JP3518923B2 - Automatic image forming equipment for confocal scanning optical microscope - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an automatic image forming apparatus for a confocal scanning optical microscope.

[0002]

2. Description of the Related Art Conventionally, a point light source illuminates an observation sample in a point shape, and transmitted light or reflected light from the illuminated sample is imaged again in a point shape, and a detector having a pinhole opening is used. There have been microscopes with confocal optics to obtain image density information.

A microscope having this confocal optical system will be described below with reference to FIG.

FIG. 7 (a) is a schematic configuration diagram thereof, in which light emitted from a point light source 31 passes through a half mirror 32,
An image is formed as a point on the observation sample 34 by the objective lens 33 whose aberration is corrected well, and the observation sample 34 is illuminated.

The light reflected by the observation sample 34 passes through the objective lens 33 again, and then is reflected by the half mirror 32 to be condensed. A pinhole plate 35 having a pinhole opening is arranged at this condensing position, and light passing through the pinhole opening is detected by a photodetector 36. Then, the two-dimensional image of the observation sample 34 is obtained by two-dimensionally scanning the observation sample 34 in the same manner as the raster scanning of the television.

By the way, the light shown by the broken line in FIG. 7A is the light from the position L deviated from the condensing position of the objective lens 33. This displaced light is not condensed at the position of the pinhole opening of the pinhole plate 35, so it cannot pass through the pinhole plate 35 and does not reach the photodetector 36. From this, with such an optical system, it is possible to obtain an image only at the focusing position of the objective lens 33, that is, the focusing position.

This is effective when the observation surface of the observation sample 34 is flat, but as shown in FIG. 7 (b), the observation surface has a stepped shape and a non-planar shape having a plurality of portions with different heights. In the case of the observation sample 34, it is difficult to obtain an image. Figure 7
The observation sample 34 shown in (b) is composed of a high-height portion B, a low-height portion C, and an intermediate height portion A. When, for example, the observation surface of A is focused, The observation surfaces of B and C are blurred with an optical microscope. Therefore, it is impossible to obtain an image focused on all the observation planes A, B, and C by a single observation operation.

Of course, when obtaining an image of the observation surface of the observation sample 34 of FIG. 7 (b) using the microscope of FIG. 7 (a), for example, an image obtained by focusing the observation surface of A is stored,
If the image of the B observation surface and the image of the C observation surface obtained in the same manner are stored and added together, an image focused on the entire surface can be obtained. In this case, in practice, the maximum brightness value may be held for each pixel.

The surface shape of the sample can be measured by utilizing the above optical characteristics. About this technology "THEORY
AND PRACTICE OF SCANNING OPTICAL MICROS COPY ''
(P.126 to P.130).

Here, the light is focused on one point of the sample, the point is scanned in the optical axis direction (Z-axis direction), and the Z position at which the brightness is maximized during scanning is detected and stored. Next, the condensed light is moved in the X direction, scanning is performed in the Z axis direction at the point position, and the Z position at which the brightness during scanning is the highest is detected and stored. By repeating the above scanning in the X and Y directions, the surface shape (concavities and convexities) of the sample can be measured.

[0011]

In the microscope having the confocal optical system described above, the image disappears when the observation sample deviates from the focus position of the objective lens. Deviating from the focus position indicates that the position of the sample at that time is past the highest position or the lowest position. Utilizing this characteristic,
The highest position and the lowest position of the sample can be set automatically.

This technique is disclosed in JP-A-6-308393. That is, the farther the sample is from the in-focus position, the darker the entire image obtained.
This indicates that the brightness level of each pixel forming an image approaches zero and the number of pixels of this brightness level increases. In Japanese Patent Laid-Open No. 6-308393,
The highest position and the lowest position of the sample are determined by setting the level of the luminance signal of the image and checking the ratio of the pixels of that level to all the pixels.

