JP4065129B2 - Color confocal microscope, measuring range selection method, and control program therefor - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a technique for selecting a predetermined region of a three-dimensional confocal microscope image having color information corresponding to a sample acquired by a color confocal microscope as a measurement range of the sample.
[0002]
[Prior art]
A confocal microscope illuminates a sample in a spot shape, collects light from the sample, for example, fluorescence, transmitted light, or reflected light, onto a pinhole, and receives the light passing through the pinhole with a photodetector. Thus, the surface information of the sample is acquired. Here, an example of the confocal microscope will be described.
[0003]
FIG. 11 is a diagram illustrating a schematic configuration example of a general confocal microscope.
In the figure, the light emitted from the light source 31 passes through the beam splitter 32 and then enters the two-dimensional scanning mechanism 33. The two-dimensional scanning mechanism 33 includes a first optical scanner (X scanner) 33a and a second optical scanner (Y scanner) 33b. The light beams are two-dimensionally scanned by scanning light with the respective optical scanners 33a and 33b. Scan. By guiding the light beam to the objective lens 34, the light beam incident on the objective lens 34 is condensed by the objective lens 34 and scanned two-dimensionally on the surface of the sample 35. The light from the surface of the sample 35 passes through the objective lens 34 again, is guided to the beam splitter 32 via the two-dimensional scanning mechanism 33, is reflected by the beam splitter 32, and is condensed on the pinhole 37 by the imaging lens 36. Is done. Since the pinhole 37 is at a position optically conjugate with the condensing position by the objective lens 34, the light from the sample 35 is collected on the pinhole 37 when the sample 35 is at the condensing position by the objective lens 34. When light passes through the pinhole 37 and the sample 35 is not located at the condensing position by the objective lens 34, the light from the sample 35 cannot be condensed on the pinhole 37 and therefore does not pass through the pinhole 37. Accordingly, the light from the sample 35 is cut by the pinhole 37 except for the focal point of the objective lens 34, and only the light passing through the pinhole 37 is detected by the photodetector 38. The sample 35 is placed on a sample table 40 corresponding to the X and Y stages, and can be moved in the optical axis direction by the Z stage 41. The two-dimensional scanning mechanism 33, the Z stage 41, and the photodetector 38 are controlled by a computer 39.
[0004]
With such a configuration, if the focal point is two-dimensionally scanned by the two-dimensional scanning mechanism 33 and the output of the photodetector 38 is imaged in synchronization with the two-dimensional scanning mechanism 33, only a specific height of the sample 35 is obtained. Is imaged, and an image (confocal microscope image) obtained by optically slicing the sample 35 is obtained. Further, the sample 35 is discretely moved in the optical axis direction by the Z stage 41, the two-dimensional scanning mechanism 33 is scanned at each position to acquire a confocal microscope image, and a photodetector 38 is obtained at each point on the sample surface. The surface shape information (height information) of the sample 35 is obtained by detecting the position of the Z stage 41 at which the maximum output is obtained, and a three-dimensional image (three-dimensional confocal microscope image) corresponding to the surface shape information is obtained. can get.
[0005]
In recent years, a color confocal microscope for colorizing the obtained three-dimensional image has been proposed in order to make it easy to understand the correspondence with an actual sample. For example, Japanese Patent Laid-Open No. 2001-82935 discloses a three-dimensional image corresponding to the surface shape information of a sample obtained by a confocal optical system, and a color image corresponding to the color information of the sample obtained by a non-confocal optical system. A color confocal microscope that obtains a color three-dimensional image (color three-dimensional confocal microscope image) by combining the above is disclosed.
[0006]
[Problems to be solved by the invention]
However, in such a color confocal microscope, a color three-dimensional image can be obtained, so that it is easy to understand the correspondence with the actual sample, but the sample is measured from the obtained color three-dimensional image. In such a case, effects such as reduction of the burden on the user (measurer, etc.) related to the measurement operation and improvement of measurement accuracy cannot be obtained. In particular, the burden on the user related to the operation when selecting the region of the obtained color three-dimensional image as the measurement range of the sample is still large.
[0007]
In addition, in a color confocal microscope, a technique for selecting a color three-dimensional image area based on color information (color) as a sample measurement range, and a technique for automatically selecting the area have not been proposed. .
In view of the above situation, an object of the present invention is to provide a color confocal microscope that reduces the burden on the user when selecting a region of a color three-dimensional image obtained from a color confocal microscope as a measurement range of a sample. It is to provide a measuring range selection method and a control program thereof.
