JP2005156651A - Scanning optical microscope - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To automatically adjust brightness of an image against the change of brightness of the image, which is caused by switching of the objective lens and the observation optical system, in a scanning optical microscope. <P>SOLUTION: In response to switching of the objective lens or the observation optical system, a CPU 102 reads information on optical characteristics, corresponding to a combination of the objective lens and the observation optical system after switching, and reference information on brightness from a memory 104 and uses these information to calculate the gain value of a light-receiving element 123 or 124 or the light quantity value of a light source 111 and sets the calculated value to the light-receiving element or the light source. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a scanning optical microscope having a plurality of objective lenses and a plurality of observation optical systems.

  The scanning optical microscope obtains a microscopic image of a sample from luminance information of the spot light reflected from the sample by XY scanning the spot light with respect to the surface of the sample. In this case, the magnification is adjusted by switching the objective lens. However, since the amount of transmitted light changes depending on the aperture angle, transmittance, focal length, and the like of the objective lens, the brightness of the image changes when the objective lens is switched.

  Therefore, in order to adjust the change in the brightness of the image caused by the switching of the objective lens, in the conventional scanning optical microscope, the brightness of the obtained image is adjusted every time the magnification is changed. It has been carried out by adjusting the level of the luminance information signal. For example, it is configured to give a signal of luminance information to the image display device through a manual adjustment type variable gain amplifier, and when the brightness of the image changes with the switching of the objective lens, the gain of the variable gain amplifier is manually adjusted. By doing so, the image has been adjusted to an optimal brightness.

  However, such manual brightness adjustment has a problem that it takes time until an appropriate brightness can be obtained. Moreover, the adjustments required for switching the objective lens include not only the brightness but also the focus adjustment.Therefore, several adjustments must be made manually each time the objective lens is switched, which is very troublesome. I was invited.

Therefore, the scanning optical microscope is equipped with a function that can automate the adjustment of the brightness change of the image caused by the switching of the objective lens and can follow the switching of the objective lens without time difference. (For example, see Patent Document 1).
FIG. 9 is a configuration diagram of the scanning optical microscope of Patent Document 1. The scanning optical microscope of FIG. 9 includes a microscope main body 1001, a CPU (Central Processing Unit) 1002, an operation unit 1003, a memory 1004, and a monitor 1005. The microscope main body 1001 includes a scanning optical microscope light source 1011, a polarizing beam splitter 1012, a two-dimensional scanning mechanism 1013, an objective lens holding revolver 1014, a beam splitter (optical path branching element) 1017, a pinhole 1018, and a light receiving element 1019.

A plurality of objective lenses 1015a, 1015b, and 1015c are attached to the revolver 1014. An observation target in the microscope main body 1001 is a sample 1016, and only one optical system including a light receiving element 1019 is provided as an observation optical system. It has been.
When exchanging the objective lens, the CPU 1002 issues an instruction to the revolver 1014, and rotates the revolver 1014 so that the objective lens to be used is arranged in the optical path. At the same time, the CPU 1002 reads the type information of the objective lens before and after the change from the memory 1004, calculates the gain after the change of the light receiving element 1019, and changes the gain of the light receiving element 1019 to the calculated value.

Further, a method for adjusting the gain of the light receiving element while observing an image (for example, see Patent Document 2), and a method for eliminating variations in the light receiving sensitivity range of a photomultiplier used as the light receiving element (for example, see Patent Document 3). ) Is also known.
Japanese Patent Application Laid-Open No. 08-021956 JP 2000-321503 A JP 2001-059939 A

However, image brightness adjustment in the above-described conventional scanning optical microscope has the following problems.
In a scanning optical microscope, an operation that causes a change in image brightness is not limited to switching of the objective lens. For example, in a scanning optical microscope having a plurality of observation optical systems, including microscope observation with a television camera, even if the observation optical system is switched, the brightness changes due to the difference in the configuration of the optical system and the light receiving element used. Occurs.

  Furthermore, even if the change in image brightness caused by switching the objective lens can be automatically adjusted to the optimum brightness, the operator can intentionally increase or decrease the brightness of the image. When it is desired to observe the image, it is necessary to adjust the brightness every time the objective lens is switched.

A first problem of the present invention is to provide a scanning optical microscope that automatically adjusts the brightness of an image in response to a change in the brightness of the image caused by switching between an objective lens and an observation optical system. is there.
A second problem of the present invention is to provide a scanning optical microscope that automatically adjusts the brightness of an image based on an arbitrary brightness set by an operator.

