JP2011027804A - Confocal microscope - Google Patents

Abstract

The image quality of a confocal microscope is improved.
An optical deflection element 25 of a confocal microscope 1 has a deflection surface 25A provided with a plurality of micromirrors arranged at a position conjugate with a predetermined observation surface of a sample S, and is condensed by an objective lens 29. Thus, the polarization direction of the observation light from the sample S incident on the deflection surface 25A is controlled for each micromirror. The control unit 41 sequentially selects the micro mirrors of the light deflection element 25 and controls the observation light incident on the selected micro mirrors to be deflected in the direction of the imaging lens 24. The observation light deflected in the direction of the imaging lens 24 is imaged on the light receiving surface 31A of the detector 31 by the imaging lens 24. A surface obtained by extending the deflection surface 25A of the optical deflection element 25, the main plane 24A of the imaging lens 24, and the light receiving surface 31A of the detector 31 intersects with one straight line passing through the point A1. The present invention can be applied to, for example, a confocal microscope.
[Selection] Figure 1

Description

  The present invention relates to a confocal microscope, and more particularly, to a confocal microscope that can improve image quality.

  Conventionally, illumination light emitted from a light source is scanned in a two-dimensional direction on a predetermined observation surface of a sample using a digital mirror device (DMD: Digital Mirror Device), and observation light from the sample is scanned by another DMD. There has been proposed a confocal microscope that reflects in the direction of the detector, detects the detector, and generates a confocal image (see, for example, Patent Document 1).

JP 2001-521205 A

  However, in the confocal microscope described in Patent Document 1, the optical path length of the observation light reflected by the micromirror provided in the DMD and incident on the detector is different for each micromirror. Therefore, the observation light incident on the micromirror near the optical axis forms an image on the light receiving surface of the detector, but the observation light incident on the micromirror far from the optical axis is combined at a position shifted from the light receiving surface of the detector. Image. Therefore, the further away from the center of the confocal image, the lower the image quality and the blurred image.

  The present invention has been made in view of such circumstances, and is intended to improve the image quality of a confocal microscope.

  A confocal microscope according to one aspect of the present invention includes an illumination optical system that condenses illumination light from a light source on a predetermined observation surface of a sample and scans a condensing position, and an observation lens from the sample. A confocal microscope including an imaging optical system that collects light and forms an image to detect the light, and an optical deflecting unit capable of controlling a deflection direction of incident light incident on a predetermined deflection surface for each predetermined region. The deflecting surface is arranged at a position conjugate with the observation surface, and the light deflecting unit controls the polarization direction of the observation light that is collected by the objective lens and enters the deflection surface for each region; The region of the light deflection unit is sequentially selected, and deflection control means for controlling the observation light incident on the selected region to be deflected in a predetermined first direction, and the light deflection unit controls the first Detecting the observation light deflected in the direction A light detection unit; and an imaging lens that forms an image of the observation light deflected in the first direction by the light deflection unit on a light receiving surface of the light detection unit, the deflection surface of the light deflection unit, The light deflecting means, the imaging lens, and the light detecting means are arranged such that a main plane of the imaging lens and a surface obtained by extending the light receiving surface of the light detecting means intersect each other with a single straight line. Is arranged.

  In one aspect of the present invention, the entire range of the sample image connected to the deflection surface of the light deflection element by the objective lens is imaged on the imaging surface of the light detection means by the imaging lens.

  According to the present invention, the image quality of a confocal microscope can be improved.

It is a figure which shows 1st Embodiment of the confocal microscope to which this invention is applied. It is a figure for demonstrating operation | movement of the micromirror which the light deflection | deviation element of the confocal microscope of FIG. 1 has. It is a figure which shows the example of the scanning by the micromirror which an optical deflection element has. It is a figure for demonstrating the distortion of the image which generate | occur | produces with the confocal microscope to which this invention is applied. It is a figure for demonstrating the distortion of the image which generate | occur | produces with the confocal microscope to which this invention is applied. It is a figure which shows 2nd Embodiment of the confocal microscope to which this invention is applied. It is a figure for demonstrating operation | movement of the micromirror which the light deflection | deviation element of the confocal microscope of FIG. 6 has. It is a figure which shows the other example of the scanning by the micromirror which an optical deflection element has.

  Hereinafter, embodiments of the present invention will be described with reference to the drawings.

  First, a first embodiment of a confocal microscope to which the present invention is applied will be described with reference to FIGS.

  FIG. 1 shows an external view of a confocal microscope 1 as a first embodiment of a confocal microscope to which the present invention is applied, as viewed from the side. In the drawing, a part of the optical system arranged inside the confocal microscope 1 is shown in cross section so that the configuration of the optical system can be seen, and the members indicated by the cross section are hatched.

  The confocal microscope 1 includes a light source 21, a collector lens 22a, a lens 22b, a half mirror 23, an imaging lens 24, a light deflection element 25, a lens 26, a light dimming member 27, a revolver 28, an objective lens 29, a stage 30, and a detection. The device 31 and the control unit 41 are included.