[0008]
[Means for Solving the Problems]
According to the first aspect of the present invention, a first step of acquiring a three-dimensional confocal microscope image having color information corresponding to a sample, a second step of receiving an instruction of color information, and receiving in the second step A third step of selecting a region of the three-dimensional confocal microscope image having the color information acquired in the first step based on an instruction of the color information as a measurement range of the sample, is there.
[0009]
According to the above method, the area as the measurement range is selected by instructing the color information (color), and the burden on the user related to the selection of the measurement range is reduced.
According to a second aspect of the present invention, in the first aspect of the invention, in the second step, at least the content of RBG is instructed numerically or via a color sample, or a color map is obtained. Or at least one of density, luminance, hue, saturation, and lightness of color information is numerically indicated, or at least one of density, luminance, hue, saturation, and lightness of color information The range is indicated by a numerical value, and an instruction for color information is accepted, and in the third step, the three-dimensional image having the same color information as the color information indicated in the second step In this method, a region of a confocal microscope image is selected as a measurement range of the sample.
[0010]
According to this method, the color information is instructed by the content of RBG, color sample, color map, color information density, luminance, hue, saturation, brightness, range thereof, etc. The area is selected, and the burden on the user related to the measurement range selection is reduced.
[0011]
According to a third aspect of the present invention, in the first aspect of the invention, in the second step, the three-dimensional confocal microscope image having the color information acquired in the first step is displayed on the display unit, and the color information is displayed. As the instruction, an instruction of a point on the three-dimensional confocal microscope image having the color information displayed on the display unit is received, and in the third step, the color information displayed on the display unit is The region of the three-dimensional confocal microscope image having the same color information as the color information of the point indicated on the three-dimensional confocal microscope image or the color information within a predetermined range including the point, or the color information A region of only the point indicated on the three-dimensional confocal microscope image having or a region of only a predetermined range including the point is selected as the measurement range of the sample.
[0012]
According to this method, an area as a measurement range is selected by instructing a point on the three-dimensional confocal microscope image having the displayed color information, and the burden on the user related to the measurement range selection. Is reduced.
According to a fourth aspect of the present invention, in the first aspect of the invention, in the second step, a three-dimensional confocal microscope image having the color information acquired in the first step is displayed on a display unit, and a color is displayed. As an instruction of information, an instruction of a range on a three-dimensional confocal microscope image having the color information displayed on the display unit is accepted, and in the third step, the color information displayed on the display unit A region of the three-dimensional confocal microscope image having the same color information as the color information within the range indicated on the three-dimensional confocal microscope image having the indication, or an indication on the three-dimensional confocal microscope image having the color information In this method, a region having only a specified range is selected as the measurement range of the sample.
[0013]
According to this method, an area as a measurement range is selected by designating a range on the three-dimensional confocal microscope image having the displayed color information, and the burden on the user related to the measurement range selection is selected. Is reduced.
A fifth aspect of the present invention is the method according to any one of the first to fourth aspects, wherein in the second step, an instruction for a plurality of color information is received.
[0014]
According to this method, a plurality of color information can be designated.
According to a sixth aspect of the present invention, there is provided an objective lens that focuses light from a light source onto a sample, and the focusing position of the objective lens and the position of the sample are relative to each other along the optical axis direction of the light focused by the objective lens. Moving means, a micro aperture arranged at a position conjugate with the focusing position of the objective lens, a photodetector for detecting the intensity of light passing through the micro aperture, and detection by the photo detector Confocal microscope image acquisition means for acquiring a three-dimensional confocal microscope image of the sample based on the intensity of the light, color information acquisition means for acquiring color information of the sample surface, and confocal microscope image acquisition means A color confocal microscope image for acquiring a three-dimensional confocal microscope image having color information based on the three-dimensional confocal microscope image acquired by the color information and the color information acquired by the color information acquisition means A three-dimensional confocal microscope image having color information acquired by the color confocal microscope image acquiring unit based on the instruction of the color information received by the receiving unit; A color confocal microscope provided with measurement range selection means for selecting the region as a measurement range of the sample.
[0015]
According to the above configuration, when the color information (color) is instructed, a region as the measurement range is selected, and the burden on the user related to the measurement range selection is reduced.