However, for example, in the observation sample 34 having the shape shown in FIG. 7B, when the steep surface between the A surface and the B surface or the B surface and the C surface has a steep inclination, The light reflected by may not return to the objective lens. Therefore, most of the image signals at such positions are black level images. This state is very similar to the state where the sample is out of focus. Therefore, in the technique of Japanese Patent Laid-Open No. 6-308393, there is always a possibility of making an erroneous judgment that the highest position or the lowest position of the sample is detected, even though the focus position is on the slope. Yes, there was a problem of poor reliability.

The present invention has been made in view of the above circumstances, and is for the case of measuring an observation sample having a plurality of planes having different heights and having a steep slope between the planes. Even if there is, it is possible to accurately set the measurement range automatically, and to prevent interference between the sample and the objective lens during automatic setting .
An object is to provide an automatic image forming apparatus .

[0015]

The invention according to claim 1 is
A point light source illuminates the observation sample placed on the stage.
An objective that refocuses the light emitted from the observation sample.
On the observation sample collected by the lens and the objective lens
Means for relatively scanning the light and the observation sample
And light detection means for detecting the reflected light from the observation sample
And a scanning microscope having moving means for moving a relative position between the focal position of the objective lens and the observation sample in the optical axis direction, wherein the number of plane regions of different heights of the observation sample is set. The plane number setting means and the detection signal of the light detecting means.
Pixels whose signal level is below a preset signal level
The pixel number counting means for counting the number and the moving means
First detecting means that is moved in the axial direction and detects whether or not the scanning position of the observation sample is a flat area by determining whether or not the number of pixels from the pixel number counting means is equal to or more than a preset number of pixels; , the detection result of the first detecting means, so as to comprise a second detection means for detecting the upper limit position and lower limit position of the observation sample based on the plane area number and set in the plane-setting means There is.

[0016] In a second aspect of the present invention is the invention of claim 1, wherein said second detection means, the upper limit position of the observation sample is detected earlier, in detecting the lower limit position of the observation sample
The moving range of the moving means is limited. In the invention of claim 3, the invention of claim 2
In the invention, the movement range is limited to the objective lens.
Try to be within the operating distance of.

[0017]

As a result, according to the invention described in claim 1, there is a case of measuring an observation sample having a plurality of planes having different heights and having a steep slope between the planes. However, the relative position between the focus position of the objective lens and the observation sample can be moved to accurately detect the upper and lower limit positions thereof, and the automatic setting of the measurement range can be surely performed.

According to the invention of claim 2, in addition to the effect of the invention of claim 1, the relative position between the focal position of the objective lens and the observation sample is moved to set the upper and lower limit positions of the observation sample. When automatically setting, the interference between the observation sample and the objective lens can be prevented in advance.

[0019]

DESCRIPTION OF THE PREFERRED EMBODIMENTS An embodiment of the present invention will be described below with reference to the drawings.

FIG. 1 shows the system configuration of the entire confocal scanning optical microscope. In the figure, 1 is a confocal scanning optical microscope, and 2 is a laser light source which is a light source of scanning light. The laser light as the spot light generated from the laser light source 2 is totally reflected by the mirror 3, and the half mirror 4
And then sent to the two-dimensional scanning mechanism 5.

The two-dimensional scanning mechanism 5 has, for example, a galvano mirror for scanning in the X-axis direction and a galvano mirror for scanning in the Y-axis direction, and the XY scanning control unit 1 to be described later.
Under the control of 6, the two galvano mirrors are swung in the X-axis direction and the Y-direction to scan the spot light in the same way as the raster scan of the television, and the spot light scanned by the two-dimensional scanning mechanism 5 is scanned. Is radiated from the objective lens 7 through the revolver 6 onto the sample 8 mounted and supported on the fine movement stage 9.

That is, the revolver 6 holds a plurality of objective lenses 7 having different magnifications, and the fine movement stage 9 holds the sample 8. The fine movement stage 9 is provided on the coarse movement stage 10.

