JPH0375508A - Incorporation of data for reconstructing stereoscopic image from continuous section - Google Patents

Abstract

PURPOSE:To make it possible to align preparations efficiently by sequentially incorporating the position data of a plurality of the preparations in a broad range as the positions on the coordinates of a world. CONSTITUTION:Preparations are set at a microscope 10. A moving stage 12 is operated. An intended image is displayed on a TV monitor 16 through a frame memory 18. A cursor is displayed on the monitor 16 with a signal from a computer 20. A suitable position is selected, and a reference point is inputted. Said reference point is made to be the original point of the X-Y plane of world. coordinates. The coordinate Z of the world coordinates is determined by the interval between the sections of the preparation. Then, the position data are inputted, and the cursor on a screen is moved with a mouse 26 and digitizer 28 and the like. Thus the intended position is selected in the image that is displayed on the monitor 16. Here, the computer 20 can grasp the position on the monitor 16 as the position on the world coordinates by the signals and the like from the stage 12.

Description

[Detailed Description of the Invention] [Industrial Application Fields] This invention relates to a method for capturing data for reconstructing a three-dimensional image from serial sections for reconstructing a three-dimensional image by observing human tissues, cells, etc. with a microscope, Especially regarding this efficiency.

[Conventional technology] Traditionally, in the field of basic medicine, human organs, bones,
Three-dimensional images of cells, etc. are being reproduced. Normally, a plurality of continuous thin slices are first formed from the object to be measured, and each slice is observed one by one using a microscope as a preparation. The observer then recognized the three-dimensional connections from the observation results of each image, sketched them, and reconstructed the three-dimensional image. On the other hand, the observation results of these microscopes are also being quantified and handled. That is, a method is used in which a two-dimensional image of a cross section is measured and interpolated into a three-dimensional quantity as a cylinder, sphere, or spheroid by circular or elliptical approximation.

It has also been proposed to reconstruct a three-dimensional image by inputting various image position data for a plurality of slices into a personal computer.

[Problem to be solved by the invention] In the above-mentioned method of constructing a three-dimensional image from the observation results of multiple slides, the recognition of three-dimensional connections is entirely left to human imagination and intuition, and the construction is difficult. The problem was that the 3D images depended on the experience and abilities of the individual observer.

Further, the conventional method using circular or ellipse approximation has a problem in that it cannot handle cases where the three-dimensional shape is complex. Furthermore, in the conventional method of quantifying using a personal computer, etc., it is possible to sequentially import position data for one field of view observed with a microscope, but when the object to be observed protrudes from the field of view, the preparation is moved and the field of view is When the materials are changed, it is not possible to grasp the corresponding materials, it is not possible to capture data from a wide field of view, and it is impossible to reconstruct a three-dimensional image.

This invention was made with the aim of solving the above-mentioned problems, and it is a three-dimensional image that can capture data for a three-dimensional image with the same standard even when the slide is moved or when the slide is replaced. The purpose of this invention is to provide a method for capturing image data.

[Means for Solving the Problems] In order to achieve the above object, the present invention includes a step of mounting and fixing a slide on a moving stage that is movable within a predetermined plane at an observation position of a microscope, and A step of displaying an image of the preparation on a screen, a step of selecting an arbitrary point from among the images displayed on this screen and determining a reference point, and a step of determining the position of the moving stage and the screen when this reference point is determined. A process of determining the origin and coordinate axes of the world coordinates that serve as the reference for all position data from the position of the reference point above, a process of outputting the position signal of the moving stage, and a process of looking at the image displayed on the screen above. The process of moving the cursor displayed on the screen to a location corresponding to the outline of the image and sequentially inputting data about that position to import positional data about the outline, and transferring the positional data on this screen to the moving stage. A step of converting the position data into world coordinates using a position signal from the controller, a step of exchanging the slide to be observed, and a step of superimposing the image of the exchanged slide and the position data input for the previous slide on the screen. and a process of matching the position and direction of the reference point for the images before and after the exchange by comparing both displayed images, and from the position of the moving stage and the position of the reference point on the screen at this time. The present invention is characterized by comprising the step of recognizing the position on the screen after exchanging the slides as a position on world coordinates, and sequentially capturing a wide range of position data regarding a plurality of slides as positions on world coordinates.

