JP2003167197A - Confocal microscope - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a confocal microscope in which the efficiency of using light can be enhanced and the shorter time for photographing can be achieved. <P>SOLUTION: This confocal microscope has a lamp 1 and multislits 2 and a plurality of linear bright-lines from the multislits 2 are scanned by a galvanometer 4. The reflected light from a sample 6 is detected by a two-dimensional array photodetector 7. An image is formed by using only the data of the pixels of an illuminated region 71 in this two-dimensional array photodetector 7. As a result, the confocal image can be formed. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a confocal microscope for imaging a sample or the like.

[0002]

2. Description of the Related Art An example of a conventional confocal microscope is disclosed in, for example, Japanese Patent Application Laid-Open No. 10-104523. In the conventional confocal microscope disclosed in the publication, the light transmitted through one slit illuminates the sample, and the reflected light from the sample is a one-dimensional CCD (Charge Coupl).
ed Device) is used to receive light.

[0003]

The conventional confocal microscope has a problem in that the efficiency of use of light is low because only the light transmitted through one slit illuminates the sample. As a result, a long exposure time is required to obtain a sufficient luminance signal, which causes a problem of a long imaging time.

The present invention has been made to solve the above problems, and an object of the present invention is to provide a confocal microscope capable of improving the efficiency of use of light and shortening the imaging time. To do.

[0005]

A confocal microscope according to the present invention comprises a light source, a bright line converting means for converting light emitted from the light source into a plurality of linear bright lines, and a plurality of bright line converting means for converting the bright line converting means. Image forming means for forming an image of the bright line on a sample installed at a position conjugate with the bright line scanning means for scanning the bright line and the bright line converting means, and forming an image of reflected light or transmitted light from the sample on an image forming surface. Means, a two-dimensional array photodetector arranged on an image plane formed by the image forming means, and a pixel of the two-dimensional array photodetector in an illumination region illuminated by the bright line. It is provided with an image forming means for forming an image using only data. With such a configuration, it is possible to improve the use efficiency of illumination light and shorten the imaging time.

Here, according to a preferred embodiment, the bright line converting means is a multi-slit having a plurality of slits.

Further, the bright line converting means may be constituted by a transmissive liquid crystal panel, and the bright line scanning means may be constituted by scanning a portion which transmits light by the transmissive liquid crystal panel.

Further, the bright line converting means may be constituted by a digital micromirror device, and the bright line scanning means may be constituted by scanning the mirror on state by the digital micromirror device.

Further, the image forming means forms a confocal image forming mode in which an image is formed using only the data of the pixels of the illumination area illuminated by the bright line among the pixels of the two-dimensional array photodetector. It is preferable to provide means for illuminating the entire surface, capturing all image data, and selecting a non-confocal image forming mode for forming an image based on the image data. With such a configuration, the confocal image and the non-confocal image can be switched extremely easily.

Further, it is preferable to provide a bright line interval control means for controlling the interval between the bright lines generated in the bright line converting means. With such a configuration, ghost can be easily suppressed.

Further, it is preferable that the bright line scanning means scans the pixels of the two-dimensional array photodetector with the bright lines so that a time difference in irradiation between adjacent pixels is reduced. Thereby, a three-dimensional clear image can be obtained.

The light source may be composed of a plurality of two-dimensionally arranged laser diodes having different oscillation phases. With such a configuration, generation of speckles (small spots) can be suppressed.

Here, when the area illuminated by the bright line extends beyond a predetermined number of pixels, the image forming means performs weighting based on the brightness of illumination of those pixels when the reference sample is imaged. Moving average processing or weighted averaging processing may be performed, and correction may be performed based on the processing result when forming an image. With such a configuration, the positional deviation can be easily adjusted without adjusting the optical system. Further, the shading can be corrected by adjusting the weighting coefficient so that the brightness of the screen becomes constant.

Further, when the two-dimensional array photodetector is composed of a color image sensor and a reference sample is imaged, the position of the illumination area is determined based on the illuminance detected by the pixel of a specific color of the color image sensor. A position shift detection unit that detects a shift may be further provided. With such a configuration, the positional deviation can be easily detected. Further, the optical system may be provided with a magnification adjusting function and an optical axis adjusting function in order to adjust an interval between a plurality of bright lines and an interval between pixels of the two-dimensional array photodetector. This allows for easy adjustment.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described below with reference to a plurality of embodiments of the invention.

First Embodiment of the Invention FIG. 1 shows a configuration example of the confocal microscope according to the first embodiment. As shown in the figure, the confocal microscope includes a lamp 1, a multi-slit 2, objective lenses 31, 32, 33,
34, 35, galvanometer 4, beam splitter 5,
The two-dimensional array photodetector 7 is provided and the sample 6 is imaged.
Although the configuration is simplified in this example for easy understanding, it goes without saying that another configuration may be provided.

