JP2001083426A - Confocal microscope - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a confocal microscope constituted so that a bright visual field can be observed without attaching/-detaching a disk. SOLUTION: This confocal microscope is provided with an optical system guiding light to a sample, a rotary body 4'w having a semi-transmission part where a part through which the light guided by the optical system is transmitted and a part shielding the light coexist and a transmission part through which the light is transmitted, a driving means 32 rotating the rotary body 4', a control means 31 controlling the driving of the driving means 32, 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' and 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 detection means 33 detecting the prescribed place of the rotary body 4'. Then, the control means controls the driving means based on the detected result of the detection means 33 and stops the rotation of the rotary body 4' so that the transmission part thereof is positioned on an observation optical path.

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 the optical path of the light emitted from the light source 1. A rotating disk 4 and a 波長 wavelength plate 12 The sample 6 is arranged via the objective lens 5.

[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 intervals as the pinhole diameter.
And an opening 4b through which light can pass freely, and shielding portions 4c and 4d for blocking light are formed between the random pinhole pattern portion 4a and the opening 4b.

The rotary disk 4 is formed of a transparent glass disk, and a low reflection film such as a Cr film is deposited on portions corresponding to the shielding portions 4c and 4d on the disk. 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. 12 can be used. In this disk, instead of the random pinhole pattern portion 4a shown in FIG. 11, a striped line pattern portion 4e in which a plurality of lines are arranged at substantially equal intervals on transparent glass.
Are formed. Then, this line pattern portion 4e
In each of the lines, for example, a Cr film is deposited,
Each line is shaded. That is, the line pattern portion 4e 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 an imaging lens 8. A computer 10 is connected to an image output terminal of the CCD camera 9, and an 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 that has passed through the random pinhole pattern portion 4a (or the line pattern portion 4e) or the opening 4b of the rotating disk 4 becomes circularly polarized by the quarter-wave plate 12, is imaged by the objective lens 5, and is imaged. Is incident on.

The light from the sample 6 forms an image with the objective lens 5, passes through the quarter-wave plate 12, becomes polarized light orthogonal to the incident light, and is again turned into a random pinhole pattern portion of the rotating disk 4. The light passes through 4a (or the line pattern portion 4e) or the opening 4b, further passes through the PBS 3, and enters 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 passing through the random pinhole pattern portion 4a (or line pattern portion 4e) and an image passing through the opening 4b. 2
One image is captured. The output image of the CCD camera 9 is stored in the computer 10. Computer 10
Performs a difference operation between confocal image data containing a non-confocal component obtained from the random pinhole pattern portion 4a (or line pattern portion 4e) and non-confocal image data obtained from the opening 4b. Thus, the confocal image data is converted into confocal image data of only the confocal component, 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]

As described above, the conventional T.I. With a Wilson-type disk scanning confocal microscope, a disk is rotated to capture a composite image obtained in a portion provided with a large number of pinholes and stripes, and a bright-field image obtained in a transmission portion. Thus, a confocal image is obtained by calculating the difference between the two. That is, in this confocal microscope, a composite image and a bright-field image are captured during one rotation of the disk.

However, when using a confocal microscope having such a configuration, it is sometimes necessary to perform not only a confocal image but also a bright-field observation. In that case,
In the conventional disc-scanning confocal microscope, it is necessary for the operator to stop the confocal observation, stop the drive of the disc, perform the operation of removing the disc from the observation optical path, and then perform the bright field observation.

As described above, the conventional confocal microscope requires a complicated operation when switching from confocal observation to bright-field observation.

An object of the present invention is to provide a confocal microscope capable of performing bright-field observation without attaching / detaching a disk.

[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 rotating unit having a transmitting unit that transmits light, a driving unit that rotates the rotating unit, a control unit that controls the driving of the driving unit, and the sample that is obtained through the semi-transmitting unit of the rotating unit. A composite image composed of a confocal image and a non-confocal image, and an imaging unit that captures a bright-field image of the sample obtained through the transmission unit, and the composite image captured by the imaging unit and the composite image. A confocal microscope comprising: a bright-field image; and a calculating means for obtaining a confocal image of the sample, wherein a detecting means for detecting a predetermined portion of the rotating body is provided, and the control means detects a detection result of the detecting means. Controlling the driving means based on Stopping the rotation of the rotating body so that the transmitting portion is positioned on the observation optical path.