Among the objective lenses 7, one having a desired magnification is switched by the revolver 6 to set the position in the observation optical path of the microscope, and the two-dimensional scanning mechanism 5 is moved from the two-dimensional scanning mechanism 5 via the objective lens 7 set in this position. It is possible to irradiate the sample 8 on the fine movement stage 9 with the spot light while scanning two-dimensionally.

The reflected light by this irradiation returns to the two-dimensional scanning mechanism 5 through the objective lens 7 and is returned from the two-dimensional scanning mechanism 5 to the half mirror 4.

The half mirror 4 is provided on the emission optical path of the laser light source 2 with respect to the two-dimensional scanning mechanism 5, and is for guiding the reflected light from the sample 8 obtained through the two-dimensional scanning mechanism 5 to the detection system. The reflected light from the sample 8 obtained by the half mirror 4 is condensed by the lens 11 and sent to the photodetector 13 via the pinhole plate 12 having an opening of a predetermined diameter.

That is, the photodetector 13 has the lens 11 and the pinhole plate 12 arranged on the front surface of its light receiving surface, and the pinhole plate 12 is located at the focal position of the lens 11.
The light obtained through the pinhole plate 12 is converted into an analog electric signal (luminance signal) corresponding to the amount of light and sent to the image processing unit 14.

The image processing unit 14 has two image memories 14A and 14B, each of which has a storage capacity for one frame. Here, for example, 512 pixels × 512 pixels × 8 bits ( (256 gradations)
The image is stored as 1 frame.

Of these, the image memory 14A stores the brightness information of the reflected light obtained by the photodetector 13 as 8-bit data at a pixel position corresponding to the current XY scanning position of the spot light, thereby generating an image signal. Is to remember.

When the current brightness information is larger (brighter) than the brightness information at the previous scan position, this memory is updated and saved as the image information at the scan pixel position, so that the height position This is so that different images can be added.

Further, the image memory 14B counts the number of times the revolver 6 has moved from the Z moving circuit 15 to the information on the Z moving direction at the pixel position corresponding to the current XY scanning position of the spot light. The value of is given.

In this case, the image processing unit 14 refers to the image data in the image memory 14A, and when the level of the luminance information of this time is higher than the luminance information of the position at the previous time, the image processing unit 14B is compared with the image memory 14B. A function of updating the value of the number of times as data and storing and storing is provided.

In this way, when the image memory 14B has information indicating the maximum brightness at each pixel position, the revolver movement count value is the information in the Z scanning direction (this information indicates the height position). Will be stored).

The current XY scanning position information of the spot light for the image processing unit 14 is the XY scanning control unit 1.
6 through the computer 17. The image processing unit 14 receives the output signal from the photodetector 13 and automatically sets the automatic range at the time of automatic image formation, transmits the set value to the computer 17, controls the Z movement circuit 15, and detects an end face described later. In addition to controlling movement of the focus position,
The data stored in the image memories 14A and 14B are read out and given to the computer 17.

The Z moving circuit 15 is a circuit controlled by the computer 17 or the image processing unit 14 so as to move the revolver 6 in the height direction, that is, the Z-axis direction in units of reference width. The count function for counting the position information each time 6 is moved in the unit of the reference width, and the count value are stored in the image memory 14
It has the function of sending to B.

However, the image processing unit 14 and Z
XY for controlling the moving circuit 15 and the two-dimensional scanning mechanism 5
A computer 17 is provided to integrally control the scan control unit 16. This computer 17
In addition to the overall control of the XY scanning control unit 16, the Z movement circuit 15 and the image processing unit 14, it is the center of overall control and processing such as saving, reproducing and editing image data, and is required. Information and images are displayed and output on the monitor display 18.

Further, a television optical system is prepared for roughly focusing on the sample 8 mounted on the fine movement stage 9 when the sample 8 is measured. This is used in consideration of the point that it is slightly difficult to use because the focal depth is extremely shallow when focusing is performed on a confocal image obtained by laser scanning.

