JP4180322B2 - Confocal microscope system and computer program for parameter setting - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a confocal microscope system for acquiring information such as a confocal image and height distribution of a sample based on light information from the sample, and more specifically, a confocal system capable of automatically setting various measurement parameters. The present invention relates to a microscope system.
[0002]
[Prior art]
In a confocal microscope, light from a sample is received by a light receiving element via a confocal optical system, and information such as a confocal image (hyperfocal depth image) and height distribution of the sample is acquired based on the amount of light received. Is done. For example, when the relative distance between the sample placed on the stage and the objective lens is changed in the optical axis direction, the amount of light incident on the light receiving element via the confocal optical system, that is, the amount of received light changes, The amount of light received is maximized when the surface is in focus. Therefore, the height information of the surface of the sample is calculated from the relative distance between the sample and the objective lens when the maximum amount of received light is obtained, and the height distribution of the surface of the sample is acquired by scanning the surface of the sample with light. be able to.
[0003]
The acquired height distribution is displayed on the screen of the display device by, for example, three-dimensional display. Alternatively, the height distribution replaced with a luminance distribution or a color distribution is displayed on the screen. A CRT (cathode ray tube) or LCD (liquid crystal display device) is used as a display device, and a confocal microscope is connected to a control controller, a display device, a console, and the like to constitute a confocal microscope system.
[0004]
Also, by combining the information on the amount of light received when each point (pixel) on the sample surface is in focus (that is, the maximum luminance information of each pixel), a black and white image of the sample surface with a very deep focal depth can be obtained. Can do. This image is a so-called confocal image (superfocus depth image).
[0005]
Furthermore, a color image of the sample surface in the same range as the confocal image can be obtained by separating a part of the light from the sample halfway from the confocal optical system and receiving it with the color imaging device. Unlike a confocal image, this color image includes a focused portion and a blurred portion according to the unevenness of the sample surface. It is also possible to obtain a color image that is substantially in focus at each pixel by performing a synthesis process that replaces the luminance signal of the color image with the luminance signal of the confocal image.
[0006]
In the confocal microscope system as described above, it is necessary to set various parameters (measurement conditions) in order to maximize the capability of the optical system. For example, if the relative distance between the sample and the objective lens is changed in the optical axis direction by moving the objective lens in the optical axis direction, it is necessary to set the movement range and movement pitch of the objective lens. Alternatively, it is necessary to set the light receiving sensitivity of the light receiving element and the attenuation amount by the filter according to the light reflectance of the surface of the sample. Furthermore, it is necessary to set the shutter speed, gain, and white balance for acquiring a color image by the color image sensor.
[0007]
When the user sets the measurement parameters as described above, a certain degree of knowledge and experience is required. Therefore, in a conventional confocal microscope system, a controller (processing device) automatically sets some parameters. The automatic setting of parameters reduces the burden on the user of the confocal microscope. For example, even a user who is not accustomed can easily acquire information such as a confocal image and height distribution of a sample using a confocal microscope.
[0008]
[Problems to be solved by the invention]
However, when the types of parameters to be automatically set increase, the time required for automatic setting becomes longer, and the operability may be worsened. For example, if all parameters are automatically set every time measurement is performed, parameters that do not need to be set are also automatically set, and time is lost until the user waits for completion of the automatic setting.
[0009]
If the automatic setting of each parameter is started individually, execution of useless automatic setting can be avoided, but the number of times that the user operates for the automatic setting of the parameter (for example, the number of button presses) increases. However, it will be annoying.
[0010]
For example, when similar samples are continuously measured, if many parameters are automatically set once, it is not necessary to automatically set each time the sample is exchanged. On the other hand, depending on the type and parameters of the sample, it may be better to perform automatic setting every time the sample is replaced.
[0011]
In view of the above-described conventional problems, the present invention provides a confocal microscope system and a parameter setting program that can flexibly and rationally set parameters as measurement conditions in a flexible and rational manner. For the purpose.
[0012]
[Means for Solving the Problems]
The confocal microscope system of the present invention is a confocal microscope system capable of automatically setting a plurality of measurement parameters, and for each of a part or all of the measurement parameters, the parameter setting is automatically set for each measurement. Automatic measurement mode and automatic measurement invalid mode that retains the setting value set by executing parameter automatic setting and uses the setting value for multiple measurements, and automatically selects one of these modes. A setting mode selection means is provided.
