternatively switchable. For example, the cylindrical lenses 23, 33, 34 are U-shaped lens holders (not shown) so that they can be manually replaced. ) May be provided. Further, for example, the cylindrical lenses 23, 33, and 34 may be switchable by an external signal.

According to the scanning microscope 20 according to the present embodiment configured as described above, the cylindrical lenses 23, 33, and 34 are switched to each other (see FIG. 8A and FIG. 8B). The irradiation range of the laser light can be changed (see FIGS. 9A and 9B). Therefore, it is possible to limit the incident range of the laser beam to a minimum region including the ON region of the DMD 7 and reduce the light amount loss.
Further, since it is not necessary to adjust the position of the cylindrical lens 23 in the optical axis direction in order to change the condensing range of the laser light, the aspect ratio conversion optical system 35 can be downsized in the optical axis direction.

In addition, in this embodiment, it can deform | transform as follows.
For example, the aspect ratio conversion optical system 35 may include a lens holder (rotating unit) 60 (see FIG. 10) in which the cylindrical lenses 23, 33, and 34 are arranged.
In this case, the lens holder 60 has a rotation axis 60a parallel to the optical axis of the laser light, and is rotatable around the optical axis of the laser light. The cylindrical lenses 23, 33, and 34 are connected to the lens holder. It suffices if they are concentrically arranged on the same distance at the same distance from the rotary shaft 60a.

  By doing so, the lens holder 60 is rotated, and any one of the cylindrical lenses 23, 33, 34 is arranged on the optical axis of the laser light, thereby changing the incident range of the laser light to the DMD 7. can do. Further, the switching of the cylindrical lenses 23, 33, and 34 can be realized by a simple operation of simply rotating the lens holder 60 without accompanying the lens replacement work.

  Further, for example, the cylindrical lenses 23, 33, and 34 have different focal lengths according to the pupil diameter of the objective lens 11, and the scanning microscope 20 detects the pupil diameter of the objective lens 11 and It is good also as providing the rotation control part 62 (refer FIG. 11) which adjusts the rotation angle of the lens holder 60 according to a pupil diameter.

In this case, when the objective lens 11 is switched to another objective lens 11, the rotation control unit 62 automatically detects the pupil diameter of the objective lens 11 and the cylindrical lenses 23 and 33 provided in the lens holder 60. , 34, the rotation angle of the lens holder 60 may be adjusted so that a lens capable of obtaining a sufficient confocal effect with respect to the pupil diameter of the objective lens 11 is arranged on the optical axis of the laser light.
Thereby, a lens suitable for obtaining a confocal effect with respect to the pupil diameter of the objective lens 11 can be automatically selected from the cylindrical lenses 23, 33, and 34.

  Further, in the present embodiment, the cylindrical lenses 23, 33, and 34 have been described as an example, but instead of this, four or more cylindrical lenses having different irradiation ranges may be provided.

[Third Embodiment]
Next, a scanning microscope 30 according to the third embodiment of the present invention will be described with reference to FIG.
The scanning microscope 30 according to the present embodiment is a transmission microscope, and is arranged to face the objective lens 11 with the laser light source 1 ′ emitting a pulse laser and the sample 19 sandwiched therebetween, and the fluorescence emitted from the sample 19. , And a CCD 45 for detecting the fluorescence focused by the second objective lens 47 and imaged by the second imaging lens 15, and the dichroic mirror 5. Is different from the first embodiment in that it is not provided.
Hereinafter, in the description of the present embodiment, portions having the same configuration as those of the scanning microscope 1 according to the first embodiment are denoted by the same reference numerals and description thereof is omitted.

  According to such a configuration, the pulse laser emitted from the laser light source 1 ′ and passing through the aspect ratio conversion optical system 3 is the DMD 7, the first relay lens 25, the galvano mirror 9, the second relay lens 27, and the second relay lens 27. The light is condensed by the first objective lens 11 through one imaging lens 13 and irradiated onto the sample 19. Then, when the sample 19 is irradiated with a pulse laser, the fluorescence generated by the multiphoton excitation on the sample-side focal plane of the first objective lens 11 is condensed by the second objective lens 47 and the second result is obtained. The image is taken by the CCD 45 through the image lens 15.

  As described above, according to the scanning microscope 30 according to the present embodiment, the fluorescence obtained through the sample 19 is detected. Therefore, for example, it is suitable when the sample 19 has a thickness and it is desired to observe a position in the sample 19 away from the first objective lens 11 for illumination. In this case, a clear fluorescent image on the sample-side focal plane of the first objective lens 11 can be acquired using the multiphoton excitation effect.

[Fourth Embodiment]
Next, a scanning microscope 40 according to the fourth embodiment of the present invention will be described with reference to FIG.
The scanning microscope 40 according to the present embodiment forms an image of the laser light source 1, the second DMD 49 disposed at the position of the CCD 45 in the third embodiment, and the fluorescence reflected by the second DMD 49. The third embodiment is different from the third embodiment in that the third imaging lens 51 is provided.
Hereinafter, in the description of the present embodiment, portions having the same configuration as those of the scanning microscope 30 according to the third embodiment described above are denoted by the same reference numerals and description thereof is omitted.