The lamp 1 is, for example, a white light source, a fluorescence excitation light source, or the like, and more specifically, various light sources such as a mercury lamp and a halogen lamp can be used. Further, a laser diode having different oscillation phases may be two-dimensionally arranged to form a light source. By oscillating laser diodes having different oscillation phases, it is possible to suppress the generation of speckles (small spots).

The multi-slit 2 is provided in the vicinity of the lamp 1 and has a plurality of slits for transmitting the illumination light emitted from the lamp 1. A specific configuration example is shown in FIG. In the figure, 21 is a slit, and 22 is a light-shielding portion that blocks the transmission of illumination light. The plurality of slits 21 are openings provided in parallel along the entire side in the H direction of the multi-slit 2. Regarding the interval of the slits 21, when the reflected light of the sample 6 is incident on the two-dimensional array photodetector 7, the reflected lights corresponding to the respective slits 21 do not interfere with each other, or are equal to or less than a predetermined value such that the interference does not cause a problem. Will be determined.

The lens 31 is provided at a position where the light transmitted through the multi-slit 2, that is, the bright line is incident.

The galvanometer 4 has a function of reflecting incident light so that the light emitted from the lamp 1 and transmitted through the multi-slit 2 scans the sample 7. The galvanometer 4 includes at least a mirror, an actuator that changes the angle of the mirror, and a control unit that controls the operation of the actuator, and realizes scanning by changing the angle of the mirror.

The relay lenses 32 and 33 relay the pupil of the objective lens 34 to the galvanometer 4.

The beam splitter 5 includes a relay lens 33.
Is provided at a position below, and at which light emitted from the objective lens 33 is incident.

The objective lens 34 is provided between the beam splitter 5 and the sample 6, and is provided at a position where the light transmitted through the beam splitter 5 is incident and a position where the light reflected from the sample 6 is incident. There is.

The imaging lens 35 is provided at a position where the reflected light from the sample 6 reflected by the beam splitter 5 is incident.

The sample 6 is arranged at a position conjugate with the multi-slit 2.

The two-dimensional array photodetector 7 is a two-dimensional multi-photodetector array, and is, for example, an area CCD. An example of the light receiving surface of the two-dimensional array photodetector 7 is shown in FIG. As described above, a large number of light receiving elements are arranged on the light receiving surface. Although it is simplified in this figure, actually, for example, a light receiving element of 1.5 million pixels is arranged. In the two-dimensional array photodetector 7 shown in FIG. 3, light is illuminated on a region 71 (white portion in the figure) at a position corresponding to the slit 21 provided in the multi-slit 2. This illumination area 71 is
It moves in the V direction according to the operation of the galvanometer 4 and is scanned by an interval corresponding to the slit 21. This area 71 is a portion conjugate with the illumination area.

In the present invention, only the data of the pixels in the illuminated area 71 illuminated by the illumination light is read, and the data of the pixels in the non-illuminated area 72 not illuminated by the illumination light is discarded and not used. By doing so, a confocal image can be acquired. When all the data of the pixels on the two-dimensional array photodetector 7 is completed by scanning the illumination area 71, these data are combined by an image forming means (not shown) to form a two-dimensional image. To do. This two-dimensional image is stored in a predetermined storage means and is displayed on the display.

Here, FIG. 4 shows a principle configuration diagram of the two-dimensional array photodetector 7. This two-dimensional array photodetector 7 is
This is an interline CCD solid-state imaging device. As shown in the figure, the two-dimensional array photodetector 7 includes a plurality of light receiving portions 701 which are pixels arranged in a horizontal and vertical direction at a predetermined pitch, and a vertical direction provided on one side of the light receiving portions 701 in each column. A vertical transfer register 702 having a CCD structure and a horizontal transfer register 703 having a CCD structure provided at one end of each vertical transfer register 702. Then, the signal charges generated in each light receiving unit 701 according to the amount of received light are transferred to the corresponding vertical transfer registers 702, and the signal charges of each vertical transfer register 702 are transferred to the horizontal transfer register 703.
And the signal charges for each horizontal line are read out.

