JP2001083427A - Confocal microscope - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a confocal microscope constituted so that the brightness of an image can be easily adjusted. SOLUTION: This confocal microscope is provided with an optical system guiding light to a sample, a rotary body 4' having a semitransmission part where a part through which the light guided by the optical system is transmitted, a part shielding the light and a transmission part through which the light is transmitted, a driving means 33 rotating the rotary body 4', an image pickup means 9 picking up a composite image consisting of the confocal image and the non-confocal image of the sample obtained through the semi-transmission part of the rotary body 4', the bright visual field image of the sample obtained through the transmission part,, and an arithmetic means 10 obtaining the confocal image of the sample from the picked-up composite image and the picked-up bright visual field image. Besides it is provided with a control means 10 controlling the driving means 33 so as to change the rotational speed the rotary body 4' and also the exposure time of the image pickup means 9 according to the change of rotational speed of the rotary body 4'.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a confocal microscope which is most suitable for observing and measuring the shape of a minute structure or a three-dimensional structure of a sample, for example.

[0002]

2. Description of the Related Art A typical conventional confocal microscope is a disk scanning type using a Nippow disk in which a large number of pinholes are spirally arranged at intervals of about 10 times the diameter of the pinholes. As an improved technique of a confocal microscope using this disc, for example, R.S. Juskait
is, T .; Wilson et al., “Efficient r.
Eal-time Confocal microsc
opy with white light sour
ces ", Nature, Vol. 383 Oct.
1996 p804-806. There is a technique described in.

[0003] FIG. FIG. 1 is a diagram showing a configuration of a disk scanning confocal microscope by Wilson et al. As the light source 1, a halogen light source, a mercury light source, or the like is used.
An optical lens 2 and a PBS (polarizing beam splitter) 3 are arranged on an optical path of light emitted from the light source 1.
A sample (specimen) 6 is disposed on the reflection optical path of the PBS 3 via a rotating disk 4, a first imaging lens 23, a 波長 wavelength plate 24, and an objective lens 5, which will be described later.

[0004] The rotating disk 4 is a random pinhole disk shown in FIG. The disk has a random pinhole pattern portion 4a in which a large number of pinholes are randomly arranged at substantially the same interval as the pinhole diameter,
An opening 4b through which light can freely pass is formed, and light blocking portions 4c and 4d for blocking light are formed between the random pinhole pattern portion 4a and the opening 4b.

The rotating disk 4 is formed of a transparent glass disk, and a low reflection film (for example, chromium oxide) such as a Cr film is deposited on portions corresponding to the light shielding portions 4c and 4d on the disk. I have. Similarly, a low-reflection film such as a Cr film is deposited on portions other than the pinholes in the random pinhole pattern portion 4a. As a result, light is shielded between the light shielding portions 4c and 4d and portions other than the pinholes on the random pinhole pattern portion 4a. The rotary disk 4 is connected to a shaft of a DC motor (not shown) via a rotary shaft 7 and rotates at a constant rotation speed.

Further, as the rotating disk 4, a line pattern disk shown in FIG. 8 can be used. In this disk, instead of the random pinhole pattern portion 4a shown in FIG. 7, a striped line pattern portion 4e in which a plurality of lines are arranged at substantially equal intervals on transparent glass is formed. Then, for example, a Cr film is vapor-deposited on each line in the line pattern section 4e, and light is shielded on each line. That is, the line pattern portion 4
e has a linear light shielding portion and a light transmitting portion alternately.

On the other hand, a CCD camera 9 is arranged on the transmission optical path of the PBS 3 via a second imaging lens 8. A computer 10 is connected to an image output terminal of the CCD camera 9, and a confocal image is displayed on a monitor 11 by image processing in the computer 10.

With such a configuration, light emitted from the light source 1 is guided to the PBS 3 via the optical lens 2. The light reflected by the PBS 3 is incident on a rotating disk 4 rotating at a constant speed. The light passing through the random pinhole pattern portion 4a (or the line pattern portion 4e) or the opening portion 4b of the rotating disk 4 passes through the first imaging lens 23 and becomes circularly polarized by the quarter-wave plate 24.
An image is formed by the objective lens 5 and is incident on the sample 6.