  First, the illumination optical system of the confocal microscope 1 will be described. The illumination optical system of the confocal microscope 1 includes a light source 21, a collector lens 22 a, a lens 22 b, a half mirror 23, an imaging lens 24, a light deflection element 25, a lens 26, and an objective lens 29. (That is, illumination light) is condensed on a predetermined observation surface of the sample S placed on the stage 30, and the condensing position is scanned.

  Specifically, the light source 21 includes, for example, a halogen lamp, a mercury lamp, or an LED (Light Emitting Diode). The illumination light emitted from the light source 21 is collected by the collector lens 22 a and the lens 22 b and guided to the half mirror 23. The illumination light that has reached the half mirror 23 is deflected in the upper left direction in the drawing, passes through the imaging lens 24, becomes a parallel light beam, and reaches the light deflection element 25.

  The light deflection element 25 has a conjugate positional relationship with a predetermined observation surface of the sample S, and is arranged at a position corresponding to the arrangement position of the field stop. The light deflection element 25 is configured by an element capable of controlling the polarization direction of incident light incident on a predetermined deflection surface 25A for each predetermined region. For example, the light deflection element 25 is provided with a plurality of minute light deflection mirrors (hereinafter referred to as minute mirrors) that can be selectively controlled for each region on the deflection surface 25A, and in accordance with control from the control unit 41, With a micromirror, light incident from one direction can be deflected in various directions for each region. For example, a DMD (Digital Micromirror Device), which is a unit having a pixel structure in which minute micromirrors are two-dimensionally arranged, is used as the light deflection element 25.

In the optical deflecting element 25 such as DMD, as shown in FIG. 2, the micromirror 51 is controlled by on / off control of a voltage applied to a holding unit that holds the micromirrors 51 two-dimensionally arranged on the deflection surface 25A. It is possible to control the angle of each of the two in two directions. Specifically, among the plurality of micromirrors 51 _{i, j} arranged in two dimensions of i × j (i, j: natural number), micromirrors 51 _{m, n} (1 ≦ m ≦ i, 1 ≦ n) ≦ j) and the minute mirror 51 _{p, q} (1 ≦ p ≦ i, 1 ≦ q ≦ j), the lower right in the figure reflected by the half mirror 23 and transmitted through the imaging lens 24. The illumination light from the direction is incident on each of the micromirrors 51 _{m, n} and the micromirrors 51 _{p, q} . At this time, since the micromirrors 51 _{p, q} are in the on state, the incident illumination light is deflected downward toward the lens 26 in the drawing. On the other hand, since the micro mirrors 51 _{m, n} are in the off state, the incident illumination light is deflected toward the light attenuating member 27 in the lower left direction in the drawing. That is, micromirrors 51 _i, by performing the voltage control of _j on / off the micromirrors 51 _i, by each of _j, the illumination light coming incident from one direction, a lower direction of the lens 26 in the drawing, It is possible to guide in two directions with the light dimming member 27 in the lower left direction in the figure.

In addition, the deflection surface 25 </ b> A of the light deflection element 25 is disposed at a position conjugate with a predetermined observation surface of the sample S placed on the stage 30. Therefore, in order to guide the light reaching the deflection surface 25A of the light deflection element 25 to a desired location on the sample S, the micromirrors 51 _{i, j} arranged in the region corresponding to the desired location are turned on. Then, the incident illumination light may be guided to the lens 26 side. For example, as shown in FIG. 2, the illumination light deflected toward the lens 26 by the micro mirrors 51 _{p and q} passes through the lens 26 and enters the objective lens 29 held by the revolver 28, and the objective lens 29, the light is condensed on the observation surface of the sample S conjugate with the deflecting surface 25A of the light deflection element 25, and is irradiated only to the portion corresponding to the micro mirrors _{51p, q} of the sample S. Thus, in the confocal microscope 1, the illumination light can be guided to a desired location on the sample S by individually controlling the on / off of each of the micromirrors 51 _{i, j} of the light deflection element 25. it can.

  On the other hand, the light deflected toward the light attenuating member 27 by the light deflecting element 25 is unnecessary light that does not contribute to the image and is attenuated by the light attenuating member 27. As the light attenuating member 27, for example, a member that absorbs light such as a neutral density (ND) filter or flocking paper is used. By disposing such a light attenuating member 27, unnecessary light is prevented from becoming stray light and affecting observation. The same effect can be obtained by painting the region in black instead of arranging the light dimming member 27, but an image with better contrast can be obtained by arranging the light dimming member 27.

Here, with reference to FIG. 3 _, the control of the micromirrors 51 _{i, j} of the light deflection element 25 will be described. FIG. 3 is a simplified representation of the two-dimensional arrangement of the micromirrors 51 _{i, j} of the light deflection element 25, and each grid cell represents the micromirrors 51 _{i, j} . In the present embodiment, in order to simplify the description, a case where 6 × 8 micromirrors 51 _{i, j} are arrayed will be described as an example of a two-dimensional array.