That is, the light generated by the white light source 19 is totally reflected by the half mirror 21 after passing through the lens 20, and is inserted into the laser optical path between the two-dimensional scanning mechanism 5 and the revolver 6 through the lens 22. The sample 8 is reflected by the mirror 23 and is radiated from the objective lens 7 through the revolver 6 to the sample 8 mounted and supported on the fine movement stage 9.

The light reflected by this irradiation passes through the objective lens 7, is reflected by the mirror 23, passes through the lens 22, and then passes through the half mirror 21, and the lens 24
The image is formed on the television camera 25 by.

The sample image taken by the television camera 25 is displayed on the monitor display 18, and the observer moves the coarse movement stage 10 while looking at the image on the monitor display 18 for focusing. At this time, the observer can know the approximate shape and in-focus position of the sample 8.

Next, as the operation of the above-mentioned embodiment, details of the case where the shape of the sample 8 is measured by the laser light generated by the laser light source 2 will be described. When measuring the sample shape, it is necessary to set the measurement range in the height direction (Z-axis direction). Here, the automatic setting of the measurement range according to the present invention is used.

In the following description, it is assumed that the sample 8 has a shape as shown in FIG. The sample 8 shown in FIG. 2 (a) includes a flat portion A having a high height, a flat portion E having a low height, a flat intermediate portion C having a height intermediate between these planes A and E, and the above A. The inclined surface portion B between the surface C and the surface C, the inclined surface portion D between the surface C and the surface E, and the inclined surface portion F between the surface E and the bottom surface of the sample 8 respectively. , F shall be sharp and steep.

Then, the focus position roughly adjusted by the television optical system is set as the focus initial position X in FIG.
In addition, as shown in FIG. 2B together with a side view of the sample 8,
It is assumed that the direction from the initial focusing position X to the surface A is the upper limit direction, and the direction from the initial focusing position X to the surface E is the lower limit direction.

FIGS. 3 and 4 mainly show the contents of the operation processing by the image processing unit 14, and at the beginning, each parameter is set (steps S1 to S7).

First, the detection reference level Vref is set (step S1).

This detection reference level Vref is the photodetector 1
It is used for comparison with the level of the image signal Vin from No. 3. As described above, the image signal Vin from the photodetector 13 is ideally zero when the sample 8 is out of the focus position of the objective lens 7 or when the reflected laser light does not return. Therefore, the value of the detection reference level Vref may be set to zero, but usually, a noise component or the like is often superposed on the image signal Vin, so that a level of zero or a level close to zero is set. is there.

Next, the detection reference pixel number Nref is set (step S2).

This detection reference pixel number Nref indicates the number of pixels when the image signal Vin is below the detection reference level Vref with respect to the total number of pixels forming the laser image. The black area on the image increases as the sample 8 moves away from the in-focus position of the objective lens 7 or the reflected laser light does not return, that is, the detection reference level V.
The proportion of pixels having the image signal Vin equal to or lower than ref increases. Based on such a property, the value of the detection reference pixel number Nref is set.

Next, the number of surfaces Nup in the upper limit direction is set (step S3).

The number of surfaces Nup in the upper limit direction indicates the number of planes located in the upper limit direction from the in-focus initial position X, and in the present embodiment, the planes in the upper limit direction from the in-focus initial position X as shown in FIG. Since there is only the A side, “Nup = 1”.

Next, the number of surfaces Ndown in the lower limit direction is set (step S4).

The number of planes Ndown in the lower limit direction indicates the number of planes located in the lower limit direction from the in-focus initial position X, and in this embodiment, the planes in the lower limit direction from the in-focus initial position X as shown in FIG. Is C-plane and E-plane, so "Ndown = 2".

Next, which of the upper limit direction and the lower limit direction is detected first is set (step S).
5).

Basically, the upper and lower limits may be moved first, but when the measurement range (shape of the sample 8) is wider than the operating distance of the objective lens 7, the sample 8 and the objective lens 7 are moved. There is a risk of physical interference between and. for that reason,
First, it is better to move the objective lens 7 and the sample 8 in such a direction that the distance between them widens, that is, the upper limit direction.

