JP4044914B2 - Scanning microscope - Google Patents

Description

  The present invention relates to a scanning microscope using a confocal principle.

  Conventionally, there are scanning microscopes using the confocal principle. The scanning microscope is provided with an objective lens and a pinhole, and when the sample is at the focal position of the objective lens, the laser beam that has passed through the pinhole is received by the first light receiving element, so that the height to be observed Only the image (confocal image) for the portion is clearly displayed (the resolution is increased). Such a confocal image is a black and white (achromatic) image. However, there is little information in such a black and white video, that is, information on the color of the sample cannot be obtained, and detailed observation such as determination of the type of scratches or deposits may be difficult. For this reason, there are color (chromatic) scanning microscopes.

  However, a conventional color scanning microscope scans the sample surface using laser beams of three primary colors from three light sources. This complicates the optical system, making the microscope expensive and large.

Accordingly, an object of the present invention is to reduce the cost and size of a color scanning microscope by including a laser optical system and an optical system for illumination light for color imaging information.
The height (Z coordinate) of the sample stage is also movable, and the in-focus color imaging information is stored for each pixel (imaging unit) so that a focused image can be obtained.

The novel scanning microscope according to the present invention includes luminance information from the first light receiving element of the first optical system using laser light and a second light receiving element of the second optical system using illumination light for color imaging information. A color video signal is obtained based on the color imaging information.
The present invention includes a sample stage capable of moving the sample three-dimensionally with respect to the objective lens, and means for storing the height of the sample stage when the first optical system is in focus.
The present invention also includes means for storing focused color imaging information from the second light receiving element at the stored height of the sample stage.
Further, according to the present invention, the second light receiving element is a color CCD, and a color video signal in focus is obtained using color imaging information when the first optical system is in focus for each pixel. Configured.
Further, according to the present invention, the second light receiving element is a solid-state image sensor, and a focused color video signal is obtained using color imaging information when the first optical system is in focus for each pixel. Configured as follows.
By the way, in the present invention, the “color image signal” means an image signal composed of intensities for the three primary colors of light (red, green, blue), a signal composed of a luminance signal and a color difference signal, a horizontal synchronization signal, and a color signal. A composite color video signal including a burst signal is a signal that can display a color video as it is or after being processed.
Further, “color imaging information” refers to information obtained by imaging and includes “luminance information” and “color information”. “Luminance information” refers to information relating to luminance that does not include color, and “color information” refers to information relating to the balance of color intensity, such as color difference signals.
In the following description, the terms “luminance information from the first light receiving element” and “color information from the second light receiving element” mean that the case of obtaining a color video signal as follows is included. Sometimes. That is, in the following description, the color video signal may include luminance information from the first light receiving element and color information of the color imaging information from the second light receiving element. . The color video signal may include corrected luminance information based on luminance information from the first light receiving element and color information in the color imaging information from the second light receiving element. Further, the color video signal may include luminance information from the first light receiving element and correction color information based on color information in the color imaging information from the second light receiving element. In addition, the color video signal may include the corrected luminance information and the corrected color information.
In addition, when the condensed laser beam is dot-like, the laser beam can be relatively scanned two-dimensionally along the sample surface. On the other hand, when the condensed laser beam is a linear line laser beam, the line laser beam can be relatively scanned along the sample surface in a direction substantially orthogonal to the condensed line laser beam. Here, “relatively scanning along the sample surface” means that the sample is stationary and scanning with laser light or line laser light, and the sample is moved without scanning with laser light or line laser light. This means that there are at least three cases: scanning with laser light and scanning the laser beam in the X direction and moving the sample in the Y direction (direction orthogonal to the X direction).

