JP2009223164A - Microscopic image photographing apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a microscopic image photographing apparatus, wherin an appropriate control range including a specimen is set through automatic focus adjustment, by automatically detecting the thickness of a slide glass specimen before the automatic focus adjustment is made, and to provide a photographing method and a program. <P>SOLUTION: In synchronization with the conveyance of a slide glass 10 to a stage 9 from a slide glass accommodation unit 16, the thickness of a slide glass specimen is detected by a thickness detection unit 60. Based on the detected thickness, the range of the focus control is determined, and the focus control is performed using this range. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a technique for taking a microscope image of a slide glass specimen.

2. Description of the Related Art Conventionally, there has been known a microscopic image photographing apparatus that magnifies a biological specimen such as a human cell sample with a microscope and photographs the microscopic image.
To observe or photograph a specimen with a microscope, first place the specimen on the slide glass on the microscope stage, set the specimen under the objective lens, and focus on the specimen (microscopic image). Observe and shoot.

  By the way, photographing of microscope images performed in a hospital or laboratory is performed on a large number of slide glass specimens, and it is desired to efficiently capture a huge amount of microscope images.

  For example, Patent Document 1 includes an automatic conveyance unit that automatically conveys a slide glass specimen to a microscope stage and captures a microscope image, and acquires a macro image and label information of the slide glass specimen. A microscope image photographing apparatus is disclosed, and a large number of slide glass specimens can be automatically captured.

Patent Document 2 discloses a technique for automatically recognizing a specimen region from an image of an entire slide glass specimen, and only a necessary area with a slide glass specimen can be automatically photographed.
JP 2003-248176 A JP 2000-295462 A JP-A-8-210818

  However, the thickness varies depending on the type of the slide glass, and the thickness of the slide glass specimen also varies. Therefore, automatic focus adjustment may not be performed correctly when observing and photographing a slide glass specimen.

  In order to prevent this, if the focus adjustment range is widened, the focus position where there is no sample is searched for focus, so that the efficiency is low, and there is a possibility that the focus is on dust that is not a sample.

To cope with this, it is only necessary to know the thickness of the specimen.
For example, Patent Document 3 discloses a technique of a microscope apparatus provided with a unit for measuring depth data of a transparent specimen on a microscope stage. It is possible to get.

However, Patent Document 3 does not describe any means for efficiently focusing the specimen image using the measured thickness data.
In view of the above problems, the present invention provides a microscope image photographing apparatus, photographing method, and program capable of automatically detecting the thickness of a slide glass specimen before performing automatic focus adjustment and setting an appropriate control range with the specimen by automatic focus adjustment. The purpose is to provide.

  The microscope image photographing apparatus of the present invention is a microscope image photographing apparatus including a microscope image photographing unit that photographs a specimen microscope image obtained by enlarging a specimen on a slide glass, and detects the thickness of the slide glass specimen to be photographed. A slide glass specimen thickness detector; and a focus control method determination unit that determines a focus control range of the automatic focus controller based on the slide glass specimen thickness data detected by the slide glass specimen thickness detector; And an automatic focus control unit for determining a focus position for photographing based on the focus control range.

  A photographing method according to the present invention is a photographing method by a microscope image photographing device provided with a microscope image photographing unit for photographing a specimen microscope image obtained by enlarging a specimen on a slide glass, and the thickness of the slide glass specimen to be photographed is determined. Detecting and determining a focus control range based on the thickness data of the slide glass specimen, and determining a focus position for photographing based on the focus control range.

  Further, the program according to the present invention is a program executed by a microscope image photographing apparatus including a microscope image photographing unit that photographs a specimen microscope image obtained by enlarging a specimen on a slide glass, and the thickness of the slide glass specimen to be photographed Determining a focus control range based on thickness data of the slide glass specimen, and causing the microscope image capturing apparatus to determine a focus position for imaging based on the focus control range. Features.

  According to the present invention, the thickness of the slide glass specimen is automatically detected before performing the automatic focus adjustment, and the automatic focus adjustment is performed based on the thickness. Therefore, the processing by automatically adjusting the focus within the minimum necessary range is performed. Time can be shortened, and errors due to the in-focus position outside the detection range of automatic focus adjustment can be prevented.