As described above, when the detection direction is always started from the upper limit direction, the internal setting may be made in advance so that the observer can save the parameter setting.

Next, it is set whether the initial focusing position X is a flat area or a slope area (step S6).

This is set because the method of judgment is different between the plane area such as the A surface and the slope area such as the B surface at the focus position of the sample 8 having the shape of FIG. 2, for example. Here, since the initial focus position X is the B surface, that is, the slope area, the slope area is set.

At the end of the parameters, the movement amount unit of the revolver 6 is set (step S7).

This is to evaluate the image signal while moving the in-focus position of the objective lens 7 in the automatic setting of the measurement range. By moving the revolver 6 up and down, the revolver 6 is moved.
The objective lens 7 attached to the lens moves up and down, and the relative position between the in-focus position and the sample 8 is in the optical axis direction (Z direction).
To change to.

If the movement amount unit of the revolver 6 is set small, the movement in the Z direction is finely performed, so that the detection accuracy is high, but there is a demerit that it takes time accordingly. On the other hand, if the movement amount unit of the revolver 6 is set to a large value, the movement in the Z direction will be rough and the detection accuracy will be low. Is set.

After automatically setting the measurement range, the revolver 6 is moved again for the actual measurement, but the moving amount unit of the revolver 6 for this measurement is set separately.

When the setting of each parameter is completed as described above, the scanning with the laser beam is started at the initial focusing position X (step S8).

The light reflected by the sample 8 by this scanning is photoelectrically converted by the photodetector 13 to obtain an image signal Vin of each pixel forming an image. The obtained image signal is obtained for each pixel at the scanning position. Based on Vin and the detection reference level Vref, it is determined whether “Vin <Vref” is satisfied (step S
9).

Then, only when "Vin <Vref", the number of pixels is counted (step S10).

Thereafter, until it is judged that the scanning of one screen is completed (step S11), the counting of the number of pixels is continued only at the time of "Vin <Vref" at steps S9 and S10 at each pixel position. When it is determined that the scanning of one screen is completed, the number N of pixels of which the image signal Vin is determined to be lower than the detection reference level Vref out of the total number of pixels forming the image at this point is found. To do.

Here, as described above, since the determination method is different between the flat area and the slope area, it is determined whether or not the current focus position set in step S6 is the flat area (step S12).

At this point in time, the initial focusing position X is on the B surface, which is a slope, and this is determined in step S12,
Next, it is determined whether or not the counted pixel number N is smaller than the preset detection reference pixel number Nref (step S13).

As shown in FIG. 5, B with the initial focusing position X
Since the surface is substantially vertical, the laser light emitted from the objective lens 7 to the sample 8 is reflected almost obliquely downward. Therefore, most of the reflected light does not return to the objective lens 7, and most of the image becomes black, and as a result, the image signal V below the detection reference level Vref.
The number of pixels of in increases.

As described above, in the slope area, "N>Nref" while scanning one screen, and this is determined in step S13, and the current position is the upper limit position in order to determine the moving direction of the revolver 6. It is determined whether or not is being set (step S15).

Since the upper limit position is currently being set,
When this is determined, the revolver 6 is moved in the upper limit direction by the Z movement circuit 15 by the movement amount unit set in step S7 (step S16), and the step S16 is performed again.
Returning to the processing from 8, the scanning for one screen is performed.

As a result, from the state shown in FIG.
As shown in (b), the focus position of the objective lens 7 moves in the upper limit direction, and the laser beam scanning is started at the new focus position and the position of the sample 8. While in the slope area, the above processing is repeatedly executed.

Next, the case where the focus position of the objective lens 7 approaches the flat area will be described. When the focus position of the objective lens 7 approaches the plane area, the reflected light from the sample 8 comes back to the objective lens 7.