The light reflected from the sample 6 passes through the objective lens 5 and becomes polarized light orthogonal to the incident light by the quarter-wave plate 24, passes through the first imaging lens 23, and returns to the rotating disk 4. Pass through the random pinhole pattern portion 4a (or line pattern portion 4e) or the opening 4b, further pass through the PBS 3, and enter the CCD camera 9 via the imaging lens 8.

The CCD camera 9 has an image pickup timing controlled in synchronization with the rotation speed of the rotary disk 4, and an image (composite image) passing through the random pinhole pattern section 4a (or line pattern section 4e) and the opening section 4b. Are captured (bright-field images). The output image of the CCD camera 9 is stored in the computer 10. The computer 10 controls the confocal image data including the non-confocal component obtained from the random pinhole pattern portion 4a (or the line pattern portion 4e) and the aperture 4b.
The confocal image data of only the confocal component is obtained by performing a difference operation with the non-confocal image data obtained from the above, and the confocal image is displayed on the monitor 11 based on the confocal image data.

The three-dimensional image in the vicinity of the surface of the sample 6 is moved in the vertical direction (a) together with the horizontal movement stage by a piezo element attached to a horizontal movement stage (not shown), so that The images are obtained by synthesizing the images with the computer 10.

Further, in the Nippow disk type confocal microscope, the reflected light from the sample that can be used with respect to the incident light from the light source is 0.5 to 1%. Wi
With an lson-type confocal microscope, the reflected light available for incident light is 25 to 50%, and it is reported that a brighter image can be obtained.

[0013]

SUMMARY OF THE INVENTION Wils
In the on-type disk scanning confocal microscope, the CCD camera 9 needs to be exposed while the random pinhole pattern portion 4a (or line pattern portion 4e) and the opening 4b of the rotating disk 4 pass through the optical path, respectively. Therefore, it is not possible to change the brightness of the image simply by changing the exposure time of the CCD camera 9 alone.

Usually, in the visible light observation, the brightness can be adjusted by adjusting the voltage of the lamp of the halogen light source. In the ultraviolet light observation, the brightness is adjusted by the lamp voltage because a mercury light source is used. Can not do.
In the case of ultraviolet light observation, it is conceivable to adjust the brightness using an ND filter in the ultraviolet region.
The expensive filters increase the cost of manufacturing the microscope.

An object of the present invention is to provide a confocal microscope capable of easily adjusting the brightness of an image.

Another object of the present invention is to make it possible to easily adjust the brightness of an image regardless of whether the observation method is confocal observation or non-confocal observation, and to improve the operability of the brightness adjustment. It is to provide a confocal microscope.

[0017]

Means for Solving the Problems In order to solve the above problems and achieve the object, a confocal microscope according to the present invention is configured as follows.

(1) The confocal microscope according to the present invention comprises: an optical system for guiding light to a sample; a semi-transmissive portion in which a portion for transmitting and shielding a light guided by the optical system is provided; A rotator having a transmission part that transmits light, a driving unit that rotates the rotator, and a confocal image and a non-confocal image of the sample obtained through the semi-transmission part of the rotator. A confocal image of the specimen, from an imager that captures an image and a bright-field image of the sample obtained through the transmission unit; and the composite image and the bright-field image captured by the imager. And a calculation means for obtaining
Control means for controlling the driving means to change the rotation speed of the rotating body and changing the exposure time of the imaging means according to the change in the rotation speed of the rotating body.

(2) The confocal microscope according to the present invention has an optical system for guiding the light emitted from the light source to the sample, and a semi-transmissive portion in which a portion for transmitting the light guided by the optical system and a portion for shielding are mixed. Unit, a transmitting unit that transmits the light, a rotator, a rotator driving unit that rotates the rotator, and a confocal image of the sample obtained through the semi-transmitting portion of the rotator. An imaging unit that captures a composite image composed of a non-confocal image and a bright field image of the sample obtained through the transmission unit, and the composite image and the bright field image captured by the imaging unit. A confocal microscope comprising: a calculating means for obtaining a confocal image of the specimen; a first polarizing member provided between the light source and the rotating body; and a light source and the first polarizing member. And a second polarizing member provided between the second polarizing member and the second polarizing member. A polarization direction rotation driving unit for rotating the polarization direction with respect to the optical axis of the light source; and a control unit for controlling the driving of the rotator driving unit and the driving of the polarization direction rotation driving unit in association with each other. .