  The scanning optical microscope according to the first aspect of the present invention includes at least a plurality of objective lenses, a revolver that switches between the objective lenses, and a light receiving element that receives reflected light from an observation target and converts it into an electrical signal. A microscope image is obtained by measuring reflected light from an observation object while two-dimensionally scanning the spot light collected on the observation object with two or more observation optical systems, storage means, and adjustment means. To do. The storage unit stores optical characteristic information corresponding to each of a plurality of combinations of the objective lens and the observation optical system, and reference information representing the brightness of the reference image. When the objective lens or the observation optical system is switched, the adjustment unit reads out the optical characteristic information and the reference information corresponding to the combination of the objective lens and the observation optical system after switching from the storage unit, and uses the read information. The brightness value of the image after switching is adjusted by calculating the gain value of the light receiving element and setting the obtained gain value in the light receiving element.

  The scanning optical microscope according to the second aspect of the present invention receives a plurality of objective lenses, a revolver that switches between the objective lenses, a light source that can adjust the amount of light, and reflected light from an observation target to generate an electrical signal. At least two or more observation optical systems each having a light receiving element to be converted, storage means, and adjustment means are provided, and the reflected light from the observation target is measured while two-dimensionally scanning the spot light condensed on the observation target. By doing so, a microscope image is acquired. The storage unit stores optical characteristic information corresponding to each of a plurality of combinations of the objective lens and the observation optical system, and reference information representing the brightness of the reference image. When the objective lens or the observation optical system is switched, the adjustment unit reads out the optical characteristic information and the reference information corresponding to the combination of the objective lens and the observation optical system after switching from the storage unit, and uses the read information. The brightness value of the image after switching is adjusted by calculating the light quantity value of the light source and setting the obtained light quantity value as the light source.

  In the third aspect of the present invention, at least two or more observation optical systems of the scanning optical microscope according to the first or second aspect are arranged at a position conjugate with the light receiving element and the imaging position on the observation target. A confocal optical system having a pinhole and an imaging lens that forms an image of reflected light from the observation object at the position of the pinhole.

In the fourth aspect of the present invention, at least two observation optical systems of the scanning optical microscope in the first or second aspect include a non-confocal optical system having a light receiving element.
In the fifth aspect of the present invention, at least two or more observation optical systems of the scanning optical microscope in the first or second aspect include an observation optical system having an image sensor as a light receiving element.

  In the sixth aspect of the present invention, the scanning optical microscope according to the first or second aspect further includes display means. The display means displays brightness information indicating the brightness of the microscope image, and even if the objective lens or the observation optical system is switched, the image is displayed with the brightness adjusted by the adjustment means. Display the same brightness information as before switching.

  In the seventh aspect of the present invention, the scanning optical microscope according to the first or second aspect further includes an operation means. The operation unit causes the adjustment unit to perform a process of adjusting the brightness of the image using the optical characteristic information and the reference information stored in the storage unit regardless of whether the objective lens or the observation optical system is switched. .

  In the eighth aspect of the present invention, the scanning optical microscope according to the first or second aspect further includes an operating means. The operating means changes the brightness adjusted by the adjusting means, and the adjusting means is changed by changing the optical characteristic information or the reference information stored in the storage means based on the changed brightness. The brightness information is reflected in the optical characteristic information or the reference information.

  In the scanning optical microscope of the present invention, when the observation condition comprising the combination of the objective lens and the observation optical system is switched, the adjustment unit is configured to display an image having a predetermined brightness based on the information stored in the storage unit. Automatically adjusts the gain value of the light receiving element or the light amount value of the light source.

  An observation optical system is an optical system that forms an optical path that guides illumination light from a light source to an observation target and guides reflected light from the observation target to a light receiving element. Two or more observation optical systems share a part of the optical path. It is possible that Therefore, an observation optical system in which some of the optical elements included in a certain observation optical system are different is an observation optical system different from the observation optical system. In general, different observation optical systems use different observation methods.

  When the observation condition is switched, the adjustment unit calculates an optimal gain value or light amount value based on the optical characteristic information and the reference information corresponding to the switched observation condition stored in the storage unit, The gain value of the light receiving element or the light amount value of the light source used after switching is set to the calculated value. The information stored in the storage means is, for example, that the operator manually adjusts the image to an optimum brightness in advance and registers information such as optical characteristics, gain values, and light quantity values at that time in the storage means. Prepared.