According to the scanning microscope of the present invention, since color imaging information is obtained by the second optical system using illumination light for color imaging different from the laser light of the first optical system, the conventional color using the three primary colors of laser light is obtained. Compared to a laser microscope, the optical system has a remarkably simple structure, so that the cost and size can be reduced.
Then, by providing a means for storing the height of the sample stage when the first optical system is in focus, the height of the sample stage at the focus can be stored.
Further, by providing means for storing in-focus color imaging information from the second light receiving element at the stored height of the sample stage, a strictly focused image can be obtained.
Further, since a color CCD is used as the light receiving element of the second optical system and the color imaging information at the focal point of the first optical system is used for each pixel, a color video signal is obtained. Matched images can be obtained.
Further, a solid-state image sensor is used as the light receiving element of the second optical system, and color imaging information at the focal point of the first optical system is used for each pixel to obtain a color video signal. Can be obtained.
In addition, although the resolution about color information is low compared with the conventional color laser microscope, when luminance information is obtained by laser light, the resolution is higher than that of a normal enlarged image, and thus the utility value is sufficiently high. That is, when luminance information from the first optical system using laser light is used, the resolution is higher than that of a simple enlarged image using white light, and a color image is generated using color information from the second light receiving element. As a result, detailed observation of scratches and deposits becomes possible.
Further, by obtaining the maximum luminance of the response light incident on the first light receiving element while changing the distance from the objective lens to the sample, and using it as luminance information of the maximum luminance, a part of the image can be obtained even in a sample with large unevenness. There is no danger of darkening.

Hereinafter, embodiments of the present invention will be described with reference to the drawings.
1 to 4 are drawings showing a first embodiment of a scanning microscope of the present invention.

FIG. 1 is a schematic configuration diagram showing the first embodiment.
In FIG. 1, the scanning microscope includes a laser optical system (first optical system) 1 and a white light optical system (second optical system) 2.

  FIG. 2 is a block diagram showing the first embodiment.

  FIG. 3 is a plan view showing an imaging region in the first embodiment.

  FIG. 4 is a flowchart regarding the first embodiment.

  First, the laser optical system 1 will be described. The laser optical system 1 is a confocal optical system that can detect information related to the depth of the sample w, and uses, for example, a He-Ne laser 10 that emits red laser light L1 as a light source. On the optical axis of the laser 10, there are a first collimating lens 11, a polarizing beam splitter 12, a quarter-wave plate 13, a two-dimensional scanning device 14, a first relay lens 15, a second relay lens 16 and an objective lens 17. Are arranged in order. A sample stage 30 is disposed in the vicinity of the focal position of the objective lens 17, and the objective lens 17 condenses the laser light L1 on the surface of the sample w. The above-described two-dimensional scanning device 14 is composed of, for example, two galvanometer mirrors, and deflects the laser light L1 so that the condensing position on the sample w is two-dimensionally (X, Y) along the surface of the sample w. Direction). The sample stage 30 is driven and controlled in the Z (vertical) direction by the stage control circuit 40, and can be moved by a manual handle in the X and Y directions.

  The laser light (response light) L1 reflected by the sample w passes through the objective lens 17, the second relay lens 16, and the first relay lens 15, and again passes through the two-dimensional scanning device 14 to the 1/4 wavelength plate 13 and The light passes through the polarization beam splitter 12 and travels toward the second imaging lens 18. The laser beam L1 is collected by the second imaging lens 18, passes through the optical aperture 19a having a pinhole, and enters the first light receiving element 19b. The first light receiving element 19b is composed of, for example, a photomultiplier or a photodiode, and photoelectrically converts the incident laser light L1 and outputs an analog light quantity signal via an output amplifier and a gain control circuit (not shown). Output to the 1 A / D converter 41.

  Next, luminance information obtained by the laser optical system 1 will be described. The optical diaphragm 19a is disposed at the focal position of the second imaging lens 18. On the other hand, since the pinhole of the optical diaphragm 19a is very small, the laser beam L1 is focused on the sample w. , The reflected light L1 forms an image at the pinhole of the optical diaphragm 19a, and the amount of received light incident on the first light receiving element 19b becomes remarkably large. Conversely, the laser light L1 is focused on the sample w. Otherwise, the reflected light L1 hardly passes through the pinhole of the optical aperture portion 19a, so that the amount of light received by the first light receiving element 19b is significantly reduced. Therefore, a bright image can be obtained for a focused portion (focused imaging unit) in an imaging region (scanning region) by the laser optical system 1, while a dark image can be obtained for other height portions. It is done. In addition, since the laser optical system 1 is a confocal optical system using the monochromatic laser beam L1, luminance information with excellent resolution can be obtained.