(2) The confocal microscope of the present invention provides the above-mentioned (1)
And the driving means comprises a pulse motor.

(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 (3)
And the material of the rotating body is synthetic quartz or fluorite.

(5) The confocal microscope according to the present invention is characterized in that (1)
(4) The microscope according to any one of (1) to (4), wherein the rotating body has a shielding part that shields the light guided by the optical system, and the shielding part and the part that the semi-transmissive part shields are: And a low reflection film.

(6) A confocal microscope according to the present invention is the microscope according to the above (1) or (2), wherein an arbitrary position in the transmission section of the rotating body is set as a stop position of the rotating body. Rotating body stop position setting means,
The stop position of the rotating body set by the rotating body stop position setting means is a position at which foreign matter on the transmission portion of the rotating body does not affect a bright field image, and the control means The rotation of the rotating body is stopped at the stop position set by the body stop position setting means.

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

(1) According to the confocal microscope of the present invention, when performing only bright-field observation from a state in which confocal observation is being performed, the rotating body can be stopped in a state where only bright-field observation is possible. Therefore, bright-field observation can be performed only by stopping the rotation of the rotating body, and there is no need to perform a complicated operation such as removing the rotating body from the observation optical path. That is, switching between confocal observation and bright field observation can be performed very easily. Further, since it is not necessary to take one bright-field image while the rotator makes one rotation, the bright-field image can be taken in a short time without wasting light.

(2) According to the confocal microscope of the present invention, the rotation of the rotator can be stopped while the transmitting portion of the rotator is securely positioned on the observation optical path.

(3) According to the confocal microscope of the present invention, a confocal image in an ultraviolet light region can be obtained.

(4) According to the confocal microscope of the present invention, a confocal image in the deep ultraviolet region can be obtained.

(5) According to the confocal microscope of the present invention, it is possible to reduce the occurrence of flare between the shielding portion of the rotating body and the shielding portion of the semi-transmissive portion.

(6) According to the confocal microscope of the present invention, even if there is foreign matter such as dust on the transmission part of the rotating body, the foreign matter affects the bright field image by the rotating body stop position setting means. Since the position where no rotation is given is set as the rotating body stop position, during bright field observation, the rotating body can be stopped at a position where foreign matter on the transmission part of the rotating body does not affect the bright field image.

[0031]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS (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. 10 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 xenon mercury 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 and a U that transmits only ultraviolet rays are provided.
A V filter 21, a polarizer 22, and a PBS (polarizing beam splitter) 3 are arranged. On the reflection optical path of the PBS 3, a sample is provided via a rotating disk 4 ′, an imaging lens 23, a 波長 wavelength plate 24, and an objective lens 5. 6 are arranged.

The rotating disk 4 'is made of synthetic quartz or fluorite and is made of a random pinhole disk shown in FIG. 2 or a line pattern disk shown in FIG. The basic configurations of these disks are shown in FIG.
12 and FIG. 11, FIG.
The same parts as those in 2 are denoted by the same reference numerals. 2 and FIG. 3, the edge 41 of the half circumference on the side of the opening 4b is painted black and the half of the circumference of the random pinhole pattern part 4a and the half circumference on the line pattern part 4e are different. The edge 42 is transparent.

In each of the disks shown in FIGS. 2 and 3, 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. A computer 10 is connected to an image output terminal of the CCD camera 9, and an operation unit 101 and a monitor 11 are connected to the computer 10.

Further, a controller 31 is connected to the computer 10, and a pulse motor 32 and a photo interrupter 33 are connected to the controller 31. The rotating shaft of the pulse motor 32 is connected to the rotating shaft 7 of the rotating disk 4 '. Photo interrupter 33
Is arranged such that the edge of the rotating disk 4 ′ is inserted into the recess and does not always contact the edge.

The controller 31 receives a detection signal from the photo-interrupter 33 in response to an instruction from the computer 10, and controls the rotation of the rotary disk 4 'by controlling the pulse motor 32 as described later.