[0013]
According to such a configuration, a parameter that is a measurement condition is automatically set in a flexible manner when measuring a plurality of samples under the same measurement conditions or when sequentially measuring a plurality of different types of samples. The operation can be performed easily and rationally. When automatic invalid mode is selected, automatic setting is not performed for each measurement when measuring multiple samples in sequence, so the entire measurement is compared to when automatic valid mode is selected. It is possible to reduce the time required for In addition, when the automatic measurement valid mode is selected, an appropriate parameter is automatically set for each measurement, and the user does not need to perform a parameter setting operation for each measurement.
[0014]
In a preferred embodiment, the apparatus further comprises a manual mode in which a manually set value is used for a plurality of measurements, and the automatic setting mode selection means is configured to perform the measurement automatic valid mode, the measurement automatic invalid mode, and the manual mode. One of the three modes is selectable. Thereby, for example, it is possible to cope with a case where a user who is accustomed to the operation wants to shorten the time required for the entire measurement by manual setting without using automatic parameter setting.
[0015]
The computer program for parameter setting in the confocal microscope system of the present invention is a program to be executed by a computer of a confocal microscope system capable of automatically setting a plurality of measurement parameters, and each of some or all of the measurement parameters A measurement automatic enable mode that automatically sets parameters for each measurement, and a measurement automatic invalid mode that retains the set values set by executing parameter automatic settings and uses the set values for multiple measurements. A step of displaying a selection screen for the user to select one of them on the display device, and when the mode selected by the user using the selection screen is the automatic measurement valid mode, a measurement start command is issued. Performing automatic measurement parameter setting before executing measurement and using the selection screen If the mode selected by the user is the automatic invalid mode during measurement, the automatic parameter setting is executed according to the automatic setting command, the automatic setting value is stored, and the measurement is executed according to the measurement start command. Includes a step of reading out an automatically set value stored without executing automatic setting and using it for measurement.
[0016]
In a preferred embodiment, the computer program described above is configured so that a part of or all of the measurement parameters are automatically set at the time of measurement for each parameter, and a setting set by executing the parameter automatic setting. The user can select one of three modes including a measurement automatic invalid mode that retains a value and uses the set value for a plurality of measurements, and a manual mode that uses a manually set value for a plurality of measurements. A step of displaying a selection screen for selection on the display device, a step of displaying a manual setting screen for manual setting of the parameter on the display device, and a mode selected by the user using the selection screen is automatically measured In the valid mode, the measurement parameter automatic setting is executed before the measurement is executed in response to the measurement start command. If the mode selected by the user using the selection screen is the automatic invalid mode during measurement, the automatic setting of parameters is executed according to the automatic setting command, the automatic setting value is stored, and the measurement is started. Reading out the stored automatic setting value when measurement is performed in accordance with the command, and using it for measurement, and when the mode selected by the user using the selection screen is the manual mode, start measurement. A step of reading a set value manually set using the manual setting screen when performing measurement in accordance with the command, and using it for the measurement.
[0017]
Such a computer program is supplied in a state of being stored in a computer-readable storage medium such as a CD-ROM, and installed in the computer from the storage medium and executed.
[0018]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, embodiments of the present invention will be described with reference to the drawings.
[0019]
FIG. 1 shows a schematic configuration of a confocal microscope system according to an embodiment of the present invention. The confocal microscope system 1 includes a confocal microscope having a confocal optical system 2 and a non-confocal optical system 3, a laser drive circuit 44 of the confocal microscope, signal processing circuits 41, 42, and 43 from a light receiving element, and an objective lens. The controller includes a moving mechanism 40, a control unit 46 using a microcomputer, and the like, and a display device 47 and an input device 48 connected to the controller.
[0020]
First, the confocal optical system 2 of the confocal microscope and its signal processing will be described. The confocal optical system 2 includes a light source 10 for irradiating a sample wk with monochromatic light (for example, laser light), a first collimating lens 11, a polarizing beam splitter 12, a quarter wavelength plate 13, a horizontal deflection device 14a, and vertical deflection. A device 14b, a first relay lens 15, a second relay lens 16, an objective lens 17, an imaging lens 18, a pinhole plate 9, a light receiving element 19 and the like are included.
