JP2003021790A - Image photographing device for microscope and image photographing method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To enable a user to easily photograph a good microscope observation image by fewer operations. SOLUTION: A specimen T is placed on a stage 1. An observation image is obtained by observing the specimen T through an objective lens 6. A stage driver 13 is capable of adjusting the focal position of the observation image by vertically moving the stage 1 across a stage rise and fall mechanism section 15. In bracketing of the observation image, the observation image is photographed by a camera head 8 and the stage 1 is moved by as much as the prescribed distance by the stage driver 13 after the photographing, by which the focal position is adjusted. The observation image is photographed again in the fresh focal position. Such operation to photograph the observation image and to move the stage 1 after the photographing is repeated a prescribed number of times until the observation images are photographed.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a technique for photographing an image observed by a microscope, and more particularly to a technique which can be suitably used for measuring fine dimensions.

[0002]

2. Description of the Related Art With the widespread use of digital cameras for general photographing, the use of digital image photographing apparatuses has become widespread also in photographing observation images of a microscope. High-definition, high-pixel count, high-priced digital image capturing devices that have been sold in some areas, but digital image capturing devices in a relatively low-pixel popular model have become particularly popular due to their price range. ing. This type of digital image capture device has
Various types of image capturing devices were proposed, such as a so-called stand-alone type that is attached to the microscope body later, a so-called integrated type that is built in the microscope body in advance, and a type that has an intermediate character between the two. There is. Further, in a digital image capturing apparatus of a popular model band, generally, an image display unit such as an LCD (Liquid Crystal Display) of several inches (1 inch corresponds to 2.54 cm) for image confirmation or focusing. have.

Compared with photography using a silver salt film, digital image photography is achieving a degree of perfection that is comparable to or superior to operability, image quality, and running cost. However, in order to obtain a high quality captured image when observing an image observed with a microscope, whether it is a digital image capture or an image capture using a silver salt film, a large amount of exposure control and focusing are required. experience,
Often requires skill. The difficulty of exposure control is represented by factors such as various special observation methods with a microscope and illumination methods.
On the other hand, the difficulty of focusing is due to a factor in the mechanical transmission mechanism that the distance handled is a unit of micron ([μm]) or less, and also the depth of focus of the objective lens and the unevenness of the sample to be observed. It is typified by factors derived from the relationship.

The unevenness of the specimen observed with a microscope is
In cells, which is the main target in the biological field, or in semiconductors, which is the main target in the industrial field,
It is about 10 microns. On the other hand, the depth of focus of the objective lens used for microscopic observation is precisely the low-magnification objective lens (4 to 10 times) used to confirm the overall image.
Although it is approximately 10 microns or more, it is approximately several microns or less in a commonly used medium-high magnification objective lens. Therefore, when observing an uneven sample using the medium-high magnification objective lens, one focus position cannot cover the unevenness of the sample observed by the microscope.

As a technique for solving the above-mentioned difficulty of exposure control in image pickup of a microscope, Japanese Patent Application Laid-Open No. 9-179038 discloses an exposure control method for a microscope photography apparatus. The invention described in this publication is capable of obtaining a plurality of photographs in which an intended difference in brightness appears when performing continuous shooting of a plurality of pieces of an object whose brightness changes by an auto bracket function. The present invention relates to an exposure control method for a microscopic photography device. Then, in the exposure control method of the microscopic photography device when continuously taking a plurality of photographs with different exposure values for the same test object by the auto bracket function, each photometry is performed immediately before each photography. There has been proposed an exposure control method for a microscopic photography apparatus, which sets an exposure value based on a value measured immediately before each photography.

[0006] On the other hand, various autofocus devices for microscopes have been proposed which overcome the difficulty of focusing in photographing an image of a microscope. An object of the invention described in Japanese Patent Laid-Open No. 11-258483 is to provide an autofocus device for a microscope which ensures high focusing accuracy and is highly reliable.

Therefore, a mounting means for mounting the observation body, an objective lens facing the observation body mounted on the mounting means, and a driving means for driving at least one of the mounting means and the objective lens. An image forming optical system for forming an image of an observation object through the objective lens; an optical path difference optical system for providing an optical path difference between a front position and a rear position conjugate with respect to an image forming point of the image forming optical system; Observation means for picking up an image of an observation object through an optical path difference optical system, and an operation of operating the driving means by obtaining the degree of focus of the observation object based on the image pickup information by the imaging means and placing it on the mounting means Control means for guiding the body to the in-focus position,
In the autofocus device for a microscope having a microscope, the control means uses an observation object search mode for searching an observation object, a rough focus mode for adjusting the observation object to a rough focus position with low focusing accuracy, and a focusing operation by the rough focus mode. With respect to the rough focus position, the front and rear defocus detection mode for obtaining the front and rear focusing degrees of at least two front and rear observation objects, and the high focusing accuracy of the observation object from at least the front and rear focusing degrees obtained by the front and rear defocus detection mode The present invention proposes an autofocus device for a microscope, which is equipped with a fine focus mode for linearly approximating the degree of focus of (1) and for adjusting an observation object to a focus position and an autofocus control mode of four modes.