In particular, in the present invention, the data is read only in the illuminated area 71 and the data in the non-illuminated area 72 is discarded.
Such processing is executed by electrical processing as described below, for example. The data of the pixels located in the non-illuminated area 72 as well as the data of the pixels located in the illuminated area 71 are once sent to the vertical transfer register 702. The data on the vertical transfer register 702 is sent to the horizontal transfer register 703 pixel by pixel. At the time of sending out, the pixel data of the non-illuminated area 72 is stored in the horizontal transfer register 703 and is not output,
Only the pixel data of the illumination area 71 is output. In addition, a gate circuit may be added to the output data of the two-dimensional array photodetector 7 to limit the output of the data of the pixels in the non-illumination area 72 and output only the data of the pixels in the illumination area 71. Good. Furthermore, the data may be temporarily stored in the frame memory and only the data of the illumination area 71 may be addressed and read. In addition, a CID (Charge Inj) having a circuit for controlling the readout of the two-dimensional array photodetector 7 for each pixel
section device), the output from the pixels in the non-illumination area 72 to the vertical transfer register 702 may be stopped, and only the pixels in the illumination area 71 may be output to the vertical transfer register 702.

Next, the image pickup operation of the confocal microscope according to the first embodiment will be described.

First, the light emitted from the lamp 1 enters the multi-slit 2. Multi slit 2 is slit 2
Since it has 1 and the non-transmissive portion 22, the incident light passes only through the slit 21. The light that has passed through the multi-slit 2, that is, the line-shaped bright line is refracted by the lens 31, and enters the galvanometer 4. The galvanometer 4 sets its angle according to the control signal and reflects the incident light. The light reflected by the galvanometer 5 is refracted by the relay lens 32. The light refracted by the relay lens 32 is refracted by the relay lens 33, then passes through the beam splitter 5, and is imaged on the sample 6 by the objective lens 34.

The reflected light of the sample 6 is again reflected by the objective lens 34.
After being refracted by, the light is reflected by the beam splitter 5. The light reflected by the beam splitter 5 is formed by the imaging lens 3
After refracting by 5, the image is formed on the two-dimensional array photodetector 7. The two-dimensional array photodetector 7 is illuminated only with the light passing through the position of the slit 21 of the multi-slit 2. The two-dimensional array photodetector 7 reads data only for pixels in the illuminated area 71 and discards data for pixels in the non-illuminated area 72. Subsequently, the galvanometer 4 is scanned, and similarly, only the data of the pixels in the illumination area 71 is read by the two-dimensional array photodetector 7. Such processing is repeated until the entire area of the sample 6 is illuminated and imaged. Then, the data output from the two-dimensional array photodetector 7 is combined to obtain 2
Form a three-dimensional image. This two-dimensional image is stored in a predetermined storage means and is displayed on the display.

As described above, according to the confocal microscope of the first embodiment, since the utilization efficiency of the illumination light is high, the illumination with high illuminance can be performed, and the S
A confocal image with a high N ratio can be captured. Moreover, since the scan angle of the galvanometer is small, a small galvanometer can be used.

On the other hand, in the conventional method using the one-dimensional CCD, the position accuracy in the V direction is determined by the angle setting accuracy of the galvanometer, whereas in this method, the position accuracy of each light receiving element on the light receiving element array is almost the same. Since it is determined, the position accuracy of the captured image or the dimension measurement accuracy is high.

The lamp 1 is illuminated with a white light source,
If a color CCD is used as the two-dimensional array photodetector 7, a color image can be captured. At this time, since RGB are arranged in two rows on the color CCD, it is preferable that the illumination area 71 is arranged in two or more rows. In particular, when the illumination is not scanned over the entire area of the sample 6, that is, when a rough image is output, there is an advantage that the image processing can be easily performed by doing this.

Further, not only the data obtained from the pixels in the illuminated area 71, but also the data obtained from the pixels in the non-illuminated area 72 which are not conjugate, integrated, and synthesized, a non-confocal image can be obtained. Specifically, of the pixels of the two-dimensional array photodetector 7, both the confocal image forming mode in which an image is formed using only the data of the pixels in the illumination area 71 and the data of the pixels in the non-illumination area 72 are used. A means for selecting a non-confocal image forming mode for forming an image based on the above is provided. By controlling the reading of data in the two-dimensional array photodetector 7, it becomes possible to switch between the confocal image and the non-confocal image extremely easily and instantaneously. When obtaining a non-confocal image, the multi-slit 2 may be removed from the optical path so that the light is irradiated to the entire area of the sample.

Embodiment 2 of the Invention In the confocal microscope according to the second embodiment, a white light source is used as a light source, a switchable color filter is used on the illumination side, and RGB are sequentially switched to illuminate and an image of each color is captured to obtain a color. The method of composing images is adopted.