[0021]
As the light source 10, for example, a semiconductor laser that emits red laser light is used. The laser light emitted from the light source 10 driven by the laser driving circuit 44 passes through the first collimating lens 11, the optical path is bent by the polarization beam splitter 12, and passes through the quarter wavelength plate 13. Thereafter, the light is deflected in the horizontal (lateral) direction and the vertical (longitudinal) direction by the horizontal deflecting device 14a and the vertical deflecting device 14b, passes through the first relay lens 15 and the second relay lens 16, and is sampled by the objective lens 17. The light is condensed on the surface of the sample wk placed on the stage 30.
[0022]
The horizontal deflection device 14a and the vertical deflection device 14b are each configured by a galvanometer mirror, and scan the surface of the sample wk with the laser light by deflecting the laser light in the horizontal and vertical directions. For convenience of explanation, the horizontal direction is referred to as the X direction, and the vertical direction is referred to as the Y direction. The objective lens 17 is driven in the Z direction (optical axis direction) by the objective lens moving mechanism 40. Thereby, the relative position of the focal point of the objective lens 17 and the sample wk in the optical axis direction can be changed.
[0023]
However, the relative position in the optical axis direction between the focal point of the objective lens 17 and the sample wk can be changed by other methods. For example, instead of driving the objective lens 17 in the Z-axis direction, the sample stage 30 may be driven in the Z-axis direction. Or the structure which moves the focus of the objective lens 17 to a Z-axis direction by inserting the lens from which a refractive index changes between the objective lens 17 and the sample wk is also possible. The sample stage 30 can be displaced in the X, Y and Z directions by manual operation.
[0024]
The laser beam reflected by the sample wk follows the above optical path in reverse. That is, it passes through the objective lens 17, the second relay lens 16, and the first relay lens 15, and again passes through the quarter wavelength plate 13 through the horizontal deflection device 14a and the vertical deflection device 14b. As a result, the laser beam passes through the polarization beam splitter 12 and is collected by the imaging lens 18. The condensed laser light passes through the pinhole of the pinhole plate 9 disposed at the focal position of the imaging lens 18 and enters the light receiving element 19. The light receiving element 19 is composed of, for example, a photomultiplier tube (photomultiplier tube) or a photodiode, and converts the amount of received light into an electric signal. An electrical signal corresponding to the amount of received light is supplied to the first AD converter 41 via an output amplifier and a gain control circuit (not shown), and converted into a digital value.
[0025]
With the confocal optical system 2 configured as described above, height (depth) information of the sample wk can be acquired. The principle will be briefly described below.
[0026]
As described above, when the objective lens 17 is driven in the Z direction (optical axis direction) by the objective lens moving mechanism 40, the relative distance between the focal point of the objective lens 17 and the sample wk in the optical axis direction changes. Then, when the focal point of the objective lens 17 is focused on the surface of the sample wk, the laser light reflected on the surface of the sample wk is condensed by the imaging lens 18 through the above optical path, and almost all the laser light is collected. Passes through the pinhole of the pinhole plate 9. Therefore, at this time, the amount of light received by the light receiving element 19 is maximized. On the contrary, in a state where the focus of the objective lens 17 is deviated from the surface of the sample wk, the laser light collected by the imaging lens 18 is focused at a position deviated from the pinhole plate 9, so that some lasers Only light can pass through the pinhole. As a result, the amount of light received by the light receiving element 19 is significantly reduced.
[0027]
Therefore, if the received light amount of the light receiving element 19 is detected while driving the objective lens 17 in the Z direction (optical axis direction) at any point on the surface of the sample wk, the objective lens 17 when the received light amount becomes maximum. The position in the Z direction (the relative position of the focal point of the objective lens 17 and the sample wk in the optical axis direction) can be uniquely determined as height information.
[0028]
Actually, each time the objective lens 17 is moved by one step (one pitch), the surface of the sample wk is scanned by the horizontal deflection device 14a and the vertical deflection device 14b to obtain the amount of light received by the light receiving element 19. When the objective lens 17 is moved in the Z direction from the lower end to the upper end of the measurement range, received light amount data that changes according to the position in the Z direction is obtained for each point (pixel) in the scanning range.
[0029]
FIG. 2 is a graph showing an example of received light amount data that changes in accordance with the position of the objective lens 17 in the Z direction. Based on such received light amount data, the maximum received light amount and the position in the Z direction at that time are obtained for each point (pixel). Therefore, the distribution of the surface height of the sample wk on the XY plane is obtained. This process is executed by the control unit 46 using a microcomputer.