A first embodiment of the present invention will be described below with reference to the drawings.
FIG. 1 is a diagram showing an overall configuration of the microscope image photographing apparatus of the present embodiment.
The microscope image photographing apparatus 1000 in FIG. 1 is roughly divided into a microscope photographing mechanism 500 for photographing a slide glass specimen and a large number of slide glass specimens, and the observation position of the microscope photographing mechanism 500 based on an instruction. And a slide glass transport mechanism 501 that transports the microscope unit 500 and a computer 502 that controls the microscope unit 500 and the slide glass transport mechanism 501.

First, the configuration of the microscope photographing mechanism 500 will be described.
The microscope photographing mechanism 500 includes a transmission illumination light source 2 made of, for example, a halogen lamp or an LED, and the transmission illumination light source 2 generates illumination light. As shown by the broken line in FIG. 1, the illumination light is first collected by the collector lens 3, and then passes through various filters 4, such as an ND filter or an LBD filter, and the field stop 5 in this order, and then by the mirror 6. It is deflected toward the stage 9.

  The illumination light deflected in the direction of the stage 9 by the mirror 6 passes through the aperture stop 7 and the condenser lens unit 8 and then passes through an illumination opening (not shown) of the stage 9, so that the slide glass on the stage 9. The whole is configured to illuminate the specimen S on 10.

  A revolver 12 holding a plurality of objective lenses 11 is disposed above the stage 9, and the revolver 12 is rotated in any direction forward and backward as indicated by an arrow A in the figure, whereby an objective with a desired magnification is obtained. The position of the lens 11 can be exchanged to the observation position. These objective lenses 11 can be detachably exchanged with respect to the revolver 12. Accordingly, not only the revolver 12 is rotated, but also the objective lens 11 itself attached to the revolver 12 can be replaced to obtain a desired magnification.

  The stage 9 is configured to be movable in the horizontal direction in order to change the observation position of the sample S. Further, the stage 9 is configured to be driven up and down in the Z direction (vertical direction) as indicated by a double arrow D in the figure so that the focus control can be performed by the focus drive unit 100.

  Note that the focus control of the microscope image capturing apparatus 1000 in FIG. 1 is not limited to that realized by the raising / lowering drive of the stage 9, and is realized by a configuration in which the revolver 12 holding the objective lens 11 is raised and lowered in the Z direction. Needless to say.

  Further, the brightness of the image may vary depending on the shooting position on the slide glass 10. The brightness control for such an image is performed by the CPU 20 of the computer 502 controlling the camera unit 14 to fix the exposure. It is possible to perform control such as making an overall adjustment by correcting the brightness level by software.

  The specimen image of the slide glass 10 incident on the objective lens 11 positioned on the optical axis in the observation optical path in this way is branched by the beam splitter 103 through the intermediate magnification lens 13, and the autofocus detection unit 101 and the camera unit 14. The entire microscope photographing mechanism 500 is configured to be guided to

The sample image captured by the camera unit 14 is compressed by the image processing board 37 according to the specification of the JPEG format and converted into digital data.
In the autofocus detection unit 101, the entire or part of the luminance distribution of the specimen image is input as an electrical signal to the autofocus circuit 102, and the contrast state is digitized as a contrast value. The autofocus circuit 102 instructs the focus drive unit 100 to maximize the contrast value and drives the specimen up and down.

  In the microscope image photographing apparatus 1000 of the present embodiment, the focus drive range for performing the autofocus control is instructed from the computer 502 based on the slide glass thickness data described later. Therefore, the focus drive range is a range based on the thickness of the slide glass, and the focus position of the sample image can be efficiently focused by quickly finding the focus position to be photographed.

  In the above description, the contrast value is obtained by the autofocus detection unit 101. However, the image processing board 37 calculates the contrast value from the sample image obtained by the camera unit 14 and performs autofocus control. It is also possible to do.

Next, the configuration of the slide glass transport mechanism 501 will be described.
The slide glass transport mechanism 501 includes a slide glass storage unit 16 that can store a plurality of slide glasses, and a slide glass transport unit that transports the slide glass between the slide glass storage unit 16 and the stage 9 based on instructions from the computer 502. 17 is provided.

  From the slide glass storage unit 16, a predetermined slide glass 10 is taken out by the slide glass carrying unit 17, and the taken slide glass 10 is carried to the stage 9.