The invention according to claim 7 is a control program for causing a computer to execute a control for selecting a measurement range of a three-dimensional confocal microscope image having color information corresponding to a sample, the three-dimensional confocal microscope image having color information. A first step of acquiring color information, a second step of receiving an instruction of color information, and the tertiary having the color information acquired in the first step based on the instruction of color information received in the second step A control program for causing the computer to execute a third step of selecting an area of the original confocal microscope image as a measurement range of the sample.
[0016]
By causing the computer to execute the above-described control program, an area as a measurement range is selected by instructing color information (color), and a user's burden related to measurement range selection is reduced.
[0017]
DETAILED DESCRIPTION OF THE INVENTION
Embodiments of the present invention will be described below with reference to the drawings.
FIG. 1 is a diagram showing a configuration example of a color confocal microscope according to the first embodiment of the present invention.
[0018]
The color confocal microscope shown in the figure is configured to acquire a computer 9, a sample surface shape information (three-dimensional confocal microscope image, height information), and a sample color information (color image). Etc.
The computer 9 includes a CPU (central processing unit) that controls the operation of the entire color confocal microscope by executing a control program stored in advance in an internal memory, a keyboard and a mouse that receive various instructions from the user, and the like. An operation unit, a display unit for displaying a GUI (Graphical User Interface), an image, and the like.
[0019]
Further, the configuration for acquiring the surface shape information of the sample includes a light source 1, a beam splitter 2, a two-dimensional scanning mechanism 3, an objective lens 4, an imaging lens 6, a pinhole 7, a photodetector 8, and the like. Yes. Here, the two-dimensional scanning mechanism 3, the photodetector 8, and the Z stage 11 are controlled by a computer 9. In the configuration for acquiring the surface shape information, light (laser light or the like) emitted from the light source 1 passes through the beam splitter 2 and then enters the two-dimensional scanning mechanism 3. The two-dimensional scanning mechanism 3 includes a first optical scanner 3 a and a second optical scanner 3 b, scans the light beam in two dimensions, and guides the light beam to the objective lens 4. Thereby, the light beam incident on the objective lens 4 is condensed by the objective lens 27 and scanned on the surface of the sample 5. Light from the surface of the sample 5, here reflected light, is guided again from the objective lens 4 to the beam splitter 2 through the two-dimensional scanning mechanism 3, reflected by the beam splitter 2, and pinned by the imaging lens 6. It is condensed on the hole 7. The pinhole 7 is at a position optically conjugate with the condensing position by the objective lens 4. When the sample 5 is at the condensing position by the objective lens 4, the reflected light from the sample 5 is collected on the pinhole 7. When light passes through the pinhole 7 and the sample 5 is not located at the condensing position by the objective lens 4, the reflected light from the sample 5 does not converge on the pinhole 7 and cannot pass through the pinhole 7. Therefore, the reflected light from the sample 5 other than the focal point is cut by the pinhole 7, and only the light passing through the pinhole 7 is detected by the photodetector 8. The sample 5 is placed on the sample table 10 and can be moved in the optical axis direction by the Z stage 11. With such a configuration, the two-dimensional scanning mechanism 3 performs two-dimensional scanning of the condensing point, and the output of the photodetector 8 is imaged in synchronization with the two-dimensional scanning mechanism 3, so that a specific height of the sample 5 is obtained. Only an image is obtained, and an image (confocal microscope image) obtained by optically slicing the sample 5 is obtained. Further, the sample 5 is discretely moved in the optical axis direction by the Z stage 11, the two-dimensional scanning mechanism 3 is scanned at each position to acquire a confocal microscope image, and the photodetector 8 is detected at each point on the sample surface. The surface shape information corresponding to the sample 5 is obtained by detecting the position of the Z stage 11 at which the maximum output is obtained, and a three-dimensional image (three-dimensional confocal microscope image) of the sample 5 corresponding to the surface shape information is obtained. can get.
[0020]
The configuration for obtaining the color information of the sample includes the white light source 12, the lens 13, the half mirrors 14 and 15, the color CCD 16, the CCD drive circuit 17, and the like. Here, the CCD drive circuit 17 is controlled by the computer 9. In the configuration for acquiring the color information, the white light emitted from the white light source 12 passes through the lens 13, the optical path is bent by the half mirror 14, and is condensed on the surface of the sample 5 on the sample stage 10 by the objective lens 4. The The white light reflected by the sample 5 passes through the objective lens 4, the half mirror 14, etc., the light path is bent by the half mirror 15, and the light is incident on the color CCD 16 to form an image. The color CCD 16 is provided at a position conjugate with the pinhole 7. An analog signal corresponding to the color image of the sample 5 imaged by the color CCD 16 is read by the CCD drive circuit 17, output to the computer 9, and converted into a digital signal by the computer 9. As a result, color information corresponding to the imaged sample 5 is obtained, and a color image of the sample 5 corresponding to the color information is obtained.