The operation of the disk scanning confocal microscope constructed as described above will be described below. When the above-described ordinary confocal observation is performed by the present confocal microscope, the rotating disk 4 'is rotating. When performing bright field observation from this state, the operator turns on the changeover switch of the operation unit 101. When the computer 10 detects that the changeover switch has been turned ON, the computer 10 outputs a signal to the controller 31 to instruct the controller 31 to stop rotating the rotating disk 4 '.

When the controller 31 receives the above signal, it inputs a detection signal from the photo interrupter 33.
The detection signal includes a first signal indicating that the edge of the rotating rotating disk 4 'has changed from blocking (41) to transmission (42), and a change from transmission (42) to blocking (41). There are two types of second signals that indicate this.

In the controller 31, two types of set pulse numbers respectively corresponding to the first signal and the second signal are set. The number of set pulses is determined by setting the center of the opening 4b in the pulse motor 32 after the first or second signal is input to the photointerrupter 33 by PB.
It shows the number of pulses required to stop on the transmission light path (observation light path) in S3.

The controller 31 outputs the number of pulses corresponding to the input first signal or second signal to the pulse motor 32. The pulse motor 32 rotates the rotary disk 4 ′ via its own shaft and the rotary shaft 7 according to the number of input pulses, and then stops. As a result, the center of the opening 4b of the rotating disk 4 'stops on the transmitted light path of the PBS 3. In this state, the CCD camera 9 captures only a non-confocal image obtained from the opening 4b, that is, a bright-field image.

When the confocal observation is performed again, the operator turns off the changeover switch of the operation unit 101. Then, the computer 10 detects that the changeover switch has been turned off, and outputs a signal to the controller 31 to instruct the controller 31 to start rotating the rotating disk 4 ′. Thus, the controller 31 controls the rotation of the rotating disk 4 'again.

As described above, according to the first embodiment, when only bright-field observation is performed from the state where confocal observation is performed in the ultraviolet region, only bright-field observation of the rotating disk 4 'is possible. Can be stopped in a normal state,
It is not necessary to perform a complicated operation such as removing the rotating disk 4 'from the observation optical path as in the related art. That is, switching between confocal observation and bright field observation can be performed very easily. In addition, since it is not necessary to capture one bright-field image while the rotating disk 4 'makes one rotation, a plurality of bright-field images can be captured in a short time without wasting light.

(Second Embodiment) FIG. 4 is a diagram showing a configuration of a disk scanning confocal microscope according to a second embodiment of the present invention. 4, the same parts as those in FIG. 1 are denoted by the same reference numerals.

The confocal microscope shown in FIG. 4 is a microscope using visible light, and a halogen light source or the like is used as the light source 1. 4 is different from FIG. 1 in that the UV filter 21 and the polarizer 22 are omitted on the optical path of the light emitted from the light source 1. In addition, Polarizer 2
Leaving 2 is effective in preventing flare. Also, PB
A half mirror 51 is disposed between the PBS 3 and the imaging lens 8 on the transmission optical path of S3. Further, an eyepiece lens 52 is arranged on the reflection optical path of the half mirror 51.

The disk scanning confocal microscope using visible light configured as described above also operates in the same manner as in the first embodiment, so that the center of the opening 4b of the rotating disk 4 'is positioned at the center of the PBS3. Stop on the transmitted light path, C
The CD camera 9 can capture only a non-confocal image in the visible light region obtained from the opening 4b, that is, only a bright-field image. Further, the bright field image transmitted through the opening 4 b and reflected by the half mirror 51 can be observed with the naked eye via the eyepiece 52.

(Third Embodiment) FIG. 5 is a diagram showing a configuration of a disk scanning confocal microscope using visible light according to a third embodiment. In FIG. 5, the same parts as those in FIG. 4 are denoted by the same reference numerals.

FIG. 5 differs from FIG. 4 in that the branch point to the eyepiece observation optical path is located closer to the sample 6 than the rotating disk 4 '.