[0008]

However, in the invention described in Japanese Patent Application Laid-Open No. 9-179038, the exposure value is set based on the value measured immediately before each photo shooting in the auto bracket shooting. It is set. For this reason, it is necessary to carefully and surely focus the observation image before starting the auto bracket photographing. If this focusing is not performed correctly, there is a problem that the result of a series of auto bracket shooting becomes useless.

Further, when the depth of focus of the objective lens to be used is small (shallow) with respect to the unevenness of the sample to be observed, the auto bracket photographing must be executed a plurality of times at a plurality of different focus positions. was there. By the way, the operator is forced to perform a troublesome microscopic image capturing operation and focusing with a skillful technique in order to obtain a desired high-quality microscope observation image.
There was a problem that the shooting environment was far from comfortable.

On the other hand, according to the invention described in Japanese Unexamined Patent Publication No. 11-258483, an autofocus device for a microscope, which ensures high focusing accuracy and is highly reliable, is provided.
Even if a microscope image is taken using this autofocus device, if the depth of focus of the objective lens to be used is small (shallow) with respect to the unevenness of the specimen to be observed, a high-quality microscope observation image There was a problem that I could not get.

In view of the above problems, an object of the present invention is to make it possible to take a high-quality observation image of a microscope easily and with few operations.

[0012]

According to one aspect of the present invention, there is provided a photographing device having a function of continuously photographing observation images obtained by observing a specimen through an objective lens,
A driving unit that adjusts the focus position of the observation image by changing the distance between the mounting unit that mounts the sample and the objective lens, and an image capturing unit that captures the observation image. The image capturing unit and the driving unit change the distance between the mounting unit and the objective lens by the driving unit after capturing the image by the image capturing unit until the observation image is captured a predetermined number of times. Characterized by repeating.

With this structure, the photographing device can photograph the observation image a predetermined number of times while automatically changing the focus position of the observation image, so that the observation image of the microscope is focused. The operator does not need to have high skill for focusing. As a result, even an inexperienced operator can obtain a high quality captured image without failure. Also, since the image is taken while changing the focus position of the observed image, even if the sample has relatively large unevenness or thickness compared to the depth of focus of the objective lens used, a good quality image can be obtained by just performing bracket shooting once. It becomes possible to obtain. Furthermore, it becomes possible to solve the above problems.

Here, in the above-mentioned photographing apparatus, at least one of the predetermined number of times and the amount of movement of the distance when changing the distance between the placing means and the objective lens depends on the performance of the objective lens. It may be decided. For example, at least one of the number of times of photographing and the amount of movement may be determined according to the depth of focus of the objective lens. Further, the number of times of photographing may be determined according to the unevenness or depth of the sample in addition to the performance of the objective lens. As a result, it is possible to perform continuous shooting a reasonable number of times and to perform a reasonable focus adjustment according to the performance of the objective lens. As a result, you can get good quality images with less number of shots and without failure.
It is possible to provide an image capturing device for a microscope with improved operation efficiency.

Further, in the above-mentioned photographing device, after changing the distance between the placing means and the objective lens by the driving means, waiting for a predetermined time until the photographing is started by the photographing means. Time may be provided. This makes it possible to eliminate the influence of the vibration generated by the driving unit even when the observation image is captured using the high-magnification objective lens. In addition, it is possible to obtain a high-quality photographed image without failure, and it is possible to provide a microscope image photographing device with improved reliability.

According to another aspect of the present invention, in a photographing method for continuously photographing observation images obtained by observing a sample through an objective lens, the observation images are photographed and the photographing is performed. After that, adjusting the focus position of the observation image by changing the distance between the sample and the objective lens is repeated until the observation image is captured a predetermined number of times. With this method as well, the same actions and effects as those of the above-described image pickup apparatus can be obtained, and the above-mentioned problems can be solved.

Even in the case of a program that causes a computer to control each means constituting the image capturing apparatus so that the image capturing apparatus can continuously capture images, the above-described problems can be solved by causing the computer to execute the program. can do. The above-mentioned problem can also be solved by causing a computer to read and execute the program from a computer-readable recording medium recording the program.

[0018]

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described below with reference to the drawings. First, a first embodiment of the present invention will be described. FIG. 1 is an overall configuration diagram of a microscope including an image capturing device according to the first embodiment. In FIG. 1, the vertical direction of the paper surface indicates the vertical direction (vertical direction), and the horizontal direction indicates the horizontal direction. The microscope includes a light source 2 for irradiating a specimen T, which is an observation object, mounted on a stage 1 that is vertically movable from below.