[0021]
Thus, by combining the color image of the sample 5 obtained by the configuration for acquiring the color information with the three-dimensional image of the sample 5 obtained by the configuration for acquiring the surface shape information described above, the color image corresponding to the sample 5 is obtained. A color three-dimensional image (color three-dimensional confocal microscope image, three-dimensional confocal microscope image having color information) is obtained.
[0022]
FIGS. 2A and 2B are examples of the color three-dimensional image obtained at this time.
Figures (a) and (b) both correspond to the sample having a plurality of colors on the sample surface. Figure (a) shows each region of the sample surface (Fig. (A) The area surrounded by the square in Fig. 4 is an example of a color three-dimensional image corresponding to a sample having a single color. Figure (b) shows each area on the surface of the sample (as shown in Figure (b)). Is an example of a color three-dimensional image corresponding to a sample having color unevenness (density difference or the like).
[0023]
Next, the measurement process performed by the CPU of the computer 9 will be described. This measurement process is realized by the CPU of the computer 9 reading and executing a control program stored in the internal memory.
FIG. 3 is a flowchart showing the processing contents of the measurement processing according to the first embodiment of the present invention.
[0024]
In the figure, first, in S301, the generated color three-dimensional image is acquired. As described above, the color three-dimensional image is generated by combining the three-dimensional image corresponding to the obtained surface shape information and the color image corresponding to the obtained color information.
[0025]
In S302, the color three-dimensional image generated in the previous step is displayed on the display unit of the computer 9, and display related to the color selection GUI is performed.
The color selection GUI according to the present embodiment provides an interface function for accepting an instruction of color information (color) from the user. For example, red (R), blue (B), green ( G) the function of selecting the corresponding color by specifying the content of the three primary colors numerically, the function of selecting the corresponding color by specifying a predetermined color on the color palette (color swatch), The specified color density, brightness, hue, saturation, brightness, etc. (this density, brightness, hue, saturation, brightness, etc., hereinafter simply referred to as density etc.) are indicated by numerical values. A function for selecting a color, a function for selecting a color within a corresponding range by specifying a numerical range of the specified color density, etc., or a predetermined position on a color map Select a color within a specified range It has a that function, and the like.
[0026]
In S303, a color instruction from the user is accepted via the color selection GUI. In this step, the content of the three primary colors of RBG described above is indicated by the user as a numerical value, and the range of the indicated color, such as the density, is indicated by a numerical value. A color included in the approximate color range is selected. As a result, a region of the color three-dimensional image having the same color as the color included in the approximate color range is selected as the measurement range of the sample 5.
[0027]
In S304, a color three-dimensional image in which a region corresponding to a color in the approximate color range selected in the previous step, that is, a region as a measurement range is distinguished from other regions is displayed on the display unit of the computer 9.
4A, 4B, and 4C are display examples of the color three-dimensional image displayed at this time, and FIG. 4A is a display example of the color three-dimensional image before the areas are distinguished. FIGS. 7B and 7C are display examples of a color three-dimensional image after the areas are distinguished. FIG. 2A is a part of the color three-dimensional image shown in FIG. The example shown in FIG. 4B is an example in which a region as a measurement range and a region in another range are distinguished by being displayed in different colors. Here, the range shown in black is shown as an area of another range. In addition, the example shown in FIG. 5C is an example in which the area as the measurement range is distinguished from the area of the other range by displaying the area as the measurement range surrounded by a conspicuous line such as a dotted line. is there. As another display example, the display examples shown in FIGS. 2B and 2C may be used in combination. Moreover, the display form which distinguishes the area | region as such a measurement range and the area | region of another range may be selected by the user.
[0028]
Returning to FIG. 3, in S <b> 305, it is determined whether or not the region as the selected measurement range is confirmed by the user. If the determination result is Yes, the process proceeds to S <b> 306, and if No, the process proceeds to S <b> 303. Returning, the above process is repeated. As a result, if this determination is not made, the color three-dimensional image displayed in S304 is displayed almost simultaneously with the color instruction in S303, and the process is repeated.
[0029]
In S306, a measurement item is selected according to a user instruction. The selected measurement items are used for existing confocal microscopes such as height measurement (step difference measurement), area measurement, roughness measurement, particle analysis, line width measurement, and film thickness measurement.