FIG. 5 shows the structure of the confocal microscope. As shown in the figure, a color filter 8 and a motor 9 for rotating the color filter 8 are provided between the white light source 1 and the multi-slit 2. The color filter 8 has a transmissive area 81, a red colored area 82, a green colored area 83, and a blue colored area 84, as shown in the figure.
When the color filter 8 is rotated by the motor 9, RGB is sequentially switched. Then, a color image can be obtained by appropriately reading the image of each color by the two-dimensional array photodetector 7. Other configurations are basically the same as those of the confocal microscope according to the first embodiment of the present invention, and therefore description thereof will be omitted.

Third Embodiment of the Invention In the confocal microscope according to the third embodiment, the transmissive liquid crystal panel 10 is used instead of the multi-slit 2.

FIG. 6 shows the structure of the confocal microscope. The transmissive liquid crystal panel 10 is provided with a plurality of slits 21 serving as transmissive regions, similarly to the multi-slit 2 shown in FIG. In the first embodiment of the invention, the galvanometer 4
However, in the third embodiment, scanning can be performed by sequentially moving the transmissive area of the transmissive liquid crystal panel 10. Therefore, it is not necessary to provide the galvanometer 4 as the scanning means. For other configurations,
The description is omitted because it is basically the same as the confocal microscope according to the first embodiment of the invention.

Fourth Embodiment of the Invention In the confocal microscope according to the fourth embodiment, the digital micromirror device 11 is used instead of the multi-slit 2.

FIG. 7 shows the configuration of the confocal microscope. The digital micromirror device 11 is also called DMD (registered trademark), has a large number of minute mirrors, and is configured to be able to tilt each mirror by approximately ± 10 degrees. When each mirror is tilted +10 degrees, it is turned on, and when it is tilted -10 degrees, it is turned off. The mirror tilts electronically at a rate of more than a thousand times per second.

This digital micromirror device 11 is
The part corresponding to the slit 21 of the multi-slit 2 shown in FIG. 2 is controlled to be in the ON state. In the first embodiment of the invention, scanning is performed by the galvanometer 4, but in the fourth embodiment, scanning can be performed by turning on the digital micromirror device 11, that is, by sequentially shifting the reflection area. Therefore, it is not necessary to provide the galvanometer 4 as the scanning means. For other configurations,
The description is omitted because it is basically the same as the confocal microscope according to the first embodiment of the invention.

In the confocal microscope having such a structure, the light emitted from the lamp 1 enters the digital micromirror device 11. Then, in the digital micromirror device 11, since the mirror of the portion corresponding to the slit 21 is in the on state, only the mirror in the on state reflects the light emitted from the lamp 1 toward the objective lens 33. The reflected light is applied to the sample 6 via the relay lens 33 and the objective lens 34, and then is imaged on the light receiving surface of the two-dimensional array photodetector 7 via the objective lens 34, the beam splitter 5, and the imaging lens 35. . After that, the digital micromirror device 11 sequentially shifts the mirrors to be turned on. When the irradiation of the entire area of the sample 6 is completed in this way, the image data is combined and output to a display (not shown).

Fifth Embodiment of the Invention Next, a case where a color CCD is used for the two-dimensional array photodetector 7 will be described. In this example, the two rows of pixels of the color CCD are controlled to be simultaneously irradiated with light. FIG. 8 shows the intensity distribution of illumination light and the point image intensity distribution on the light receiving surface. As shown in the figure, the intensity distribution of the illumination light is a superposition of the point image intensity distributions. Further, when the illumination light intensity distribution between the adjacent illumination areas is seen, it is found that the pixels in the illumination area are irradiated with light of sufficient intensity.

Subsequently, movement control of the illumination area on the color CCD will be described with reference to FIGS. 9, 10, 11 and 12. The illumination area shown in FIG. 9 is sequentially shown in FIGS.
1, FIG. 12 and the illumination area has moved. Focusing on the illumination area located at the top of the drawing, in FIG.
The pixels in the second and second columns are illuminated. Next, FIG.
In, the pixels in the fifth and sixth columns are illuminated. Further, in FIG. 11, the pixels in the third and fourth columns are illuminated. Finally, in FIG. 12, the pixels in the seventh and eighth columns are illuminated. That is, the illumination area is shifted as follows. 1st and 2nd row → 5th and 6th row → 3rd and 4th row →
7th and 8th rows

When the illumination area is shifted in this manner, the time difference between adjacent pixels is reduced as compared with the case where the first and second columns are shifted by two columns, so that a clear 3
There is an effect that a three-dimensional image can be obtained.

In order to reduce a ghost when capturing an image of a sample having a high contrast, it is advisable to sufficiently separate the illumination areas as shown in FIGS. 13 and 14. When the illumination areas are close to each other, light from adjacent illuminations enters and a ghost occurs. Therefore, in such a case, it is possible to suppress the occurrence of the ghost by widening the interval between the illumination areas. For example, one illumination area for 4 lines is 8
Change to one illuminated area on the line. Particularly, when the digital micromirror device 11 is used, such control can be easily performed.