As described above, since the deflection surface 25A of the light deflection element 25 and the observation surface of the sample S placed on the stage 30 have a conjugate relationship, a plurality of micromirrors 51 included in the light deflection element 25 are provided. _i, of the _j, the micromirrors 51 i which is turned _on, only the portion of the sample S corresponding to _j is illuminated. FIG. 3A shows a state in which only the first micro mirror 51 _1,1 from the left in the uppermost stage among the micro mirrors 51 _{i, j} of the light deflecting element 25 is turned on. This means that only the corresponding part of is illuminated. Similar to FIG. 3a, FIG. 3b shows a state in which only the micro mirrors 51 _1,2 are in an on state, and FIG. 3c shows a state in which only the micro mirrors 51 _1,3 are in an on state. In the cases of FIGS. 3b and 3c as well, only corresponding portions of the sample S are illuminated at the same time as in FIG. 3a.

Of the micro mirrors 51 _{i, j} of the light deflection element 25, the control unit 41 sequentially turns on the micro mirror 51 that is turned on from the state shown in FIG. 3a, as shown in FIGS. 3b, 3c,. I will make a transition. That is, the control unit 41 sequentially selects the uppermost micromirrors 51 _1,4 to 51 _1,8 and sequentially turns them on, and similarly, the micromirrors 51 _{2, 2nd and} subsequent stages are similarly turned on _{. 1} to 51 _2,8 ,..., By sequentially selecting the micro mirrors 51 _7,1 to 51 _7,8 and turning them on, finally, the 8th from the left of the bottom (sixth stage) The micro mirrors _{516 and 8} are turned on.

In this way, the region where the sample S is irradiated with the illumination light is scanned. That is, as shown in FIGS. 3a, 3b, 3c,..., 3d, the micromirrors 51 _{i, j} of the light deflection element 25 are sequentially driven one by one to illuminate the entire range of the sample S. By scanning the light, it is possible to scan the entire observation site of the sample S.

As shown in FIG. 3, not only scanning is performed by sequentially driving one minute mirror 51 _{i, j} (one pixel) one by one, but also, for example, a plurality of adjacent minute mirrors 51 _{i, j} ( A plurality of pixels) may be used as one region, and the micromirrors 51 _{i, j} may be sequentially driven for each of the plurality of pixels to perform scanning.

  A driving device (not shown) is connected to the stage 30 and relatively moves the objective lens 29 and the stage 30 in the Z direction. Further, the stage 30 itself is configured to be freely movable in the XY directions, and the spectroscope can observe a desired part of the sample S. Further, the revolver 28 is configured to be rotatable manually or electrically, and a desired objective lens 29 can be selectively used from among a plurality of objective lenses 29 attached to the revolver 28.

  Next, the imaging optical system of the confocal microscope 1 will be described. The imaging optical system of the confocal microscope 1 includes an objective lens 29, a lens 26, a light deflecting element 25, an imaging lens 24, a half mirror 23, and a detector 31, and the observation light from the sample S is converted into the objective lens. The light is condensed by 29, condensed by the imaging lens 24 on the light receiving surface 31 </ b> A of the detector 31, and detected by the detector 31.

Specifically, the light (that is, observation light) from the sample S that is illuminated only at the portion corresponding to the micromirrors 51 _{i, j} in the on state is condensed by the objective lens 29 and then transmitted through the lens 26. Then, the light is guided to the light deflection element 25. At this time, the objective lens 29 and the lens 26 form an image of the sample S on the deflection surface 25A of the light deflection element 25. As a matter of course, in the micro mirror 51 _{i, j} that has guided the illumination light, an image of the illumination region of the sample S with respect to the micro mirror 51 _{i, j} is formed, and therefore the micro mirror 51 _i that is in the on state. _{, j} deflects the image light to the imaging lens 24. For example, the micromirrors 51 _1,1 that are turned on in FIG. 3a, with the illumination light at positions corresponding to the micromirrors 51 _1,1 of the sample S is irradiated, the light image of the illuminated region, The light enters the minute mirror 51 _1,1 and is reflected so as to follow the same optical path as the illumination light in the reverse direction.

  The observation light deflected to the imaging lens 24 passes through the imaging lens 24 and reaches the half mirror 23, and the observation light that has passed through the half mirror 23 reaches the light receiving surface 31 </ b> A of the detector 31. Here, the imaging lens 24 is configured to cause all observation light incident from the light deflection element 25 to be incident on the light receiving surface 31A of the detector 31, and the sample imaged on the deflection surface 25A of the light deflection element 25. The light flux in the S illumination region is condensed on the light receiving surface 31 </ b> A of the detector 31 by the imaging lens 24.

  The detector 31 is configured by a two-dimensional image sensor that detects a two-dimensional image, such as a CCD (Charge Coupled Device) image sensor or a CMOS (Complementary Metal Oxide Semiconductor) image sensor. The detector 31 detects the intensity of light incident on each pixel, and outputs an image signal indicating the result (ie, the confocal image of the sample S) to the control unit 41. That is, the detector 31 captures an image of the sample S with the observation light.