  The slide glass storage unit 16 includes a thickness detection unit 60 that detects the thickness of the slide glass being conveyed. When the slide glass 10 is taken out by the slide glass transport unit 17, the thickness detection unit 60 measures the thickness of the slide glass specimen.

  As the thickness detection unit 60, for example, an ultrasonic measuring device as disclosed in JP-A-3-262909 or an air servo as disclosed in JP-A-10-160420 can be considered. Further, a glass boundary surface using an optical distance measuring method applying a triangulation method as disclosed in JP-A-6-011341 or a confocal optical system as disclosed in JP-A-2005-98833. If the thickness detection unit 60 by the method of measuring the difference in the distance of the laser light reflected by the lens is used, the thickness of each of the slide glass part, specimen part, and cover glass part constituting each layer of the slide glass specimen is measured. Is possible.

  Further, as shown in FIG. 2, the reflected light sensor is used as the thickness detection unit 60, and the thickness of the slide glass is adjusted in conjunction with the operation of mapping the position of the slide glass stored in the slide glass storage unit 16. It may be detected.

  The slide glass photographed by the microscope photographing mechanism 500 is taken out from the stage 9 by the slide glass transport unit 17 and stored in a predetermined position of the slide glass storage unit 16.

Next, the configuration of the computer 502 will be described.
Transport of the slide glass 10 by the slide glass transport unit 17, measurement of the slide glass specimen thickness by the thickness detection unit 60, taking a microscope image by the camera unit 14, other movements of the stage 9 left and right and up and down, automatic focusing, etc. The operation of the microscope is performed under the control of the CPU 20 of the computer 502.

  The control program recorded on the recording medium is read by the medium access device 22 and loaded into the memory 21 or the large-capacity recording medium 23, and the CPU 20 controls each unit according to the loaded control program. An operation control screen display program for displaying an operation control screen on the operation monitor 53 is also installed in the control program so that the operator can easily control the microscope image photographing apparatus 1000.

  The operator operates the keyboard 55 or a pointing device 56 such as a mouse to input necessary instructions, thereby operating the microscope photographing mechanism 500, transporting a slide glass, taking a microscope image, and capturing a wide-field image. Execution such as imaging can be instructed. Each unit described above and each unit to be described are connected to the CPU 20 via an interface circuit (hereinafter referred to as I / F circuit) and a dedicated driver for each unit, and further via a CPU bus 49.

  The CPU 20 issues a control signal to each unit via each interface circuit connected to the CPU bus 49 to control each unit constituting the microscope image photographing apparatus 1000. For example, taking an example of the illumination light control of the microscope photographing mechanism 500, the analog voltage value changing circuit 44 is controlled by the CPU 20 via the light source control I / F 32, and the analog voltage value changing circuit 44 is illuminated under this control. The illumination light is controlled by changing the voltage of the lamp.

  Further, the CPU bus 49 includes a memory 21, a medium access device 22 for reading and writing programs and data from a portable storage medium such as a CD and a DVD, a large-capacity recording medium 23 such as a hard disk, a screen display memory 52, and an operation monitor 53. And an input control I / F circuit 54 for controlling an event of a pointing device 56 such as a keyboard 55 or a mouse.

  The microscope photographing mechanism 500 and the slide glass transport mechanism 501 are each provided with a dedicated driver (drive mechanism) for electrically driving each unit. These dedicated drivers are composed of, for example, a unit dedicated motor, a drive driver for the motor, a drive transmission system, and the like. These dedicated drivers are connected to the CPU 20 via a dedicated control I / F circuit and a CPU bus 49.

  For example, the revolver 12 is provided with a revolver rotation driver 38 including a revolver rotation motor, a motor driver, and a drive transmission system. Similarly, a condenser lens drive driver 41, an aperture stop driver 42, a field stop driver 46, various filter control drivers 45, a stage transfer unit driver 47, a slide glass transfer driver 48, and the like are connected to corresponding units.

  In addition, in the microscope image photographing apparatus 1000 of the present embodiment, a minimum number of sensors (not shown) are provided at various locations so that each unit can correctly perform operations such as movement and rotation to a predetermined position. Is provided. For example, the revolver 12 is provided with an objective optical axis position sensor so that it can be rotated and stopped correctly at the objective optical axis position.

