JP2009294465A - Imaging device for microscope and objective micrometer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an imaging device for a microscope and an objective micrometer capable of reducing troublesomeness of calibration operation. <P>SOLUTION: This imaging device 2 for the microscope has an imaging means 21 for imaging a sample image observed by an observing means and outputting the observed image, a dimension information acquiring means 23 for selecting at least one pattern for calibrating dimension positioned close to the outermost peripheral part in an imaging range of the imaging means from among one or a plurality of patterns for calibrating dimension included in the imaging range and dimension information patterns arranged close to one or a plurality of patterns for calibrating dimension, respectively, and acquiring the dimension information of the pattern for calibrating dimension from the dimension information pattern corresponding to the selected pattern for calibrating dimension, and a dimension calibrating means 23 for calculating actual dimension per pixel from the acquired dimension information and the number of pixels of the pattern for calibrating dimension corresponding to the selected dimension information pattern. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a microscope imaging apparatus and an objective micrometer.

There is known an apparatus that takes an image of a sample observed with a microscope and measures an actual size of an arbitrary portion on the observed image. In this device, after taking an image of a ruler serving as a reference for length such as an objective micrometer for measurement, the user inputs two points on the image and inputs the length between the two points by the user. A calibration operation is performed to calculate the length per pixel from the number of pixels between two points in the captured image, and then the actual size between any two specified points on the observation image is used using the value per pixel. The actual dimensions are converted (for example, see Patent Document 1).
JP 2004-258495 A

  In the conventional apparatus, at the time of the calibration operation, the user needs to specify two points on the image and input the length and unit between the two points. When the objective lens is replaced and the magnification of the microscope is changed, the user needs to select an optimum scale according to the magnification of the image of the ruled image to be captured. For this reason, the calibration operation is troublesome for the user, and there is a problem that calibration errors frequently occur due to input errors.

  In order to solve the above-mentioned problems, the present invention captures a specimen image observed by an observation unit and outputs an observation image, and one or a plurality of dimension calibrations included in the imaging range of the imaging unit. Selecting at least one of the dimension calibration patterns located in the vicinity of the outermost peripheral portion in the imaging range from the pattern and the dimension information patterns respectively arranged in the vicinity of the one or more dimension calibration patterns; Dimension information acquisition means for acquiring dimensional information of the dimensional calibration pattern from the dimensional information pattern corresponding to the selected dimensional calibration pattern, and the dimensional calibration corresponding to the acquired dimensional information and the selected dimensional information pattern And a calibrating means for calculating an actual size per pixel from the number of pixels of the pattern image for use. That.

  Further, the present invention includes one or more dimension calibration patterns formed on the substrate, and dimension information patterns arranged in the vicinity of the one or more dimension calibration patterns, respectively. An objective micrometer is provided.

  ADVANTAGE OF THE INVENTION According to this invention, the imaging device for microscopes and objective micrometer which can reduce the troublesome of calibration operation can be provided.

  Hereinafter, an imaging device for a microscope and an objective micrometer according to an embodiment of the present invention will be described with reference to the drawings. The following embodiments are merely for facilitating understanding of the invention, and excluding additions and substitutions that can be performed by those skilled in the art without departing from the technical idea of the present invention. It is not intended.

  FIG. 1 is a schematic configuration diagram of a microscope photographing apparatus according to an embodiment.

  In FIG. 1, an imaging device 2 connected to the microscope 1 captures an observation image, which will be described later, of a sample 13 placed on the stage 12 of the microscope 1 with a camera unit 21. At this time, the objective lens 16 to be used is replaced by rotating the revolver 15, and the magnification of the observation image of the sample 13 is changed. An observation image captured by the camera unit 21 is sent from the camera unit 21 to the CCU 23 (Camera Control Unit) via the cable 22.

  The CCU 23 can display the observation image sent from the camera unit 21 on the monitor 3 and perform various image processing and measurement processing instructed by the user. At this time, the user operates the mouse 5 to instruct the image processing contents to be executed by the CCU 23.

  FIG. 2 is a functional block diagram of the CCU of the microscope imaging apparatus according to the embodiment.

  In FIG. 2, the observation image sent from the camera unit 21 via the cable 22 is input to the image processing IC 2301 of the CCU 23 and recorded in the image memory 2302. The image processing IC 2301 executes various kinds of image processing using the image information recorded in the image memory 2302, converts the image into a display image on the monitor 3 by the image composition unit 2303, and outputs the image to the monitor 3.