  The speed at which the illumination light is scanned on the sample S by the light deflection element 25 is sufficiently faster than the frame rate at which the detector 31 captures an image of the sample S, and the detector 31 captures a confocal image for one frame. In the meantime, the illumination light is scanned on the sample S at least once.

  Further, the number of pixels of the detector 31 is larger than the number of micromirrors 51 of the light deflection element 25, and the light beam reflected by one micromirror 51 is incident on a plurality of pixels of the detector 31 and detected.

Here, the light deflection element 25, the imaging lens 24, and the detector 31 are the deflection surface 25A of the light deflection element 25, the main plane 24A including the principal point of the imaging lens 24, and the light receiving surface 31A of the detector 31. Are extended so as to intersect each other by a single straight line passing through the point A1 and perpendicular to the paper surface. That is, the deflection surface 25A of the light deflection element 25, the principal plane 24A including the principal point of the imaging lens 24, and the light receiving surface 31A of the detector 31 have a condition that satisfies the Scheinproof law (hereinafter referred to as the Scheinproof condition). Meet). Accordingly, the entire range of the image of the sample S connected on the deflection surface 25A of the light deflection element 25 is formed on the light receiving surface 31A of the detector 31. That is, all the light beams of the observation light deflected in the direction of the imaging lens 24 by the respective micro mirrors 51 _{i, j} are imaged on the light receiving surface 31A of the detector 31. Therefore, a high quality image of the sample S can be obtained with less blur in all ranges.

  However, the vertical and horizontal scales of the image formed on the light receiving surface 31 </ b> A of the detector 31 are changed in comparison with the image formed on the deflecting surface 25 </ b> A of the light deflection element 25 when the Scheinproof condition is satisfied. . For example, an image that was square on the deflection surface 25A of the light deflection element 25 becomes an isosceles trapezoid (trapezoid) on the light receiving surface 31A of the detector 31. For example, as shown in FIG. 4, on the deflection surface 25A of the optical deflection element 25, a square 61 whose sides are parallel to the X axis and Y axis and whose center (intersection of diagonal lines) is on the optical axis is a detector. On the 31 light receiving surface 31A, an isosceles trapezoid 71 as shown in FIG. Note that the distortion of the image varies depending on the position and orientation of the image.

Therefore, for example, a CPU (Central Processing Unit) (not shown) acquires the image signal obtained by the detector 31 from the control unit 41 and corrects the distortion of the image of the sample S. Since the amount of image distortion is determined by the positional relationship between the deflection surface 25A of the light deflection element 25, the main plane 24A of the imaging lens 24, and the light receiving surface 31A of the detector 31, it can be easily corrected by a predetermined calculation. Is possible. Then, for example, the CPU supplies the corrected image signal to a monitor (not shown), and the monitor displays an image of the sample S with almost no blur and distortion. Note that the image at this time is a confocal image with a very shallow depth of focus because the micromirrors 51 _{i, j} play the role of a pinhole.

  As described above, the confocal microscope 1 can obtain a high-quality image of the sample S with less blur.

  Further, a two-dimensional image of the sample S can be obtained almost in real time by the detector 31 without performing image processing such as arranging light intensity signals at each position of the sample S as in a conventional confocal microscope. Can do.

  Further, by changing the relative position of the objective lens 29 and the stage 30 in the Z direction, a two-dimensional image of each observation plane is acquired while shifting the focal plane of the objective lens 29 in the sample S, that is, the observation plane, in the Z direction. The spectrographer can easily and finely observe the desired surface of the sample S. In addition, a high-definition and high-quality three-dimensional image of the sample S can be obtained by correcting and superimposing a plurality of acquired two-dimensional images as described above with reference to FIGS. 4 and 5. Can do.

Further, in the confocal microscope 1, when the two-dimensional scanning is performed by the deflection control of the micro mirrors 51 _{i, j} of the light deflection element 25, unnecessary light is attenuated by the light attenuating member 27. Therefore, it is possible to detect light from the observation object with high sensitivity. This makes it possible to provide a confocal microscope with good detection sensitivity.

Furthermore, in the confocal microscope 1, the illumination optical system that guides the illumination light from the light source 21 to the sample S, and the image plane of the sample S illuminated by the illumination light are displayed on the spectroscopic side, that is, on the detector 31 side. The light deflection element 25 is disposed on both optical paths of the imaging optical system that guides light to the light source, and the illumination optical system and the imaging optical system are separated on the optical path between the light source 21 and the light deflection element 25. A half mirror 23 is arranged, and both the optical system from the half mirror 23 to the objective lens 29 is used in common. Then, the illumination position on the sample S and the observation position of the sample S can be selected by the same light deflection element 25. By performing deflection control of the micromirrors 51 _{i, j} of the light deflection element 25, 2 of the sample S is controlled. Dimension scanning is performed. Therefore, the imaging optical system and the illumination optical system are different from each other in the light deflection element in comparison with the confocal microscope including the light deflection element for selecting the illumination position and the light deflection element for selecting the observation position. It becomes unnecessary to adjust the corresponding position of the micromirror, and the illumination position and the observation position can be easily synchronized with the light deflection element.