[0030]
The obtained surface height distribution information can be displayed on the monitor screen of the display device 47 by several methods. For example, the height distribution (surface shape) of the sample can be displayed three-dimensionally by three-dimensional display. Alternatively, it can be displayed as a two-dimensional distribution of brightness by converting the height data into luminance data. By converting the height data into color difference data, the height distribution can also be displayed as a color distribution.
[0031]
Further, a surface image (black and white image) of the sample wk is obtained from a luminance signal having the received light amount obtained for each point (pixel) in the XY scanning range as luminance data. If a luminance signal is generated using the maximum amount of received light in each pixel as luminance data, a confocal image with a very deep focal depth in focus at each point having a different surface height can be obtained. In addition, if the height (Z-direction position) at which the maximum received light amount is obtained at any target pixel is fixed, the received light amount of the pixel of the target pixel portion and the portion where the height difference is large is significantly reduced. A bright image is obtained only at the same height.
[0032]
Next, the non-confocal optical system 3 provided in the confocal microscope and its signal processing will be described. The non-confocal optical system 3 includes a white light source 20, a second collimating lens 21, a first half mirror 22, a second half mirror 23, and a color for irradiating the sample wk with white light (illumination light for photographing a color image). A CCD (image sensor) 24 and the like are included. Further, the non-confocal optical system 3 shares the objective lens 17 of the confocal optical system 2, and the optical axes of the two optical systems 1 and 2 partially coincide.
[0033]
For example, a white lamp is used as the white light source 20, but a dedicated light source may not be provided, and natural light or room light may be used. White light emitted from the white light source 20 passes through the second collimating lens 21, the optical path is bent by the first half mirror 22, and is condensed on the surface of the sample wk placed on the sample stage 30 by the objective lens 17. .
[0034]
The white light reflected by the sample wk passes through the objective lens 17, the first half mirror 22, and the second relay lens 16, is reflected by the second half mirror 23, and enters the color CCD 24 to form an image. The color CCD 24 is provided at a position conjugate to or close to the conjugate with the pinhole of the pinhole plate 9 of the confocal optical system 2. The color image picked up by the color CCD 24 is read out by the CCD drive circuit 43, and the analog output signal is given to the second AD converter 42 and converted into a digital value. The color image thus obtained is displayed on the monitor screen of the display device 47 as an enlarged color image for observing the sample wk.
[0035]
In addition, a combination of a confocal image having a deep focal depth obtained by the confocal optical system 2 and a normal color image obtained by the non-confocal optical system 3 can be used to obtain a focal depth that is substantially in focus at all pixels. Deep color confocal images can also be generated and displayed. For example, a color confocal image can be easily generated by replacing the luminance signal constituting the color image obtained by the non-confocal optical system 3 with the luminance signal of the confocal image obtained by the confocal optical system 2. be able to.
[0036]
The controller including the control unit 46 also controls the processing related to the color image as described above. An input device 48 such as a console (console console) and a display device 47 such as a CRT (cathode ray tube) or LCD (liquid crystal display device) are connected to the controller. A pointing device such as a mouse is also connected as the input device 48.
[0037]
The user can set various measurement parameters using the input device 48 in accordance with the guidance displayed on the screen of the display device 47. For example, the Z direction movement range (measurement range) and movement pitch of the objective lens 17 are set. Alternatively, the light receiving sensitivity (PMT gain) of the light receiving element 19 and the attenuation amount by the ND filter are set according to the light reflectance of the surface of the sample wk, so that the brightness of the confocal image becomes appropriate. . The shutter speed, gain, and white balance for obtaining a color image by the color CCD 24 are set.
[0038]
Many of these parameters can be set automatically (automatic adjustment), and even unskilled users can easily obtain confocal images, height distribution information, etc. of the sample wk by using automatic parameter settings. can do. In addition, manual setting of measurement parameters is possible, and for users who are accustomed to operation, manual setting may be more suitable for quick measurement and fine measurement than automatic setting.
[0039]
Further, the confocal microscope system 1 (controller) of the present embodiment is also provided with an interface for connecting an external computer system such as a personal computer. By connecting an external computer system in which dedicated software for controlling the confocal microscope is installed to the confocal microscope system 1, it is possible to seamlessly process the image information and height distribution information of the acquired sample wk. Is possible. Furthermore, by including a parameter setting program as described below in the dedicated software, the measurement parameters of the confocal microscope can be easily and rationally set using an external computer system.