[Function] In the present invention, an image of a slide obtained by a microscope is displayed on a screen.

Even when the slide is moved, the position on the screen can always be taken in as position data in world coordinates, which is the reference for all position data, using position data from the moving stage.

Furthermore, when replacing the slide, the position data obtained from the previous observation is displayed superimposed on the current image. Therefore, it is possible to align the replaced slides while displaying both.

Therefore, position data for a stereoscopic image can be captured in a wide field of view as position data based on the same standard in world coordinates for each of a plurality of slides.

[Example 1] FIG. 1 is a block diagram showing the configuration of a system to which the stereoscopic image data acquisition method according to the present invention is applied.

In this system, the object to be measured is divided into a predetermined number of slices, preparations are prepared for each slice, and the plurality of preparations are sequentially observed using a microscope 10, as in the conventional example.

Here, the position of the stage used for mounting the preparation in this microscope 10 can be detected by a moving stage 12.

Then, as shown in FIG. 1A, the slide 40 is
It is rotatably attached to a moving stage 12 by a sample holder 9. Therefore, by rotating the sample holder 9, the image observed by the microscope 10 can be rotated, and the x and y positions of the moving stage 12 can be rotated.
The observation position can be adjusted by moving in the direction. Also,
Movement of the moving stage 12 is detected by a sensor 13. Further, the microscope 10 can change the display magnification using a revolver 11.

The image observed by the microscope 10 is photographed by a television camera 14 and displayed on a television monitor 16. Here, the moving stage 12 and the television camera 14 are mounted parallel to each other so that when the moving stage 12 moves in the X direction, the image moves horizontally on the television monitor 16, and the coordinate axes of the world coordinates are determined. .

Note that the RGB signals, which are color signals obtained by the television camera 14, are once stored in the color frame memory 18, and then supplied to the television monitor 16 and displayed there.

A computer 20 is connected to the color frame memory 18 so that data from the computer 20 can be displayed on the television monitor 16. Furthermore, this computer 20 has a hard disk 22 and a floppy disk 24 as external storage devices.
In addition to signals from the movable stage 12, the human power means include a mouse 26, a digitizer 28, and so on. It has a keyboard 30.

The computer 20 is a personal computer with a memory capacity of about 512 kilobytes, the hard disk 22 is 40 megabytes, and the color frame memory 18 is 1600 megabytes.
About a thousand colors are used.

When entering data in such a system,
First, a slide is set on the microscope 10. and,
The moving stage 12 is operated to display a desired image on the television monitor 16, and a cursor is displayed on the television monitor 16 in response to a signal from the computer 20. Then, select an appropriate position and input the reference point. This reference point serves as the reference for all position data to be input from now on, and normally this point is set as the origin (0, O) of the XY plane of the world coordinates. All data entered manually from now on will be recognized as relative coordinates to this origin (0, O).

The Z coordinate of the world coordinates is determined from the intercept interval of the slide. In other words, the interval between the intercept of one slide and the intercept of the next slide is 2 in world coordinates.
This is the difference in coordinates on the Z axis obtained from the two slides.

To input position data, move the cursor on the screen using the mouse 26,
This is done by selecting a desired position from among the images displayed on the television monitor 16 while moving the image using the digitizer 28 or the like. That is, mouse 26. Digitizer 28 or keyboard 30 input signals provide signals to computer 20 about the current cursor position. This position data is normally contour data for the displayed image, and by inputting the position data while sequentially moving the cursor along the contour, the contour is stored in the computer 20 as point sequence data. Note that on the screen, each manual force of the position data is displayed as a polygonal line connecting those points.

Here, the data regarding the cursor position is regarding the position on the screen of the image at that time, that is, the position of the pixels forming the screen. This is then converted into relative coordinates from the world coordinate origin mentioned above.

Therefore, it is difficult to operate the moving stage 12 or use the revolver.
11 to change the display magnification, the cursor position data will no longer correspond to the position on the slide.