Further, in the confocal microscope 1, since the two-dimensional scanning is performed by the deflection control of the micro mirrors 51 _{i, j} of the light deflecting element 25, the scanning mechanism has a scanning mechanism as compared with the case where a scanning mirror such as a galvanometer mirror is used. The number of parts can be reduced, and the configuration can be simplified. Moreover, since the number of parts can be reduced, the probability of failure can be lowered and the reliability of the product can be improved. Furthermore, the manufacturing cost can be reduced as compared with the case of using a scanning mirror such as a galvanometer mirror.

  Next, a second embodiment of a confocal microscope to which the present invention is applied will be described with reference to FIGS.

  FIG. 6 shows an external view of a confocal microscope 101, which is a second embodiment of a confocal microscope to which the present invention is applied, as viewed from the side. In the drawing, a part of the optical system disposed inside the confocal microscope 101 is shown in cross section so that the configuration of the optical system can be seen, and the members indicated by the cross section are hatched. Further, in the figure, the same reference numerals are given to the lower two digits in the parts corresponding to those in FIG.

  The confocal microscope 101 includes an optical system that is almost the same as that of the confocal microscope 1 of FIG. 1, and is provided with a total reflection mirror 132 that is configured by two units of the microscope 111 and the lens barrel unit 112. The point and the installation position of each optical element are different from those of the confocal microscope 1. Hereinafter, description of the same parts as the confocal microscope 1 will be omitted or simplified as appropriate.

  The microscope 111 of the confocal microscope 101 is configured to include a revolver 128, an objective lens 129, and a stage 130. The lens barrel unit 112 is detachable from the microscope 111, and includes a light source 121, a collector lens 122a, a lens 122b, a half mirror 123, an imaging lens 124, a light deflection element 125, a lens 126, and a light dimming member 127. The detector 131, the total reflection mirror 132, and the control unit 141 are stored.

  First, the illumination optical system of the confocal microscope 101 will be described. The illumination optical system of the confocal microscope 101 includes a light source 121, a collector lens 122a, a lens 122b, a half mirror 123, an imaging lens 124, a light deflection element 125, a lens 126, a total reflection mirror 132, and an objective lens 129. . That is, the illumination optical system of the confocal microscope 101 is different from the illumination optical system of the confocal microscope 1 in that a total reflection mirror 132 is provided between the lens 126 and the objective lens 129.

  The illumination light emitted from the light source 121 is condensed by the collector lens 122 a and the lens 122 b and guided to the half mirror 23. The illumination light that has reached the half mirror 23 is deflected in the lower right direction in the drawing, passes through the imaging lens 124, becomes a parallel light beam, and reaches the light deflection element 125.

  The light deflection element 125 has a deflection surface 125A having a positional relationship conjugate with a predetermined observation surface of the sample S, and is arranged at a position corresponding to the arrangement position of the field stop. The light deflection element 125 is configured by, for example, DMD, and can control the polarization direction of incident light incident on the deflection surface 125A for each predetermined region.

In the optical deflecting element 125 such as DMD, as shown in FIG. 7, the micromirror 151 is controlled by on / off control of a voltage applied to a holding unit that holds the micromirrors 151 two-dimensionally arranged on the deflecting surface 125A. It is possible to control the angle of each of the two in two directions. Specifically, among the plurality of micromirrors 151 _{i, j} arranged in two dimensions of i × j (i, j: natural number), micromirrors 151 _{m, n} (1 ≦ m ≦ i, 1 ≦ n) ≦ j) and the minute mirror 151 _{p, q} (1 ≦ p ≦ i, 1 ≦ q ≦ j), the upper left direction in the figure reflected by the half mirror 123 and transmitted through the imaging lens 124 will be described. Illumination light is incident on each of the micromirrors 151 _{m, n} and the micromirrors 151 _{p, q} . At this time, since the micromirrors 151 _{p, q} are in the on state, the incident illumination light is deflected to the lens 126 side in the left direction in the figure. On the other hand, since the micromirrors 51 _{m, n} are in the off state, the incident illumination light is deflected toward the light attenuating member 127 in the lower left direction in the figure. That is, the micro mirrors 151 i, by performing the voltage control of _j on / off, the micromirrors 151 i, by each of _j, the illumination light coming incident from one direction, the leftward in the drawing of the lens 126, It is possible to guide in two directions with the light dimming member 127 in the lower left direction in the figure.