  The CCU 23 has a built-in flash memory 2304. When the user operates the mouse 5, the CPU 2305 reads the menu image component from the flash memory 2304 and sends it to the menu screen generation unit 2306. The menu screen generation city 2306 generates a menu image by combining menu image components, and sends the menu image to the image composition unit 2303. The image combining unit 2303 combines the observation image stored in the image memory 2302 and the menu image, and displays the combined image on the monitor 3 (see FIG. 3).

  The user operates the mouse 5 while viewing the observation image and the menu image displayed on the monitor 3, so that the CPU 2305 changes image processing parameters or performs measurement processing.

  For example, when the user operates the mouse 5 to perform the measurement processing of the observation image and selects the measurement menu, the CPU 2305 reads the calibration determination flag from the memory 2307 and determines whether the calibration determination flag is set. to decide. If the calibration determination flag is cleared, it is determined that calibration for setting an actual dimension per pixel has not been performed, and the CPU 2305 executes calibration for the user as shown in FIG. An item indicating this is displayed on the monitor 3. Here, FIG. 3 shows an example of a screen for instructing the user to perform calibration.

  When calibration is performed according to the procedure to be described later, a calibration determination flag is set and stored in the memory 2307, and the CPU 2305 displays on the monitor 3 a screen for instructing execution of the calibration result item and the observation image as shown in FIG. indicate. Here, FIG. 5 shows an example of a measurement execution instruction screen.

  Next, the calibration operation will be described with reference to FIG. FIG. 4 shows a state in which an objective micrometer for calibration is imaged by the camera unit 21 and displayed on the monitor 3.

  When the calibration execution instruction screen shown in FIG. 3 is displayed, the user replaces the sample 13 on the stage 12 of the microscope 1 with the objective micrometer 100 (the ruler) on which the dimension calibration pattern is formed. The objective micrometer 100 may be arranged in advance at a predetermined position on the stage 12 to drive the stage 12 and position it in the optical path of the objective lens 16 to acquire an observation image. The observation image may be acquired by exchanging with the above.

  The image of the objective micrometer 100 shown in FIG. 4 has a plurality of concentric patterns a, b, c and d. The concentric patterns a, b, c, d are such that the pattern a close to the center has a small diameter D so that it can correspond to the objective lens 16 having various magnifications, and the radius D increases as the distance from the center increases. It is formed to become. That is, since the field diameter is small when the magnification of the objective lens 16 is high, the pattern a having a small diameter D is used. When the magnification of the objective lens 16 is low, the field diameter is large. Use as Thus, the accuracy of the actual dimension per pixel is increased by selecting the dimension calibration pattern corresponding to the field diameter.

  A two-dimensional code (also referred to as a QR code) including dimensional information such as the value of the diameter D of each of the patterns a, b, c and d and the unit thereof is provided in the vicinity of the dimension calibration patterns a, b, c and d. Patterns ap, bp, cp, dp are arranged.

  In the observation image shown in FIG. 4, the CPU 2305 shown in FIG. 2 instructs the scale detection unit 2308 to detect a reference scale. The scale detection unit 2308 is the largest (outside) pattern (of FIG. 4) among the images of the patterns a to d in the imaging range of the camera unit 21 from the observation image of the objective micrometer 100 stored in the image memory 2302. In this case, the pattern d, which will be described below using the pattern d), is detected as a dimension calibration pattern d, and the number of pixels having a diameter D of the pattern d is sent to the CPU 2305. At this time, a colored display (for example, a red or red broken line) is displayed so that the user can easily recognize the selected pattern d.

  Next, the CPU 2305 instructs the two-dimensional code detection unit 2309 to detect the two-dimensional code dp. The two-dimensional code detection unit 2309 detects the two-dimensional code dp inside (center side) near the outermost pattern d of the observation image, and sends it to the two-dimensional code decoding unit 2310. The two-dimensional code decoding unit 2310 sends the result of decoding the two-dimensional code dp to the CPU 2305. The CPU 2305 calculates an actual dimension per pixel from the diameter D value and unit of the pattern d acquired from the two-dimensional code dp and the number of pixels corresponding to the diameter of the pattern d, and the actual dimension is displayed on the menu image shown in FIG. L value and unit are displayed.