  Next, the white light optical system 2 will be described. The white light optical system 2 uses a white light source 20 that emits white light (illumination light for color information) L2 as a light source. On the optical axis of the white light source 20, a second collimating lens 21, a first half mirror 22, and the objective lens 17 are disposed. In the first half mirror 22, two optical systems 1 and 1 are disposed. The white light optical system 2 is arranged so that the two optical axes coincide. Therefore, the white light L2 is condensed at the same location as the scanning region of the laser light L1. White light (response light) L2 reflected by the sample w passes through the objective lens 17, the first half mirror 22, and the second relay lens 16, and is further reflected by the second half mirror 23 to be a color CCD ( The image is formed on the surface of the second light receiving element 24). In other words, the color CCD 24 is disposed at a position conjugate to or close to the conjugate with the optical diaphragm 19a. The image captured by the color CCD 24 is read out as analog color imaging information to the CCD drive circuit 43 and output to the second A / D converter 42.

  Next, the color video signal generating means 5 of FIG. 2 that operates in the color confocal image mode described later will be described. The color video signal creation means 5 creates color video digital signals ro, go, bo by combining the luminance information from the first light receiving element 19 b and the color information from the color CCD 24. The color video signal generating means 5 includes first and second area circuits 51 and 52, a luminance conversion circuit 53, and the like.

  As shown in FIG. 3, the first and second area circuits 51 and 52 select predetermined common portions of the imaging areas A1 and A2 of the laser optical system 1 and the white light optical system 2 as an image area A0, respectively. The digital signal is output for the selected part. That is, the first area circuit 51 of FIG. 2 stores the luminance signal i in the luminance memory Mi with the resolution corresponding to each pixel of the color CCD 24 for the video area A0. On the other hand, the second area circuit 52 stores red, green, and blue color intensity signals rm, gm, and bm for each pixel in the first color intensity memory Mr1, Mg1, Mb1 for the video area A0. The color intensity signal is a signal including luminance (intensity) for the three primary colors.

The luminance conversion circuit 53 converts the luminance information of the color intensity signals rm, gm, bm for each pixel into luminance information of the luminance signal i according to the following arithmetic expressions (1), (2), (3). The converted color intensity signals ro, go, bo are obtained by replacement, and the signals are stored in the second color intensity memories Mr2, Mg2, Mb2.
Ro = I · Rm / (Rm + Gm + Bm) (1)
Go = I · Gm / (Rm + Gm + Bm) (2)
Bo = I · Bm / (Rm + Gm + Bm) (3)
I: luminance of the luminance signal i Rm, Gm, Bm: luminance (intensity) of the color intensity signal rm, gm, bm
Ro, Go, Bo: Brightness (intensity) of converted color intensity signals ro, go, bo
The first and second color intensity memories Mr1 to Mb1 and Mr2 to Mb2 have storage units corresponding to the pixels of the above-described video area A0 in the color CCD 24.

  The converted color intensity signals ro, go, and bo thus obtained are signals obtained by replacing the luminance information in the color imaging information from the color CCD 24 with the luminance information from the first light receiving element 19b. The converted color intensity signals ro, go, bo are read from the second color intensity memories Mr2, Mg2, Mb2 and output to the D / A converter 60. Further, the adder 61 adds the synchronization signal a. Thus, an analog composite color video signal c is obtained. The composite color video signal c is output to the monitor 62, and an image of the sample w is displayed.

  Next, how to use the present scanning microscope will be described. The present scanning microscope selects and uses one of three modes: an area search mode, a monochrome (achromatic) confocal image mode, and a color confocal image mode. These modes are set by operating the operation unit 63.

  When the area search mode is selected, the color video signal generating means 5 stops the laser drive circuit 44 of FIG. 1 and operates the CCD drive circuit 43 to cause the color CCD 24 to take an image. In this area search mode, the color intensity signals rm, gm, bm stored in the first color intensity memories Mr1, Mg1, Mb1 from the second area circuit 52 in FIG. 2 are output to the D / A converter 60 as they are. A normal enlarged image having a deep depth of field is displayed on the monitor 62. Therefore, by moving the sample stage 30 in FIG. 1 in the X and Y directions, it is possible to find an area to be imaged.