Next, a configuration for conveying the slide glass will be described.
The stage 9 can be driven to slide in at least two directions of X and Y (left and right direction and depth direction in FIG. 1), and the slide glass is transferred between the slide glass transport unit 17 and the stage 9. It has a mechanism for glass slide delivery.

FIG. 3 is a perspective view schematically showing a detailed configuration of the slide glass transport mechanism 501.
As shown in the figure, the slide glass storage unit 16 is configured to store a plurality of slide glasses 10 (10-1, 10-2,..., 10-n).

A conveying suction device 86 is provided in the slide glass storage unit 16.
The conveying suction device 86 takes out the fried glass 10 accommodated in the slide glass storage unit 16 one by one by the suction nozzle and the suction pad at the tip thereof, and reaches the delivery position to the stage of the slide glass conveyance unit 17. Then, the slide glass 10 is moved as indicated by the arrow F in FIG.

  In this suction device for conveyance 86, a suction nozzle is connected to a vacuum pump (not shown) via a tube (not shown), and sucks and holds at least one slide glass 10 and moves to a position where it is delivered to the stage. It has an adsorption power to the slide glass 10 as much as possible. The stage 9 shown in FIG. 1 transfers the slide glass 10 to the optical axis position below the objective lens 11 on the right side of the figure.

  In addition, a thickness detection unit 60 including a condenser lens 84 and a wide-field imaging camera 85 shown in FIG. 1 is disposed in the slide glass storage unit 16. Is connected to an analog / digital signal processing device 61.

  As described above, when the slide glass 10 is taken out from the slide glass storage unit 16 by the conveying suction device 86, the thickness of the slide glass specimen is detected by the thickness detection unit 60. The position where the thickness detection unit 60 measures the thickness of the slide glass can be moved by the conveying suction device 86, and an arbitrary location on the slide glass can be measured. The CPU 20 stores and saves the measured slide glass specimen thickness data in a predetermined storage area of the large-capacity recording device 23.

Next, a first embodiment of an image capturing method by the microscope image capturing apparatus 1000 having the basic configuration as described above will be described.
FIG. 4 is a flowchart showing the processing operation in the first embodiment. The processing in the figure is realized by the CPU 20 of the computer 502 executing a control program.

In the figure, first, the CPU 20 causes the slide glass transport mechanism 501 to transport the slide glass onto the stage 9.
While viewing the control screen displayed on the operation monitor 53, the operator changes the pointing device 56 such as a keyboard 55 or a mouse, and takes a microscope image so as to take out an arbitrary slide glass 10 from the slide glass storage unit 16. Instruct the device. Based on this instruction, under the control of the CPU 20, the designated slide glass 10 is taken out from the slide glass storage unit 16 and is placed under the objective lens 11 on the optical axis by the slide glass transport unit 17 and the stage 9. It is transferred (step S1).

  Subsequently, the CPU 20 causes the thickness detection unit 60 to detect the thickness of the slide glass specimen (S02). This process is performed in conjunction with the removal of the slide glass 10 in step S1. That is, the CPU 20 controls the thickness detection unit 60 shown in FIG. 2 simultaneously with the conveyance control of the slide glass 10 to detect the thickness of the slide glass specimen, and the thickness data (= TS) is recorded on the recording medium 22. Save to. Thus, the entire processing time until the sample image is captured is not increased by the thickness measurement processing of the slide glass sample in step S2.

Next, in step S3, the CPU 20 calculates a search range for automatic focus adjustment (autofocus) for the slide glass specimen.
Here, it is assumed that the in-focus position (= F0) at the reference slide glass thickness (= T0) is detected in advance during calibration of the apparatus and stored in the large-capacity recording medium 23. The CPU 20 reads T0 and F0 from the large-capacity recording medium 23 and the thickness data TS detected in step S2, and obtains the in-focus position FS of the sample from the following equation 1.

(Formula 1) FS = F0 + (TS−T0)
When the thickness data TS is obtained, the CPU 20 sets the focus width set in advance by the operator as the search range with respect to the focus driving unit 100 with the focus position FS of the sample as the center. In step S4, the CPU 20 searches for a focus position where the contrast value of the autofocus detection unit 101 is maximized while driving the stage 9 in this search range.

  At this time, it is necessary to automatically recognize the position of the specimen on the slide glass specimen before the automatic focus adjustment, and to move the stage 9 so that the position is below the objective lens 11, but this specimen is automatically recognized. For example, the technique disclosed in Japanese Patent Laid-Open No. 2000-295462 is used.