(3) The confocal microscope of the present invention provides the above-mentioned (1)
Alternatively, in the microscope according to (2), the material of the rotating body and each member of the optical system are optical members that transmit ultraviolet light.

(4) The confocal microscope of the present invention provides the above-mentioned (1)
The microscope described above, and capable of switching to non-confocal observation, a brightness change instructing means for instructing a change in image brightness, and an irradiation for changing an irradiation light amount to a sample guided by the optical system. A light amount changing unit, an exposure time changing unit for changing an exposure time of imaging by the imaging unit, an observation method detecting unit for detecting whether a currently performed observation method is confocal observation or non-confocal observation, At the time of observation and at the time of non-confocal observation, the change value of the rotation speed of the rotating body, the change value of the irradiation light amount by the irradiation light amount change unit, and the exposure when there is a brightness change instruction from the brightness change instruction unit. Setting change value calculating means for obtaining at least one of the change values of the exposure time of the imaging by the time changing means.

As a result of taking the above-described measures, the following effects are obtained.

(1) According to the confocal microscope of the present invention, the brightness of an image is changed by changing the rotation speed of the rotator in accordance with the exposure time of the image pickup device whose exposure time can be controlled by an external signal input. Adjustments can be made. This makes it possible to easily adjust the brightness of the image.

(2) According to the confocal microscope of the present invention, the brightness of the image can be adjusted by changing the rotation speed of the rotator in accordance with the exposure time of the image pickup device. The amount of light can be adjusted by rotating the polarization direction by the polarizing member to change the angle of the polarized light and extracting only one direction of the polarized light by the first polarizing member. This makes it possible to easily and widely adjust the brightness of the image.

(3) According to the confocal microscope of the present invention, a confocal image in an ultraviolet light region can be obtained, and light can be modulated even when a mercury light source is used or in a deep ultraviolet light region.

(4) According to the confocal microscope of the present invention, the current observation method is examined when a brightness change instruction is input, and during the confocal observation, the rotation speed of the rotating body or irradiation of the sample is performed. The brightness can be changed by changing the amount of light, and in non-confocal observation, the brightness can be changed by changing the exposure time of imaging or the amount of light applied to the sample. This makes it possible to easily adjust the brightness by using only one brightness change instruction input unit regardless of the observation method.

[0027]

(First Embodiment) FIG. 1 is a diagram showing a configuration of a disk scanning confocal microscope according to a first embodiment of the present invention. In FIG. 1, the same parts as those in FIG. 6 are denoted by the same reference numerals.

The confocal microscope shown in FIG. 1 is a microscope that projects ultraviolet light, and a mercury light source or a xenomercury light source is used as the light source 1. On the optical path of the light emitted from the light source 1, an optical lens 2, a UV transmitting only the ultraviolet rays,
Filter 21, PBS (or polarizing plate, hereinafter referred to as PO) 2
2, PBS3 is arranged, and on the reflected light path of PBS3,
Rotating disk 4 ', imaging lens 23, quarter-wave plate 2
4. A sample 6 is arranged via an objective lens 5.

The rotating disk 4 'is an optical member made of synthetic quartz, fluorite, etc., which transmits ultraviolet light, and has the same basic structure as that shown in FIGS. In the rotating disk 4 ', a low-reflection film made of chromium oxide or the like is deposited on the shielding portions 4c and 4d. Similarly, a low reflection film made of chromium oxide or the like is deposited on portions other than the pinholes in the random pinhole pattern portion 4a and on the line portions in the line pattern portion 4e. Further, the occurrence of flare is reduced by applying an anti-reflection coating to each disk as a whole.

On the other hand, a CCD camera 9 is arranged on the transmission optical path of the PBS 3 via an imaging lens 8. The CCD camera 9 has an image sensor capable of controlling the exposure time by inputting an external signal. A computer 10 is connected to an image output terminal and an input terminal. And monitor 11
Is connected.

The computer 10 has a first controller 3
1 is connected, and a first pulse motor 33 is connected to the first controller 31 via a first driver 32. The shaft (not shown) of the first pulse motor 33 is connected to the rotating shaft 7 of the rotating disk 4 '. Further, the computer 10 has a compound image (CF) bright field image (NCF) (hereinafter, CF / NCF) reading sensor 34 for identifying an image type arranged near the side of the rotating disk 4 ′.
Is connected.