[0040]
FIG. 3 is a block diagram illustrating a hardware configuration example in which the external computer system 50 is connected to the controller of the confocal microscope system 1. The external computer system 50 includes a display device 51 such as a CRT or LCD, a keyboard 52, a mouse (may be another pointing device) 53, a communication interface 54 such as RS232C or USB (Universal Serial Bus), a processing device (CPU) 55, A main memory 56 that is a semiconductor storage medium, a fixed disk device 57 that is an auxiliary storage device, and a removable disk device 58 are provided.
[0041]
Dedicated software (hereinafter referred to as observation application) for controlling the confocal microscope is supplied in a state of being stored in a storage medium 59 such as a CD-ROM, and a removable disk device 58 such as a CD-ROM drive device. Is read from the storage medium 59 and installed in the fixed disk device 57. The program installed in the fixed disk device 57 is loaded into the main memory 56 and executed by the processing device 55.
[0042]
FIG. 4 is a diagram illustrating an example of a screen displayed on the display device 51 when the observation application is activated. A large rectangular result display area 61 on the left side of the screen is a confocal image obtained from the color image obtained from the color CCD 24 of the non-confocal optical system 3 of the confocal microscope system 1 or the light receiving element 19 of the confocal optical system 2. This is an area for displaying an image or a measurement result of the height distribution. The vertically long operation unit area 62 on the right side is an area for setting various measurement parameters.
[0043]
FIG. 5 is an enlarged view of the operation unit area 62 of FIG. Manual operation of each measurement parameter can be performed by operating the push button and slide bar of the operation unit area 62 displayed on the screen of the display device 51 and selecting each pull-down menu using the mouse 53. . For example, the brightness of the display of the color image obtained from the color CCD 24 can be adjusted by moving the camera gain adjustment slide lever 63 up and down by dragging the mouse. The brightness of the color image display can also be adjusted by moving the adjacent ND filter adjusting slide lever 64 up and down.
[0044]
Alternatively, the Z-direction movement range (measurement range) of the objective lens 17 can be set by selecting an appropriate value from a pull-down menu that appears when the triangle mark 65 on the right side of the distance is clicked with the mouse 53. Similarly, an appropriate Z-direction movement pitch can be set for the pitch below that by selecting an appropriate numerical value from a pull-down menu that appears when the triangle mark 66 is clicked.
[0045]
The “measurement” button 67 is a push button for instructing the start of measurement. The “auto set” button 68 is used in conjunction with the selection of “invalid during measurement” described later, and is a push button for automatically setting parameters. An “Auto focus” button 69 is a push button for activating autofocus, and is provided on the main screen of the operation unit area 62 so that a frequently used autofocus function can be activated with few operations.
[0046]
In addition, an item “auto setting” is included in a pull-down menu that appears when “option” on the menu bar is clicked with the mouse 53. By selecting this “auto setting”, an automatic setting dialog shown in FIG. 6 appears, and automatic setting of main measurement parameters can be performed easily and efficiently.
[0047]
FIG. 6 is a diagram illustrating an example of an auto setting dialog for easily and efficiently performing automatic setting of main measurement parameters. The auto setting dialog 70 includes three screens that can be switched by three tabs. FIG. 6 shows a screen when the “Pitch & Distance” tag 71 is selected. FIG. 7 shows a screen when the “auto focus & PMT gain” tag 72 is selected, and FIG. 8 shows a screen when the “camera” tag 73 is selected. In each screen of FIGS. 6 to 8, the lower half is used for setting common items. This will be described later.
[0048]
In the screen of FIG. 6, the pitch (Z-direction movement pitch) and the distance (Z-direction movement range) can be automatically set. Any one of “valid at measurement”, “invalid at measurement”, and “manual” included in the automatic setting mode selection means 74 can be selected. These three modes correspond to an automatic valid mode at measurement, an automatic invalid mode at measurement, and a manual mode, respectively.
[0049]
When “valid at measurement” is selected, automatic setting is performed prior to measurement processing at the timing when the start of measurement is instructed. When “invalid at measurement” is selected, automatic setting is executed when the “auto set” button 68 in the operation unit area 62 shown in FIG. 5 is pressed, and the automatically set value is held. That is, automatic setting is not performed every time measurement is started. When “Manual” is selected, the parameter is not automatically set. That is, the value manually set using the distance triangle mark 65 and the pitch triangle mark 66 in the operation unit area 62 shown in FIG. 5 is effective.