  Furthermore, according to the operation means of the eighth aspect, it is possible to manually change the brightness again after switching, and the changed brightness can be reflected in the information stored in the storage means. For example, if you want to reflect the changed brightness only in the current observation condition, you only need to update the information of the optical characteristics corresponding to the observation condition at that time, if you want to reflect in all the observation conditions, Information representing the brightness at that time may be stored as new reference information.

  As described above, the brightness of the microscope image of the scanning optical microscope is adjusted together with the switching of the observation condition, and an image having the optimum brightness for the operator can be obtained immediately after the switching. Therefore, it is possible to automate adjustments to changes in image brightness accompanying switching of the objective lens and the observation optical system.

In addition, manual brightness adjustment after automatic adjustment and the adjustment result can be reflected in specific observation conditions or all observation conditions. The brightness manually set by the operator can be used in subsequent automatic adjustments. It can be used as a reference.
Therefore, in the operation of a scanning optical microscope having a plurality of objective lenses and a plurality of observation optical systems, an optimal observation image can be obtained in a short time after switching between the objective lens and the observation optical system.

  The storage means and the adjustment means correspond to, for example, a memory 104 and a CPU 102, which will be described later, the optical characteristic information corresponds to, for example, matrix data described later, and the reference information includes, for example, a gain of a light receiving element described later or a light source. This corresponds to the reference value of the amount of light and the reference value of the matrix data.

  The imaging element corresponds to, for example, a CCD (Charge Coupled Device) camera 515 described later, and the display unit corresponds to, for example, a monitor 105 and an operation unit 103 described later. Brightness information displayed by the display unit is, for example, This corresponds to a brightness display bar 903 described later. The operation means corresponds to, for example, an operation unit 103 described later and a graphical user interface (GUI) screen shown in FIG.

According to the present invention, in the operation of a scanning optical microscope having a plurality of objective lenses and a plurality of observation optical systems, a change in brightness caused by switching between the objective lens and the observation optical system is automatically adjusted, and Observation images having the same brightness can be obtained.
In addition, when the brightness is manually adjusted again after the automatic brightness adjustment, the adjustment result is recorded in correspondence with the specific observation condition including the combination of the objective lens and the observation optical system at that time. When switching to the same observation condition again, it is possible to reproduce the manually adjusted brightness. In addition, it is possible to reflect the result of the brightness adjustment performed manually not only for the observation condition at that time but also for all the observation conditions.

  Therefore, in the operation of a scanning optical microscope having a plurality of objective lenses and a plurality of observation optical systems, the operability after switching between the objective lens and the observation optical system is improved.

The best mode for carrying out the present invention will be described below in detail with reference to the drawings.
FIG. 1 is a configuration diagram of a scanning optical microscope according to an embodiment. This scanning optical microscope includes a microscope main body 101, a CPU 102, an operation unit 103, a memory 104, and a monitor 105. The microscope main body 101 includes a scanning optical microscope light source 111, a polarizing beam splitter 112, a ¼ wavelength plate 113, a two-dimensional scanning mechanism 114, a pupil projection lens 115, imaging lenses 116, 121, and 125, an objective lens holding revolver 117, A beam splitter 120, a pinhole 122, and light receiving elements 123 and 124 are included.

  A plurality of objective lenses 118a, 118b, and 118c are attached to the revolver 117. Here, three objective lenses are shown for convenience, but the number of objective lenses attached to the revolver 117 is generally arbitrary. The observation object in the microscope body 101 is a sample 119, and the light source 111 is a light emission source for illuminating the sample 119. For example, a laser light source is used as the light source 111.

  In this case, as the observation optical system, the sample 119 is irradiated with illumination light from the light source 111 and the reflected light from the sample 119 is guided to the light receiving element 123. Similarly, the illumination light from the light source 111 is sample 119. Two optical systems, a non-confocal optical system that guides the reflected light from the sample 119 to the light receiving element 124, are provided. The light receiving unit of the confocal optical system includes an imaging lens 121, a pinhole 122, and a light receiving element 123. The pinhole 122 is disposed at a position optically conjugate with the imaging position on the sample 119. On the other hand, the light receiving unit of the non-confocal optical system includes an imaging lens 125 and a light receiving element 124.

  The illumination light emitted from the light source 111 passes through the polarization beam splitter 112, passes through the quarter wavelength plate 113, rotates the polarization direction by 90 °, and is guided to the two-dimensional scanning mechanism 114. The two-dimensional scanning mechanism 114 scans the spot light on the sample 119 in the XY direction by two-dimensionally scanning the illumination light. As the two-dimensional scanning mechanism 114, for example, a galvanometer mirror is used.