When the color CCD is used as described above, the positional deviation between the illumination area and the pixel is based on the data of the pixel of the specific color obtained by photographing the reference sample, that is, the data of the illuminance. Can be detected. For example, the color CCD shown in FIG. 9 has a configuration in which the green (G) pixel data output from the respective illumination areas are equal, but if they are not equal, such a positional deviation has occurred. You can judge. On the contrary, if the position of the illumination area is adjusted so that the data of the pixels of the specific color output from each illumination area become equal, the positional deviation can be eliminated.

In such a color CCD,
It is also possible to adjust the white balance of RGB by changing the positional relationship between the illumination light and the imaging element and changing the position of the illumination area.

Sixth Embodiment of the Invention Next, a specific configuration example of the confocal microscope will be described with reference to FIG. In this example, the mercury lamp 1 is provided as the illumination means. Then, the light emitted from the mercury lamp 1 is
It enters the cone lens 12. The cone lens 12 refracts the incident light so as to prevent the hollow portion of the far field pattern. The light refracted by the cone lens 12 enters the incident end of the bundle fiber 13. Then, light is emitted from the emission end of the handle fiber 13 and reflected by the mirror 14 at a predetermined angle. The light reflected by the mirror 14 enters the objective lens 32.

The light that has entered the objective lens 32 is tropic,
The light enters the digital micromirror device 11. At this time, the digital micromirror device 11 is controlled so that the mirror at a place corresponding to the slit 21 of the multi-slit 2 as shown in FIG. 2 is turned on. The digital micromirror device 11 emits the incident light to the lens 33 by the mirror in the ON state. On the other hand, the mirror in the off state in the digital micromirror device 11 reflects the incident light to the optical trap 15. The light emitted from the digital micromirror device 11 becomes equivalent to the light transmitted through the multi-slit 2 as shown in FIG. The emitted light is refracted by the lens 33 and passes through the beam splitter 5. The light transmitted through the beam splitter 5 is refracted by the objective lens 34 and condensed on the sample 6.

The reflected light of the sample 6 is refracted again by the objective lens 34 and enters the beam splitter 5. And
The light that has entered the beam splitter 5 is reflected by the beam splitter 5 and enters the imaging lens 35. The light that has entered the imaging lens 35 is refracted and enters the color CCD 7. The color CCD 7 is illuminated only with the light reflected by the mirrors of the digital micromirror device 11 in the ON state. The color CCD 7 reads the data of only the pixels in the illuminated area 71 and discards the data of the pixels in the non-illuminated area 72. Then, the reflected light is sequentially scanned by the digital micromirror device 11, and the entire predetermined region of the sample 6 is imaged. A two-dimensional image is formed by combining the data output from the color CCD 7. This two-dimensional image is stored in a predetermined storage means and is displayed on the display.

As described above, according to the confocal microscope of the sixth embodiment, since the utilization efficiency of the illumination light is high, the illumination with high illuminance can be performed, and the S
A confocal image with a high N ratio can be captured.

Seventh Embodiment of the Invention The configuration of the confocal microscope according to the seventh embodiment is shown in FIG. As shown in the figure, this confocal microscope is configured by combining a personal computer 200 and a projector 100. Personal computer 20
The S-VGA line of the 0 video board 201 is connected to the projector 100. The RS422 line of the frame grabber is connected to the CCD driver 73. The line of the DAC is connected to the fine movement stage driver 18. Therefore, the digital micromirror device 11 can be controlled by the output signal from the video board 201 of the personal computer 200. Further, the CCD driver 73 can be controlled by the output signal from the frame grabber 202. Then, the fine movement stage driver 18 can be controlled by the output signal from the DAC. The fine movement stage driver 18 can drive the operation of the fine movement stage 17. The personal computer 200 stores a control program for controlling the confocal microscope in a memory. This control program includes, for example, multi-line illumination, multi-line image acquisition, alignment spatial filter function creation, alignment / spatial filtering processing, non-interlaced image formation, Z
Includes control programs for axis drive, afocal depth imaging, surface topography imaging, and color image capture / display.

The projector 100 includes a mercury lamp 1, a color filter 8, a motor 9, an objective lens 31, and a digital micromirror device 11.