[0050]
FIG. 9 is a flowchart showing an outline of processing relating to parameter setting and measurement execution depending on which of the measurement automatic valid mode, the measurement automatic invalid mode, and the manual mode is selected. Based on this flowchart, the difference in processing by the above three types of selection will be described. In step # 101, it is checked whether the selection contents of the automatic setting, that is, “valid at measurement”, “invalid at measurement”, or “manual” is selected.
[0051]
If “valid at measurement” is selected (Yes in step # 102), the process jumps to step # 109. If neither “valid during measurement” nor “invalid during measurement” is selected (No in step # 103), that is, if “manual” is selected, the manual setting value is read in step # 104, and step # 109.
[0052]
When “invalid at measurement” is selected (Yes in step # 103), automatic setting is executed (step # 105) when the “auto set” button 68 in the operation unit area 62 is pressed (step # 105). 106) After the automatic setting value is stored (step # 107), the process proceeds to step # 109. In other cases, the automatic setting value is read (step # 108), and the process proceeds to step # 109.
[0053]
In step # 109, a measurement start command is checked. That is, when the “measurement” button 67 in the operation unit area 62 is pressed (Yes in step # 109), it is checked again in step # 110 whether “valid at measurement” is selected, and “valid at measurement”. Is selected (Yes in Step # 110), automatic parameter setting is executed here (Step # 111). When “valid at measurement” is not selected (No in step # 110), the measurement automatic disable mode or manual mode is set, and parameter setting values used for measurement are already prepared as described above. Step # 111 is skipped. In the subsequent step # 112, measurement is performed.
[0054]
Therefore, the user selects “invalid at measurement” (automatic invalid mode at measurement) when performing measurement of a plurality of samples under the same measurement conditions, sets the first sample on the stage 30 and performs “auto set”. By pressing the button 68, each parameter is automatically set and stored, and then the “measurement” button 67 is pressed. Since automatic setting is not executed when measuring a plurality of samples sequentially, the time required for the entire measurement can be shortened as compared with the case where “valid at measurement” (automatically valid mode at measurement) is selected.
[0055]
Also, when there is a possibility that the appropriate measurement conditions (parameters) may change, such as when measuring multiple samples of different types, select “Valid at measurement” (automatically valid at measurement mode). Good. In this case, since an appropriate parameter is automatically set for each measurement, the user does not need to perform a parameter setting operation for each measurement.
[0056]
Note that the flowchart of FIG. 9 is simplified, and in actuality, depending on the measurement parameters, different automatic setting selections may be performed as described later. Further, the parameter setting values automatically set in step # 106 or step # 111 are temporarily stored, and the display positions and triangles of the adjustment slide levers 63 and 64 for manual setting in the operation section area 62 shown in FIG. This is reflected in the setting value displayed on the left side of the marks 65 and 66. If the user is not satisfied with the automatic setting value, the setting value can be changed (finely adjusted) by manual setting as described above. That is, even if the manual mode is not changed, the set value can be changed (finely adjusted) in the measurement automatic invalid mode.
[0057]
In the screen of FIG. 7 in which the “auto focus & PMT gain” tag 72 is selected, auto focus and PMT gain can be automatically set. The PMT gain is linked to the attenuation factor by the ND filter, and the brightness of the display of the color image obtained from the color CCD 24 can be automatically adjusted. With respect to this parameter, any one of “valid at measurement”, “invalid at measurement”, and “manual” included in the automatic setting mode selection means 74 can be selected. The difference in the parameter setting operation when each is selected is as described above. When “Manual” is selected, values manually set using the camera gain adjusting slide lever 63 and the ND filter adjusting slide lever 64 in the operation section area 62 shown in FIG. 5 are valid.
[0058]
In addition, it is possible to select whether to perform standard detection or high-speed detection. When “high-speed detection” is selected, automatic setting of the PMT gain is completed in a short time without moving the objective lens 17 up and down. For example, when a user who is familiar with the operation performs the automatic setting of the PMT gain after manually focusing, if this “high speed detection” is selected, the objective lens 17 is not moved up and down. Automatic setting can be completed in a short time.
[0059]
If a “target range designation” button is pressed on the screen of FIG. 7, it is possible to designate a range (range on the XY plane) to be automatically set for autofocus and PMT gain. For example, the range of the rectangle can be specified by dragging the mouse along the diagonal line of the rectangle. Measurement of the measurement target location when the light reflectance is significantly different due to the difference in material, such as the soldered part or bonding part of the printed wiring board and its peripheral area, or the resin metal plating part and other parts. It is necessary to prevent the conditions from being automatically set inappropriately due to the influence of surrounding parts. In such a case, by using the above-described target range specification, the measurement condition of the measurement target portion is automatically set appropriately.