The illumination light from the light source 2 is ND for dimming.
(Neutral Density) The light enters the sample T from below through the filter 3, the field stop 4, the aperture stop 100, and the condenser lens 5. The light flux from the sample T passes through the objective lens 6 arranged in the revolver type lens barrel portion 16, a part of the light flux is guided to the eyepiece lens 7, and the other light flux passes through the imaging lens 200. Incident on the camera head 8. The light flux incident on the camera head 8 passes through the shutter 300 and forms an image on the incident surface of the image sensor 400. The structure of the camera head 8 will be described later (see FIG. 2).

The stage driving device 13 includes a controller 5
When the electric signal is received from 00, for example, the stage up-and-down mechanism unit 15 is driven to move the stage 1 in the up-down direction.
When receiving the electric signal from the controller 500, the revolver driving device 800 drives the revolver type lens barrel portion 16 to expose the various objective lenses 6 to the sample T on the stage 1. Further, the revolver type lens barrel section 16 has a built-in detection section for detecting the revolver position. The detection unit detects which position (which objective lens 6) of the revolver is currently inserted in the optical path facing the sample T, and sends the detection result to the controller 500. Detailed description of the detection unit is omitted. Upon receiving an electric signal from the controller 500, the aperture stop driving device 110 drives the aperture stop adjusting mechanism unit 120 to adjust the aperture stop 100 to an arbitrary aperture diameter.

The controller 500 includes an operation input section 510.
And a display portion 520. In addition, the controller 50
Reference numeral 0 is electrically connected to the camera head 8, the stage driving device 13, the revolver driving device 800, and the aperture stop driving device 110, and controls each device. The structure of the controller will be described later (see FIG. 3).

The structure of the camera head will be described below with reference to FIG. As shown in FIG. 2, the camera head 8
Is a sampling circuit 21 for sampling the electric signal supplied from the image sensor 400, and an A / D converter 22 for converting the obtained analog electric signal into a digital signal.
And an image processing unit 23 that performs a process for reproduction based on the converted digital signal, and the image processing unit 23 is connected to the controller 500. The shutter 300 that blocks the observation image incident on the image sensor 400 when desired is connected to the controller 500 via a shutter drive circuit 310.

The configuration of the controller will be described below with reference to FIG. As shown in FIG. 3, the controller 5
00 is a CPU (Central Processing Unit) 501,
Operation input unit 510, display unit 520, display memory 53
0, a display driver 540, and a memory device 550.

The CPU 501 has a ROM (Read Only Memory) 501a for storing a control program and a RAM (Random Access Mem) for securing variable data of the control program.
ory) 501b, a non-volatile memory 501c that stores various setting information, and a frame memory 5 that holds image data.
01d and 01d are provided inside (both not shown), and control overall control of this device.

The CPU 501 is connected to the operation input unit 510, and in response to an input from the operation input unit 510, the shutter drive circuit 310 of the camera head 8 and the stage drive device 1 are connected.
3, revolver driving device 800, aperture stop driving device 110
Send operation instructions to. The CPU 501 also stores the image data input from the camera head 8 in the display memory 53.
0, and display on the display unit 520 via the display unit driver 540. As the display unit 520, for example, a TFT (Thin
LCD etc. which are represented by Film Transistor) are mentioned.

The memory device 550 stores image data. The memory device 550 includes a removable medium 551 such as a flexible disk or a magneto-optical disk, which is widely used in PCs, and a memory reading / writing device 552 for writing and reading image data on the removable medium 551. .
As a result, the CPU 501 causes the memory read / write device 55 to read the image data input from the camera head 8.
2 to the removable medium 551. Further, the CPU 501 can also read, for example, a directory entry from the removable medium 551 via the memory reading / writing device 552 to obtain information on the image data file written in the removable medium 551. Further, the CPU 501 reads out arbitrary image data from this information, displays the display memory 530, the display driver 540.
It is also possible to display it on the display unit 520 via.

Next, the operation input section of the controller will be described in more detail with reference to FIG. FIG. 4 is an example of a front view of the operation input unit 510. As shown in FIG. 4, the operation input unit 510 includes at least the release button 511,
It has mode selection buttons 512a, 512b, 512c for selecting the operation mode of this device. These mode selection buttons are, for example, LED (Light Emitting Dio)
This is a so-called illuminated button switch in which a light emitting device such as de) is incorporated, and the mode selection status can be easily recognized by turning on or off the LED or the like. Each operation mode will be described later.