Therefore, in this invention, when one breparat is set and the above-mentioned reference point is determined, the position of the reference point is set in world coordinates based on the position of the moving stage at that time, the position of the reference point on the screen, and the display magnification. It is recognized as the origin (0,0). Then, as the moving stage 12 moves, it outputs position data at that time, but this position data corresponds to the position relative to the origin of the world coordinates, that is, the coordinates relative to the above-mentioned reference point on an l-to-l basis. . Therefore, the computer 20 can always grasp the position of the image displayed on the television monitor 16 as the position on the world coordinates based on the signal from the moving stage 12 and the display magnification regardless of its movement or change in magnification.

In particular, in the present invention, the image is magnified by a microscope 10. Therefore, the computer 20 correlates both the position data regarding the image displayed on the television monitor 16 and the above-mentioned position data from the moving stage, taking into account the presence of the microscope filo.

Therefore, this will be explained based on FIG. 2.

In the figure, the slide 40 is, for example, 8 cm wide and 2 cm tall.
．． It has a size of about 5 cm.

A sample area 42 several cm square is located near the center.
is set. Here, the size of this sample area 42 is assumed to be LXLmm. The field of view of the microscope 10 is a specific field of view 44 within this sample area 42.
becomes.

This visual field range 44 is magnified by the microscope 10, and the image is obtained as a pixel signal by the television camera 14. That is, the horizontal direction 512 . of the television camera 14 corresponds to this viewing range 44 . 480 pixel signals in the vertical direction are obtained. Then, this (7) 512×480 pixel signal is stored in the frame memory 18.

The data about the pixels stored in frame memory 18 is supplied to computer 20 where it is converted into coordinates within the entire sample area 42 and stored. Therefore, according to the apparatus of the present invention, the sample area 42
It is possible to grasp the position data at any point within.

That is, the computer 20 has world coordinates corresponding to the entire sample area 42, and data regarding each position in the viewing range 44 is recognized as coordinates within this world coordinate.

Here, this association will be explained in more detail.

First, the positions of predetermined alignment reference points in the slide 40 are assumed to be (X oo, Y oo).

Further, the center position of the objective lens of the microscope at the same coordinates is assumed to be (xo, yo). Then, the center position of this objective lens is set at the center of the television monitor 16.

Since the size of the screen is (512,480), the center position (Xo, Yo) of the screen is (25
6,240).

Here, if the position signal of the moving stage is regarded as the coordinates of the center point of the screen, (X 00' y DO) is the position signal of the moving stage when the reference point is brought to the center of the screen. Also, if the size of one pixel determined by the microscope magnification is Aμm, any point on the slide (x,
y) is the coordinate on the screen (X.

Y), the following equation is obtained.

(X, Y)-” (Xo, Yo) +A (X-256, Y-240>
Then, the alignment reference point is set to (0.
0), the length Lav+ is 20.00 in world coordinates
If set to 0, any point (X, Y) on the screen becomes a point (X, Y) on the world coordinates using the following equation.

y). Here, the coordinate system of the slide is μ
The unit is m.

20.000 0 ”” ((Xo -Xoo-”10 Yoo
) +A (X-258, Y-240) If the Z coordinate of the 111th slide is 0, then the Z coordinate of the second slide is 0 2 !, if the interval between slides is ΔZ in the world coordinate system. ΔZ Generally, if the Z coordinate of the immediately preceding slide is Zn-1 and the interval between slides is 628 m, then 0 z −2+ ΔZ n n-1 is obtained.

The length Lmm is set to 20,000 in the world coordinate system because when a 16-bit computer is used, the range of single-precision integers that can be handled is approximately ±32,000.

In this way, according to this embodiment, position data can be inputted not only for the viewing range 44 of the microscope 10 but also for the entire sample area 42 as position data based on the same reference. Therefore, data acquisition for one slide 40 can be completed in one observation,
Data capture efficiency can be dramatically increased.

Next, when the preparation 40 is replaced, consistency is maintained as follows. That is, the slide 40 is observed in order for each slice. Therefore, the distance between the previous measurement object and the next measurement object is input from the keyboard to obtain the Z coordinate of the new measurement object.

Then, the observer can match the coordinates before and after the exchange by searching for the reference point in the current image with reference to the position data manually entered last time and matching the reference points.