[0060]
In the example of the present embodiment, the “target range designation” button is provided only for the automatic setting of autofocus and PMT gain. However, the “target range designation” button is provided for other parameters as necessary, and the target of automatic setting is set. A range may be designated. Also, in principle, all parameters may be automatically set according to information obtained from the specified range, and parameters that should not be specified for range specification, such as pitch, may be specified.
[0061]
On the screen shown in FIG. 8 when the “camera” tag 73 is selected, automatic setting of parameters relating to the color CCD camera can be designated. That is, automatic setting of “white balance”, “camera gain”, and “camera brightness” can be designated. White balance is a function that automatically adjusts the color temperature of a color image obtained from the color CCD 24. The camera gain is a function for automatically adjusting the brightness (amplification amount) of the color image. The camera brightness is a function for automatically adjusting the brightness (light receiving amount) of a color image by changing the shutter speed.
[0062]
As shown in FIG. 8, for “camera gain” and “camera brightness”, either “always enabled” or “always disabled” can be selected. When “always enabled” is selected, the automatic adjustment function provided in the color CCD camera is utilized. When “always disabled” is set, the automatic adjustment function provided in the color CCD camera is set to be disabled. With respect to “white balance”, one can be selected from three types obtained by adding “invalid at measurement” to “always valid” and “always invalid”. “Invalid at measurement” is a function that sets a white color as a reference and determines a white balance in advance for the set white color, and is realized by using a function called “one push”. The “always enabled” function in white balance is a function that disables the auto adjustment function provided in the color CCD camera only during measurement.
[0063]
Next, setting of common items displayed in the lower half of each screen of FIGS. 6 to 8 will be described. As a first common item, it is possible to set whether or not to perform autofocus interlock. When the above-mentioned “invalid at measurement” is set, if autofocus is disabled while the focus is not in focus, other parameters may not be set to the optimum conditions. Therefore, if this “auto focus interlock” is checked, items that are selected to be processed automatically (automatic) among items such as pitch, distance, PMT gain, white balance, camera gain, and camera brightness. On the other hand, before performing the auto process, the auto focus is executed only once, and the selected item is auto-processed in a focused state.
[0064]
Further, as a second common item, it is possible to set a thinning amount in order to shorten the detection time of “distance” and “PMT gain”. For parameters that are automatically set while moving the objective lens 17 in the Z direction, such as “Distance” and “PMT gain”, the setting is faster if the processing is performed manually or at a pitch larger (coarse) than the automatically set pitch. Processing is complete. Therefore, the thinning amount can be set. The detection accuracy becomes worse as the thinning amount is set larger, but the detection time becomes shorter.
[0065]
In the above embodiment, the case where dedicated software for controlling the confocal microscope is installed in the external computer system 50 and measurement parameters are set on the computer screen has been described. However, the present invention includes such an embodiment. It is not limited. For example, a measurement parameter automatic setting program as in the above embodiment is incorporated into a control program stored in a storage medium such as a ROM built in the controller of the confocal microscope system 1, and the confocal microscope system 1 has a standard display device. 47 and the input device 48 may be used to automatically set the measurement parameters as in the above embodiment.
[0066]
Further, in order to acquire the height information of the surface of the sample wk by the confocal optical system 2, the stage 30 may be moved up and down instead of moving the objective lens 17 in the Z direction (up and down movement). In addition, the present invention can be implemented in various forms.
[0067]
For example, scanning of a sample by light is not limited to two-dimensional scanning by horizontal deflection and vertical deflection, and various scanning methods can be considered. For example, if a cylindrical lens is used to generate light that is elongated in the X direction (slit light) and is deflected in the Y direction, two-dimensional scanning is possible.
[0068]
The confocal microscope of the above embodiment is a reflection type microscope, but the present invention can be applied to a transmission type confocal microscope. In the case of a transmission type microscope, laser light from a confocal optical system and white light from a non-confocal optical system are irradiated from the back surface of the sample. The light source of the confocal optical system may include not only a monochromatic light source including a laser light source but also a plurality of wavelengths. The light source of the non-confocal optical system can be replaced with natural light or room light.