The operation input unit 510 further includes at least
Objective lens switching buttons 513a and 513b, aperture stop adjusting buttons 514a and 514b, index buttons 515a and 515b for specifying a file of image data written in the removable medium 551, a memory button 516 for recording various setting information, and,
It has a jog dial 517. The jog dial 517 is mechanically connected to a rotary encoder (not shown), and the rotary encoder outputs a pulse corresponding to the rotating operation of the jog dial 517. Further, the operation input section 510 is provided with an index display section 518 such as a 7-segment LED, and is configured to display the number of files of image data written in the removable medium 551.

The basic operation of this apparatus will be described below. In the following description, there are three operation modes, that is, a reproduction mode, a recording mode and a bracket recording mode, and mode selection buttons 512a, 512b and 512c are assigned to select each mode. I assume.

First, the operation in the reproduction mode will be described. The reproduction mode is an operation mode in which the image data written in the removable medium 551 is displayed on the display unit 520. First, when the mode selection button 512a is pressed, the CPU 501 detects a signal from the operation input unit 510, lights the LED of the mode selection button 512a, and turns on the LED of the mode selection button 512b and the mode selection button 512c. The LED is turned off and the operation mode is switched to the reproduction mode.

Next, the CPU 501 causes the removable medium 55 to be read via the memory read / write device 552.
The directory entry of No. 1 is read, the information of the image data file written in the removable medium 551 is obtained, and the number of written files is displayed on the index display unit 518. For example, if there are 10 image data files on the removable medium 551, the CPU 501 displays “10” on the index display unit 518.

Subsequently, the CPU 501 reads the tenth image data file of the directory entry from the removable medium 551 via the memory reading / writing device 552, expands it in the frame memory 501d, and displays this expanded image data for display. Transfer to the memory 530. The image data transferred to the display memory 530 is immediately transmitted to the display unit 5 via the display unit driver 540.
20 is displayed. In this way, the 10th image data file written in the removable medium 551 is displayed on the display unit 520.

The index button 515a designates an image data file in the order immediately preceding the currently designated image data file. Therefore, while the 10th image data file is displayed on the display unit 520,
When the index button 515a is pressed, the ninth image data file written in the removable medium 551 is displayed on the display unit 5 in the same process flow as above.
20 is displayed.

That is, the CPU 501 displays "9" on the index display portion 518. Then, the CPU 50
1 reads the ninth image data file of the directory entry from the removable medium 551 via the memory read / write device 552, expands it in the frame memory 501d, and transfers this expanded image data to the display memory 530. The image data transferred to the display memory 530 is immediately displayed on the display unit 520 via the display unit driver 540. In this way, the ninth image data file written in the removable medium 551 is displayed on the display unit 520.

Contrary to the index button 515a, the index button 515b designates an image data file which is one sequence behind the currently designated image data file. Now, the 10th image data file written in the removable medium 551 is the display unit 5
20 is displayed. In this case, even if the index button 515b is pressed, the 11th image data file does not exist. Therefore, the removable media 551 is not the 11th image data file.
The first image data file written in is displayed on the display unit 520.

On the other hand, when the index button 515a is pressed while the first image data file written in the removable medium 551 is displayed on the display section 520, the 0th image data file is Since it does not exist, the 10th image data file written in the removable medium 551 is displayed on the display unit 520.

That is, the CPU 501 displays "10" on the index display portion 518. Then, CPU5
01 reads the tenth image data file of the directory entry from the removable medium 551 via the memory reading / writing device 552, expands it in the frame memory 501d, and transfers this expanded image data to the display memory 530. The image data transferred to the display memory 530 is immediately transferred to the display driver 54.
It is displayed on the display unit 520 via 0. In this way, the 1 written in the removable medium 551
The 0th image data file is displayed on the display unit 520.

In this way, according to this apparatus, the operator operates the index button 515 in the reproduction mode.
By operating a and the index button 515b, it is possible to reproduce and display the image data file desired by the operator among the image data files written in the removable medium 551 on the display unit 520.

Next, the operation in the recording mode will be described. When the mode selection button 512b is pressed, CP
U501 detects the signal from the operation input unit 510,
The LED of the mode selection button 512b is turned on, and the LED of the mode selection button 512a and the mode selection button 512 are turned on.
The LED of c is turned off, and the operation mode is switched to the recording mode.

In the recording mode, a microscope observation image formed on the incident surface of the image pickup device 400 is displayed on the display unit 520 at a real time / full frame rate. That is, the observation image from the microscope body is imaged on the incident surface of the image sensor 400 via the imaging lens 500. This allows
The observed image of the sample T is converted into an electric signal by the image sensor 400. This electrical signal representing the observed image is spatially and temporally sampled by the sampling circuit 21. The sampled electrical signal is digitized by the A / D converter 22, and then image-processed by the image processing unit 23 by a predetermined method based on the sampling component. As a result, a reproducible digital image data signal of the sample is generated. This digital image data signal is sent to the display memory 530 via the CPU 501.
Transferred to. The image data transferred to the display memory 530 is immediately displayed on the display unit 520 via the display unit driver 540.