Therefore, the number of pixels that have obtained the image signal Vin higher than the detection reference level Vref increases.
As a result, the number of count pixels N decreases and “N <Nref
Is determined in step S13, and processing for declaring that the in-focus position has entered the plane area is performed (step S14).

Then, after confirming that the upper limit is being set (step S15), the revolver 6 is further moved in the upper limit direction by the movement amount unit (step S16), and the process returns from the step S8 to 1 Scan the screen.

After the scanning for one screen is completed, the count pixel number N and the detection reference pixel number Nref similar to the above are compared. However, this is the step because the focusing position of the objective lens 7 is in the plane area. The determination is made in S12, and subsequently in step S20, it is determined whether or not “N> Nref”.

When the in-focus position of the objective lens 7 is in the plane area, "N <Nref" is satisfied as described above.
This is determined in step S20, and the above step S15
In step S16, the revolver 6 is further moved in the upper limit direction by the movement amount unit, and the process returns from step S8 to scan one screen.

Due to the movement of the revolver 6 in the upper limit direction, the image signal Vin becomes smaller and the image of the sample 8 disappears as the focus position of the objective lens 7 moves out of the plane area and approaches the upper limit position. Therefore, the count pixel number N becomes large, and finally "N>Nref". If this is determined in step S20, that is, it means that the plane area has been detected, so that the number of plane areas is counted and the count value is set to M (step S21).

Then, it is judged whether or not the detected plane area is the final detection surface (step S22).

This is determined by whether or not the count value M of the detection plane and the value of the number Nup of faces in the upper limit direction or the number Ndown of faces in the lower limit direction set in steps S3 and S4 match. If it is done, it is judged as the final side,
If they do not match, a process of declaring that the focus position of the objective lens 7 has entered the slope area is performed (step S2).
3).

Here, since the count value M of the detected plane is "1" and the number Nup of faces in the upper limit direction set in step S3 is "1", it is determined that the detected plane is the final face. Then, it is determined whether the final surface is the final surface at the upper limit position or the final surface at the lower limit position (step S22).

Since this is the last surface of the upper limit position, this is determined in step S24, and the position of the revolver 6 at this time is stored as the upper limit position of the measurement range (step S25).

After that, in order to detect the lower limit position of the measurement range, the focus position of the objective lens 7 is set to the above-mentioned focus initial position X.
Then, the revolver 6 is moved by the Z moving circuit 15 (step S26), and the detection direction is set to the lower limit direction (step S27). Then, the process returns from step S8 to the upper limit direction. Similarly to the case, scanning is executed while moving the revolver 6 in the lower limit direction.

However, when moving the revolver 6 in the lower limit direction, a process partially different from the above-described movement in the upper limit direction is performed.

This is done in order to prevent the objective lens 7 and the sample 8 from physically interfering with each other. After it is determined in step S15 that the upper limit position is not currently set, It is determined whether the operating distance WD of the objective lens 7 has exceeded the movement amount L of the revolver 6 from the upper limit position (step S17).

If “WD> L”, then the revolver 6
Is moved in the lower limit direction by the movement amount unit by the Z moving circuit 15 (step S18), and the process is returned from step S8 again to scan one screen.

However, when “WD <L”, that is, when the moving amount L of the revolver 6 from the upper limit position exceeds the operating distance WD of the objective lens 7, the sample 8
Since the highest portion of the sample 8 will hit the objective lens 7 before the in-focus position of the objective lens 7 reaches the lowest surface of the objective lens 7, a warning is issued and, for example, it is displayed on the monitor display 18 (step S19), Revolver 6
3 is stopped, or the processing of FIGS. 3 and 4 is ended as shown in the figure.

Since there is a possibility that the operating distance WD of the objective lens 7 may vary, a value slightly smaller than the standard value may be set when setting the value in consideration of the margin.

However, the processing of steps S17 to S19 is effective only when the processing procedure of first detecting the upper limit position and then detecting the lower limit position is performed, and almost effective when first detecting the lower limit position. I can't get it.