In S307, the measurement corresponding to the measurement item selected in the previous step is performed on the region of the color three-dimensional image selected as the measurement range of the sample 5 in S303.
[0030]
In S308, the result of the measurement performed in the previous step is displayed on the display unit of the computer 9, and this flow ends.
The above processing is the measurement processing according to the first embodiment, and when this processing is performed by the CPU of the computer 9, the region of the color three-dimensional image having the same color as the color according to the user's instruction is obtained. The measurement range of the sample 5 is selected, and the measurement for the measurement range is performed.
[0031]
In S303 of this flow, if an instruction for a range such as a density is not given, an area of a color three-dimensional image having the same color as the color according to the instruction related to the RBG is used as the measurement range. It may be selected.
Further, in S303 of this flow, an instruction for only the density or the like may be given instead of an instruction for the range of the density or the like. In this case, a region of the color three-dimensional image having the same color as the color according to the instruction such as the density is selected as the measurement range.
[0032]
Further, the approximate color range determined in S303 of the present flow may be displayed on the display unit of the computer 9 as color information, for example, by a color bar or the like.
In S303 of the present flow, the ratio of the three primary colors of RBG is digitized, and the range of the ratio is indicated by a numerical value, so that the color included in the corresponding range, that is, the color included in the approximate color range is displayed. It may be selected.
[0033]
Also, in S303 of this flow, the RBG content is indicated numerically for each of the plurality of colors, the approximate color range is indicated for each of the indicated plurality of colors, and the indicated plurality of approximate color ranges is indicated. All of the colors included in the image may be selected.
[0034]
In S303 of the present flow, the color instruction is performed by specifying the numerical value of the RBG content. However, the color instruction may be performed via the above-described color palette. FIG. 5 is an example of a color palette displayed on the display unit of the computer 9 in this case. A color is designated by instructing a predetermined color from the color palette shown in FIG. A plurality of colors may be designated.
[0035]
Further, in S303 of this flow, a color in the approximate color range is selected by instructing a range such as the density of the instructed color, but this instruction is performed via the color map. Anyway. FIG. 6 is an example of a color map displayed on the display unit of the computer 9 in this case. When a predetermined point on the color map shown in the figure is designated by the user, a color within a predetermined range centered on the point is selected as a color within the approximate color range. A plurality of points may be indicated. Further, only the color of the point designated by the user may be selected.
[0036]
In S307 of this flow, for example, when measurement related to area measurement, particle analysis, or the like is performed, not only the region as the measurement range selected in the above-described processing of S303 but also the height range is instructed and selected. Measurement of a portion corresponding to the specified measurement range and the designated height range is performed. FIG. 7 is a diagram showing an example of the area as the selected measurement range and the designated height range. The example shown in the figure shows an example in which h1 to h2 are designated as the height range, and the area indicated by the bold line is selected as the area as the measurement range. As a result, the measurement is performed on the region within the measurement range indicated by the bold line and the portion within the height range (h1 to h2).
[0037]
As described above, according to the first embodiment, a user can easily select a region as a measurement range simply by specifying a color, and the burden on the user related to the measurement range selection operation can be reduced. Can be made.
Next, a second embodiment of the present invention will be described.
[0038]
This embodiment is different from the above-described first embodiment in measurement processing. Therefore, here, the measurement process will be mainly described.
FIG. 8 is a flowchart showing the processing contents of the measurement processing according to the second embodiment of the present invention. Note that the measurement processing according to this embodiment is also realized by the CPU of the computer 9 reading and executing a control program stored in the internal memory.
[0039]
In the figure, in S801, the generated color three-dimensional image is acquired in the same manner as in the process of S301 in FIG.
In S802, the color three-dimensional image acquired in the previous step is displayed on the display unit of the computer 9, and the display related to the color selection GUI is performed.
[0040]
Note that the color selection GUI according to the present embodiment has a function of selecting a color included in a predetermined range centered on the point when a point on the displayed color three-dimensional image is designated. It has.
In step S803, a point on the color three-dimensional image is instructed by the user via the color selection GUI and is included in a predetermined range (within an approximate color range) centered on the point on the designated color three-dimensional image. The color to be selected is selected. In this step, a color included in the approximate color range may be selected by designating numerically the range such as the color density of the point on the designated color three-dimensional image.