Here, when the release button 511 is pressed, the CPU 501 starts the recording operation. That is, in response to an instruction from the CPU 501, the shutter 300 via the shutter drive circuit 310 or an electronic shutter (not shown) opens and closes in accordance with an appropriate exposure time, and the camera head 8 observes the stopped state. You can take a picture. This image data is transferred to the display memory 530 and displayed on the display unit 520 as captured image data. On the other hand, the CPU 501 writes this image data in the removable medium 551 via the memory reading / writing device 552, and the removable medium 5
Add one file to the 51 directory entry.

In this way, this apparatus writes and records a desired observation image in the removable medium 551 in response to the operation of the release button 511 in the recording mode. The operation of the bracket recording mode according to the first embodiment will be described below with reference to the flowchart shown in FIG.

First, when the mode selection button 512c is pressed, the CPU 501 detects a signal from the operation input section 510, turns on the LED of the mode selection button 512c, and the LED of the mode selection button 512a and the mode selection button 512c. The LED of the button 512b is turned off, and the operation mode is switched to the bracket recording mode.

Even in the bracket recording mode, the observation image of the microscope imaged on the incident surface of the image pickup device 400 is displayed on the display unit 520 at the real time / full frame rate, as in the recording mode. is there. That is, the observation image from the microscope body is the image forming lens 500.
Then, an image is formed on the incident surface of the image sensor 400. As a result, the observed image of the sample T is converted into an electric signal by the image sensor 400. This electrical signal representing the observed image is spatially and temporally sampled by the sampling circuit 21. The sampled electrical signal is digitized by the A / D converter 22, and then image-processed by the image processing unit 23 by a predetermined method based on the sampling component. As a result, a reproducible digital image data signal of the sample is generated. This digital image data signal is sent to the display memory 530 via the CPU 501.
Transferred to. The image data transferred to the display memory 530 is immediately displayed on the display unit 520 via the display unit driver 540.

When the release button 511 is pressed (step S500), the CPU 501 starts the bracket recording operation. The CPU 501 reads the previously stored movement amount from the non-volatile memory 501c and substitutes it into the variable Z secured in the RAM 501b (step S501). Similarly, the CPU 501 reads the previously stored number of shots (the number of shots) from the non-volatile memory 501c and substitutes it into the variable N secured in the RAM 501b (step S502).

Here, as an example, it is assumed that the movement amount Z and the number N of shots are Z: 4 [μm] and N: 3, respectively.

The CPU 501 clears a counter C (not shown) for counting the number of shots to 0, and starts shooting (step S503). Here, the stage position is X. However, in FIG. 1, the upward direction is positive and the downward direction is negative.
Based on an instruction from the CPU 501, the shutter 300 via the shutter drive circuit 310 or an electronic shutter (not shown) opens and closes in accordance with an appropriate exposure time, so that the camera head 8 displays an observation image in a stopped state. You can shoot. This image data is transferred to the display memory 530 and displayed on the display unit 520 as captured image data.
On the other hand, the CPU 501 writes this image data in the removable medium 551 via the memory reading / writing device 552, and the removable medium 551.
Add one file to the directory entry of
The shooting of the first image is finished (step S505). CPU5
01 increments the counter C by 1 (step S506). That is, C: 1.

Subsequently, the CPU 501 compares the number of shots N with the counter C to determine whether or not a predetermined number of shots have been finished (step S507). According to the series of description so far, the number of captured images is N: 3 and the counter is C: 1. Comparing the two, since N ≠ C,
The result of the determination in step S507 branches to No.

The CPU 501 drives the stage up-and-down mechanism section 15 via the stage drive unit 13, and the stage 1
In the downward direction in FIG. 1, the stage movement is started by the movement amount Z: 4 [μm] (step S508).
The CPU 501 waits for a stage movement end signal from the stage driving device 13 (step S509). When the stage movement is completed and a stage movement end signal is input from the stage driving device 13 to the CPU 501, step S50
The end determination of 9 branches to Yes. The process is step S
Returning to 504, the CPU 501 starts shooting again. Since the stage has moved by the amount of movement Z: 4 [μm] in step S508, the stage position here is
It is X-4 [μm].

Thereafter, the above sequence is repeated until the counter C: 3 is reached. Since the stage is moved twice until the observation image is captured three times, the counter C:
The stage position at time 3 is X-8 [μm]. When the counter C is 3, the end determination in step S507 is branched to Yes, and a series of bracket shooting in the bracket recording mode ends.