  When the black and white confocal image mode is selected, the color video signal generating means 5 (FIG. 2) operates the laser drive circuit 44 and the two-dimensional scanning device 14 of the laser optical system 1 and causes the laser optical system 1 to take an image. . In the black and white confocal image mode, the luminance signal i stored in the luminance memory Mi from the first area circuit 51 in FIG. 2 is output to the D / A converter 60 as it is, and has a high resolution black and white (achromatic color). Is displayed on the monitor 62.

  When the color confocal image mode is selected, the laser drive circuit 44 and the CCD drive circuit 43 are alternately driven as described below. That is, when the mode is selected in step S1 of FIG. 4, the process proceeds to step S2, and one screen is scanned with the laser light L1, and then the process proceeds to step S3. In step S3, the laser drive circuit 44 in FIG. 1 is stopped, and the laser beam L1 is not emitted from the laser 10. In this state, the process proceeds to step S4 in FIG. In the color intensity signals rm, gm, bm of FIG. 2 obtained in step S4, the luminance information contained in the signals is replaced with the luminance information of the luminance signal i obtained in step S2, and the converted color intensity signal ro. , Go, bo. The converted color intensity signals ro, go, and bo are stored in the second color intensity memories Mr2, Mg2, and Mb2, respectively, and then output to the D / A converter 60 to display an enlarged color image on the monitor 62. Note that the process proceeds to step S5 after step S4 in FIG. 4, and scanning of the laser beam L1 and accumulation and reading of charges by the CCD drive circuit 43 are repeated.

  Since the color confocal image thus obtained has a low resolution for color information, the resolution is slightly lower than that of a conventional color laser microscope using three primary colors of laser light, but the resolution is higher than that of a normal enlarged image. A video with a sufficiently high utility value can be obtained.

  Further, since the color information is obtained by the white light optical system 2 using the white light source 20 and the color CCD 24 of FIG. 1, the optical system has a remarkably simple structure as compared with a conventional color laser microscope using three primary color laser beams. Therefore, the cost and size of the scanning microscope can be reduced.

  In the case of the present embodiment, when imaging is performed by the color CCD 24 of FIG. 1 (charge is accumulated in the color CCD 24), the laser drive circuit 44 is stopped so that the laser light L1 does not enter the color CCD 24. . Therefore, an image having a color close to the actual color of the sample w can be obtained without forming an image having the color of the laser beam L1.

  As means for preventing the laser light L1 from entering the color CCD 24, various means such as using a shutter for shielding the laser light L1 or setting the scanning range of the laser light L1 outside the imaging region of the color CCD 24 are used. The method can be adopted. Further, even if the laser beam L1 is incident on the color CCD 24 and takes on the color of the laser beam L1, a color image can be obtained, which is included in the scope of the present invention.

  By the way, if the color of the sample w in FIG. 1 and the color of the laser beam L1 are similar colors, the amount of reflected light L1 on the sample w (the luminance I in the equations (1) to (3)) increases. The amount of light incident on the light receiving element 19b increases. Therefore, in such a case, the image becomes whitish than the actual color of the sample w. Therefore, the second embodiment shown in FIG. 5 has a function of obtaining a color close to the color of the actual sample even in the case of such a similar color.

The second embodiment shown in FIG. 5 differs from the first embodiment in that the luminance conversion circuit 53 includes a determination unit 53a and a correction variable storage unit 53b that stores three correction variables. In the present embodiment, the luminance conversion circuit 53 corrects the luminances Rm, Gm, and Bm of the color intensity signals rm, gm, and bm according to the following arithmetic expressions (11) to (13) to convert the converted color intensity signals ro, go. , Bo brightness (intensity) Ro, Go, Bo.
Ro = Kr · I · Rm / (Rm + Gm + Bm) (11)
Go = Kg · I · Gm / (Rm + Gm + Bm) (12)
Bo = Kb · I · Bm / (Rm + Gm + Bm) (13)
Kr, Kg, Kb: correction variables

The discriminating means 53a of the luminance conversion circuit 53 compares the red ratio Rc shown in the following equation (14) with a predetermined threshold Sr for each imaging unit (each pixel of the color CCD 24) in the imaging area of the sample w. In addition, it is determined whether or not the portion of each imaging unit is more reddish than the predetermined threshold value Sr.
Rc = Rm / (Rm + Gm + Bm) (14)
That is, when the red luminance Rm received by each pixel of the color CCD 24 is larger than the total value (Rm + Gm + Bm) of the luminances of the three colors, the determination unit 53a determines that the imaging unit portion is reddish. To do.