Further, a second controller 35 is connected to the computer 10.
5 has a second pulse motor 37 via a second driver 36.
Is connected. The shaft of the second pulse motor 37 is connected to a gear (not shown), and the rotation of the shaft causes the PBS 22 to rotate around the optical axis of the light source 1 via the gear. Computer 1
0 is connected to an origin detection sensor 38 arranged near the PBS 22.

When the CCD camera 9 receives the operation start signal from the computer 10, it opens the shutter and starts exposure, and when it receives the shutter close signal from the computer 10, closes the shutter and ends the exposure, and reads out the accumulated charges. Is completed, the shutter is opened again.

The computer 10 includes a first controller 3
1. The rotation speed of the rotating disk 4 'is controlled via the first driver 32 and the first pulse motor 33. This rotation speed is recognized by the computer itself based on a motor drive command (command speed) output by the computer 10. At this time, the first controller 31 outputs a motor drive pulse signal to the first driver 32 according to a motor drive command (drive amount, drive speed) input from the computer 10.
The first driver 32 amplifies the drive pulse signal from the first controller 31 and sends it to the first pulse motor 33, and rotates the first pulse motor 33 according to the drive pulse signal.

Note that the computer 10 controls the exposure time of the CCD camera 9 to be extended correspondingly when the rotation speed of the rotary disk 4 'is reduced, and to be reduced when the rotation speed is increased.

Further, the computer 10 receives the origin identification signal of the PBS (PO) 22 from the origin detection sensor 38, and transmits the polarization of the PBS (PO) 22 via the second controller 35, the second driver 36, and the second pulse motor 37. To control the orientation. At this time, the second controller 35 outputs a motor drive command (drive amount,
A driving pulse signal corresponding to the driving speed is output to the second driver 36. The second driver 36 amplifies the drive pulse signal from the second controller 35, sends the amplified signal to the second pulse motor 37, and rotates the second pulse motor 37 according to the drive pulse signal.

As a result, the PBS (PO) 22 is rotated to change its direction, and the polarization direction of the illumination light incident from the PBS (PO) 22 to the PBS 3 for branching the illumination / imaging optical path is changed. As a result, the amount of illumination light guided toward the sample 6 changes.

The computer 10 has a PBS (PO) 22
Is detected by inputting an origin identification signal generated by the origin detection sensor 38. Origin detection sensor 3
Reference numeral 8 denotes a direction in which the PBS (PO) 22 has a direction that maximizes the amount of illumination light, that is, a direction in which the PBS 3 for light branching matches the direction of polarized light that reflects light toward the sample 6.
An origin identification signal indicating that the direction of the PBS (PO) 22 is at the origin is generated. The rotation amount of the PBS (PO) 22 is recognized by the computer 10 as a motor drive command (rotation amount) output by the computer 10.

The computer 10 reads an image type identification signal from a CF / NCF reading sensor 34 for reading a bright field image (NCF image) identification mark and a composite image (CF image) identification mark provided on the side of the rotating disk 4 '. Receive. In the bright-field image identification mark, the opening 4b of the rotating disk 4 'is P
To be able to detect that it is located on the transmission light path of BS3,
It is provided on the side of the rotating disk 4 '. Similarly, in the composite image identification mark, the random pinhole pattern portion 4a (or the line pattern portion 4e) of the rotating disk 4 'is P
To be able to detect that it is located on the transmission light path of BS3,
It is provided on the side of the rotating disk 4 '.

Then, the computer 10 executes the CF / NCF
When the reading sensor 34 notifies that the image type has been changed by the image type identification signal, it outputs an instruction to the CCD camera 9 to close the shutter.

The computer 10 reads the instruction from the brightness instruction input unit 101 and changes the PBS (P) according to the content of the instruction, that is, whether the image is to be brightened or darkened.
O) It is determined whether to change the direction of 22 or the rotation speed of the disk 4 '.

When brightening an image, the computer 10
Is the PBS (P) input from the origin detection sensor 38 first.
O) Confirm the origin identification signal of the PBS (PO) 22
Is not at the origin, the PBS (PO) 22 is controlled to rotate gradually toward the origin to brighten the image.
If an instruction to further brighten the image is given after the PBS returns to the origin, the computer 10 controls to reduce the rotation speed of the disk 4 '.