[0041]
In S804, the area selected as the measurement range of the sample 5 is set to an area only within the approximate color range centered on the point on the color three-dimensional image specified in the previous step, or the approximation selected in the previous step. The user designates whether or not to set the area on the color three-dimensional image having the same color as the color within the color range.
[0042]
In step S <b> 805, a color three-dimensional image in which a region as a measurement range and other regions are distinguished according to the region instructed in the previous step is displayed on the display unit of the computer 9. The display example at this time is as shown in FIGS. 5A, 5B, and 5C, for example.
[0043]
In the subsequent processes in S806 to S809, the same processes as those in S305 to S308 in FIG. 3 described above are performed.
The above processing is the measurement processing according to the second embodiment, and this processing is performed by the CPU of the computer 9 so that the region within the approximate color range centered on the point designated by the user or An area of the color three-dimensional image having the same color as the color included in the approximate color range is selected as the measurement range of the sample 5, and measurement for the measurement range is performed.
[0044]
Note that the approximate color range determined in step S803 of this flow may be displayed on the display unit of the computer 9 as color information, for example, by a color bar or the like.
In S803 of this flow, a plurality of points are designated on the displayed color three-dimensional image, and within a predetermined range (within an approximate color range) centered on each of the designated plurality of points. All of the included colors may be selected.
[0045]
Further, in step S803 of the present flow, only a point color may be selected by designating a point on the displayed color three-dimensional image.
As described above, according to the second embodiment, the user can easily select an area as a measurement range simply by indicating a point on a color three-dimensional image obtained from a color confocal microscope. Thus, it is possible to reduce the burden on the user related to the measurement range selection operation.
[0046]
Next, a third embodiment of the present invention will be described.
This embodiment is different from the above-described first embodiment in measurement processing. Therefore, here, the measurement process will be mainly described.
FIG. 9 is a flowchart showing the processing contents of the measurement processing according to the third embodiment of the present invention. Note that the measurement processing according to this embodiment is also realized by the CPU of the computer 9 reading and executing a control program stored in the internal memory.
[0047]
In the figure, in S901, the generated color three-dimensional image is acquired in the same manner as in the process of S301 in FIG.
In step S <b> 902, the color three-dimensional image acquired in the previous step is displayed on the display unit of the computer 9, and display related to the color selection GUI is performed.
[0048]
Note that the color selection GUI according to the present embodiment has a function of selecting a color included in the predetermined range when a predetermined range on the displayed color three-dimensional image is designated. . The predetermined range is instructed, for example, by the user surrounding a predetermined area on the color three-dimensional image with a straight line or a curve.
[0049]
In step S903, a predetermined range (approximate color range) on the color three-dimensional image is instructed by the user via the color selection GUI, and colors included in the instructed approximate color range are selected. Further, when a range such as a density of a color included in the designated approximate color range is designated numerically, a color contained in a new approximate color range corresponding to the instruction is selected.
[0050]
In S904, the area selected as the measurement range of the sample 5 is set to an area only in the approximate color range on the color three-dimensional image instructed in the previous step, or the same color as the color included in the approximate color range. It is a new area by specifying a range such as a density of a color included in the approximate color range on the color 3D image specified in the previous step, or an area on the color 3D image having color. The user instructs whether the area is only within the approximate color range or the area on the color three-dimensional image having the same color as the color included in the new approximate color range.
[0051]
In step S <b> 905, a color three-dimensional image in which a region as a measurement range is distinguished from other regions according to the region designated in the previous step is displayed on the display unit of the computer 9. The display example at this time is as shown in FIGS. 5A, 5B, and 5C, for example.
[0052]
In the subsequent processes in S906 to S909, the same processes as those in S305 to S308 in FIG. 3 described above are performed.
The above processing is the measurement processing according to the third embodiment, and this processing is performed by the CPU of the computer 9 so that the region within the approximate color range on the color three-dimensional image instructed by the user or The numerical value indicates the area on the color 3D image that has the same color as the color included in the approximate color range, or the range of the color density included in the approximate color range on the specified color 3D image. A region only in the new approximate color range or a region on the color three-dimensional image having the same color as the color included in the new approximate color range is selected as the measurement range of the sample 5. Measurement is performed on the measurement range.
[0053]
In the present flow, the approximate color range determined in S903 may be displayed on the display unit of the computer 9 as color information by a color bar or the like, for example.
As described above, according to the third embodiment, the user can easily select a region as a measurement range simply by designating a predetermined range on a color three-dimensional image obtained from a color confocal microscope. Thus, the burden on the user related to the selection operation of the measurement range can be reduced.