As described above, according to this apparatus, it is possible to photograph a predetermined number of observation images while automatically shifting the focus position with respect to the observation image by a predetermined amount by one release operation. Therefore, it becomes relatively easy and easy to focus the observation image prior to the start of photographing. Since the operator who focuses the observation image of the microscope does not require much experience and high skill based on heat training, even an inexperienced operator can obtain a high-quality photographed image without failure. Further, it becomes possible to provide a more comfortable shooting operation even when focusing on an image display unit such as an LCD of several inches which is widely used. Furthermore, even if the sample has relatively large unevenness and thickness with respect to the depth of focus of the objective lens used, a high quality photographed image can be obtained by one release operation.

Next, an image capturing apparatus for a microscope according to the second embodiment of the present invention will be described. Since the device configuration is the same as that of the first embodiment, the description of the device configuration according to the second embodiment will be omitted. Since the operation is basically the same as that of the first embodiment, the different operation between the first embodiment and the second embodiment is emphasized.
explain.

The operation of the bracket recording mode according to the second embodiment will be described below with reference to the flow chart shown in FIG. First, when the release button 511 is pressed (step S600), the CPU 501 starts the bracket recording operation.

The CPU 501 is a revolver type lens barrel unit 1.
The revolver position of the objective lens currently used for observation is read from a detection unit (not shown) provided in 6 and the read revolver position is substituted for the variable P secured in the RAM 501b (step S601). Here, it is assumed that the revolver type lens barrel portion 16 is a five-hole revolver, and an objective lens is attached to each revolver position.

FIG. 7 shows an example of a table showing the magnification and the depth of focus of the objective lens mounted at each revolver position.
As shown in FIG. 7, an objective lens having a magnification of 5 and a depth of focus of 12 [μm] is attached to the revolver position of 1. Similarly, the revolver position: 2 has an objective lens with a magnification of 10 and a focal depth of 3 [μm], and the revolver position: 3 has a magnification of 20 and a focal depth of 1.3 [μm].
Objective lens, revolver position: 4, magnification: 50
And an objective lens having a focal depth of 0.4 [μm] and a revolver position of 5, a magnification of 100 and a focal depth of 0.3.
Objective lenses each having a size of [μm] are mounted. Hereinafter, the bracket recording operation will be described on the assumption that an objective lens as shown in FIG. 7 is attached to the revolver.

First, assume that the current revolver position P is P: 2. In addition, the nonvolatile memory 50
It is assumed that the depth of the unevenness or the depth of the observation target stored in 1c, that is, the target depth D is 10 [μm].

It can be read from the table of the objective lens shown in FIG. 7 that the depth of focus FD (P) of the objective lens 6 mounted at the revolver position P: 2 is 3 [μm]. In this case, the target depth D is 10 [μm], whereas the focal depth FD (P) of the objective lens 6 is 3
Since it is [μm], it is difficult to cover the unevenness of the observation target at one focus position.

The CPU 501 substitutes FD (P) into the variable Z secured in the RAM 501b (step S60).
2). Thereby, the CPU 501 sets the depth of focus FD (P) of the objective lens 6 used for observation as the movement amount. That is, the movement amount is Z: 3 [μm].

Next, the CPU 501 calculates the number of shots N by adding 1 to the integer part of the value obtained by dividing the target depth D by the depth of focus FD (P). The calculation formula is as follows. N = 1 + D / FD (P) (integer operation) ... Equation 1 Here, when D: 10 and FD (P): 3 are actually substituted into Equation 1, N = 4. The CPU 501 is the RAM 5
The calculation result is substituted into the variable N secured in 01b (step S603). The sequence after step S604 is the same as that after step S503 of the first embodiment, and thus the description thereof is omitted.

As described above, according to the second embodiment, when continuously photographing a predetermined number of observation images while shifting the focus position with respect to the observation image by a predetermined amount, the depth of focus of the objective lens and the unevenness of the sample to be observed are obtained. Alternatively, it is possible to rationally determine at least one of the suitable amount of movement of the stage and the number of shots based on the depth. As a result, it is possible to obtain a high-quality photographed image without making a mistake with a smaller number of operations and a smaller number of photographed images.

In the second embodiment, the movement amount Z is set as the depth of focus of the objective lens attached to the revolver, and the number N of shots is calculated by the above equation 1. By the way, once various objective lenses are attached to the revolver, it is usually necessary to replace these objective lenses.
Very rare. Therefore, it is also preferable to store in the non-volatile memory 501c a table that uniquely determines the movement amount Z and the number of captured images N according to the revolver position P.
Alternatively, even when the objective lens is replaced, the same effect can be obtained by simply re-registering the movement amount Z and the number of captured images N in the nonvolatile memory 501c. By storing the movement amount Z and the number of captured images N in the non-volatile memory 501c, the logic becomes simpler, and further improvement in the reliability and processing speed of the device is expected.