The imaging operation of the confocal microscope having such a configuration will be briefly described. The illumination light of the mercury lamp 1 is colored by the color filter 8 and then enters the digital micromirror device 11 by the mirror 14. At this time, the digital micromirror device 11 is controlled so that the mirror at a place corresponding to the slit 21 of the multi-slit 2 as shown in FIG. 2 is turned on. The digital micromirror device 11 emits the incident light to the objective lens 32 installed outside the projector 100 by the mirror in the ON state. On the other hand, the mirror in the off state in the digital micromirror device 11 reflects incident light to an optical trap (not shown). The light emitted from the digital micromirror device 11 is as shown in FIG.
The light becomes equivalent to the light transmitted through the multi-slit 2 as shown in FIG.

The light incident on the lens 32 is refracted and enters the beam splitter 5. Also, the beam splitter 5
A part of the light incident on is transmitted through the beam splitter 5, refracted by the objective lens 33, and then focused on the sample on the XYZ stage 16. The light reflected from the sample is refracted by the objective lens 33 and reflected by the beam splitter 5. This reflected light is refracted by the imaging lens 35 and is condensed on the CCD 7. The CCD 7 reads only the data of the pixels in the illuminated area 71 illuminated by the illumination light, discards the data of the pixels in the non-illuminated area 72 not illuminated by the illumination light, and does not use them. By doing so, a confocal image can be acquired.

Further, the digital micromirror device 11 is controlled to scan the illumination area 71. When the acquisition of all the data of the pixels on the two-dimensional array photodetector 7 is completed, these data are combined to form a two-dimensional image. This two-dimensional image is stored in a predetermined storage means and is displayed on the display.

Further, the confocal microscope according to the seventh embodiment includes the digital micromirror device 11 and C.
It has a function of correcting the positional deviation of the pixels of the CD 7.
The positional deviation correction process may be performed when the lens is replaced, or may be performed each time a new image is captured. This function will be described in detail below.

FIG. 17 is a flow chart showing a process of creating an alignment spatial filter function necessary for realizing the position shift correction function. First, the reference sample is attached on the XYZ stage 16 of the confocal microscope (S101). The reference sample is a reflector that reflects incident light uniformly and uniformly, and is, for example, a mirror or a silicon wafer. Next, the digital micromirror device 11 of the projector 100 is controlled to perform multi-line illumination (S102). Then, the reflected light from the reference sample corresponding to the multi-line illumination is read by the CCD 7 and the multi-line image is captured (S10).
3). An alignment spatial filter function is created based on the last-acquired multiline image (S104).

Further, the creation of the alignment spatial filter function will be described with reference to FIG. In FIG. 18, the point image intensity distribution on the light receiving surface is shown together with the pixels of the CCD 7. Normally, a digital micromirror device 1
By controlling 1, the light emitted to the CCD 7 is
The optical system is designed so that a plurality of line-shaped bright lines are formed and the illumination area 71 formed by the bright lines coincides with one row of pixels of the CCD 7. However, in the vicinity of the edge of the CCD 7, the illumination area 71 may not coincide with one row of pixels of the CCD 7. The white part at the bottom of FIG. 17 is
The case where the illumination area 71 is aligned with one row of pixels of the CCD 7 is shown. In this case, a confocal image can be finally obtained by reading the data only from the pixels in which the illumination areas 71 match.

In the upper white portion in FIG. 17, the CCD 7
The illumination area extends over two columns of pixels. in this case,
Data is read from these two pixel columns and a coefficient for weighting each luminance is determined. In the example shown in this figure, since the light line is located at a substantially intermediate position between the two columns of pixels of the CCD 7, the data of the pixels in the two columns are weighted equally. And
In consideration of such weighting coefficient, the luminance is calculated by performing the weighted moving average processing or the weighted average processing.

In order to perform such processing, when the reference plane is observed and the illumination in the illumination area 71 covers not only one pixel column but also a plurality of pixel columns, the center of gravity of the local luminance in the V direction is set. Ask. Then, by weighted moving average processing or weighted averaging processing for two pixel rows close to the center of gravity,
The displacement of the pixel between the digital micromirror device 11 and the CCD 7 is corrected. When the number of pixels in the V direction is processed by the number of pixels of the CCD 7, weighted moving average processing is performed, and when the number of pixels of the digital micromirror device 11 is matched, weighted average processing is performed. The luminance is set to 0 for the pixels other than the illumination area 71.

If weighted moving average processing is performed for the n-th and n + 1-th pixel columns, the luminance ln
Can be expressed as: ln = aln + bl (n + 1) where a + b = 1.

On the other hand, when the weighted averaging process is performed, the brightness ln can be expressed as follows. ln = al2n + bl (2n + 1) where a + b = 1.

Further, the illumination area 71 may not be uniformly displaced in the pixel row in the V direction, but may be obliquely displaced due to distortion of the optical system. Also in this case, the gravity center of brightness is obtained for each pixel, and the weighted moving average processing or the weighted average processing is performed. For this reason, for example, processing is performed on different pixel rows, such as processing on the n-th and n + 1-th pixel rows at one pixel and processing on the n-th and n-1 th pixel rows at other pixels. Will be done.