[0054]
In the first to third embodiments, the measurement process for the color three-dimensional image has been described. Needless to say, the measurement process can be similarly performed for the color two-dimensional image. .
In the first to third embodiments, what is displayed on the display unit of the computer 9 may be displayed on, for example, an external display device.
[0055]
In the first to third embodiments, the area as the measurement range of the color three-dimensional image and the other areas are inverted, and the area that was the other area is set as the area as the measurement range. May be.
In the first to third embodiments, the area of the measurement range of the color three-dimensional image may be enlarged or reduced by several pixels, for example.
[0056]
In the first to third embodiments, the above-described measurement process may be automatically performed by instructing the color, measurement items, and the like in advance.
In the first to third embodiments, when a region that is not a measurement range is included in a region surrounded by a region as a measurement range of a color three-dimensional image, the measurement range is Only when the region that is not the region is very small, the region that is not the region as the measurement range may be used as the region as the measurement range.
[0057]
In the first to third embodiments, the configuration shown in FIG. 1 is applied as an example of a color confocal microscope. However, a color confocal microscope having another configuration capable of acquiring a color three-dimensional image is used. You may make it apply. For example, an XY stage that moves the sample 5 perpendicular to the optical axis may be used as a scanning mechanism that relatively scans the focused light from the objective lens 4 along the surface of the sample 5. Further, as a moving mechanism for moving the focusing position of the objective lens 4 and the position of the sample 5 in place, a mechanism for moving the objective lens 7 instead of the Z stage 11 for moving the position of the sample 5 may be used. good. Further, the stage on which the sample 5 or the like is installed may be manually operated. Further, a manual sample transport device may be provided in the color confocal microscope, or an automatic sample transport device may be provided so that the above-described measurement processing is automatically performed on a plurality of samples. However, in this case, the color and the measurement item are instructed in advance. In this case, if the display on the display unit during the measurement process described above is not performed, the time required for the measurement process for one sample can be shortened. Further, other means may be used in place of the color CCD 16 to obtain a color image.
[0058]
In the first to third embodiments, the control process performed by the CPU of the computer 9 of the color confocal microscope can be executed by a computer as shown in FIG. 10, for example. In this case, the control program stored in the internal memory of the computer 9 of the color confocal microscope is a CD-ROM, floppy (registered trademark) disk (or MO, DVD, CD-R, A portable storage medium 21 such as a CD-RW or a removable hard disk), the portable storage medium 21 is read by the medium driving device 23 of the computer 22, and the read control program is read by the computer 22. May be stored in the internal memory (RAM or hard disk) 24 and the computer 22 may execute the control program. Alternatively, the control program is stored in storage means (database or the like) 25 in an external device (server or the like) of the information provider, transferred to the computer 22 by communication, and stored in the internal memory 24, and the control is performed. The program may be executed by the computer 22. Note that the control program stored in these may execute only a part of the control processing performed by the CPU of the computer 9 of the color confocal microscope.
[0059]
As described above, the color confocal microscope, the measurement range selection method, and the control program thereof according to the present invention have been described in detail. However, the present invention is not limited to the above embodiment, and various improvements can be made without departing from the gist of the present invention. Of course, changes may be made.
[0060]
【The invention's effect】
As described above in detail, according to the present invention, the user can easily select the region of the confocal microscope image having the color information as the measurement range of the sample only by indicating the color information, It is possible to reduce the burden on the user related to the selection operation of the area as the measurement range. If color information is instructed in advance, the measurement process can be automatically performed.
[Brief description of the drawings]
FIG. 1 is a diagram showing a configuration example of a color confocal microscope according to a first embodiment of the present invention.
FIGS. 2A and 2B are examples of color three-dimensional images. FIG.
FIG. 3 is a flowchart showing processing contents of measurement processing according to the first embodiment of the present invention.
4 (a), (b), and (c) are display examples in the processing of S304, (a) is a display example of a color three-dimensional image before the region is distinguished, and (b), (c ) Is a display example of a color three-dimensional image after the regions are distinguished.
FIG. 5 is an example of a color palette displayed on a display unit of a computer.
FIG. 6 is an example of a color map displayed on a display unit of a computer.
FIG. 7 is a diagram showing an example of a region as a selected measurement range and an indicated height range.
FIG. 8 is a flowchart showing the processing contents of measurement processing according to the second embodiment of the present invention.