A half mirror 64 for guiding light from the sample 6 to the eyepiece 52 is provided between the quarter-wave plate 12 and the imaging lens 23. The half mirror 64
Alternatively, a switching cube in which three types of prisms of 100% transmission prism, 100% reflection prism, and 50% reflection prism can be mechanically switched can be used.

Further, another half mirror 63 is arranged between the half mirror 64 and the quarter-wave plate 12. The half mirror 63 is for guiding light from a visible light source (hereinafter, referred to as a light source) 61 to the sample 6 via a condenser lens 62. When the light source 61 is turned on, it is assumed that a light source 1 described later is turned off or light from the light source 1 is blocked.

To perform bright field observation with this confocal microscope, the light source 61 is turned on. When a switching cube is used instead of the half mirror 64, a 100% reflection or 50% reflection prism is selected so that light is guided to the eyepiece 52.

In this case, the illumination light from the light source 61 is guided to the observation optical path via the condenser lens 62 and the half mirror 63, and illuminates the sample 6. Observation light from the sample 6 passes through the objective lens 5 and the half mirror 63 and is guided to the eyepiece 52 by the half mirror 64, thereby enabling bright field observation with the naked eye.

At this time, since the rotating disk 4 'has nothing to do with observation, the rotating disk may be rotated or stopped. When stopping, first and second
Unlike the first embodiment, the stop position of the rotating disk 4 ′ may be any position. When a bright-field image is captured by the CCD camera 9, the rotating disk 4 ′ is similar to the first and second embodiments. It is necessary to stop the rotating disk 4 ′ so that the center position of the opening 4 b is located on the optical path.
If another CCD camera is provided in the optical path of the eyepiece 52,
A bright-field image can be electrically captured without controlling the rotating disk 4 '.

On the other hand, when performing confocal observation, the light source 1 is turned on and the rotating disk 4 'is rotated. When the light source 1 is turned on, the light source 61 is turned off or the light from the light source 61 is blocked as in the case of turning on the light source 61 described above. In this case, the illumination light from the light source 1 passes through the condenser lens 2, the PBS 3, the rotating disk 4 ', the imaging lens 23, the half mirrors 64 and 63, the quarter wavelength plate 12, and the objective lens 5 in order, and passes through the sample 6. Light up. Since the observation light of the sample 6 passes through the quarter-wave plate 12 again and changes the polarization direction by 90 degrees, it passes through the PBS 3 and passes through the imaging lens 8 to the CCD.
An image is formed on the camera 9 and an observation image is captured. A confocal image is generated from the captured composite image and the bright field image, and the monitor 1
1 is displayed.

In the present embodiment, since the rotating disk is not involved in the bright-field image at all, the bright-field image can be observed only by switching the light source to be used without controlling the rotating disk.

The confocal microscope according to the present embodiment has the following configuration.

(1) A first illumination optical system that has a first light source and guides light to a sample, and a portion that transmits and blocks light guided by the first illumination optical system is mixed. A rotating body having a semi-transmissive part and a transmissive part for transmitting the light, a driving means for rotating the rotating body, a control means for controlling the driving of the driving means, and Imaging means for capturing a composite image composed of a confocal image and a non-confocal image of the specimen, and a bright-field image of the specimen obtained through the transmission unit, and the composite image captured by the imaging means And a calculating means for obtaining a confocal image of the specimen from the bright-field image and a second means provided between the specimen and the rotating body and different from the first light source. A second illumination optical system for guiding light from the light source to the sample, Confocal microscope for the optical branching means for branching the observation light from the at the specimen side of the rotating body, and observation means for observing the observation light which is branched by the optical branching unit, characterized by comprising a.

(2) The confocal microscope according to the above (1), wherein the observation means is an eyepiece.

(Fourth Embodiment) The hardware configuration of a confocal microscope according to the fourth embodiment is the same as the configuration of the disk scanning confocal microscope shown in the first embodiment. The description is omitted. When referring to the hardware configuration, reference numerals shown in FIGS. 1, 2 and 3 are used.

When a confocal image or a bright field image is obtained by the confocal microscope of the fourth embodiment, an instruction for switching between confocal observation and bright field observation is received from an operator. ′ Is stopped and rotated, and 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 opening portion 4 by the CCD camera 9.
Operations such as performing a difference calculation with the non-confocal image data obtained from b or displaying the observed images on the monitor 11 are performed by software running on the computer 10.