In addition, the deflection surface 125A of the light deflection element 125 is disposed at a position conjugate with a predetermined observation surface of the sample S placed on the stage 130. Therefore, in order to guide the light reaching the deflection surface 125A of the light deflection element 125 to a desired location on the sample S, the micromirrors 151 _{i, j} arranged in the region corresponding to the desired location are turned on. Thus, the incident illumination light may be guided to the lens 126 side. For example, as shown in FIG. 7, the illumination light deflected to the lens 126 side by the micromirrors 151 _{p, q} is transmitted through the lens 126, is deflected downward in the figure by the total reflection mirror 132, and is applied to the revolver 128. A portion incident on the held objective lens 129, focused on the observation surface of the sample S conjugate with the deflection surface 125 A of the light deflection element 125 by the objective lens 129 _, and corresponding to the micromirrors 151 _{p and q} of the sample S Only irradiated. Thus, in the confocal microscope 101, the illumination light can be guided to a desired position on the sample S by individually controlling the on / off of each of the micro mirrors 151 _{i, j} of the light deflection element 125. it can.

  On the other hand, the light deflected toward the light attenuating member 127 by the light deflecting element 125 is unnecessary light that does not contribute to the image and is attenuated by the light attenuating member 127 similar to the light attenuating member 27 of the confocal microscope 1. To be lighted.

Then, the illumination light can be scanned over the entire range of the sample S by controlling the micromirrors 151 _{i, j} of the light deflection element 125 in the same manner as described above with reference to FIG.

  A driving device (not shown) is connected to the stage 130 and relatively moves the objective lens 129 and the stage 130 in the Z direction. Further, the stage 130 itself is configured to be freely movable in the XY directions, and the spectroscope can observe a desired part of the sample S. Further, the revolver 128 is configured to be rotatable manually or electrically, and a desired objective lens 129 can be selectively used from among a plurality of objective lenses 129 attached to the revolver 128.

  Next, the imaging optical system of the confocal microscope 101 will be described. The imaging optical system of the confocal microscope 101 includes an objective lens 129, a total reflection mirror 132, a lens 126, a light deflection element 125, an imaging lens 124, a half mirror 123, and a detector 131. That is, the imaging optical system of the confocal microscope 101 is different from the imaging optical system of the confocal microscope 1 in that a total reflection mirror 132 is provided between the lens 126 and the objective lens 129. .

Light from the sample S that is illuminated only at the position corresponding to the micromirrors 151 _{i, j} in the on state (that is, observation light) is collected by the objective lens 129 and then rightward in the figure by the total reflection mirror 132. And is transmitted through the lens 126 and guided to the light deflection element 125. At this time, the objective lens 129 and the lens 126 form an image of the sample S on the deflection surface 125A of the light deflection element 125. Then, the micromirrors 151 i Led illumination light, at _j, the micromirrors 151 i, since the image of the illuminated region of the sample S with respect to _j is formed, the micro mirrors 151 i where in the ON state, _j is The light of the image is deflected to the imaging lens 124.

  The observation light deflected to the imaging lens 124 passes through the imaging lens 124 and reaches the half mirror 123, and the observation light that has passed through the half mirror 123 reaches the light receiving surface 131A of the detector 131. Here, the imaging lens 124 is configured to cause all the observation light incident from the light deflection element 125 to be incident on the light receiving surface 131A of the detector 131, and the sample imaged on the deflection surface 125A of the light deflection element 125. The luminous flux in the S illumination region is condensed on the light receiving surface 131A of the detector 131 by the imaging lens 124.

  Similarly to the detector 31 of the confocal microscope 1, the detector 131 is constituted by a two-dimensional image sensor, detects the intensity of light incident on each pixel, and obtains the result (ie, the confocal image of the sample S). The image signal shown is output to the control unit 141. That is, the detector 131 captures an image of the sample S using observation light.

Here, as for the deflection surface 125A of the light deflection element 125, the main plane 124A including the main point of the imaging lens 124, and the light receiving surface 131A of the detector 131, the extended surfaces are perpendicular to the paper surface through the point A11. It intersects with a single straight line and satisfies the Scheinproof condition. Accordingly, the entire range of the image of the sample S connected on the deflection surface 125A of the light deflection element 125 is formed on the light receiving surface 131A of the detector 131. That is, all the light beams of the observation light deflected in the direction of the imaging lens 124 by the respective micromirrors 151 _{i, j} are imaged on the light receiving surface 131A of the detector 131. Therefore, a high quality image of the sample S can be obtained with less blur in all ranges.

  A CPU (Central Processing Unit) (not shown) acquires the image signal obtained by the detector 131 from the control unit 141 and corrects the distortion of the image of the sample S. Further, the CPU supplies, for example, the corrected image signal to a monitor (not shown), and the monitor displays an image of the sample S with almost no blur and distortion.

  As described above, the confocal microscope 101 can obtain a high-quality image of the sample S with less blur as with the confocal microscope 1.

  Further, the confocal microscope 101 has a total reflection mirror 132 disposed between the lens 126 and the objective lens 129, and the arrangement of each optical element is devised to make the entire apparatus compact compared to the confocal microscope 1. Can be configured.