When darkening an image, the computer 10
First, the rotation speed of the disk 4 'is the normal speed (maximum speed)
Is checked, and if it is determined that the rotational speed of the disk 4 'is not the maximum speed, the speed is increased to control to darken the image. If an instruction to darken the image is given after the rotation speed of the disk 4 'has reached the normal speed, the computer 10 controls the PBS (PO) 22 to change the direction to darken the image.

FIG. 2 is a flowchart showing the operation procedure of the disk scanning confocal microscope having the above-described configuration. Hereinafter, an operation procedure of the present disk scanning confocal microscope will be described with reference to FIG.

First, in step S1, when the operator inputs a brightness change instruction from the brightness instruction input unit 101,
In step S2, the computer 10 determines whether the direction of changing the content of the instruction is a direction for brightening or darkening the image. If it is determined that the image is to be brightened, the computer 10 determines in step S3 whether or not the direction of the PBS (PO) 22 is the direction of the maximum light amount based on the origin identification signal input from the origin detection sensor 38.

If the polarization direction of the PBS (PO) 22 is the direction of the maximum light quantity, the computer 10 lowers the rotation speed of the first pulse motor 33 to lower the rotation speed of the disk 4 'in step S4. In S5, the shutter control of the CCD camera 9 is performed to lengthen the exposure time. If it is determined in step S3 that the direction is not the maximum light amount,
In step S6, the computer 10 is rotated by the second pulse motor 37 in a direction in which the amount of light from the PBS (PO) 22 is increased, that is, in the direction of the origin.

If it is determined in step S2 that the image is to be darkened, in step S7, the computer 10 determines whether or not the rotation speed of the disk 4 'is the maximum. Here, when it is determined that the rotation speed is the maximum, in step S8,
The computer 10 uses the second pulse motor 37 to
It rotates in a direction to decrease the amount of light from the S (PO) 22, that is, in a direction away from the origin. In the above step S7,
If it is determined that the rotation speed of the disk 4 'is not the maximum,
In step S9, the computer 10 increases the rotation speed of the first pulse motor 33 to increase the rotation speed of the disk 4 '. In step S10, the computer 10 controls the shutter of the CCD camera 9 to shorten the exposure time.

In the confocal microscope, there is a limit in adjusting the brightness of an image only by changing the rotation speed of the disk and the exposure time of the image sensor. That is, when the image is darkened, there is a problem that the rotation speed of the disk cannot be increased to a predetermined maximum speed or more, and the exposure time of the image sensor cannot be shorter than the predetermined time.

However, according to the first embodiment, when the rotation speed of the disk 4 'is not the maximum, the rotation speed of the disk 4' is increased. Control is performed to reduce the amount of light from (PO) 22. Thus, the brightness of the image can be adjusted in a wide light amount range.

As the polarization direction rotation driving means, as in the first embodiment, the polarization beam splitter or the polarization plate as the second polarization member is mechanically rotated to move the polarization plane around the optical axis. In addition to the above, an element that can electrically rotate the plane of polarization by changing the applied voltage may be used.

(Second Embodiment) FIG. 3 is a diagram showing a configuration of a disk scanning confocal microscope according to a second embodiment of the present invention. 3, the same parts as those in FIG. 1 are denoted by the same reference numerals.

The confocal microscope shown in FIG. 3 is a microscope capable of switching between confocal observation and non-confocal observation. The rotating disk 4 ′ can be moved in and out of the optical path by moving the rotating shaft 7. Has become.

FIGS. 4A and 4B are diagrams showing the structure of the moving operation of the rotating disk 4 'and the rotating shaft 7. FIG. FIG.
In (a) and (b), the same parts as those in FIG. 3 are denoted by the same reference numerals. When the rotating disk 4 'is in the optical path as shown in FIG. 4A, it is possible to obtain a confocal image by the method of the first embodiment. When the rotating disk 4 'is out of the optical path as shown in FIG. 4B, non-confocal observation becomes possible.

The computer 10 is connected to a rotary disk position detecting sensor 39 arranged near the rotary shaft 7 of the rotary disk 4 '.
9, it is possible to detect whether or not the rotating disk 4 'is in the optical path.