  The illumination light that is XY scanned by the two-dimensional scanning mechanism 114 passes through the pupil projection lens 115 and the imaging lens 116 and is guided to the objective lens 118b. The objective lens is switched by rotating the revolver 117 and changing the objective lens arranged on the optical path of the illumination light. The revolver 117 may be rotated electrically or manually.

  At this time, the illumination light XY-scanned by the two-dimensional scanning mechanism 114 is configured to always pass through the pupil position of the objective lens 118b. The illumination light that has passed through the objective lens 118b is condensed on the sample 119 by the objective lens 118b to become spot light, and is scanned on the sample 119 in the XY direction.

  The illumination light is reflected on the sample 119 and guided as reflected light to the objective lens 118b, the imaging lens 116, the pupil projection lens 115, the two-dimensional scanning mechanism 114, and the quarter wavelength plate 113. The reflected light that has passed through the quarter-wave plate 113 is rotated again by 90 ° in the polarization direction and reaches the polarization beam splitter 112. At this time, the reflected light from the polarizing beam splitter 112 is guided not to the light source 111 but to the beam splitter 120 because the direction of polarization is 180 ° different from that of the illumination light.

  The reflected light guided to the beam splitter 120 has a constant branching ratio at the beam splitter 120 and a light receiving unit of a confocal optical system including the imaging lens 121, the pinhole 122, and the light receiving element 123, and the imaging lens. The light is guided to a light receiving portion of a non-confocal optical system composed of 125 and a light receiving element 124.

  The light receiving elements 123 and 124 are elements that detect luminance information of the reflected light, convert it into an electrical signal, and output it. As these light receiving elements, for example, a photomultiplier, a photodiode or the like is used. In the case of a photomultiplier, the gain of an electrical signal to be output is determined by adjusting sensitivity according to the applied voltage. In the case of a photodiode, the gain of an electric signal to be output is determined by adjusting an internal amplifier circuit.

  The electrical signals output from the light receiving elements 123 and 124 are converted into image data by data processing, and are displayed on the screen of the monitor 105 as a microscope image. At this time, the brightness of the displayed microscope image is determined by the magnitude of the electrical signal output from the light receiving elements 123 and 124.

  The CPU 102 and the memory 104 correspond to an information processing apparatus (computer) such as a personal computer, for example, and the memory 104 includes a ROM (read only memory) and a RAM (random access memory). The CPU 102 executes a control program using the memory 104 to control the microscope main body 101 and perform processing necessary for image brightness adjustment. As a result, the revolver 117 and the light receiving elements 123 and 124 are controlled by the CPU 102.

  Programs and data used by the CPU 102 are loaded into the memory 104 via an arbitrary computer-readable recording medium such as a memory card, a flexible disk, a CD-ROM (compact disk-read only memory), an optical disk, or a magneto-optical disk. be able to. Also, these programs and data can be loaded into the memory 104 from an external device via a communication network. In this case, the external device generates a carrier signal for carrying the program and data and transmits it to the scanning optical microscope via the communication network.

  The operation unit 103 is, for example, an input device such as a keyboard, a pointing device such as a mouse, or a touch panel, and inputs instructions and information from the operator to the CPU 102 via the GUI. The monitor 105 is a display device that displays a microscope image, and displays a GUI operation screen together with the image.

Next, the operation procedure of the operator and the control processing procedure by the CPU 102 will be described with reference to the flowchart of FIG.
The operator first sets the sample 119 on the microscope body 101, activates the scanning optical microscope, and starts scanning (step 301). Next, an objective lens and an observation optical system used for observation are set (step 302). Then, focus adjustment, zoom adjustment of the microscope image, and the like are performed, and the microscope image to be observed is displayed on the monitor 105 (step 303). In this case, as shown in FIG. 1, setting and adjustment are performed by the CPU 102 receiving an instruction from the operation unit 103 controlling the microscope body 101.

  Next, the operator adjusts the brightness of the image (step 304). This adjustment is manually performed via the operation unit 103. The brightness value adjusted by the operation unit 103 is instructed to the light receiving element of the currently selected observation optical system via the CPU 102. For example, when the confocal optical system is selected, the brightness adjustment result is instructed to the light receiving element 123, and when the non-confocal optical system is selected, the brightness adjustment result is input to the light receiving element 124. Instructed.

  When the microscope image is adjusted to an optimum brightness, the CPU 102 registers the gain value and optical characteristic matrix data of the light receiving element at that time in the memory 104 from the operation unit 103 (step 305). The gain value and matrix data are defined as H_0 and T_0, respectively, and when performing automatic brightness adjustment thereafter, the CPU 102 performs calculations using these values as reference values.