Next, an image capturing apparatus for a microscope according to the third embodiment of the present invention will be described. Since the device configuration is the same as that of the first embodiment, the description of the device configuration according to the third embodiment will be omitted. Since the operation is basically the same as that of the first embodiment, the operation different from that of the first embodiment and that of the third embodiment will be emphasized.
explain.

The operation of the bracket recording mode according to the third embodiment will be described below with reference to the flow chart shown in FIG. The sequence from steps S800 to S806 is the same as steps S500 to S of the first embodiment.
The same as 506. First, when the release button 511 is pressed (step S800), the CPU 501 starts the bracket recording operation. The CPU 501 reads the previously stored movement amount from the nonvolatile memory 501c and substitutes it into the variable Z secured in the RAM 501b (step S801). Similarly, the CPU 501
The number of shots stored in advance is stored in the nonvolatile memory 5
It is read from 01c and is substituted into the variable N secured in the RAM 501b (step S802).

Here, as an example, it is assumed that the movement amount Z and the number of captured images N are Z: 4 [μm] and N: 3, respectively.

The CPU 501 clears the counter C for counting the number of shots to 0 (step S803), and starts shooting (step S804). Hereinafter, for the sake of explanation, it is assumed that the stage position at this point is X. As in the first embodiment, the upward direction is positive and the downward direction is negative in FIG.

Based on an instruction from the CPU 501, the shutter 300 via the shutter drive circuit 310 or an electronic shutter (not shown) opens and closes in accordance with an appropriate exposure time, so that the camera head 8 is in a stopped state. An image of the observation image can be taken. This image data is transferred to the display memory 530 and displayed on the display unit 520 as captured image data.

On the other hand, the CPU 501 writes this image data in the removable medium 551 via the memory reading / writing device 552, adds one file to the directory entry of the removable medium 551, and takes the first image. finish. (Step S80
5). Subsequently, the CPU 501 increments the counter C by 1 (step S806). This gives C: 1.

The CPU 501 controls the number of shots N and the counter C.
Compare with. According to the explanations so far, N: 3,
Since C: 1, N ≠ C, and as a result of the end determination in step S806, the process branches to No. Then the CPU
501 drives the stage up-and-down mechanism part 15 via the stage drive device 13 to start moving the stage 1 downward in FIG. 1 by a movement amount of N: 4 [μm] (step S808). ). The CPU 501 waits for a stage movement end signal from the stage driving device 13 (step S809). When the stage movement is completed and a stage movement end signal is input from the stage drive device 13 to the CPU 501, the end determination in step S809 is Y.
Branch to es.

Next, the CPU 501 controls the nonvolatile memory 5
01c or the standby time T stored in advance in the ROM 501a, the timer is set based on the acquired standby time T, and this timer is started (step S810). This standby time may be set in consideration of the time required until the vibration of the mechanical transmission mechanism caused by the movement of the stage is attenuated to the extent that it does not affect the desired quality of the captured image. good. Here, the waiting time T is, for example, T: 100 [m
s].

The CPU 501 waits for the waiting time T, that is,
Wait for 100 [ms] to elapse (step S81
1). When the waiting time T has elapsed, the time-up determination of step S811 branches to Yes, and the CPU 501
Photographing is started again (step S804). Since the stage is moved by the movement amount in step S808, the stage position in this case is X-4 [μm].
Has become. After that, the above sequence is repeated until the counter C: 3 is reached. When the counter C is 3, the end determination in step S807 branches to Yes, and a series of bracket shooting in the bracket recording mode ends.

As described above, according to the third embodiment, when a predetermined number of observation images are photographed while shifting the focus position with respect to the observation image by a predetermined amount by one release operation, the vibration accompanying the movement of the stage is generated. Do not start the next shot until it has decayed. As a result, even when the observation image is captured using the high-magnification objective lens, the effect of vibration of the mechanical transmission mechanism can be removed, and a high-quality captured image can be obtained without failure.

Modifications of each embodiment will be described below. According to the first, second and third embodiments of the present invention described above, when the focus position is shifted by a predetermined amount, the stage movement is performed as shown in FIG.
In the above, the direction is downward, but is not limited to this. Further, when the focus position is shifted by a predetermined amount, the observation images may be sequentially captured at positions moved in both up and down directions around the stage position X in the first image capturing. For example,
The observation image may be sequentially captured by controlling the stage to move to a position X + 4 [μm] moved upward and a position X-4 [μm] moved downward. Here, when the movement of the stage is in the upward direction, so-called limit limiting control may be performed to limit the moving position of the stage. As a result, it is possible to realize a safer image capturing apparatus for a microscope, which can prevent accidental destruction and failure of the objective lens and the specimen.