The software in the confocal microscope according to the fourth embodiment has a menu form as shown in FIG. 6, for example.

When performing image observation, the operator operates the monitor 1
1 from the observation image selection menu 91 displayed on
A desired observation image of the confocal image 91a, the composite image 91b, and the bright field image 91c is selected via the operation unit 101. Then, when obtaining a confocal image, the computer 10 outputs an instruction to rotate the rotating disk 4 ′ to the controller 31, and calculates a difference between the confocal image from the composite image obtained from the CCD camera 9 and the bright field image. And displays it on the monitor 11. When obtaining a bright-field image, the computer 10 outputs to the controller 31 an instruction to stop the rotating disk 4 'so that the center position of the opening 4b of the rotating disk 4' is located on the observation optical path, A bright field image obtained from the CCD camera 9 is displayed on the monitor 11.

Here, as shown in the first embodiment, in order to obtain a bright-field image, a predetermined position (here, the center position) of the opening 4b of the rotating disk 4 'is set. When the rotating disk 4 'was stopped so as to be located on the observation optical path, dust was attached to the center position of the opening 4b as shown in FIG. 7A, and as shown in FIG. 7B. It is assumed that dust is reflected in the bright field image.

In this case, in the confocal microscope of the fourth embodiment, in addition to the observation image selection menu 91,
A disc stop position setting menu 92 is provided.
This menu is selected via the operation unit 101. This menu is for the operator to set a stop position when stopping the rotating disk 4 'to obtain a bright field image.

Hereinafter, the disc stop position setting menu will be described. When the disc stop position setting menu is selected, the processing shown in the flowchart of FIG. 8 is performed.

The process of setting the stop position of the rotating disk 4 'during bright-field observation can be roughly divided into a process of freely rotating the rotating disk 4' within a range where the opening 4b falls within the observation range (step S111).・ S111 ′ ・
S112) and a process of determining and determining whether or not the position where the rotating disk 4 'is freely rotated and stopped is regarded as the disk stopping position during bright field observation (step S11).
3) calculating the amount of rotation of the rotating disk 4 'from the disk stop position before this menu is selected to the disk stop position determined and determined as the disk stop position for bright field observation in this menu; The processing comprises a step of notifying the controller 31 (step S114).

First, in step S111, the computer 10 waits for an input of the number of drive pulses. The number of drive pulses is the number of drive pulses of the pulse motor 32 corresponding to the rotation amount of the rotating disk 4 ′ desired by the operator, and is input to the computer 10 via the operation unit 101 by the operator. The number of drive pulses input by the operator can be arbitrarily input within the range from the current stop position of the rotating disk 4 '(here, the center position of the opening 4b) to the opening 4b. Here, for example, the number of drive pulses corresponding to the rotation amount of the distance corresponding to the observation range is input.

When the computer 10 detects that the number of driving pulses has been input through the operation unit 101, the computer 10 proceeds to step S1.
In step 12, the number of drive pulses is stored in a built-in memory, and an instruction to rotate the rotary disk 4 'by the number of input drive pulses is output to the controller 31.

Then, the controller 31 outputs pulse signals for the number of drive pulses to the pulse motor 32, and the rotating disk 4 'rotates by a predetermined amount of rotation (here, the distance for the observation range), and the opening portion is opened. Since the position of the observation light path 4b also changes, the monitor 11 is located at the center of the opening 4b.
The position of the dust reflected in the image moves as shown in FIG. As a result of rotating the rotating disk 4 'by a predetermined amount, dust disappears from the bright-field image as shown in FIG. 9B, and the position is regarded as a disk stop position during bright-field image observation. The operator presses the disc stop position determination switch of the operation unit 101 during bright field observation.

The computer 10 proceeds to step S113
If it is not detected that the disc stop position determination switch is pressed during the bright field observation of the operation unit 101, the process proceeds to step S
At 111 ', the apparatus again waits for the input of the number of drive pulses.
If it is detected in step S113 that the disc stop position determination switch during bright field observation has been pressed, the processing in step S114 is executed.