  As shown in FIG. 3, optical characteristic matrix data is registered in advance in the memory 104 in accordance with the combination of the type of objective lens and the type of observation optical system. For example, the matrix data corresponding to the combination of the objective lens of 5 × magnification and the confocal optical system is T_C5, and the matrix data corresponding to the combination of the objective lens of 100 × magnification and the non-confocal optical system is T_N100.

  These matrix data are determined from the relative value of the brightness of the microscope image when the standard sample is observed in advance as the sample 119 for each combination of the objective lens and the observation optical system. For example, if the observation optical system is common, the matrix data is proportional to the square of the transmittance of each objective lens with respect to the illumination light.

Next, the operator switches the objective lens or the observation optical system (step 306). In this case, as shown in FIG. 1, the CPU 102 that receives an instruction from the operation unit 103 controls the microscope main body 101 to switch observation conditions.
If the revolver 117 is an electric type, the CPU 102 controls to rotate the revolver 117 so that the designated objective lens is arranged on the optical path. If the revolver 117 is a manual type, the operator rotates the revolver 117 by itself. . In any case, the CPU 102 knows from the rotational position of the revolver 117 which objective lens is currently arranged on the optical path. Further, the CPU 102 determines which of the electrical signals output from the light receiving elements 123 and 124 is to be displayed on the monitor 105 as a microscope image, and switches the signal input to the monitor 105.

  During this switching, the CPU 102 reads out the gain value H_0 and matrix data T_0 registered in the memory 104 and matrix data corresponding to the observation condition after switching, and uses these data to determine the light receiving element after switching. A gain value is calculated (step 307). In this case, the CPU 102 multiplies H_0 by a correction term basically obtained from the ratio of T_0 and the matrix data after switching to calculate the gain value after switching.

For example, when photomultipliers are used as the light receiving elements 123 and 124, T is the matrix data after switching, and m is a unique parameter determined uniquely for each photomultiplier. H is calculated.

H = H_0 * (T / T_0) ^m

The value of the parameter m may be obtained experimentally or may be calculated from the data sheet of the photomultiplier manufacturer. This value is registered in advance in the memory 104 for each photomultiplier, and is read out by the CPU 102 in accordance with the observation optical system after switching.

The gain value calculated in this way is instructed from the CPU 102 to the light receiving element 123 or 124 as shown in FIG. 1, and the automatic brightness adjustment is completed (step 308).
Here, the operator can manually adjust the brightness or replace the sample 119 as necessary. For example, when it is desired to make the image brighter or darker intentionally, the gain value is manually adjusted via the operation unit 103.

  The operator first determines whether or not manual brightness readjustment is necessary (step 309). If readjustment is not required, it is next determined whether or not the sample 119 is to be replaced (step 309). 313). When the sample 119 is not exchanged, the procedure after step 306 is repeated. As a result, the objective lens or the observation optical system is switched again as necessary, and automatic brightness adjustment is repeated in the same procedure.

In step 309, if manual brightness readjustment is necessary, the same brightness adjustment as in step 304 is performed (step 310). At this time, the CPU 102 reflects the readjusted brightness in the memory 104 in accordance with an instruction from the operation unit 103.
If the operator wants to reproduce the brightness state after readjustment only under the observation conditions consisting of the current objective lens and observation optical system, he / she inputs an instruction to reflect only the current observation conditions and performs all observations. If you want to reproduce under conditions, enter an instruction to reflect as an offset in all observation conditions.

When an instruction to reflect the readjusted brightness only in the current observation condition is input from the operation unit 103, the CPU 102 updates and registers only the matrix data corresponding to the current observation condition in the memory 104 (step 311). ), The processing after step 306 is repeated.
At this time, matrix data that can reproduce the current gain value without changing H_0 and T_0 is calculated, and the value is registered in the memory 104 as new matrix data corresponding to the current observation condition. As a result, when the current observation condition is switched again in the future, the newly updated and registered matrix data can be read from the memory 104 and the switched gain value can be calculated.

  On the other hand, when an instruction for reflecting the readjusted brightness in all observation conditions is input from the operation unit 103, the processing after step 305 is repeated and the readjusted gain value is transferred from the operation unit 103 to the memory 104. Re-register as a new reference value H_0. Thereby, the state of the brightness after readjustment is reflected on all the observation conditions to be switched in the future.