Thus, the detection of the slope area of the B surface is completed, and then the detection of the flat area of the C surface is performed. Since the C surface is not the final surface, the slope area of the D surface is continuously detected, and the flat area of the E surface is further detected.

When the detection of the plane area of the E-plane is completed, the count value M of the detection plane becomes "2", and the number N of planes in the lower limit direction is reached.
Since the values of down match, the final surface is determined in step S22.

Therefore, it is determined that the plane area of the E surface is the final surface in the lower limit direction (step S24), and the position of the revolver 6 at this time is stored as the lower limit position of the measurement range (step S28). Thus, the processing of FIGS. 3 and 4 is completed.

As described above, the upper limit position and the lower limit position of the measurement range of the sample 8 are automatically set. If each parameter used for automatic setting can be stored in the memory in the computer 17, it can be set according to various samples, and by reading the stored parameters according to the sample, The labor required for measurement can be further simplified.

In the process of counting the plane area (step S21), the following problem is taken into consideration.

That is, the plane area has a certain width as shown in FIG. Therefore, depending on the value of the movement unit amount, the focus position may cross the width twice or more.
In this case, the judgment of "N>Nref" is also performed twice or more,
Even if the same plane area is used, it will be determined as a different plane area.

Therefore, if "N>Nref" is once obtained in the process of step S21, then "N>Nref" is set.
"Do not count. However, if this is left as it is, it will not be possible to count the next plane area. For this reason, the processing is performed such that when the slope area is passed past the flat area and "N>Nref" is satisfied, the count is again performed in step S21 in preparation for the detection of the next flat area.

Next, when the final plane area is detected, the following processing is also performed in the processing of storing the position of the revolver at that time as the upper limit position or the lower limit position (steps S27 and S28).

The upper limit position and the lower limit position correspond to the measurement range. At the time of measurement, the revolver is moved again within this range to perform measurement. Here, since the revolver is moved mechanically, some movement error occurs. For this reason,
If the measurement range is set to the minimum, the movement error may cause the measurement range to be different from the preset range.

For this reason, the processing in steps S27 and S28 incorporates processing for setting the measurement range to a larger value in consideration of the above error.

The following methods can be considered as means for setting a large measurement range.

(1) A preset amount is added to the position where the plane area is detected, and the position is set as the upper limit or the lower limit position.

The preset amount is a value obtained by multiplying the moving unit amount by several times. The observer can set the amount of this several times from a computer or the like.

(2) Nothing exists in the area beyond the final plane area. Therefore, in this area, "N <Nref". Therefore, the revolver is moved even past the final plane area, and the position where “N <Nref” is satisfied is set as the upper limit or the lower limit position.

In the above embodiment, the Z moving circuit 15 moves the revolver 6 in the Z axis direction to move the in-focus position, but the relative Z between the objective lens 7 and the sample 8 is described. Since it suffices to move the position in the axial direction, for example, the fine movement stage 9 may be moved in the Z-axis direction instead of the revolver 6.

The above description is based on the embodiments, but the present invention includes the following inventions.

(1) An objective lens that collects light emitted from a point light source on an observation sample, a photodetector that detects light from the observation sample, and the light condensed on the observation sample and the observation sample are relative to each other. In the confocal scanning optical microscope having a scanning mechanism for two-dimensionally scanning and a moving mechanism for moving the relative position of the focus position of the objective lens and the observation sample in the optical axis direction, the detection of the detector is performed. Level setting means for setting an arbitrary signal level for the signal, plane number setting means for setting the number of plane regions of different heights of the observation sample, and the level of the detection signal of the detector is set by the level setting means. Pixel number counting means for counting the number of pixels below the signal level, and whether the number of pixels from the pixel number counting means is equal to or greater than a preset number of pixels while moving in the optical axis direction by the moving mechanism. or not On the basis of the first detection means for detecting whether or not the scanning position of the observation sample is a flat area, and the detection result of the first detection means and the number of flat areas set by the flat surface number setting means. An automatic image forming apparatus for a confocal scanning optical microscope, comprising: a second detecting means for detecting an upper limit position and a lower limit position of an observation sample.