Further, in order to correct the overall variation in luminance called shading, the coefficient by which the luminance is multiplied may be changed for each pixel column. In particular,
In the above formula of the luminance ln, a + b is not set to 1 and is set to an arbitrary value of a + b> 1 in a dark pixel column of small luminance,
For a bright and high-luminance pixel column, an arbitrary value of a + b <1 is set.

In the confocal microscope according to this embodiment, the illumination area 71 is aligned with one row of pixels of the CCD 7. However, the present invention is not limited to this, and the same applies when a plurality of rows are aligned. It is possible to correct the positional deviation. For example, in the case of matching in two columns, the center of gravity of the local luminance in the V direction is calculated, and the center of gravity is 3
A weighted moving average process or a weighted average process is performed on a column.

After the correction processing is performed in this way, an image of the sample 6 is photographed according to the obtained alignment spatial filter function.

Eighth Embodiment of the Invention The confocal microscope according to the eighth embodiment has a configuration particularly suitable for fluorescence observation.

First, when observing fluorescence, a CCD with an image intensifier may be used as the image pickup device. Further, the beam splitter is a dichroic mirror, which transmits excitation light having a short wavelength and reflects fluorescent light having a wavelength longer than that of the excitation light so as to enter the image intensifier. As a result, the loss of the amount of light in the beam splitter can be minimized.

Furthermore, as shown in FIG. 19, a switchable dichroic filter 8 (color filter) may be arranged in front of the CCD camera 7 in order to obtain a color fluorescence image. Here, as the excitation light source 1, for the mercury lamp, it is preferable to use a laser diode or the like having an oscillation wavelength of 405 nm.

Ninth Embodiment of the Invention The confocal microscope according to the ninth embodiment has a function of pixel misalignment. FIG. 20 shows a configuration example of the confocal microscope.

In order to adjust the pixel shift, either the illumination side lens or the image formation side lens is used as a magnification adjustment lens in order to match the pitch of the illumination slit image and the light receiving element. In this example, magnification adjustment lenses 361 and 362 are provided.

Further, in order to adjust the positional deviation between the illumination slit image and the light receiving element, an optical axis adjusting mechanism (beam positioner) is provided on either the illumination side or the image forming side optical axis. In this example, beam positioners 161 and 1
62 is provided.

Tenth Embodiment of the Invention The tenth embodiment
The confocal microscope according to the present invention has a particularly high light utilization efficiency. FIG. 21 shows a configuration example of the confocal microscope.

A polarized beam splitter can be used as the beam splitter 52. In this case, the quarter wavelength plate 17 is arranged on the objective lens 34. This will minimize the loss of light. That is, in the case of a beam splitter using a metal thin film, the transmittance is about 30% and the reflectance is about 30%, so that only 9% of light can be used for round trip. On the other hand, when the polarization beam splitter 52 is used, only the P-polarized component of the light from the lamp 1 is transmitted, and the quarter-wave plate 17 becomes circularly polarized light, which is incident on the objective lens 34. The reflected light from the sample 6 is transmitted through the quarter-wave plate 17 from the opposite direction again to S
Since it becomes polarized light, it is reflected by the polarization beam splitter 52 toward the imaging lens 35 side. Therefore, about 50% of the light can be used.

Other Embodiments. In the above-mentioned example, the reflection microscope is taken as an example, but the present invention can be applied to a transmission microscope.

[0080]

According to the present invention, the use efficiency of light is increased,
It is possible to provide a confocal microscope capable of achieving reduction in imaging time.

[Brief description of drawings]

FIG. 1 is a diagram showing a configuration of a confocal microscope according to a first embodiment of the present invention.

FIG. 2 is a diagram showing a configuration of a multi-slit according to the present invention.

FIG. 3 is a diagram showing a light receiving surface of a two-dimensional array photodetector according to the present invention.

FIG. 4 is a block diagram of a two-dimensional array photodetector according to the present invention.

FIG. 5 is a diagram showing a configuration of a confocal microscope according to a second embodiment of the present invention.

FIG. 6 is a diagram showing a configuration of a confocal microscope according to a third embodiment of the present invention.

FIG. 7 is a diagram showing a configuration of a confocal microscope according to a fourth embodiment of the present invention.

FIG. 8 is a diagram showing an intensity distribution of illumination light and a point image intensity distribution on a light receiving surface according to a fifth embodiment of the present invention.

FIG. 9 is a diagram for explaining movement control of an illumination area on a color CCD according to a fifth embodiment of the present invention.