The computer 10 has a brightness instruction input unit 1
When the instruction from 01 is read, the position of the rotating disk 4 'is read by the rotating disk position detection sensor 39 to determine whether the current observation method is confocal observation or non-confocal observation.
Further, the computer 10 changes which part of the direction of the PBS 22, the rotation speed of the disk 4 ', and the exposure time of the CCD camera 9 by the shutter control according to the current observation method and the instruction content from the brightness instruction input unit 101. Judge whether to do.

A storage unit (not shown) inside the computer 10 stores a PB at which the amount of light reflected by the PBS 3 becomes maximum.
A data table showing the amount of rotation of the PBS (PO) 22 in which the light quantity changes stepwise every 0.7 times from the direction of the origin of step S22, and the brightness of the image changes stepwise every 0.7 times A data table indicating the rotation speed of the rotating disk 4 'for confocal observation and a data table indicating the exposure time of the CCD camera 9 for non-confocal observation are stored in advance. Brightness instruction input unit 101
When the computer 10 receives an instruction from the computer, the computer 10 reads out each data table described above, and
The direction of S22, the rotation speed of the disk 4 ', and the change amount of the exposure time of the CCD camera 9 by the shutter control are determined, and the brightness of the image is changed stepwise by changing each part.

FIG. 5 is a flow chart showing the operation procedure of the disk scanning confocal microscope having the above configuration. Hereinafter, an operation procedure of the present disk scanning confocal microscope will be described with reference to FIG.

First, in step S20, when a brightness change instruction is input from the brightness instruction input unit 101 by the operator, in step S21, the computer 10 controls the rotation disk position signal input from the rotation disk position detection sensor 39. As a result, it is detected whether or not the rotating disk 4 'is in the optical path. Here, it is determined that confocal observation is being performed when the rotating disk 4 'is in the optical path, and non-confocal observation is being performed when the rotating disk 4' is out of the optical path. Here, when it is determined that the confocal observation is being performed, the processing from step S22 to step S30 is performed from step S2 to step S2 in the first embodiment.
Since the processing is the same as the processing up to 10, the description is omitted.

If it is determined in step S21 that non-confocal observation is being performed, step S21 is executed.
At 31, the computer 10 determines whether the instruction content from the brightness instruction input unit 101 is a direction for brightening or darkening the image. Here, in the case where the image is to be brightened, in step S32, the computer 10 determines whether or not the exposure time of the CCD camera 9 is the longest.

If the exposure time of the CCD camera 9 is the longest, the computer 10 changes the direction of the PBS (PO) 22 by the second pulse motor 37 in the direction of increasing the amount of light, that is, in the direction of the origin, in step S33. (P
O) Rotate 22. In step S32, CC
If the exposure time of the D camera 9 is not the longest, step S3
At 4, the computer 10 controls the shutter of the CCD camera 9 to extend the exposure time.

If it is determined in step S31 that the image is to be darkened, in step S35, the computer 10 uses the origin identification signal input from the origin detection sensor 38 to output the PBS.
It is determined whether the direction of (PO) 22 is the direction of the minimum light amount. Here, when the direction of the PBS (PO) 22 is the direction of the minimum light amount, in step S36, the computer 10
The shutter control of the D camera 9 is performed to shorten the exposure time of the CCD camera 9. In step S35, PB
If the direction of the S (PO) 22 is not the direction of the minimum light amount, the computer 10 causes the second pulse motor 37 to reduce the light amount from the PBS (PO) 22, that is, the direction of the PBS ( PO)
22 is rotated.

The switching to the non-confocal observation can also be performed by stopping the rotation of the rotary disk 4 'at a position where the opening 4b of the rotary disk 4' enters the optical path. Also in such a confocal microscope, it is possible to determine the observation method by detecting whether or not the rotation of the rotating disk 4 'is stopped. In this case, non-confocal observation is determined when the opening 4b of the rotating disk 4 'is positioned in the optical path and rotation is stopped, and confocal observation is performed when rotation is not stopped.

According to the second embodiment, the brightness can be adjusted by one brightness instruction input unit 101 regardless of whether the observation method is confocal observation or non-confocal observation. Operability of image brightness adjustment is improved.

The present invention is not limited to the above embodiments, but can be implemented with appropriate modifications without departing from the scope of the invention.

[0065]

According to the present invention, a confocal microscope capable of easily adjusting the brightness of an image can be provided.