  When the red ratio Rc in the equation (14) is smaller than the threshold value Sr, the luminance conversion circuit 53 sets Kr = Kg = Kb = 1 and converts the converted color intensity signal as in the first embodiment. Get ro, go, bo. On the other hand, when the red ratio Rc is greater than or equal to the threshold value Sr, the luminance conversion circuit 53 reads the correction variables Kr, Kg, and Kb from the correction variable storage unit 53b, and converts the converted color intensity according to the equations (11) to (13). Signals ro, go, and bo are obtained.

  FIG. 5B is a graph showing the correction variables Kr, Kg, and Kb. Each correction variable Kr, Kg, Kb is set to be smaller than “1.0” as the red ratio Rc of the imaging unit of the sample w is larger, for example. Therefore, since the brightness is reduced by multiplying the correction variables Kr, Kg, and Kb as in the above equations (11) to (13), the image does not become whitish even when the red ratio Rc is large. An image having a color close to the color of the sample w is obtained.

  By the way, when the uneven sample w is observed in the color confocal image mode described above, the image of the portion out of focus (height) is darkened. Therefore, the third embodiment shown in FIG. 6 and FIG. 7 includes an uneven color confocal image mode suitable for observation of the uneven sample w.

  Next, a configuration of the third embodiment provided with the uneven color confocal image mode will be described. As shown in FIG. 6, in the third embodiment, the color video signal generating means 5 includes a memory including a maximum luminance storage unit Mmax and a Z coordinate storage unit Mz. The maximum luminance storage unit Mmax stores the maximum luminance Imax received by the first light receiving element 19b (FIG. 5) for each imaging unit (unit corresponding to each pixel of the color CCD 24) in the scanning region. The Z coordinate storage unit Mz stores the Z coordinate (the height of the sample stage 30) when the luminance I reaches the maximum luminance Imax for each imaging unit. The second color intensity memories Mr2, Mg2, and Mb2 constitute a composite imaging information storage unit, and store color imaging information received by each pixel of the color CCD 24 at the Z coordinate stored in the Z coordinate storage unit Mz. . In other words, the second color intensity memory Mr2, Mg2, Mb2 displays the color imaging information when the height of the sample stage 30 in FIG. 1 is changed and the first optical system 1 is in focus for each pixel (imaging unit). Remember every time.

  The luminance conversion circuit 53 includes luminance comparison means 53c. The luminance comparison means 53c compares the maximum luminance Imax stored in the maximum luminance storage unit Mmax with the luminance I newly stored in the luminance memory Mi for each imaging unit. As a result of the comparison, if the luminance I newly stored in the luminance memory Mi is larger than the maximum luminance Imax as a result of the comparison, the luminance conversion circuit 53 sets the luminance I as the maximum luminance and sets the maximum luminance storage unit Mmax. Remember me. Other configurations are the same as those of the second embodiment shown in FIG. 5, and detailed description and illustration thereof are omitted.

  Next, the color confocal image mode for unevenness will be described. In FIG. 7, first, in step S11, the sample stage 30 of FIG. 1 is moved to the rising end. Next, in step S12, imaging is performed by the second optical system 2, and charges are accumulated in the color CCD 24 to obtain color imaging information. Subsequently, the process proceeds to step S13 in FIG. 7, and the luminance information is obtained by scanning the first optical system 1 with the laser beam L1 to pick up an image, and this content is stored in the maximum luminance storage unit Mmax in FIG. Each unit (unit corresponding to the pixel of the color CCD 24) is stored, and the Z coordinate at the time of imaging is stored in the Z coordinate storage unit Mz, and the converted color intensity signals ro, go, and bo subjected to luminance conversion are stored in the second color. Store in intensity memories Mr2, Mg2, and Mb2.

  Subsequently, the process proceeds to step S14, the sample stage 30 (FIG. 1) is lowered by one step, and then the process proceeds to step S15 in FIG. In step S15, color imaging information is obtained again by the second optical system 2, and the contents are stored for each pixel in the first color intensity memories Mr1, Mg1, Mb1 in FIG. 6, and the process proceeds to step S16 in FIG.