In step S114, the computer 10
Accumulates all the driving pulses required for the rotation of the rotating disk 4 'performed a plurality of times stored in the memory,
A process of notifying the controller 31 of the number of offset pulses is performed. That is, as shown in FIG. 9A, when performing bright field observation, the number of offset pulses is determined at any position from the predetermined position (in this case, the center position) of the opening 4b of the rotating disk 4 '. This indicates whether to stop the disk 4 '.

In this way, the disk stop position during bright field observation is set to a position rotated by the number of offset pulses from the predetermined position of the opening 4b of the rotary disk 4 'by using the disk stop position setting menu 92. Will be able to do it.

The switching operation from the confocal observation to the bright field observation when the disc stop position is newly set for the bright field observation as described above will be described below.

When the bright-field image 91c is selected in the observation image selection menu 91 while the confocal observation is being performed, the computer 10 issues a disk stop instruction to the controller 31.

As shown in the first embodiment, the controller 31 receives the first signal or the second signal from the photo interrupter 33 and then sets the predetermined position of the opening 4b (here, the center position). ) Are set on the observation optical path, and the number of pulses required for setting the number of pulses (set pulse number 1 and set pulse number 2 respectively) are set. When the controller 31 receives the number of offset pulses as the disk stop position information from the computer 10, the controller 31 calculates a new set number of pulses by the following equation,
Update the (old) set pulse count.

(New) Set pulse number 1 ← (Old) Set pulse number 1 + Offset pulse number (New) Set pulse number 2 ← (Old) Set pulse number 2 + Offset pulse number The updated set pulse numbers 1 and 2 are: These are the number of drive pulses required from the input of the first and second signals from the photo interrupter 33 to the stop of the rotating disk 4 'at the disk stop position set in the disk stop position setting menu 92, respectively.

When the controller 31 receives the disc stop instruction from the computer 10 and receives the first signal or the second signal from the photo interrupter 33, the controller 31 responds to the input first signal or second signal. The set number of pulses is output to the pulse motor 32. The pulse motor 32 rotates the rotating disk 4 ′ via its own shaft and the rotating shaft 7 in accordance with the number of inputted pulses,
Stop.

Thus, during bright field observation, the rotation of the rotary disk 4 'can be stopped at the disk stop position set in the disk stop position setting menu 92. As described above, even if dust adheres to the opening 4b of the rotary disk 4 ', if the position where no dust adheres is set as the disk stop position in the disk stop position setting menu 92, the light can be cleared. At the time of visual field observation,
It is possible to prevent dust from being reflected in the bright field image.

In the fourth embodiment, the position where dust is present is the center position of the opening 4b, but is not limited to this. For example, even if the rotary disk 4 'is stopped at a position other than the center position of the opening 4b and dust is present at that position, the disk stop position setting menu shown in the fourth embodiment is used, The disc stop position during bright field observation may be set to a position where dust does not exist on the opening 4b (or a position where dust does not affect the bright field observation image as much as possible). Also,
In the fourth embodiment, dust or the like adhering to the opening 4b is regarded as a foreign substance, but is not limited thereto.
For example, a scratch on the opening 4b may be treated as a foreign matter.

In the fourth embodiment, steps S111 and S1 of the disk stop position setting process are performed.
At 11 ', the operator inputs the number of drive pulses of the pulse motor as the desired disk rotation amount to the computer 10 via the operation unit 101, but is not limited to this, and inputs the disk rotation angle. Alternatively, the desired disk rotation amount may be input as a percentage of the size of the observation visual field. In this case, the computer 10 performs a process of converting each into the number of drive pulses of the pulse motor using a preset conversion coefficient.

In the fourth embodiment, in order to freely rotate and stop the rotary disk 4 'within the range of the opening 4b in the disk stop position setting process, the number of drive pulses is controlled by the operation unit. Although the input is made from the input unit 101, the present invention is not limited to this. For example, the operation unit 101 is provided with a right rotation switch and a left rotation switch, and while the operator presses the right rotation switch or the left rotation switch, 4 'may be rotated clockwise or counterclockwise, and the rotary disk 4' may be stopped when the switch is released.