When the focused sample image is obtained as a result of the automatic focus adjustment process in step S4, the CPU 20 causes the camera unit 14 to execute image capture.
When capturing images at a plurality of positions on the slide glass specimen, after moving the stage 9, the focus automatic adjustment in step S4 is performed again within the search range obtained in step S3, and the image capturing in step S5 is repeated. . When all the images have been captured, the CPU 20 instructs the stage 9 to move the slide glass specimen to the delivery position, and instructs the slide glass transport unit 17 to take out the slide glass specimen from the slide glass storage unit 16. Transport to the original position.

  In this way, automatic capture of a microscope image of one slide glass specimen is completed. In step S7, the CPU 20 determines whether or not there is a slide glass specimen that captures an image next. If there is a slide glass specimen that captures an image (Yes in step S7), the process returns to step S1. The above-described processing of steps S1 to S6 is similarly performed on the next slide glass specimen, and is repeated until all of the slide glass specimens to be captured are finished. When the image capture of all the slide glass specimens is completed (No at Step S7), this process is finished.

  As described above, according to the first embodiment, the thickness of the slide glass specimen is detected before the automatic focus adjustment for the specimen, and the search range for the automatic focus adjustment is determined based on the thickness. The efficiency and accuracy of adjustment can be improved.

Next, a second embodiment will be described.
In the second embodiment, a thickness detection sensor capable of measuring the thickness of a plurality of layers of a transparent body is used to measure the thickness of each slide glass, specimen portion, and cover glass of a slide glass specimen. A microscope image photographing apparatus capable of automatically setting the center and width of a search range for focus adjustment is realized.

  In the second embodiment, as the thickness detection sensor 60, a sensor capable of detecting each thickness from reflected light from each layer of a transparent body made of a plurality of layers is used. As such a thickness detection sensor 60, for example, there is an optical distance measurement method applying a triangulation method as disclosed in the above-mentioned Japanese Patent Laid-Open No. 6-011341, and Japanese Patent Laid-Open No. 2005-98833. It is conceivable to use a method for measuring the difference in the distance of the laser light reflected from the glass interface using such a confocal optical system.

  FIG. 5 is a flowchart showing the processing operation in the second embodiment. The processing in the figure is also realized by the CPU 20 of the computer 502 executing the control program.

  In the process shown in FIG. 4, the slide glass 10 is first transported onto the stage 9 (step S11) in the same manner as in step S1 of the first embodiment shown in FIG. 4, and the thickness of the slide glass specimen is interlocked with this transport. Is detected (step S12).

  In step S <b> 12, the CPU 20 reads the thicknesses TP, TA, and TC of the slide glass, the specimen portion, and the cover glass of the slide glass specimen from the thickness detection sensor 60 and stores them in the large-capacity recording medium 23.

  Next, the CPU 20 reads the data T0, F0 stored in advance from the large-capacity recording medium 23 and the data TP, TA, TC detected in step S12, and the center FS of automatic focus adjustment from the following formulas 2 and 3. And the width FW of the search range is calculated.

(Formula 2) FS = F0 + (TP + TA + TC-T0)
(Formula 3) FW = TA
Then, the CPU 20 instructs the focus drive unit 100 to set the range of the focus width FW around the in-focus position FS of the sample as the search range (step S13).

  In step S14, the focus position where the contrast value of the autofocus detection unit 101 is maximized is searched while driving the focus in the search range set in step S13, and the focus position is determined.

  Since the subsequent steps S15 to S17 for capturing an image and storing the slide glass sample in the slide glass storage unit 16 are the same as the processes in steps S5 to S7 in FIG.

  As described above, according to the second embodiment, the thickness of the specimen is detected in addition to the thickness of the slide glass, and the center position and the search width of the search range for automatic focus adjustment can be automatically set by taking these into account. In addition, the efficiency and accuracy in automatic focus adjustment can be further improved by matching the search width with the sample.

Next, a third embodiment will be described.
In the third embodiment, when the thickness of the specimen is larger than the depth of focus of the objective lens, it is automatically determined to capture an image at a plurality of focus positions, and the focus position and the number of images can be automatically set. The microscope image photographing device is realized.