Further, according to the present invention, the brightness of an image can be easily adjusted by one brightness instruction input unit regardless of whether the observation method is confocal observation or non-confocal observation. A confocal microscope with improved operability of adjustment can be provided.

[Brief description of the drawings]

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

FIG. 2 is a flowchart showing an operation procedure of the disk scanning confocal microscope according to the first embodiment of the present invention.

FIG. 3 is a diagram showing a configuration of a disk scanning confocal microscope according to a second embodiment of the present invention.

FIG. 4 is a diagram showing a configuration of a moving operation of a rotating disk and a rotating shaft according to a second embodiment of the present invention.

FIG. 5 is a flowchart showing an operation procedure of the disk scanning confocal microscope according to the second embodiment of the present invention.

FIG. 6 is a diagram showing a configuration of a disk scanning confocal microscope according to a conventional example.

FIG. 7 is a diagram showing a configuration of a random pinhole disk according to a conventional example.

FIG. 8 is a diagram showing a configuration of a line pattern disk according to a conventional example.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 ... Light source 2 ... Optical lens 3 ... PBS 4, 4 '... Rotating disk 4 4a ... Random pinhole pattern part 4b ... Opening 4c, 4d ... Light shielding part 4e ... Line pattern part 5 ... Objective lens 6 ... Sample 7 ... Rotation Axis 8 Second imaging lens 9 CCD camera 10 Computer 101 Brightness input unit 11 Monitor 21 UV filter 22 PBS 23 First imaging lens 24 1/4 wavelength plate 31 First controller 32 First driver 33 First pulse motor 34 CF / NCF reading sensor 35 Second controller 36 Second driver 37 Second pulse motor 38 Origin detection sensor 39 Rotating disk position detection sensor

Claims (4)

[Claims] An optical system for guiding light to a sample, a semi-transmissive portion in which a portion for transmitting light guided by the optical system and a portion for shielding are mixed, a transmitting portion for transmitting the light,
A rotating body having: a driving unit for rotating the rotating body; a composite image including a confocal image and a non-confocal image of the sample obtained through the semi-transmitting portion of the rotating body; Imaging means for capturing a bright-field image of the sample obtained through the imaging means; and computing means for obtaining a confocal image of the sample from the composite image and the bright-field image captured by the imaging means. A confocal microscope provided with control means for controlling the driving means to change the rotation speed of the rotator, and changing the exposure time of the imaging means according to the change in the rotation speed of the rotator. A confocal microscope characterized in that: 2. An optical system for guiding light emitted from a light source to a sample, a semi-transmissive portion in which a portion for transmitting the light guided by the optical system and a portion for shielding are mixed, and a transmission for transmitting the light. Department and
A rotating body driving means for rotating the rotating body; a composite image composed of a confocal image and a non-confocal image of the specimen obtained through the semi-transmissive portion of the rotating body; Imaging means for imaging a bright-field image of the specimen obtained through a section; and computing means for obtaining a confocal image of the specimen from the composite image and the bright-field image captured by the imaging means. A first polarizing member provided between the light source and the rotator, and a second polarizing member provided between the light source and the first polarizing member.
A polarizing member, a polarization direction rotation driving unit for rotating the polarization direction with respect to the optical axis of the light source by the second polarization member, and a driving of the rotator driving unit and a driving of the polarization direction rotation driving unit. A confocal microscope, comprising: control means for controlling in association with each other. 3. The confocal microscope according to claim 1, wherein the material of the rotating body and each member of the optical system are optical members that transmit ultraviolet light. 4. A brightness change instructing means capable of switching to non-confocal observation, instructing a change in brightness of an image, and changing an irradiation light amount for changing an irradiation light amount to a sample guided by the optical system. Means, an exposure time changing means for changing an exposure time of imaging in the imaging means, an observation method detecting means for detecting whether the currently performed observation method is confocal observation or non-confocal observation, and at the time of confocal observation And at the time of non-confocal observation, a change value of the rotation speed of the rotating body, a change value of the irradiation light amount by the irradiation light amount changing unit, and a change of the exposure time when there is a brightness change instruction from the brightness change instruction unit. 2. A confocal microscope according to claim 1, further comprising: setting change value calculating means for obtaining at least one of a change value of an exposure time of imaging by said means.