Further, the moving stage 12 in this embodiment is capable of not only moving in the X and Y directions but also rotating. Therefore, the observer searches for the reference point in the observed image and
Match this with the previous reference point. By rotating the moving stage 12, even that direction can be matched.

When the moving stage 12 is rotated, the subsequent movement of the moving stage 12 will be oblique on the screen. That is, when rotated by an angle θ, the moving stage 12 moves in a direction shifted by θ as the XSY axis. Therefore, the subsequent position data from the movable stage also needs to be converted in consideration of this angle θ, but this can be processed by the computer 20.

Here, in order to perform this conversion in the computer 20, the angle θ must be recognized.

This angle θ can be used if the moving stage 12 can output an angle.
The movement of the moving stage 12 after rotation may be recognized by how it moves on the screen. That is, the movement of the moving stage 12 in the X direction after rotation by angle θ is expressed as (h cos θ, hs1n
This is because the movement is θ). Therefore, by such an operation, the computer can recognize the rotation angle θ, and can convert the subsequent movement data of the moving stage 12.

Note that since rotating the moving stage 12 and rotating the television camera 14 mean the same thing, the television camera 14 may also be rotated. Furthermore, line drawings based on position data that have already been input can be rotated through processing by the computer 20.

Therefore, the line drawing may be rotated and oriented by human power at the rotation angle θ from the keyboard 30 or the like.

Furthermore, when the moving stage 12 is moved, the subsequent movement becomes oblique, resulting in poor operability, and there is a problem in that the angle must be estimated when moving the line drawing. For this reason,
After aligning the orientation by rotating the movable stage 12 and recognizing the rotation angle by the operation described above, the movable stage 12 may be returned and the line drawing may be rotated by the recognized angle θ.

Further, positioning can be performed by rotating the preparation 40. That is, the preparation is set free on the sample holder, rotated, and locked after positioning. According to this, the computer 20
There is no need to change the processing in .

Further, in the apparatus of the present invention, the previously input position data can be displayed on the television monitor 16 as a broken line connecting the input points. Therefore, an image of this broken line will be displayed superimposed on the image you are currently observing, so
By adjusting the moving stage 12 while looking at this, it is possible to match the reference point and direction of both.

Here, a plurality of position data regarding the previously input contour are normally scattered throughout the sample area 42.

Therefore, very highly accurate alignment can be achieved by aligning all of these manual data. Note that the reference points may be aligned after the direction is aligned by rotating the moving stage 12.

Next, when changing the magnification of the objective lens, do so as follows. First, the moving stage 12 is adjusted so that the location on the screen to be enlarged is centered. Next, change the objective lens and display the image with a different magnification on the TV monitor 16.
to be displayed.

Normally, this center point is a fixed point that does not move at all even when the objective lens is changed, so the position of this center point in the world coordinates is not changed when the objective lens is changed. For this reason, if you input the fact that the magnification has been changed into the computer 20 after changing the magnification, the subsequent data input by moving the point on the screen will not process the fact that the distance for each pixel has been changed. This can be done by the computer 20, and the position in world coordinates can be input as correct data.

Furthermore, when displaying the position data for each slide 40 stored in the computer 20 on the screen, the magnification on the screen can be arbitrarily changed by a predetermined calculation. Therefore, when observing at a predetermined magnification, position data that has already been input can be displayed at the magnification at that time.

Furthermore, when aligning when changing the slide 40, it is more efficient to display a plurality of already input position data at a low magnification. Therefore, preparation 40
When replacing the image, it is best to first perform alignment before and after replacement using a low-magnification screen, and then perform high-magnification observation.

Furthermore, it is preferable to display the inputted data by changing the color, such as blue for the outline of the two previous slides, yellow for the previous slide, and red for the outline of the manual slide. In this way, a plurality of pieces of slide data that have already been input can be displayed, and changes in the position of the measurement object whose outline has been input can be easily recognized.

On the other hand, in the apparatus of this embodiment, when inputting a contour line, the outer contour is manually input as a series of points around the right. That is, by arranging the position data manually, it is possible to specify the side with the most content within the outline.