For example, when the number of pixels corresponding to the diameter of the pattern d is φ, the diameter D value of the decoding result of the two-dimensional pattern pd is N, and the unit (μm), the calibration result L per pixel is as follows.
L (μm / pixel) = N / φ

  Note that the pattern d and the two-dimensional code dp may be determined by color, or may be determined by luminance by embedding substances having different reflectivities.

  The calibration is completed as described above, and the CPU 2305 displays the calibration result and measurement items as shown in FIG. Also, a calibration determination flag is set and stored in the memory 2307.

  Next, the user returns the sample 13 to the stage 12 of the microscope 1 and displays an observation image of the sample 13 on the monitor 3. Then, the user designates two points for measuring the distance with the mouse 5 on the observation image. The CPU 2305 reads the pixel coordinates of the two points, converts them into actual distances (actual dimensions) using the calibration result L, and displays the results on the monitor 3 (see FIG. 6).

For example, assuming that the pixel coordinates of two points designated by the mouse 5 are (x1, y1) and (x2, y2) and the calibration result is L (μm / pixel), the actual distance d between the designated two points d (Μm) is
d (μm) = ((x2−x1) ² + (y2−y1) ² ) ^1/2 × L
It becomes.

  FIG. 6 shows an image display on the monitor 3 after calculation of a measurement result between two designated points (indicated by a solid line in the figure). In order to display the measurement result on the menu, the CPU 2305 reads out the measurement result and the parts for creating the measurement result screen from the flash memory 2304 and sends them to the measurement result screen creation city 2311. The measurement result screen creation unit 2311 generates a measurement result screen by combining components, and sends the measurement result image to the image composition unit 2303. The image synthesis unit 2303 synthesizes the observation image stored in the image memory 2302, the measurement result image, and the menu screen creation image, and displays the synthesized image on the monitor 3.

  When it is desired to save the observation image and the measurement result, the observation image and the measurement result information are linked and saved in the memory 2307 by clicking the image saving button with the mouse 5. This makes it possible to search for a stored image from a menu screen (not shown) and redisplay it.

  7A and 7B show a modification of the objective micrometer. FIG. 7A shows an example in which the dimension calibration pattern is formed in a linear shape, and FIG. 7B shows an example in which the dimension calibration pattern is formed in a square shape. Respectively.

  FIG. 7A shows linear patterns a, b, c, d4 having different distances from the origin O, and two-dimensional codes ap, bp, cp, dp including distance and unit information in the vicinity of each pattern. Is a pattern for dimensional calibration. The two-dimensional code of the origin O can be acquired by op. At the time of calibration, an observation image is displayed on the monitor 3 as shown in FIG. 4, and the outermost pattern (for example, pattern d) among the dimensional calibration patterns a to d and the two-dimensional code dp of this pattern are used. The actual size per pixel is determined according to the above. At this time, the distance between d and d may be used as the dimension of the pattern, or the distance from the origin O to the pattern d may be obtained from the two-dimensional code op and dp. It is also possible to obtain the actual dimension per pixel using distance information other than between d and d using the respective two-dimensional code information.

  Further, FIG. 7B has quadrangular patterns a, b, c, and d4 having different distances from the origin O, and two-dimensional codes ap, bp, cp, and dp including respective distances and unit information. This is a dimensional calibration pattern. The two-dimensional codes of these patterns include information on the length of one side of the rectangle and the length of the diagonal of the rectangle as distance information. The two-dimensional code of the origin O can be acquired by op. At the time of calibration, an observation image is displayed on the monitor 3 as shown in FIG. 4, and the outermost pattern (for example, pattern d) among the dimensional calibration patterns a to d and the two-dimensional code dp of this pattern are used. The actual size per pixel is determined according to the above. At this time, the distance between d and d may be used as the dimension of the pattern, or the distance from the origin O to the pattern d may be obtained from the two-dimensional code op and dp. It is also possible to obtain the actual dimension per pixel using distance information other than between d and d using the respective two-dimensional code information.

  Needless to say, patterns having various shapes other than the above can be formed and used for the dimension calibration pattern. In the above embodiment, the case where the calibration operation is performed at the time of observing the sample 13 has been described. However, the calibration operation is performed in advance on the objective lens 16 disposed on the revolver 15 of the microscope 1, and as a result, Is stored in the memory 2307, the objective lens 16 inserted in the optical path is detected from the rotational position of the revolver 15 with the microscope 1, and the CPU 2305 uses the calibration result corresponding to the objective lens 16. Also good. Thereby, it is possible to omit the calibration operation when the objective lens 16 is replaced.