  FIG. 6 is a flowchart showing the processing operation in the third embodiment. The processing in the figure is also realized by the CPU 20 of the computer 502 executing the control program.

  The process of steps S21 and S22 for transporting the slide glass specimen to the stage 9 and acquiring the thickness data of the slide glass, specimen portion, and cover glass of the slide glass specimen, which is first performed in the process of FIG. 6, is the step of FIG. This is the same as the processing in S11 and S12.

  Next, the CPU 20 obtains the focal depth data (= FD) of the objective lens 11 stored in advance in the large-capacity recording medium 23 as step S23 and the specimen portion thickness data TA acquired in step S22 from the large-capacity recording medium 23. Read and compare the magnitude relationship between them.

  If the thickness TA of the specimen portion is smaller than the focal depth FD in step S23 (step S23, Yes), the CPU 20 determines that the specimen is sufficiently thin and that it is sufficient to capture a microscope image at only one focus position for focusing. In steps S24 to S29, automatic focus adjustment processing, image capture processing, and slide glass storage processing are performed. The processing in steps S24 to S29 is the same as the processing in steps S14 to S17 in FIG.

  In step S23, if the specimen portion thickness TA> focus depth FD (step S23, No), the CPU 20 determines that it is necessary to capture a plurality of microscope images in the focus direction because the specimen is thick. In step S26, the number of microscope images and the focus position are calculated.

  In this calculation, the CPU 20 reads the in-focus position F0 from the large-capacity recording medium 23 in addition to the thickness TA and the focal depth FD of the sample portion, and calculates the number NS and the focus position F (N) for taking a microscope image below. It calculates with the formulas 4 and 5 of these.

(Formula 4) NS = TA div FD + 1
(Formula 5) F (N) = F0 + ((NS-N) +1/2) * FD (N = 1, 2,..., NS)
Note that div in Equation 4 represents a division for rounding down values after the decimal point.

  Next, the CPU 20 controls the focus driving unit 100 as step S27, moves it to the focus position F (N) obtained in step S26, and captures a microscope image. This image capture is repeated until the image is captured at the NS focus position. Then, the slide glass specimen after image capture is transported to the slide glass storage unit 16 in step S28, and the same processing is performed on the subsequent slide glass specimen and the subsequent.

  As described above, according to the third embodiment, when the sample is thin, the image is captured at the in-focus position, and when the sample is thick, the image is automatically captured at a plurality of focus positions at the focal depth interval of the objective lens. A plurality of microscope images can be captured at every interval of the focal depth of the objective lens within the sample thickness, and the operability of the automatic focus adjustment setting is improved.

Next, a fourth embodiment will be described.
In the fourth embodiment, the thickness of the specimen is detected at a plurality of points on the slide glass specimen, the specimen thickness distribution of the whole slide glass is obtained based on the data, and appropriate automatic focus adjustment is performed at the shooting point. This makes it possible to realize a microscope image photographing apparatus that makes it possible to perform the above settings.

  FIG. 7 is a flowchart showing the processing operation in the fourth embodiment. The processing in the figure is also realized by the CPU 20 of the computer 502 executing the control program.

  In the process shown in the figure, first, the slide glass specimen in step S41 is transported to the stage 9, and when the thickness of the slide glass specimen is measured by the thickness detection unit 60 during the transport in step S42, a plurality of slide glass specimens are measured. The thickness data is acquired at the position of the position, and the coordinates (X, Y) (X, Y: horizontal coordinates) of the position and the thickness data are stored in the large-capacity recording medium 23.

FIG. 8 is a diagram showing an example of the position where the thickness data is measured in step S42.
In step S42, the coordinates (X, Y) and thickness data of a plurality of locations on the cover glass 71 of the slide glass specimen are acquired and stored in the large-capacity recording medium 23 as indicated by P1, P2, and P3 in FIG. .

Next, CPU20 calculates | requires the thickness distribution of the sample with respect to the horizontal position of a slide glass sample as step S43.
If the slide glass 10 and the cover glass 71 are sufficiently flat, the thickness data TS (X, Y) for an arbitrary position P (X, Y) of the slide glass specimen can be expressed by the following Expression 6.