  Further, when the operator wants to observe a new sample without reflecting the readjusted brightness in any observation condition, the procedure after step 302 is repeated after replacing the sample 119 (step 312). . Thereby, it is possible to reset the sample 119, start scanning, set the objective lens and the observation optical system, perform various adjustments, and reset H_0 and T_0. When it is desired to replace the sample 119 in step 313, the operation after step 312 is similarly performed.

In the embodiment of FIG. 1, two optical systems, a confocal optical system and a non-confocal optical system, are provided as the observation optical system of the scanning optical microscope, but a television observation optical system can be further added. It is.
FIG. 4 is a configuration diagram of a scanning optical microscope equipped with such a television observation optical system. The scanning optical microscope of FIG. 4 has a configuration in which the microscope main body 101 of FIG. 1 is replaced with a microscope main body 501. The microscope main body 501 includes beam splitters 511 and 512, television observation in addition to the configuration of the microscope main body 101. A television observation optical system including a light source for light 513, a slider 514, and a CCD camera 515 which is a light receiving element for television observation.

  For example, a white light source is used as the light source 513, and illumination light from the light source 513 passes through the slider 514 and the beam splitter 512, is guided to the objective lens 118 b by the beam splitter 511, and is imaged on the sample 119. The The reflected light from the sample 119 is guided toward the television observation optical system by the beam splitter 511 and guided to the CCD camera 515 by the beam splitter 512. Then, the light is received by the CCD camera 515.

  In this television observation optical system, for example, a plurality of observation methods such as bright field observation, dark field observation, and Nomarski differential interference observation can be used, and the optical system differs depending on each observation method. The plurality of observation optical systems in the television observation optical system are switched by switching a slider 514 equipped with a filter. This switching may be performed by the CPU 102 according to an instruction from the operation unit 103 or may be performed manually by the operator himself.

  The CCD camera 515 is controlled by the CPU 102 in the same manner as the light receiving elements 123 and 124, and the gain value is adjusted automatically and manually according to the same procedure as in FIG. Therefore, the matrix data registered in the memory 104 includes matrix data used for controlling the television observation optical system in addition to the matrix data shown in FIG. For example, when the observation optical systems for bright field observation, dark field observation, and Nomarski differential interference observation are mounted, matrix data as shown in FIG.

In the above embodiment, the brightness of the microscope image is adjusted by adjusting the gain of the light receiving element. However, it is also possible to adjust the brightness by adjusting the amount of light from the light source.
FIG. 6 shows control for adjusting the light amount of the light source 111 in the scanning optical microscope of FIG. In this case, the light source 111 is provided with a mechanism capable of continuously adjusting the amount of light. For example, when the light source 111 is a laser light source, although not shown, an ND (Neutral Density) filter whose density is continuously variable is installed at the emission position, and by changing the position of the ND filter on the laser optical axis, The amount of light transmitted through the ND filter can be continuously changed. This makes it possible to adjust the amount of light transmitted through the ND filter according to the combination of the objective lens and the observation optical system, and to illuminate the sample with the optimum amount of light.

In the above-described embodiment, the light amount adjustment is realized by using the ND filter. However, the present invention is not limited to this, and the light amount can be adjusted by voltage control to the laser light source.
At the time of automatic and manual brightness adjustment, the CPU 102 instructs the light source 111 on the light amount value instead of instructing the light receiving elements 123 and 124 on the gain value, and outputs the instructed light amount. Adjust the position. The light quantity value instructed at this time is calculated by the CPU 102 using matrix data as shown in FIG. 3 in the same manner as in the case of the gain value. However, a light intensity reference value is used instead of the gain reference value H_0.

  FIG. 7 shows control for adjusting the light amounts of the light sources 111 and 513 in the scanning optical microscope of FIG. In this case, not only the light source 111 but also the light source 513 is provided with a mechanism capable of continuously adjusting the amount of light. For example, when the light source 513 is a white light source, although not shown, an ND filter whose density is continuously variable is installed at the emission position, and the amount of transmitted light is adjusted by changing the position of the ND filter on the optical axis.

The light source 513 is controlled by the CPU 102 in the same manner as the light source 111, and the amount of light is adjusted automatically and manually. The light quantity value instructed at this time is calculated by the CPU 102 using matrix data as shown in FIG. 5 in the same manner as in the case of the gain value.
Next, an operation / display method for manually adjusting the brightness on the GUI screen will be described with reference to FIG.

  FIG. 8 shows an example of a GUI displayed on the screen of the monitor 105. The GUI screen in FIG. 8 includes adjustment buttons 901 and 902, a brightness display bar 903, and a reset button 904. When the adjustment button 901 is clicked, the microscope image is displayed brighter, and when the adjustment button 902 is clicked, the microscope image is displayed darker. The brightness display bar 903 moves up and down with manual adjustment of brightness using the adjustment buttons 901 and 902, and the current brightness state is represented by the position. The brightness adjustment for all viewing conditions is performed through such a common GUI.