If there is a hole inside, input the outline of the hole as a counterclockwise point sequence following the outside outline. This allows you to input objects such as a tissue that is filled with something, an object that has a hole inside, or an abstracted cortex that is empty inside. Note that it is best to input only one side of the contour to the cortex, and automatically generate and input a sequence of input points in the opposite direction for the other side. In addition, if there is a filling in a hole, it is recommended to automatically generate the same dot sequence as the hole in a reverse layer to create the outer outline of the filling.

Further, the data regarding each pre-parameter) 40 input in this manner is held in the hard disk 22, etc., which is an external storage device of the computer 2θ. Disk capacity f
In the case of f140 megabytes, the number of slices can be about 1000 without any manual effort.

Then, a three-dimensional image is constructed from these data according to a predetermined program, and in this case, the following modes can be selected.

(a) Network model mode Displaying the contour line and connecting representative points (centroids) of the contour line to display connection relationships, branches, etc. of tissues.

(b) Attach a vertical wall surface with a height equal to the slice interval to the cylindrical model mode contour line,
This is for displaying shading. This mode is effective for displaying a measurement target where the slice interval is thinner than the cross section.

(c) Solid model mode A wall surface is generated from the contour line, and shading is displayed using Gouraud shading. Usually, the light source and viewpoint are set at an infinity point in the Z direction. Then, high-speed display is performed by parallel projection.

(d) Transparent display mode Transmittance can be set for each object, allowing you to see parts hidden inside or behind the object.

Furthermore, according to this embodiment, various measurements can be performed simultaneously.

That is, it is possible to perform three-dimensional measurements of the volume of the body with the hole removed, the volume of the hole, the surface area, the length, etc.

In addition, the position on the object coordinates, the distance between two points, the cross-sectional area,
It is also possible to measure the peripheral length, average wall thickness, elliptical contour long axis diameter, contour elliptical short axis diameter, etc.

[Effects of the Invention] As explained above, according to the 3D image data acquisition method according to the present invention, the observation magnification can be increased for the entire surface of the sample, detailed contour input can be performed, and the previous contour can be displayed at the position where the slide was replaced. The data can be displayed together on the screen, and the positioning and orientation of the slides before and after replacement can be very efficiently performed.

[Brief explanation of drawings]

FIG. 1 is a block diagram showing a configuration example of a system to which the stereoscopic image data acquisition method according to the present invention is applied; FIG. 1A is a schematic diagram showing the configuration of a slide mounted on the microscope 10 of FIG. 1; The figure is an explanatory diagram showing the correspondence of position data in the same embodiment. 10... Microscope 11... Revolver 12... Moving stage 14... Television camera 16... Television monitor 18... Frame memory 20... Computer 22... Hard disk 26... Mouse 28 ... Digitizer 30 ... Keyboard 40 ... Preparation system and true vertical column Fig. 1 Fig. 1A

Claims (1)

[Claims] Three-dimensional image data acquisition in which a plurality of preparations of continuous slices of a measurement object are formed into a plurality of thin slices, and a three-dimensional image is reconstructed from the observation results of the plurality of preparations with a microscope. The method includes a step of mounting and fixing the slide on a moving stage that is movable within a predetermined plane at an observation position of the microscope, a step of displaying an image of the slide obtained by the microscope on a screen, and a step of displaying the image on the screen. The process of selecting an arbitrary point from among the images and determining the reference point, and determining the world that becomes the reference for all position data from the position of the moving stage and the position of the reference point on the screen when this reference point is determined. The process of determining the coordinate origin and coordinate axes, the process of outputting the position signal of the moving stage, and the process of moving the cursor displayed on the screen to the location corresponding to the outline of the image while looking at the image displayed on the screen above. a step of capturing positional data about the contour by sequentially inputting data about the position; a step of converting the positional data on this screen into positional data in world coordinates using a positional signal from the moving stage; and observation. The process of exchanging the slides to be replaced, the process of superimposing the image of the replaced slide and the input position data of the previous slide on the screen, and comparing the two displayed images to determine the before and after exchange. A process of matching the position and direction of the reference point for the image, and a process of recognizing the position on the screen after slide exchange as the position on the world coordinates from the position of the moving stage at this time and the position of the reference point on the screen. A data acquisition method for reconstructing a three-dimensional image from continuous slices, comprising: sequentially acquiring a wide range of position data for a plurality of slides as positions on world coordinates.