(Formula 6) TS (X, Y) = A * X + B * Y + C (A, B, C: constant)
Here, the CPU 20 reads out the coordinates of P1, P2, and P3 obtained in step S42 and the thickness data of the slide glass specimen from the recording medium 22, and substitutes them into the equation 6 for simultaneous equations relating to the constants A, B, and C. By solving the above, Equation 6 for obtaining the thickness for an arbitrary position P can be obtained.

  Next, the CPU 20 moves the stage 9 to a position where a microscope image is photographed on the slide glass specimen, obtains the thickness TS (x, y) of the slide glass specimen relative to this position from Equation 6, and as step S44, The thickness TS (x, y) is compared with the focal depth data (FD) of the objective lens 11 recorded on the large-capacity recording medium 23 in magnitude relation.

  The relationship between the horizontal coordinate system (X, Y) on the slide glass transport unit 17 and the horizontal coordinate system (x, y) on the stage 9 where the thickness detection is performed by the thickness detection unit 60 is a microscope image photographing apparatus. Assume that it is obtained in advance at the time of adjustment of 1000.

  If the comparison result of step S44 indicates that the thickness TS (x, y) of the slide glass sample <the focal depth data (FD) of the objective lens 11 (step S44, Yes), after performing automatic focus adjustment as steps S45 and S46 Capture microscopic images. If the slide glass specimen thickness TS (x, y) <the focal depth data (FD) of the objective lens 11 is not satisfied (step S44, No), the number and position of images taken from the specimen thickness as steps S47 and S48. Is calculated, and images are captured at the number and position. Since the processes in steps S45 to S48 are the same as those in steps S24 to S27 of the third embodiment in FIG. 6, detailed description thereof is omitted.

  When the capture of the microscope image is completed, the slide glass 10 is stored in the slide glass storage unit 16 in step S49. Then, it is determined whether or not there is a slide glass specimen that captures an image next. If there is a slide glass specimen that captures an image (step S50, Yes), the process returns to step S41, and the next slide glass specimen is set. The processes in steps S41 to S49 described above are performed in the same manner, and are repeated until all the slide glass specimens to be captured are finished. When the image capture of all the slide glass specimens is completed (No at Step S50), this process is terminated.

  As described above, according to the fourth embodiment, an appropriate automatic focus adjustment can be set for an arbitrary horizontal position of a specimen, and the efficiency of automatic focus adjustment at an arbitrary position even when the thickness varies depending on the position of the specimen. And accuracy can be improved.

  As described above, according to the microscope image photographing apparatus 1000 of the present embodiment, the thickness of the slide glass specimen is automatically detected before performing the automatic focus adjustment, and the automatic focus adjustment is performed based on the thickness. It is possible to shorten the processing time by automatically adjusting the focus in the minimum range, and it is possible to prevent errors due to the in-focus position outside the detection range of the automatic focus adjustment.

  In the above example, the thickness measurement of the slide glass specimen is performed in conjunction with the conveyance of the slide glass 10. However, the timing for measuring the thickness of the slide glass specimen is not limited to this time. The measurement may be performed when the specimen is on the stage 9 or when the specimen is mounted on the slide glass storage unit 16.

It is a figure which shows typically the whole structure of the microscope image imaging device in this embodiment. It is a figure which shows the thickness detection unit which detects the thickness of a slide glass in response to the operation | movement which maps the position of the slide glass accommodated in the slide glass storage unit. It is a perspective view which shows typically the detailed structure of a slide glass conveyance mechanism. It is a flowchart which shows the processing operation in 1st Embodiment. It is a flowchart which shows the processing operation in 2nd Embodiment. It is a flowchart which shows the processing operation in 3rd Embodiment. It is a flowchart which shows the processing operation in 4th Embodiment. It is a figure which shows the example of the position where thickness data is measured.