  In step S16, as described below, while scanning the laser beam L1 with the first optical system 1, the maximum luminance Imax of the maximum luminance storage unit Mmax is updated and stored, and the imaging unit with the updated maximum luminance Imax is updated. The conversion color intensity signals ro, go, bo stored in the second color intensity memories Mr2, Mg2, Mb2 are updated and stored. That is, the luminance I newly stored in the luminance memory Mi of FIG. 6 by scanning the laser beam L1 and the maximum luminance Imax stored in the maximum luminance storage unit Mmax are compared for each imaging unit. Means 53c compares. As a result of the comparison, if I> Imax, the luminance conversion circuit 53 stores the luminance I as the new maximum luminance Imax in the maximum luminance storage unit Mmax for the imaging unit, and the first color intensity memory Mr1, The color intensity signals rm, gm, bm stored in Mg1, Mb1, that is, the color imaging information is subjected to luminance conversion, and converted color intensity signals ro, go, bo in the second color intensity memories Mr2, Mg2, Mb2 Update and store the address in the imaging unit. On the other hand, as a result of the comparison, if the luminance I is less than or equal to the maximum luminance Imax, the maximum luminance Imax and the converted color intensity signals ro, go, and bo are not updated for the imaging unit.

Subsequently, the process proceeds to step S17 in FIG. 7. If the sample stage 30 is not at the lower end, the process returns to step S14. On the other hand, if the sample stage 30 is at the lower end, the process proceeds to step S18 and the converted color intensity signal ro, After go and bo are output to the D / A converter 60 (FIG. 5), a composite color video signal c (FIG. 5) is obtained. That is, by repeating steps S14 to S17 in FIG. 7, the updating of the maximum luminance Imax of the maximum luminance storage unit Mmax and the converted color intensity signals ro, go, and bo in FIG. 6 is repeated. Therefore, the luminances of the converted color intensity signals ro, go, and bo output in step S18 are expressed by the following equations (21) to (23) similar to the aforementioned equations (11) to (13).
Ro = Kr · Imax · Rm / (Rm + Gm + Bm) (21)
Go = Kg · Imax · Gm / (Rm + Gm + Bm) (22)
Bo = Kb · Imax · Bm / (Rm + Gm + Bm) (23)
If the mode is OFF in step S19 of FIG. 7, the mode is terminated.

  As described above, in this embodiment, the color imaging information about the Z coordinate when the first optical system 1 is in focus is used for each pixel of the color CCD 24 in FIG. Since an in-focus image is obtained for each pixel, an in-focus image can be obtained rather than a normal color enlarged image.

By the way, although luminance conversion was performed in each said embodiment, in this invention, it is not necessary to perform luminance conversion. That is, the color imaging information stored in the first color intensity memory Mr1, Mg1, Mb1 is stored in the second color intensity memory Mr2, Mg2, Mb2 without performing the luminance conversion in Step S13 and Step S16 of FIG. In other words, the composite color video signal c may be obtained according to the following equations (31) to (33).
Ro = Rm (31)
Go = Gm (32)
Bo = Bm (33)

  Also in this case, as in the third embodiment, the color imaging information about the Z coordinate when the first optical system 1 is in focus is used for each pixel of the color CCD 24 in FIG. Unlike images, you can get images that are strictly in focus.

  By the way, in each said embodiment, although the laser beam L1 condensed in a dot shape on the surface of the sample w and the 1st light receiving element 19b was used, in this invention, it is linear at the surface of the sample w and the 1st light receiving element 19b. It is also possible to use a line laser beam L1 that is focused on the laser beam. That is, as shown in FIG. 8, a line laser beam L1 that is long in the Y direction is used instead of the laser beam L1, and a one-dimensional CCD 19A that is long in the Y direction is used instead of the dotted first light receiving element 19b. A one-dimensional scanning device 14A is used instead of the one-dimensional scanning device 14. In this case, as shown in FIG. 8B, the line laser beam L1 is scanned in a direction orthogonal to the longitudinal direction when the line laser beam L1 is collected on the surface of the sample w. The light diaphragm portion 19a has a slit shape (groove shape).