[0069]
【The invention's effect】
As described above, according to the confocal microscope system and its parameter setting program of the present invention, when measuring a plurality of samples under the same measurement conditions or sequentially measuring a plurality of different types of samples. An operation for automatically setting a parameter as a measurement condition can be easily and rationally performed in a flexible manner corresponding to such a measurement mode.
[Brief description of the drawings]
FIG. 1 shows a schematic configuration of a confocal microscope system according to an embodiment of the present invention.
FIG. 2 is a graph showing an example of received light amount data that changes in accordance with the position of the objective lens in the Z direction.
FIG. 3 is a block diagram illustrating a hardware configuration example in which an external computer system is connected to a controller of a confocal microscope system.
FIG. 4 is a diagram illustrating an example of a screen displayed on a display device when an observation application is activated.
FIG. 5 is an enlarged view of an operation unit area in FIG. 4;
FIG. 6 is a diagram showing an example when the first tag of an auto setting dialog for easily and efficiently performing automatic setting of main measurement parameters is selected.
FIG. 7 is a diagram illustrating an example when a second tag of an auto setting dialog for easily and efficiently performing automatic setting of main measurement parameters is selected.
FIG. 8 is a diagram illustrating an example when a third tag of an auto setting dialog for easily and efficiently performing automatic setting of main measurement parameters is selected.
FIG. 9 is a flowchart showing an outline of processing relating to parameter setting and measurement execution depending on which of the measurement automatic valid mode, the measurement automatic invalid mode, and the manual mode is selected.
[Explanation of symbols]
1 Confocal microscope system
50 External computer system
59 Storage media
62 Operation area
74 Automatic setting mode selection means

Claims (4)

This is a confocal microscope system that can automatically set multiple measurement parameters. For each or all of the measurement parameters, automatic measurement mode that automatically sets parameters for each measurement and automatic parameter setting The automatic setting mode selection means for selecting any one mode is provided, which includes a measurement automatic invalid mode that retains the setting value set by execution of and uses the setting value for a plurality of measurements. A confocal microscope system characterized by that. A manual mode that uses manually set values for a plurality of measurements is further provided, and the automatic setting mode selection means includes three modes including the measurement automatic valid mode, the measurement automatic invalid mode, and the manual mode. The confocal microscope system according to claim 1, wherein one of the two is selectable. A computer program for parameter setting to be executed by a computer of a confocal microscope system capable of automatically setting a plurality of measurement parameters,
For each part or all of the measurement parameters, the automatic setting mode for measurement that automatically sets the parameter for each measurement and the setting value set by executing the parameter automatic setting is retained, and the setting value is set for multiple measurements. Displaying on the display device a selection screen for the user to alternatively select the automatic invalid mode during measurement using
When the mode selected by the user using the selection screen is the automatic measurement valid mode at the time of measurement, the step of performing automatic setting of measurement parameters before performing measurement in accordance with the measurement start command;
When the mode selected by the user using the selection screen is the automatic invalid mode at the time of measurement, the automatic setting of the parameter is executed in accordance with the automatic setting command, the automatic setting value is stored, and the measurement start command is A parameter setting computer program comprising: a step of reading a stored automatic setting value and using it for measurement without executing automatic setting when performing measurement. A computer program for parameter setting to be executed by a computer of a confocal microscope system capable of automatically setting a plurality of measurement parameters,
For each part or all of the measurement parameters, the automatic setting mode for measurement that automatically sets the parameter for each measurement and the setting value set by executing the parameter automatic setting is retained, and the setting value is set for multiple measurements. The display device displays a selection screen for the user to select one of three modes, including an automatic invalid mode during measurement that uses and a manual mode that uses manually set values for multiple measurements. Step to
Displaying a manual setting screen for manual setting of the parameter on a display device;
When the mode selected by the user using the selection screen is the automatic measurement valid mode at the time of measurement, the step of performing automatic setting of measurement parameters before performing measurement in accordance with the measurement start command;
When the mode selected by the user using the selection screen is the automatic invalid mode at the time of measurement, the automatic setting of the parameter is executed in accordance with the automatic setting command, the automatic setting value is stored, and the measurement start command is Reading out the stored automatic setting value when performing measurement and using it for measurement; and
When the mode selected by the user using the selection screen is the manual mode, when the measurement is executed in accordance with the measurement start command, the setting value manually set using the manual setting screen is read and measured. And a parameter setting computer program.

Images