As the second DMD 49, it is desirable to use one in which micromirrors are two-dimensionally arranged.
Further, the micromirror (not shown) of the second DMD 49 can be switched in the same direction as the switching direction of the micromirror 29 of the DMD 7.

  According to such a configuration, the fluorescence obtained by transmitting through the sample 19 is collected by the second objective lens 47 and is incident on the second DMD 49 via the second imaging lens 15. In this case, since the micromirror 29 of the DMD 7 and the micromirror of the second DMD 49 are switched in the same direction, the second DMD 49 is scanned in synchronism with the DMD 7 and the galvanometer mirror 9, whereby the second DMD 49 Can function as a confocal pinhole. Thereby, according to the scanning microscope 40 which concerns on this embodiment, even if it is a transmission type microscope, a clear fluorescence image in the sample side focal plane of the 1st objective lens 11 is used using a normal laser beam. Can be acquired.

  As the second DMD 49, as in the DMD 7, a DMD (for example, see FIG. 2) in which the array length in the switching direction of the micromirrors is longer than the array length in the direction orthogonal to the switching direction is used. A two-dimensional image of the sample 19 is acquired by providing a second galvanometer mirror (not shown) that scans the fluorescence obtained through the sample 19 in a direction orthogonal to the switching direction of the second DMD 49. It is good as well.

As mentioned above, although each embodiment of the present invention has been described with reference to the drawings, the specific configuration is not limited to this embodiment.
For example, the photodetector 17 may employ a two-dimensional imaging means such as a CCD or CMOS instead of the line sensor.
In this way, for example, when the ON region of the DMD 7 is increased in the scanning direction of the galvanometer mirror, a two-dimensional image of the sample 19 can be effectively acquired.

  As shown in FIG. 14, the scanning microscope 10D forms an image of another galvanometer mirror (another scanning means) 53 that scans the fluorescence emitted from the sample 19 and returned through the DMD 7 in one direction, and the fluorescence. The third imaging lens 55 may be further provided, and a photodetector 17 ′ including a two-dimensional imaging unit such as a CCD may be employed instead of the line sensor.

In this way, the galvanometer mirror 53 scans the fluorescence reflected by the DMD 7 and passed through the second imaging lens 15 in the same direction in synchronization with the galvanometer mirror 9.
The third imaging lens 55 collects the fluorescence scanned by the galvanometer mirror 53 and makes it incident on the photodetector 17 '. Thereby, the fluorescence image of the sample 19 is scanned in the two-dimensional region of the photodetector 17 ′.

  That is, since a two-dimensional fluorescent image of the sample 19 is picked up by the photodetector 17 ′ comprising a two-dimensional image pickup means, when a two-dimensional image is constructed after acquiring a line scan image. In comparison, the image acquisition speed can be increased.

  Further, as shown in FIG. 15, the scanning microscope 10 </ b> E may include, for example, a light amount equalizing unit 61 in which a rod lens 57 and a diffuser 59 are disposed in front of the aspect ratio conversion optical system 3.

  By doing so, the laser light emitted from the laser light source 1 is dispersed by passing through the rod lens 57 and the diffuser 59 so that the distribution of the amount of light on and off the optical axis becomes a uniform light flux. It is corrected.

  As a result, laser light having a uniform light amount distribution is incident on the ON region of the DMD 7, so that the sample 19 along the sample-side focal plane of the objective lens 11 can be uniformly irradiated with laser light, and unevenness is caused. A clear image with little image can be acquired.

In each of the above embodiments, the dichroic mirror 5 has been described as an example. However, instead of this, for example, a half mirror may be employed. By doing so, the laser light is collected by the objective lens 11 and applied to the sample 19, while the reflected light reflected by the sample 19 is collected by the objective lens 11 and transmitted through the half mirror. Then, the light is guided to the photodetectors 17, 17 ′ or the CCD 45. Thereby, it can replace with fluorescence and can detect reflected light.
In each of the above embodiments, the cylindrical lens 23 has been described as an example. However, instead of this, an anamorphic lens may be employed.