  As described above, according to the embodiment, the dimensional calibration pattern is automatically selected by the imaging device, and the dimension, unit information, and the like of the pattern are read from the two-dimensional code. It is not necessary to perform complicated operations such as specifying two points each time as a reference, inputting a distance between two points, and a unit.

  In addition, since the objective micrometer according to the embodiment has a plurality of dimensional calibration patterns having different dimensions, it can easily cope with a magnification change such as replacement of an objective lens.

  In addition, since the imaging apparatus can read the actual dimension two-dimensional code of the dimension calibration pattern, the imaging apparatus can be applied to an optical apparatus to which the imaging apparatus can be attached regardless of the microscope.

  Further, the dimension information of the dimension calibration pattern is set as character information, and the two-dimensional code detection unit 2309 and the two-dimensional code decoding unit 2310 in the CCU 2305 are changed to a character information detection unit and a character information decoding unit, respectively. It is also possible to determine the actual dimensions.

FIG. 1 is a schematic configuration diagram of a microscope photographing apparatus according to an embodiment. FIG. 2 is a functional block diagram of the CCU of the microscope imaging apparatus according to the embodiment. An example of the screen which instruct | indicates to perform a calibration to a user is shown. The state which imaged the objective micrometer for calibration with the camera part, and was displayed on the monitor is shown. An example of a measurement execution instruction screen is shown. An image display of a monitor after calculation of a measurement result between two designated points (indicated by a solid line in the figure) is shown. The modification of an objective micrometer is shown, (a) shows an example in which the pattern for dimension calibration is formed in linear form, (b) shows an example in which the pattern for dimension calibration is formed in quadrilateral form, respectively.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Microscope 2 Imaging device 3 Monitor 5 Mouse 12 Stage 13 Sample 15 Revolver 16 Objective lens 21 Camera part 22 Cable 23 Imaging device 100 Objective micrometer 2301 Image processing part 2302 Image memory 2303 Image composition part 2304 Flash memory 2305 CPU
2306 Menu screen creation unit 2307 Memory 2308 Scale detection unit 2309 Two-dimensional code detection unit 2310 Two-dimensional code decoding unit 2311 Measurement result screen creation unit O Origin a, b, c, d Dimensional calibration patterns ap, bp, cp, dp, op 2D code pattern

Claims (9)

An imaging means for capturing a specimen image observed by the observation means and outputting an observation image;
Among the one or more dimension calibration patterns included in the imaging range of the imaging means and the dimension information pattern arranged in the vicinity of the one or more dimension calibration patterns, the most within the imaging range. Dimension information acquisition means for selecting at least one of the dimension calibration patterns located in the vicinity of the outer periphery and acquiring the dimension information of the dimension calibration pattern from the dimension information pattern corresponding to the selected dimension calibration pattern;
Dimensional calibration means for calculating an actual dimension per pixel from the acquired dimension information and the number of pixels of the dimension calibration pattern image corresponding to the selected dimension information pattern. apparatus.   2. The microscope imaging apparatus according to claim 1, wherein the dimension calibration pattern is formed as a concentric pattern, and the dimension information pattern includes diameter information of the concentric pattern. The dimension calibration pattern is formed from an origin pattern and a plurality of rule patterns based on the origin pattern,
The microscope imaging apparatus according to claim 1, wherein the dimension information pattern of each ruler pattern includes distance information from the origin pattern.   The imaging apparatus for a microscope according to any one of claims 1 to 3, wherein the dimension information pattern includes a two-dimensional code. Display means for displaying the observed image;
Position selection means for designating two points in the observation image displayed on the display means;
5. The apparatus according to claim 1, further comprising arithmetic means for calculating an actual size between the two points from the number of pixels between the two points designated by the position selection unit and the actual size. 2. An imaging apparatus for a microscope according to item 1. One or more dimensional calibration patterns formed on the substrate;
An objective micrometer comprising: a dimension information pattern arranged in the vicinity of the one or more dimension calibration patterns.   The objective micrometer according to claim 6, wherein the dimension calibration pattern is formed as a concentric pattern, and the dimension information pattern includes diameter information of the concentric pattern. The dimension calibration pattern is formed from an origin pattern and a plurality of rule patterns based on the origin pattern,
The objective micrometer according to claim 6, wherein the dimension information pattern of each ruler pattern includes distance information from the origin pattern.   The objective micrometer according to claim 6, wherein the dimension information pattern includes a two-dimensional code.