By doing so, even when measuring an observation sample having a plurality of planes of different heights and a steep slope between the planes, the focus of the objective lens can be measured. By moving the relative position between the position and the observation sample, the upper and lower limit positions thereof can be accurately detected, and the automatic setting of the measurement range can be reliably performed.

(2) In the automatic image forming apparatus for a confocal scanning optical microscope described in (1), the second detecting means first detects the upper limit position of the observation sample and detects the lower limit position of the observation sample. In this case, the moving range of the moving mechanism is limited.

By doing so, when the relative position between the focal point of the objective lens and the observation sample is moved to automatically set the vertical source position of the observation sample, interference between the observation sample and the objective lens is obviated. Can be prevented.

[0108]

As described above, according to the present invention, it is possible to measure an observation sample having a plurality of planes having different heights and having a steep slope between the planes. Even if there is, the relative position between the focus position of the objective lens and the observation sample is moved to accurately detect the upper and lower limit positions,
It is possible to provide an automatic image forming apparatus for a confocal scanning optical microscope, which can accurately perform automatic setting of a measurement range and can also prevent interference between a sample and an objective lens during automatic setting. .

[Brief description of drawings]

FIG. 1 is a block diagram showing the overall configuration of a system according to an embodiment of the present invention.

FIG. 2 is a diagram illustrating the shape of an observation sample according to the same example.

FIG. 3 is a flowchart showing the processing contents according to the embodiment.

FIG. 4 is a flowchart showing the processing content according to the embodiment.

FIG. 5 is a view for explaining the operation according to the embodiment.

FIG. 6 is a view for explaining the operation according to the embodiment.

FIG. 7 is a diagram for explaining a conventional scanning microscope having a confocal optical system.

[Explanation of symbols]

1 ... Confocal scanning optical microscope 2 ... Laser light source 3,23 ... Mirror 4, 21 ... Half mirror 5 ... Two-dimensional scanning mechanism 6 ... Revolver 7 ... Objective lens 8 ... Sample 9 ... Fine movement stage 10 ... Coarse stage 11, 20, 22, 24 ... Lens 12 ... Pinhole plate 13 ... Photodetector 14 ... Image processing unit 14A, 14B ... Image memory 15 ... Z movement circuit 16 ... XY scanning control unit 17 ... Computer 18 ... Monitor display 19 ... White light source 25 ... Television camera (TVC)

Claims (3)

(57) [Claims] 1. An observation sample placed on a stage is spotted.
Irradiate with the light source and re-illuminate the light with which this observation sample is irradiated.
And an objective lens that collects light, and the light on the observation sample that is collected by the objective lens
Moving said observation sample and scanning means for relatively scanning light detecting means for detecting the reflected light from the observation sample, the relative position between the focus position and the observation sample of the objective lens in the optical axis direction a scanning microscope having a moving means, the plane number setting means for setting the number of planar regions of different heights the observation sample, the level of the detection signal of the light detecting means is preset signal to
Pixel number counting means for counting the number of pixels below the signal level
And whether the scanning position of the observation sample is a flat area by detecting whether or not the number of pixels from the pixel number counting means exceeds a preset number of pixels by moving the moving means in the optical axis direction. to the first detection means, said detection result of the first detecting means, a second detection for detecting the upper limit position and lower limit position of the observation sample on the basis of the planar region at a set number of said plane number setting means And an automatic image forming apparatus for a scanning microscope . Wherein said second detection means detects the upper limit position of the observation sample before, in detecting the lower limit position of the observation sample
The automatic image forming apparatus of the scanning microscope according to claim 1, wherein the moving range of the moving unit is limited.
Place 3. The objective lens is limited to the moving range.
It is within the operating distance of.
Automatic image forming device for scanning microscope.