  Furthermore, by storing optical elements other than the objective lens 129 in the lens barrel unit 112 and making it detachable from the microscope 111, the lens barrel unit 112 can be used by being mounted on a conventional microscope, for example. become. In addition, the center of gravity of the lens barrel unit 112 can be lowered by placing each optical element in the lens barrel unit 112 so that the optical path travels in the horizontal direction or the direction close to the horizontal direction as much as possible, and is attached to the microscope 111. Thus, the stability of the lens barrel unit 112 can be improved.

  Hereinafter, modifications of the embodiment of the present invention will be described.

  In the above description, an example in which the micro mirror 51 of the light deflection element 25 and the micro mirror 151 of the light deflection element 125 are selected and driven one by one to scan illumination light has been described. The illumination light may be scanned while being selected and driven.

For example, with reference to FIG. 8, a case where a plurality of micromirrors 51 of the light deflection element 25 are driven simultaneously will be described. FIG. 8 is a simplified representation of the two-dimensional arrangement of the micromirrors 51 _{i, j} of the light deflection element 25, as in FIG.

FIG. 8a shows the first micro mirror 51 _1,1 from the left at the top of the micro mirrors 51 _{i, j} of the light deflector 25, the fifth micro mirror 51 _1,5 from the left at the top, and from the top. This shows the case where the fourth micro mirrors 51 _{4,1 in} the fourth stage and the fifth micro mirrors 51 _4,5 in the fourth stage from the top are turned on. This means that only the location is illuminated. Figure 8a and in the same manner, FIG. 8b micromirrors 51 _1,2, 51 _1,6, 51 _4,2, 51 _4,6 only on state, Figure 8c micromirrors 51 _1,3, 51 _1,7 , _514,3 and _514,7 only show the state when turned on. In the cases of FIGS. 8b and 8c as well, only the corresponding four portions of the sample S are illuminated at the same time as in FIG. 8a.

The control unit 41 moves the four micromirrors 51 that are turned on one by one in the right direction among the micromirrors 51 _{i, j} of the light deflecting element 25, and moves them down one step when moving to the right end, starting from the left end. By moving in the right direction, finally, as shown in FIG. 8d, only the micromirrors 51 _3,4 , 51 _3,8 , 51 _6,4 , ₅₁₆ , ₈ are turned on.

That is, as shown in FIGS. 8a, 8b, 8c,..., 8d, the micromirrors 51 _{i, j} of the light deflection element 25 are sequentially driven four by four to illuminate the entire range of the sample S. By scanning the light, it is possible to scan the entire observation site of the sample S.

  It should be noted that the interval between the four micromirrors 51 that are simultaneously turned on is set to a distance that does not optically affect each of them. Specifically, the Airy disk of the light beam reflected by the minute mirror 51 that is in the on state at the same time is set to a distance greater than the length that does not affect each other. This distance is set to the same value as the distance between the pinholes arranged on the nipou disk in a conventional nippo disk type confocal microscope, for example.

  As described above, the scanning time of the illumination light can be shortened by simultaneously driving the plurality of minute mirrors 51 and scanning the illumination light as compared with the case of driving the minute mirrors 51 one by one. Become.

  The number of micromirrors 51 that are driven simultaneously is not limited to this example, and can be arbitrarily set within a range in which the interval between micromirrors 51 that are simultaneously turned on can be set to a distance that does not optically affect each of them. It is possible to set the number of

  In the above description, the light source 21 is built in the confocal microscope 1. However, in consideration of the influence of heat from the light source 21, the light source 21 is arranged in the lamp house and the confocal microscope 1 is used. It may be configured to be attachable to and detachable from. Similarly, in the above description, in the confocal microscope 101, the light source 121 is built in the lens barrel unit 112. However, the light source 121 is disposed in the lamp house, and the microscope 111 or the lens barrel unit 112 is attached. On the other hand, it may be configured to be detachable.

  Furthermore, in the above description, the case where epi-illumination is employed as an illumination method has been described as an example. However, the present invention can also be applied to the case where transmitted illumination that irradiates illumination light from behind the sample S is employed.

  In the above description, the configuration in which one light deflection element 25 or light deflection element 125 is disposed on the optical paths of both the illumination optical system and the imaging optical system has been described. It is also possible to arrange separate light deflection elements in each of the optical systems. In this case, the selection of the micromirrors that contribute to the optical system of these light deflection elements has a corresponding relationship. In that case, the light deflection element of the illumination optical system and the light deflection element of the imaging optical system are arranged at positions where the micromirrors having the corresponding relationship are optically coincident with each other, and the micromirror When displacing, control may be performed so as to displace in synchronization.

  In the above description, the detector 31 and the detector 131 are configured by two-dimensional image sensors. However, the detector 31 and the detector 131 are configured by a device that detects light intensity such as a PMT (Photomultiplier). You may make it do.