Explanation of symbols

2 Light source for transmitted illumination 3 Collector lens 4 Filter 5 Field stop 6 Mirror 7 Brightness stop 8 Condenser lens unit 9 Stage 10 Slide glass 11 Objective lens 12 Revolver 13 Intermediate magnification lens 14 Camera unit 16 Slide glass storage unit 17 Slide glass transport unit 20 CPU
21 Memory 22 Medium Access Device 23 Large Capacity Recording Medium 24 I / F Circuit with Image Processing Board 25 Revolver Control I / F Circuit 26 Slide Glass Storage Control I / F Circuit 27 Slide Glass Conveyance Control I / F Circuit 29 Capacitor Unit Control I / F circuit 30 Brightness stop control I / F circuit 32 Light source control I / F
33 Filter control I / F circuit 34 Field stop control I / F circuit 35 Slide glass conveyance control I / F circuit 36 Slide glass storage control I / F circuit 37 Image processing board 38 Revolver rotation driver 39 Slide glass storage unit driver 40 Slide Glass transport unit driver 41 Condenser lens drive driver 42 Brightness stop driver 44 Analog voltage value change circuit 45 Various filter control drivers 46 Field stop driver 47 Slide glass transport unit driver 48 Slide glass storage unit driver 49 CPU bus 50 Camera unit control I / F circuit 52 Screen display memory 53 Monitor 54 Input control I / F circuit 55 Keyboard 56 Pointing device 60 Thickness detection unit 81 Slide glass tray 8 Suction device for moving in storage unit 83 Wide-field image shooting stage 84 Condensing lens 85 Wide-field image shooting camera 86 Transport suction device 100 Focus drive unit 101 Autofocus detection unit 102 Autofocus circuit 103 Beam splitter 500 Microscope imaging unit 501 Slide glass conveyance unit 502 Computer 1000 Microscope image photographing device

Claims (10)

A microscope image photographing device having a microscope image photographing unit for photographing a slide glass specimen,
A slide glass specimen thickness information detection unit for detecting thickness information of the slide glass specimen;
A focus control method determination unit for determining a focus control range of the automatic focus control unit based on the slide glass sample thickness information detected by the slide glass sample thickness information detection unit;
An automatic focus control unit for determining a focus position for shooting based on the focus control range;
A microscope image photographing device characterized by comprising: A slide glass specimen storage section for storing a plurality of slide glass specimens;
A slide glass sample transport unit that transports the slide glass sample from the slide glass sample storage unit under the microscope objective lens,
2. The microscope image photographing according to claim 1, wherein the slide glass specimen thickness information detecting unit detects thickness information of the slide glass specimen in conjunction with the slide glass specimen transport by the slide glass specimen transport unit. apparatus. A slide glass specimen storage section for storing a plurality of the slide glass specimens;
The microscope image photographing apparatus according to claim 1, wherein the slide glass specimen thickness information detection unit detects the thickness of the slide glass specimen in the slide glass specimen storage unit.   The slide glass specimen thickness information detection unit detects the thickness of the slide glass of the slide glass specimen, and the focus control method determination unit performs focus control of the focus automatic control unit based on the thickness of the slide glass. The microscope image photographing apparatus according to claim 1, wherein a range of the image is determined. The slide glass specimen thickness information detection unit detects the slide glass of the slide glass specimen, the cover glass and the thickness of the specimen therebetween,
2. The microscope image according to claim 1, wherein the focus control method determination unit determines a focus control range of the automatic focus control unit based on the thickness of the slide glass, the cover glass, and the specimen therebetween. Shooting device.   The focus control method determination unit compares the thickness of the specimen with the depth of focus of the objective lens imaged by the microscope imaging unit, determines whether to capture an image at a plurality of focus positions based on the comparison result, and performs focus control The microscope image photographing apparatus according to claim 5, wherein a focus position for capturing an image within the range is obtained.   The focus control method determination unit is configured to determine the number and position of microscope images to be acquired from the thickness of the specimen and the focal depth of the objective lens when the thickness of the specimen <the depth of focus of the objective lens. The microscope image photographing device according to claim 6.   The slide glass specimen thickness detection unit detects the thickness at three or more positions on the slide glass specimen, and the focus control method determination unit obtains the thickness distribution of the entire slide glass specimen from the detection result, 2. The microscope image photographing apparatus according to claim 1, wherein a focus control method at an arbitrary specimen position is determined. It is a photographing method by a microscope image photographing device equipped with a microscope image photographing unit for photographing a slide glass specimen,
Detect the thickness information of the slide glass specimen to be photographed,
Determine the range of focus control based on the thickness information of the slide glass specimen,
A focus position for shooting is determined based on the focus control range. A program executed by a microscope image photographing apparatus having a microscope image photographing unit for photographing a slide glass specimen,
Detect the thickness information of the slide glass specimen to be photographed,
Determine the focus control range based on the thickness information of the slide glass specimen,
A program for causing the microscope image photographing apparatus to execute determination of a focus position for photographing based on the focus control range.