Further, for example, in the above description, in order to shift the focus of the observed image, the stage is moved via the stage up-and-down mechanism. However, instead of moving the stage, a portion on which the objective lens is mounted may be moved by a mechanism unit (not shown).

[0074]

According to the present invention, since the photographing device automatically photographs the observation image a predetermined number of times while changing the focus position of the observation image, many operators who focus the observation image of the microscope are many. There is no need to have a high level of skill based on the experience and skill. As a result, even an inexperienced operator can obtain a good quality captured image without failure.

Even when the depth of focus of the objective lens to be used is small (shallow) with respect to the unevenness of the specimen to be observed, the operator only needs to perform bracket photographing once to obtain a good quality photographed image. Can be obtained. Also,
At least one of the number of times of shooting and the amount of changing the focus position can be reasonably determined according to the performance of the objective lens, so that the operator can obtain a high-quality shot image with less number of times of shooting without failure. It becomes possible to acquire.

In shooting, by providing a waiting time between changing the focus position and shooting, it is possible to eliminate the influence of the vibration of the mechanical transmission mechanism caused by changing the focus position. Therefore, the operator can obtain a high-quality captured image without making a mistake.

[Brief description of drawings]

FIG. 1 is an overall configuration diagram of a microscope including an image capturing device according to a first embodiment of the present invention.

FIG. 2 is a configuration diagram of a camera head.

FIG. 3 is a configuration diagram of a controller.

FIG. 4 is an example of a front view of an operation input unit.

FIG. 5 is a flowchart showing processing in a bracket recording mode according to the first embodiment of the present invention.

FIG. 6 is a flowchart showing processing in a bracket recording mode according to the second embodiment of the present invention.

FIG. 7 is an example of a table showing the performance of the objective lens mounted at each position of the revolver.

FIG. 8 is a flowchart showing processing in a bracket recording mode according to the third embodiment of the present invention.

[Explanation of symbols]

1 stage 2 light sources 3 ND filter 4 Field stop 5 condenser lens 6 Objective lens 7 eyepiece 8 camera head 13 Stage drive 15 Stage up / down mechanism 16 Lens barrel 21 Sampling circuit 22 A / D converter 23 Image Processor 100 aperture stop 110 Aperture diaphragm drive 120 Aperture stop adjustment mechanism 200 Imaging lens 300 shutters 310 shutter drive circuit 400 image sensor 500 controller 501 CPU 510 Operation input section 511 Release button 512a, 512b, 512c Mode selection buttons 513a, 513b Objective lens switching button 514a, 514b Aperture diaphragm adjustment button 515a, 515b Index button 516 memory button 517 Jog dial 518 Index display 520 display 530 Display memory 540 Display driver 550 memory device 551 removable media 552 memory read / write device 800 revolver drive T specimen S step

Front page continuation (51) Int.Cl. ⁷ Identification code FI theme code (reference) G03B 3/12 G03B 3/12 13/34

Claims (5)

[Claims] 1. A photographing apparatus having a function of continuously photographing observation images obtained by observing a sample through an objective lens, wherein the photographing device is provided between a mounting means for mounting the sample and the objective lens. The driving means for adjusting the focus position of the observation image by changing the distance of the observation image, and the image photographing means for photographing the observation image, the image photographing means and the driving means, during the continuous photographing, Changing the focus position by changing the distance between the placing means and the objective lens by the driving means after photographing by the photographing means is repeated until the observation image is photographed a predetermined number of times. Shooting device. 2. At least one of the predetermined number of times and the amount of movement when changing the distance between the placing means and the objective lens is determined according to the performance of the objective lens. The imaging device according to claim 1. 3. A predetermined waiting time is provided after the distance between the placing means and the objective lens is changed by the driving means and before the image is taken by the image taking means. The imaging device according to claim 1 or 2. 4. A photographing method for continuously photographing observation images obtained by observing a specimen through an objective lens, wherein the observation image is photographed, and the specimen and the objective lens are photographed. Changing the focus position of the observation image by changing the distance between the observation image and the observation image until the observation image is captured a predetermined number of times. 5. An image photographing means for photographing an observation image obtained by observing a specimen through an objective lens, and a focus position of the observation image is adjusted by changing a distance between the specimen and the objective lens. In order to perform continuous photographing of the observation image by a device provided with a driving means,
A program that causes a computer to control each unit, outputs a signal instructing to capture the observation image to the image capturing unit, increments the number of times the capturing is performed by 1, Outputting a signal instructing to change the focus position of the observation image to the driving unit by changing the distance from the objective lens, is repeated until the number of times reaches a predetermined number, A program characterized by being executed by a computer.