  When manually adjusting the brightness of the image, the operator does not directly input the gain value or the light amount value, but inputs an instruction to select one of brighter or darker. On the GUI of FIG. 8, an instruction is input by clicking the adjustment button 901 or 902, and the current image can be brightened or darkened by the amount corresponding to the number of clicks. At this time, as shown in (1) of FIG. 8, the position of the brightness display bar 903 changes.

In the above-described embodiment, the brightness of the image is adjusted with the brightness display bar. However, an input screen on which a numerical value can be directly input may be displayed on the screen of the monitor 105.
Actually, as shown in FIG. 1 or FIG. 6, the CPU 102 determines the gain value of the light receiving element or the light amount value of the light source 111 included in the current observation optical system according to the number of clicks by the operation unit 103. The amount of change is determined, and the amount of change is instructed to the microscope body 101 to change the gain value or the light amount value.

  When observing with the brightness calculated from the reference value of gain or light quantity registered in the memory 104, the reference value of matrix data, and matrix data corresponding to the current observation condition, the brightness display bar 903 is displayed. Are displayed at the position of the reset button 904. Therefore, immediately after the observation conditions are switched and the automatic brightness adjustment is performed, the display is always displayed at the position of the reset button 904. Even after manual brightness adjustment, when the adjustment result is registered in the memory 104 as a reference value, the brightness display bar 903 is positioned at the position of the reset button 904 as shown in FIG. Return to.

Thus, since the position of the reset button 904 is the reference position for brightness adjustment, basically, it is optimal to perform observation with the brightness display bar 903 always displayed at the position of the reset button 904. A bright image can be obtained.
Further, according to the GUI of FIG. 8, it is possible to forcibly automatically adjust the brightness without switching the observation conditions of the scanning optical microscope. In this case, when the reset button 904 is clicked by the operation unit 103, the CPU 102 is instructed to automatically adjust the brightness.

  In response to this, the CPU 102 reads the reference value of gain or light amount, the reference value of matrix data, and matrix data corresponding to the current observation condition from the memory 104, and performs the same calculation as when the observation condition is switched. To obtain the adjusted gain value or light intensity value. Then, the gain value of the light receiving element or the light amount value of the light source 111 included in the current observation optical system is changed to the obtained value. As a result, the brightness display bar 903 returns to the position of the reset button 904 as shown in (2) of FIG.

When this function is used, a microscope image of the brightness registered in the memory 104 can be immediately displayed on the screen of the monitor 105 during manual brightness adjustment.
In the embodiments described above, the confocal optical system, the non-confocal optical system, and the television observation optical system (the bright field observation optical system, the dark field observation optical system, and the Nomarski differential interference observation optical system) are the observation optics. Although listed as a representative example of the system, the present invention is not limited to this, and can also be applied to observation optical systems based on other observation methods.

It is a block diagram of the 1st scanning optical microscope of this invention. It is a flowchart of the control processing in a 1st scanning optical microscope. It is a figure which shows the matrix data in a 1st scanning optical microscope. It is a block diagram of the 2nd scanning optical microscope of this invention. It is a figure which shows the matrix data in a 2nd scanning optical microscope. It is a figure which shows adjustment of the light quantity in a 1st scanning optical microscope. It is a figure which shows adjustment of the light quantity in a 2nd scanning optical microscope. It is a figure which shows the GUI screen of brightness adjustment. It is a block diagram of the conventional scanning optical microscope.

Explanation of symbols

101, 1001 Microscope body 102, 1002 CPU
103, 1003 Operation unit 104, 1004 Memory 105, 1005 Monitor 111, 1011 Scanning optical microscope light source 112, 1012 Polarization beam splitter 113 1/4 wavelength plate 114, 1013 Two-dimensional scanning mechanism 115 Pupil projection lens 116, 121, 125 Connection Image lens 117, 1014 Objective lens holding revolver 118a, 118b, 118c, 1015a, 1015b, 1015c Objective lens 119 Sample 120, 511, 512, 1017 Beam splitter 122, 1018 Pinhole 123, 124, 1019 Light receiving element 513 For TV observation Light source 514 Slider 515 CCD camera 901, 902 Adjustment button 903 Brightness display bar 904 Reset button

Claims (10)