FIG. 9 is a flowchart showing the processing contents of measurement processing according to the third embodiment of the present invention.
FIG. 10 is a diagram illustrating an example of a storage medium in which a control program is stored.
FIG. 11 is a diagram showing a schematic configuration example of a general conventional confocal microscope.
[Explanation of symbols]
1 Light source
2 Beam splitter
3 Two-dimensional scanning mechanism
3a First optical scanner
3b Second optical scanner
4 Objective lens
5 samples
6 Imaging lens
7 pinhole
8 Photodetector
9 Computer
10 Sample stage
11 Z stage
12 White light source
13 Lens
14, 15 half mirror
16 color CCD
17 CCD drive circuit
21 Portable storage media
22 Computer
23 Medium drive device
24 memory
25 Memory means
31 Light source
32 Beam splitter
33 Two-dimensional scanning mechanism
33a First optical scanner
33b Second optical scanner
34 Objective lens
35 samples
36 Imaging lens
37 pinhole
38 photodetectors
39 computers

Claims (7)

A first step of acquiring a three-dimensional confocal microscope image having color information according to the sample;
A second step of accepting color information instructions;
A third step of selecting a region of the three-dimensional confocal microscope image having the color information acquired in the first step based on the color information instruction received in the second step as the measurement range of the sample;
A measuring range selection method characterized by comprising: In the second step, at least the content of RBG is indicated by a numerical value, specified via a color sample, specified via a color map, or at least the density, luminance, and hue of color information. , Saturation, and lightness are indicated by numerical values, or at least one of the ranges of density, luminance, hue, saturation, and lightness of color information is indicated by numerical values. Accept information instructions,
In the third step, the region of the three-dimensional confocal microscope image having the same color information as the color information instructed in the second step is selected as the measurement range of the sample.
The measurement range selection method according to claim 1. In the second step, a three-dimensional confocal microscope image having the color information acquired in the first step is displayed on a display unit, and the color information displayed on the display unit is displayed as an instruction for color information. Accepts an indication of a point on a 3D confocal microscope image,
In the third step, the same color information as the point indicated on the three-dimensional confocal microscope image having the color information displayed on the display unit or the same color information within a predetermined range including the point A region of the three-dimensional confocal microscope image having the color information, a region of only the point indicated on the three-dimensional confocal microscope image having the color information, or a region of only a predetermined range including the point. Select as the measurement range of
The measurement range selection method according to claim 1. In the second step, the three-dimensional confocal microscope image having the color information acquired in the first step is displayed on a display unit, and the color information displayed on the display unit as an instruction of color information Accept range instructions on the 3D confocal microscope image with
In the third step, the three-dimensional confocal microscope image having the same color information as the color information in the range indicated on the three-dimensional confocal microscope image having the color information displayed on the display unit Or a region only in the range indicated on the three-dimensional confocal microscope image having the color information is selected as the measurement range of the sample.
The measurement range selection method according to claim 1. In the second step,
Accept multiple color information instructions,
The measurement range selection method according to claim 1, wherein the measurement range is selected. An objective lens that focuses the light from the light source onto the sample;
Moving means for relatively moving the focusing position of the objective lens and the position of the sample along the optical axis direction of the light focused by the objective lens;
A microscopic aperture disposed at a position conjugate with the focusing position of the objective lens;
A photodetector for detecting the intensity of light passing through the minute aperture;
Confocal microscope image acquisition means for acquiring a three-dimensional confocal microscope image of the sample based on the intensity of light detected by the photodetector;
Color information acquisition means for acquiring color information of the sample surface;
A color confocal microscope that acquires a three-dimensional confocal microscope image having color information based on the three-dimensional confocal microscope image acquired by the confocal microscope image acquisition means and the color information acquired by the color information acquisition means Image acquisition means;
Receiving means for receiving instructions for color information;
Measurement range selection for selecting a region of a three-dimensional confocal microscope image having color information acquired by the color confocal microscope image acquisition unit as a measurement range of the sample based on an instruction of color information received by the reception unit Means,
A color confocal microscope characterized by comprising: A control program for causing a computer to execute control for selecting a measurement range of a three-dimensional confocal microscope image having color information corresponding to a sample,
A first step of acquiring a three-dimensional confocal microscope image having color information;
A second step of accepting color information instructions;
A third step of selecting a region of the three-dimensional confocal microscope image having the color information acquired in the first step as a measurement range of the sample based on the color information instruction received in the second step; ,
A control program for causing the computer to execute.

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