1 is a block diagram showing a schematic configuration of a scanning microscope according to a first embodiment of the present invention. It is a figure which shows the micromirror act | operated by the ON state in DMD of FIG. It is a block diagram which shows schematic structure of the scanning microscope which concerns on the modification of the 1st Embodiment of this invention. It is a block diagram which shows schematic structure of the scanning microscope which concerns on the other modification of the 1st Embodiment of this invention. It is a block diagram which shows schematic structure around the light source of the scanning microscope which concerns on the other modification of the 1st Embodiment of this invention. It is a block diagram which shows schematic structure of the scanning microscope which concerns on the other modification of the 1st Embodiment of this invention. It is a block diagram which shows schematic structure of the scanning microscope which concerns on the 2nd Embodiment of this invention. (A) And (b) is a figure which shows schematic structure of the scanning microscope which concerns on the 2nd Embodiment of this invention which provided the cylindrical lens from which irradiation width differs, respectively. (A) And (b) is a figure which shows distribution of the light on DMD of the scanning microscope which concerns on the 2nd Embodiment of this invention, respectively. It is a figure which shows the lens holder of the scanning microscope which concerns on the modification of the 2nd Embodiment of this invention. It is a figure which shows the lens holder and rotation control part of the scanning microscope which concern on the other modification of the 2nd Embodiment of this invention. It is a block diagram which shows schematic structure of the scanning microscope which concerns on the 3rd Embodiment of this invention. It is a block diagram which shows schematic structure of the scanning microscope which concerns on the 4th Embodiment of this invention. It is a block diagram which shows schematic structure of a scanning microscope provided with the other galvanometer mirror which concerns on the modification of each embodiment of this invention. It is a block diagram which shows schematic structure of a scanning microscope provided with the light quantity equalization means which concerns on the other modification of each embodiment of this invention.

Explanation of symbols

1 Laser light source
3 Aspect ratio conversion optical system 7 DMD (micro light modulation element array, digital micro mirror array)
9 Galvano mirror (scanning means)
DESCRIPTION OF SYMBOLS 10 Scanning microscope 11 Objective lens 17 Photo detector 19 Sample 23 Cylindrical lens (curvature lens)
29 Micromirror (micro light modulation element)

Claims (10)

A light source;
A micro light modulation element array comprising a plurality of micro light modulation elements arranged two-dimensionally and switching the micro light modulation elements activated in an ON state in one direction;
An objective lens that collects the illumination light emitted from the light source and passed through the micro light modulation element array and irradiates the sample;
A photodetector having a plurality of pixels arranged at least one-dimensionally and detecting light from the sample;
It is disposed between the light source and the minute light modulation element array, has a different curvature in two axial directions perpendicular to the optical axis, and restricts an irradiation width in a direction perpendicular to the switching direction of the minute light modulation elements. An aspect ratio conversion optical system that includes an irregular curvature lens that irradiates illumination light onto the micro light modulation element array, and the irregular curvature lens is configured to be movable in the optical axis direction;
The micro light modulation element array is disposed at a position optically conjugate with a sample-side focal position of the objective lens;
The irregular curvature lens is provided on the minute light modulation element array side of the aspect ratio conversion optical system, and irradiates the illumination light to the minute light modulation element array while keeping an angle with respect to the optical axis of a light beam constant. microscope.   The scanning microscope according to claim 1, further comprising a control unit that moves the position of the different curvature lens in the optical axis direction according to the pupil diameter of the objective lens.   The scanning microscope according to claim 2, wherein the control unit moves the irregular curvature lens to a predetermined position on an optical axis associated with a pupil diameter of the objective lens. A light source;
A micro light modulation element array comprising a plurality of micro light modulation elements arranged two-dimensionally and switching the micro light modulation elements activated in an ON state in one direction;
An objective lens that collects the illumination light emitted from the light source and passed through the micro light modulation element array and irradiates the sample;
A photodetector having a plurality of pixels arranged at least one-dimensionally and detecting light from the sample;
It is disposed between the light source and the minute light modulation element array, has a different curvature in two axial directions perpendicular to the optical axis, and restricts an irradiation width in a direction perpendicular to the switching direction of the minute light modulation elements. An aspect ratio conversion optical system having a plurality of different curvature lenses with different irradiation widths for irradiating illumination light onto the minute light modulation element array, and configured to selectively switch the plurality of different curvature lenses. ,
The micro light modulation element array is disposed at a position optically conjugate with a sample-side focal position of the objective lens;
Wherein the plurality of different curvatures lens, before SL provided in the micro-optical modulation element array side of the aspect ratio conversion optical system, irradiating the illumination light while maintaining its angle to the optical axis of the light beam constant at the micro-optical modulation element array Scanning microscope. The aspect ratio conversion optical system includes a rotation unit having a rotation axis parallel to the optical axis of the light beam,
The scanning microscope according to claim 4, wherein the plurality of different curvature lenses having different irradiation widths are arranged at an equal distance from the rotation axis in the rotation unit.   The scanning microscope according to claim 5, further comprising a rotation control unit that detects information on a pupil diameter of the objective lens and adjusts a rotation angle of the rotation unit according to the pupil diameter of the objective lens.   7. The scanning device according to claim 1, further comprising a scanning unit that is disposed between the minute light modulation element array and the objective lens and scans the illumination light in a direction orthogonal to a switching direction of the minute light modulation element. A scanning microscope according to 1. The objective lens further collects light returning from the sample,
8. The apparatus according to claim 7, further comprising: another scanning unit that scans light returning through the micro light modulation element array in the same direction in synchronization with the scanning unit between the micro light modulation element array and the photodetector. The scanning microscope described.   The scanning microscope according to any one of claims 1 to 8, wherein the light source is a laser light source.   The scanning microscope according to any one of claims 1 to 9, wherein the minute light modulation element is a micromirror.

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