  Incidentally, in each of the above embodiments, the optical aperture 19a is provided in front of the first light receiving element 19b of the laser optical system 1 in FIG. 1, but the optical aperture 19a is not necessarily provided. For example, as shown in FIG. 9, a one-dimensional CCD 19A for black and white may be provided at the focal position of the second imaging lens 18. In this case, the first one-dimensional scanning device 14A is provided between the first collimating lens 11 and the polarization beam splitter 12, and the second one-dimensional scanning device 14B is provided with the quarter-wave plate 13 and the first relay lens 15. Provide between.

  In each of the above embodiments, the color CCD 24 is used as the second light receiving element of the second optical system 2 in FIG. 1, but other light receiving elements may be used. For example, the reflected light L2 may be separated into three primary colors using a dichroic mirror, and the reflected lights of these three primary colors may be incident on three black and white video two-dimensional CCDs. Although the optical system is different, a color line CCD may be used as the second light receiving element, and a one-dimensional scanning device that scans white response light in a one-dimensional manner may be provided. Further, as the second light receiving element, three monochrome CCD line CCDs (for R, G, B) can be used, and three point light receiving elements (for R, G, B) can be used. it can. In these cases, the scanning device that scans the white response light can also be used as the scanning device that scans the laser light L1.

  Further, as the second light receiving element, in addition to the color CCD, other solid-state imaging elements such as a MOS type, a television camera in which a plurality of imaging tubes are combined, or the like can be used.

  By the way, in the embodiment of FIG. 1, the reflected light of the laser light L1 and the white light L2 is received by the first light receiving element 19b and the color CCD 24, respectively. It may receive light or fluorescence obtained by replacing the reflected light.

  In the above embodiment, the colors are separated into the three primary colors of light. However, in the present invention, the colors may be separated into complementary colors (yellow, cyan, and green). Further, a color difference signal may be used as the color information.

  In each of the above embodiments, the laser relay optical system 1 and the white light optical system 2 are provided with the second relay lens 16 and the infinite correction system is employed. However, the second relay lens 16 is not provided and the finite correction system is employed. May be.

It is a schematic block diagram which shows 1st Embodiment of the scanning microscope of this invention. It is a block diagram of a 1st embodiment. It is a top view which shows the imaging area of 1st Embodiment. It is a flowchart of a 1st embodiment. It is a figure which shows the block diagram concerning 2nd Embodiment, and a correction variable. It is a block diagram of the color video signal preparation means concerning 3rd Embodiment. It is a flowchart of a 3rd embodiment. It is a schematic block diagram which shows 4th Embodiment of a scanning microscope. It is a schematic block diagram which shows 5th Embodiment of a scanning microscope.

Explanation of symbols

1: first optical system 17: objective lens 19b: first light receiving element 19A: first light receiving element 2: second optical system 20: illumination light for color information (white light source)
24: second light receiving element L1: laser light L2: illumination light ro, go, bo: signal for color video c: signal for color video (composite color video signal)

Claims (4)

A first optical system for condensing laser light onto a sample by an objective lens, causing the laser light to relatively scan along the sample surface, and causing the first light receiving element to receive response light;
A second optical system that irradiates the sample with illumination light for color imaging information different from the laser light, and causes the second light receiving element to receive the response light;
A sample stage capable of moving the sample in three dimensions relative to the objective lens;
Means for storing the height of the sample stage when the first optical system is in focus;
Based on luminance information from the first light receiving element of the first optical system using the laser light and color imaging information from the second light receiving element of the second optical system using the illumination light, A scanning microscope comprising a configuration for obtaining a color video signal. The scanning microscope according to claim 1, wherein
A scanning microscope comprising: means for storing focused color imaging information from the second light receiving element at the stored height of the sample stage. The scanning microscope according to claim 1 or 2,
The second light receiving element is a color CCD, and is configured to obtain a focused color video signal using color imaging information when the first optical system is in focus for each pixel. A scanning microscope characterized by this. The scanning microscope according to claim 1 or 2,
The second light receiving element is a solid-state imaging element, and is configured to obtain a focused color video signal using color imaging information when the first optical system is in focus for each pixel. A scanning microscope characterized by that.

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