A scanning optical microscope that acquires a microscope image by measuring reflected light from the observation object while two-dimensionally scanning the spot light condensed on the observation object,
A plurality of objective lenses;
A revolver for switching the plurality of objective lenses;
At least two observation optical systems each having a light receiving element that receives reflected light from the observation object and converts it into an electrical signal;
Storage means for storing optical characteristic information corresponding to each of a plurality of combinations of the objective lens and the observation optical system, and reference information representing the brightness of a reference image;
When the objective lens or the observation optical system is switched, information on the optical characteristics corresponding to the combination of the objective lens and the observation optical system after switching and the reference information are read from the storage means, and the light reception is performed using the read information. A scanning optical microscope comprising: an adjusting unit that calculates a gain value of an element and adjusts the brightness of an image after switching by setting the obtained gain value in the light receiving element. A scanning optical microscope that acquires a microscope image by measuring reflected light from the observation object while two-dimensionally scanning the spot light condensed on the observation object,
A plurality of objective lenses;
A revolver for switching the plurality of objective lenses;
A light source with adjustable light intensity;
At least two observation optical systems each having a light receiving element that receives reflected light from the observation object and converts it into an electrical signal;
Storage means for storing optical characteristic information corresponding to each of a plurality of combinations of the objective lens and the observation optical system, and reference information representing the brightness of a reference image;
When the objective lens or the observation optical system is switched, information on optical characteristics corresponding to the combination of the objective lens and the observation optical system after switching and the reference information are read from the storage means, and the light source is read using the read information. And an adjusting means for adjusting the brightness of the image after switching by setting the obtained light amount value to the light source.   The at least two observation optical systems include the light receiving element, a pinhole arranged at a position conjugate with the imaging position on the observation target, and reflected light from the observation target at the position of the pinhole. The scanning optical microscope according to claim 1, further comprising a confocal optical system having an imaging lens for imaging.   The scanning optical microscope according to claim 1, wherein the at least two observation optical systems include a non-confocal optical system having the light receiving element.   The scanning optical microscope according to claim 1, wherein the at least two or more observation optical systems include an observation optical system having an imaging element as the light receiving element.   Display brightness information indicating the brightness of the microscope image, even if the objective lens or the observation optical system is switched, if the image is displayed with the brightness adjusted by the adjustment means, 3. The scanning optical microscope according to claim 1, further comprising display means for displaying the same brightness information as before switching.   Regardless of whether the objective lens or the observation optical system is switched or not, the adjustment unit performs a process of adjusting the brightness of the image using the optical characteristic information and the reference information stored in the storage unit. The scanning optical microscope according to claim 1, further comprising operating means for causing the scanning optical microscope to operate.   The apparatus further includes an operation unit that changes the brightness adjusted by the adjustment unit, and the adjustment unit changes the optical characteristic information or the reference information stored in the storage unit based on the changed brightness. 3. The scanning optical microscope according to claim 1, wherein the changed brightness information is reflected in the optical characteristic information or reference information. A plurality of objective lenses, a revolver that switches between the plurality of objective lenses, and at least two observation optical systems each having a light receiving element that receives reflected light from an observation object and converts it into an electrical signal, A program for a computer that controls a scanning optical microscope that acquires a microscope image by measuring reflected light from the observation object while two-dimensionally scanning spot light collected on the object,
Storage means for storing optical characteristic information corresponding to each of a plurality of combinations of the objective lens and the observation optical system and reference information representing the brightness of the reference image when the objective lens or the observation optical system is switched. From the information of the optical characteristics and the reference information corresponding to the combination of the objective lens and the observation optical system after switching,
Calculate the gain value of the light receiving element using the read information,
A program that causes the computer to execute a process of adjusting the brightness of an image after switching by setting the obtained gain value in the light receiving element. At least two or more observations each having a plurality of objective lenses, a revolver for switching the plurality of objective lenses, a light source capable of adjusting the amount of light, and a light receiving element that receives reflected light from an observation target and converts it into an electrical signal. And a computer that controls a scanning optical microscope that acquires a microscope image by measuring reflected light from the observation object while two-dimensionally scanning the spot light collected on the observation object. The program of
Storage means for storing optical characteristic information corresponding to each of a plurality of combinations of the objective lens and the observation optical system and reference information representing the brightness of the reference image when the objective lens or the observation optical system is switched. From the information of the optical characteristics and the reference information corresponding to the combination of the objective lens and the observation optical system after switching,
Calculate the light quantity value of the light source using the read information,
A program for causing the computer to execute a process of adjusting the brightness of an image after switching by setting the obtained light quantity value in the light source.

Images