FIG. 10 is a color CCD according to a fifth embodiment of the present invention.
It is a figure for demonstrating movement control of an upper illumination area.

FIG. 11 is a color CCD according to a fifth embodiment of the present invention.
It is a figure for demonstrating movement control of an upper illumination area.

FIG. 12 is a color CCD according to a fifth embodiment of the present invention.
It is a figure for demonstrating movement control of an upper illumination area.

FIG. 13 is a color CCD according to a fifth embodiment of the present invention.
It is a figure for demonstrating movement control of an upper illumination area.

FIG. 14 is a color CCD according to a fifth embodiment of the present invention.
It is a figure for demonstrating movement control of an upper illumination area.

FIG. 15 is a diagram showing a configuration of a confocal microscope according to a sixth embodiment of the present invention.

FIG. 16 is a diagram showing a configuration of a confocal microscope according to a seventh embodiment of the present invention.

FIG. 17 is a flowchart showing a process of creating an alignment spatial filter function according to the seventh embodiment of the present invention.

FIG. 18 is a diagram showing an intensity distribution of illumination light on a light receiving surface in the seventh embodiment of the present invention.

FIG. 19 is a diagram showing a configuration of a confocal microscope according to an eighth embodiment of the present invention.

FIG. 20 is a diagram showing a configuration of a confocal microscope according to a ninth embodiment of the present invention.

FIG. 21 is a diagram showing a configuration of a confocal microscope according to a tenth embodiment of the present invention.

[Explanation of symbols]

1 Lamp 2 Multi-slit 3 Objective lens 4
Galvanometer 5 Beam splitter 7 Two-dimensional array photodetector 8
Color filter 10 Transmission type liquid crystal panel 11 Digital micro mirror device

─────────────────────────────────────────────────── ─── Continued Front Page (51) Int.Cl. ⁷ Identification Code FI Theme Coat (Reference) H04N 9/04 H04N 9/04 B

Claims (11)

[Claims] 1. A light source, bright line conversion means for converting light emitted from the light source into a plurality of line-shaped bright lines, bright line scanning means for scanning a plurality of bright lines converted by the bright line conversion means, and the bright line conversion means. An image forming unit that forms an image of the bright line on a sample installed at a position conjugate with the image forming unit and forms an image of reflected light or transmitted light from the sample on an image forming surface; A two-dimensional array photodetector arranged on the image plane, and an image forming unit for forming an image using only data of pixels of an illumination region illuminated by the bright line among pixels of the two-dimensional array photodetector. A confocal microscope equipped with. 2. The confocal microscope according to claim 1, wherein the bright line converting means is a multi-slit having a plurality of slits. 3. The bright line converting means is constituted by a transmission type liquid crystal panel, and the bright line scanning means is also constituted by scanning a portion for transmitting light by the transmission type liquid crystal panel. Item 1. The confocal microscope according to item 1. 4. The bright line converting means is constituted by a digital micromirror device, and the bright line scanning means is also constituted by scanning the ON state of a mirror by the digital micromirror device. The confocal microscope according to 1. 5. A confocal image forming mode in which the image forming means forms an image using only data of pixels in an illumination region illuminated by the bright line among pixels of the two-dimensional array photodetector. , Illuminating the entire surface with the bright line and capturing all image data of the illuminated area,
The confocal microscope according to claim 1, further comprising means for selecting a non-confocal image forming mode for forming an image based on the captured image data. 6. The confocal microscope according to claim 1, further comprising bright line spacing control means for controlling a spacing between bright lines generated in the bright line converting means. 7. The bright line scanning means scans the pixels of the two-dimensional array photodetector with bright lines so that the time difference of irradiation between adjacent pixels is reduced. Confocal microscope. 8. The confocal microscope according to claim 1, wherein the light source is composed of a plurality of two-dimensionally arranged laser diodes having different oscillation phases. 9. The image forming means, when the illumination area by the bright line extends over a predetermined pixel or more, weighted movement based on the luminance of illumination for those pixels when the reference sample is imaged. The confocal microscope according to claim 1, wherein an averaging process or a weighted averaging process is performed, and correction is performed based on the processing result at the time of image formation. 10. The position of an illumination area based on the illuminance detected by a pixel of a specific color of the color image sensor when the two-dimensional array photodetector is composed of a color image sensor and a reference sample is imaged. The confocal microscope according to claim 1, further comprising a position shift detection unit that detects a shift. 11. The optical system is provided with a magnification adjusting function and an optical axis adjusting function for adjusting an interval between a plurality of bright lines and an interval between pixels of the two-dimensional array photodetector. Confocal microscope.