  For example, when PMT is used as the detector 31 of the confocal microscope 1, the detector 31 detects the intensity of the light collected from the imaging lens 24 and outputs it to the control unit 41. The control unit 41 associates the positional information of the micromirror 51 in the on state among the micromirrors 51 of the light deflecting element 25 with the detection signal value from the previous detector 31, and stores the memory (not shown). Store in the department. This is performed while sequentially changing the micromirrors 51 to be turned on, and the position information of the micromirrors 51 corresponding to the detection signals of the entire observation region of the sample S is acquired. As a result, signals indicating the position and the light intensity are obtained, and a two-dimensional image of the sample S is obtained by subjecting these signals to predetermined image processing by a CPU (not shown) or the like.

  At this time, as described above, the observation light reflected by the micromirrors 51 in the direction of the imaging lens 24 according to Scheinproof's law, regardless of the position of the micromirrors 51, is the light receiving surface of the detector 31. The image is formed at. Therefore, the intensity of the observation light reflected by each micromirror 51 can be detected more accurately, and the image quality of the sample S is improved.

  Further, in the embodiment of the present invention, instead of the light deflection element 25 that totally reflects light, for example, a liquid crystal element that reflects light in a certain state and transmits light in a certain state, or light diffraction is used. An element capable of changing the distribution in which light is reflected two-dimensionally in a desired direction by using GLV (Grating Light Valve (registered trademark)) or the like can also be used.

  Furthermore, the present invention can be applied not only when scanning spot-shaped illumination light in a two-dimensional direction but also when scanning linear (linear) illumination light in a one-dimensional direction. is there.

  The embodiment of the present invention is not limited to the above-described embodiment, and various modifications can be made without departing from the gist of the present invention.

  1 confocal microscope, 21 light source, 22a collector lens, 22b lens, 23 half mirror, 24 imaging lens, 24A main plane, 25 light deflection element, 25A deflection surface, 26 lens, 29 objective lens, 31 detector, 31A light reception Surface, 41 control unit, 51 micromirror, 101 confocal microscope, 111 microscope, 112 lens barrel unit, 121 light source, 122a collector lens, 122b lens, 123 half mirror, 124 imaging lens, 124A main plane, 125 light deflection element , 125A deflection surface, 126 lens, 129 objective lens, 131 detector, 131A light receiving surface, 132 total reflection mirror, 141 control unit, 151 micro mirror, S sample

Claims (8)

The illumination light from the light source is condensed on a predetermined observation surface of the sample, the illumination optical system that scans the condensing position, and the observation light from the sample is condensed by the objective lens, imaged, and detected. In a confocal microscope equipped with an imaging optical system,
Light deflecting means capable of controlling the deflection direction of incident light incident on a predetermined deflecting surface for each predetermined region, wherein the deflecting surface is disposed at a position conjugate with the observation surface and is condensed by the objective lens. Light deflecting means for controlling the polarization direction of the observation light incident on the deflection surface for each region;
Deflection control means for sequentially selecting the areas of the light deflection means and controlling the observation light incident on the selected areas to be deflected in a predetermined first direction;
Light detection means for detecting the observation light deflected in the first direction by the light deflection means;
An imaging lens that forms an image of the observation light deflected in the first direction by the light deflection unit on a light receiving surface of the light detection unit;
The light deflection means, the imaging, and the light deflection means, the main plane of the imaging lens, and a surface obtained by extending the light receiving surface of the light detection means intersect with each other by a single straight line. A confocal microscope in which a lens and the light detection means are arranged. The light deflection means further controls the polarization direction of the illumination light incident on the deflection surface for each region,
The deflection control means controls the illumination light incident on the selected region to deflect in a predetermined second direction and to deflect the observation light in the first direction;
The confocal microscope according to claim 1, wherein the objective lens further condenses the illumination light deflected in the second direction by the light deflecting unit on the observation surface of the sample. The confocal microscope according to claim 2, further comprising a first optical element that is disposed on an optical path between the light source and the light deflecting unit and separates the illumination optical system and the imaging optical system. . It is disposed on the optical path between the light deflecting means and the objective lens, deflects the illumination light from the light source in the direction of the objective lens, and directs the observation light from the sample in the direction of the light deflecting means. The confocal microscope according to claim 2 or 3, further comprising a second optical element for deflecting. The confocal microscope according to any one of claims 1 to 4, wherein the light detection unit includes a two-dimensional image sensor. The light detection means is constituted by a two-dimensional image sensor,
The deflection control means simultaneously selects a plurality of the regions separated by a predetermined distance or more, deflects the illumination light incident on the selected plurality of regions in a predetermined second direction, and converts the observation light into the first light The confocal microscope according to claim 2, wherein the confocal microscope is controlled to be deflected in a direction of 1. The confocal according to any one of claims 1 to 6, wherein the light deflecting unit includes a plurality of micromirrors provided on the deflection surface, and the micromirrors can control the deflection direction of incident light for each region. microscope. A microscope comprising the objective lens;
The confocal microscope according to any one of claims 1 to 7, comprising: a unit that stores the light source, the light deflecting unit, the light detecting unit, and the imaging lens, and is detachable from the microscope. .

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