Further, in the fourth embodiment, the number of offset pulses is applied as the disk stop position information, but the rotation angle from the predetermined position of the disk opening to the new disk stop position may be used.

The confocal microscope according to the fourth embodiment is a microscope which projects the ultraviolet light shown in the first embodiment, but is not limited to this. May be a microscope that projects visible light as shown in FIG.

In the confocal microscope according to the fourth embodiment having the structure and operation described above, even if foreign matter such as dust adheres to the opening 4b of the rotating disk 4 ', the foreign matter is reflected in the bright field image. Since a clear bright-field observation image can be obtained at any time, the trouble of cleaning attached foreign matter can be omitted.

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.

[0086]

According to the present invention, it is possible to provide a confocal microscope capable of performing bright-field observation without mounting / dismounting a disk.

[Brief description of the drawings]

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

FIG. 2 is a diagram showing a configuration of a random pinhole disk according to the embodiment of the present invention.

FIG. 3 is a diagram showing a configuration of a line pattern disk according to the embodiment of the present invention.

FIG. 4 is a diagram showing a configuration of a disk scanning confocal microscope according to the embodiment of the present invention.

FIG. 5 is a diagram showing a configuration of a disk scanning confocal microscope using visible light according to the embodiment of the present invention.

FIG. 6 is an exemplary view showing a menu configuration of software operating on the computer according to the embodiment of the present invention.

FIG. 7 is a view showing a state of the disk and a bright-field image when the disk is stopped at the center position of the disk opening according to the embodiment of the present invention.

FIG. 8 is a flowchart showing the flow of a disk stop position setting process according to the embodiment of the present invention.

FIG. 9 is a diagram showing a disk state and a bright-field image when the disk is stopped at the disk stop position set in the disk stop position setting process according to the embodiment of the present invention.

FIG. 10 is a diagram showing a configuration of a disk scanning confocal microscope according to a conventional example.

FIG. 11 is a diagram showing a configuration of a random pinhole disk according to a conventional example.

FIG. 12 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 (polarization beam splitter) 4, 4 '... Rotating disk 4a ... Random pinhole pattern part 4b ... Opening 4c, 4d ... Light shielding part 4e ... Line pattern part 5 ... Objective lens 6 ... Sample 7 ... Rotating axis 8 ... Imaging lens 9 ... CCD camera 10 ... Computer 101 ... Operation unit 11 ... Monitor 21 ... UV filter 22 ... Polarizer 23 ... Imaging lens 24 ... 1/4 wavelength plate 31 ... Controller 32 ... Pulse motor 33 photointerrupters 41 and 42 edge 51 half mirror 52 eyepiece 61 light source 62 optical lens 63 PBS 64 half mirror

Claims (6)

[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 unit having: a driving unit for rotating the rotating unit; a control unit for controlling the driving of the driving unit; and a non-confocal image of the specimen obtained through the semi-transmissive portion of the rotating body. An imaging unit that captures a composite image composed of a focus image and a bright field image of the sample obtained through the transmission unit; andthe 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; and a detecting means for detecting a predetermined portion of the rotating body, wherein the controlling means is configured to control the driving means based on a detection result of the detecting means. A confocal microscope, wherein the rotation of the rotator is stopped so that the transmission portion of the rotator is positioned on an observation optical path. 2. The confocal microscope according to claim 1, wherein said driving means comprises a pulse motor. 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. The confocal microscope according to claim 3, wherein a material of the rotating body is synthetic quartz or fluorite. 5. The rotator has a shielding part for shielding the light guided by the optical system, and the shielding part and the shielding part of the semi-transmission part are made of a low reflection film. The confocal microscope according to any one of claims 1 to 4. 6. A rotating body stop position setting means for setting an arbitrary position of the rotating body in the transmission section as a stop position of the rotating body, wherein the rotating body set by the rotating body stop position setting means is provided. Is a position at which foreign matter on the transmission portion of the rotating body does not affect the bright field image. The control means controls the rotation at the stop position set by the rotating body stop position setting means. 3. The confocal microscope according to claim 1, wherein rotation of the body is stopped.