JPH0821956A - Scanning type optical microscope - Google Patents

Abstract

PURPOSE:To automatize adjustment for the change of the brightness of an image accompanying the switching of an objective lens and to follow up the switching of the objective lens without a time difference. CONSTITUTION:In this scanning type optical microscope; a sample is irradiated with spot light to perform two-dimensional scanning through the objective lens of the optical micorscope and light quantity from the sample is measured so as to obtain the microscopic image of the sample. The microscope is provided with a means 11 giving information on the kinds of the objective lens in the optical path of the spot light at present and the objective lens after it is changed in the case of changing the objective lens 7, and a storage means 13 storing information on the optical characteristic of the objective lens corresponding to the information on the kind of the objective lens. Furthermore, the microscope is equipped with an arithmetic means 12 calculating the difference of the light quantity between the objective lens before it is changed and the objective lens after it is changed based on the information on the optical characteristic corresponding to the information on the kinds of the objective lens at present in the optical path of the spot light and the objective lens after it is changed obtained from the storage means 13 and a means 10 adjusting sensitivity based on output from the arithmetic means 12.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a scanning optical microscope for observing an image by scanning a point light source on a sample.

[0002]

2. Description of the Related Art In an optical microscope, a sample on a slide placed on a stage is magnified and observed with an objective lens. Generally, the sample is illuminated by condensing a light from a light source such as a lamp with a condenser lens. Was used to make it uniform over the entire observation area of the sample.

However, when such a structure is adopted as an illumination system, there are problems such as flare, and it is difficult to observe a low-contrast sample. A light projection type (spot light projection type) optical microscope has been proposed. This optical microscope irradiates an observation sample in a point shape with a point light source,
Thereby, the light transmitted through the observation sample (transmitted light) is imaged again in a point shape, and this is detected by a detector having a pinhole opening to obtain density information of the image. However,
Since this alone can obtain only the density of the point illuminated by the point light source, the XY scanning method is adopted in which the sample is moved in the X-axis and Y-axis directions and mechanically moved in the two-dimensional plane.
CRT while XY scanning is synchronized with this XY scanning
An image is displayed on an image display device such as a display corresponding to the signal of the density information so that the image can be observed.

However, in the case of this apparatus, problems such as flare can be solved and a high-resolution image can be observed, but on the other hand, the sample is moved in the directions of the X-axis and the Y-axis to be two-dimensional. Since the scanning method that mechanically moves in the plane is adopted, only light samples can be targeted, and non-fixed samples such as culture samples in petri dishes could not be observed. .

Therefore, as the next stage of such a scanning type microscope apparatus (scanning optical microscope), spot-like light for irradiating the sample is applied to the surface of the sample in the X-axis and Y-axis.
A configuration in which scanning is performed in the axial direction has been proposed. With this configuration, the problems of the conventional scanning microscope apparatus have been solved, but the following problems have newly arisen.

That is, the scanning optical microscope obtains concentration information by scanning spot light on the surface of the sample by XY scanning, but in principle, it is the optical microscope itself, and the magnification adjustment is performed by the objective. This is done by switching the lens.
Since the amount of transmitted light of the optical lens changes depending on the aperture angle, the transmittance, the focal length, etc., the brightness of the image changes when the objective lens is switched.

Therefore, it becomes necessary to adjust the change in the brightness of the image due to such switching of the objective lens.
In the conventional scanning optical microscope, the brightness of the obtained image is adjusted by adjusting the level of the signal of the density information. For example, a signal of density information is provided to an image display device through a manually adjustable variable gain amplifier, and when the image brightness changes as the objective lens is switched, the gain of the variable gain amplifier is manually adjusted. By doing so, an image with optimum brightness is adjusted.

[0008]

A scanning optical microscope irradiates a sample with a spot light through an objective lens facing the sample and scans the sample with the spot light by XY scanning. The density information of the surface is obtained, and the density information is given to the image display device as brightness information so that it is displayed as an image and the microscopic image of the sample is observed. Since the microscope itself is used, when observing the sample, the magnification of the objective lens is changed as necessary.

However, since the amount of transmitted light of the optical lens changes depending on the aperture angle, the transmittance, the focal length, etc., the brightness of the image changes when the objective lens is switched.

Therefore, in the conventional scanning optical microscope,
The sensitivity adjustment for determining the brightness of the obtained image is a manual adjustment method of an observer, which is performed by adjusting the level of the signal of the density information using a manually adjustable variable gain amplifier.

As described above, conventionally, a method has been used in which the observer manually adjusts the brightness of the obtained image so as to obtain the optimum brightness. Therefore, the objective lens may be replaced.
If there is a sudden change in brightness, there is a problem that it takes time to obtain appropriate brightness with manual adjustment, and the adjustments required when switching the objective lens Not only the brightness but also the focus and the like require that many adjustments be performed manually each time the objective lens is switched, which is very cumbersome and causes poor operability.

Therefore, the object of the present invention is to
An object of the present invention is to provide a scanning optical microscope that can automatically adjust the change in the brightness of an image due to the switching of the objective lens and can follow the switching of the objective lens without any time difference.

[0013]

In order to achieve the above object, the present invention is configured as follows. That is, in a scanning optical microscope, a sample image is obtained by irradiating a sample with spot light that is two-dimensionally scanned through an objective lens of the optical microscope and measuring the amount of light from the sample obtained by this. Means for giving the type information of the current objective lens in the optical path of the spot light and the objective lens after the exchange, and the memory for storing the optical characteristic information of each objective lens in correspondence with the type information of the objective lens. Means and the amount of light of the objective lens before the exchange and the objective lens after the exchange from the optical characteristic information corresponding to the type information of the objective lens after the exchange with the current objective lens in the optical path of the spot light obtained from the storage means. The calculation means for calculating the difference and the means for adjusting the sensitivity by the output of the calculation means are provided.

[0014]

The scanning optical microscope irradiates a sample with spot light that is two-dimensionally scanned through the objective lens of the optical microscope, and the light quantity from the sample obtained by this is obtained, for example, reflected light from the sample, transmitted light, and fluorescence. The microscope image of the sample is obtained by measuring the amount of light such as, but in the scanning optical microscope of the present invention, when the objective lens is replaced, the means for giving the type information of the objective lens is currently in the optical path. The type information of the objective lens after replacement with the objective lens in (1) is given to the calculation means. Then, the calculation means obtains optical characteristic information corresponding to the type information of the objective lens from the storage means for storing the characteristic of each objective lens, and based on this, the light amount difference between the objective lens before replacement and the objective lens after replacement is obtained. calculate. Then, the sensitivity of the scanning optical microscope is adjusted by the sensitivity adjusting means in accordance with the light amount difference output obtained by the calculating means.

As described above, the sensitivity of the scanning optical microscope is adjusted as the objective lens is replaced, and an image having the same brightness can be obtained before and after the replacement. Therefore, according to the present invention, it becomes possible to automatically adjust the change in the brightness of the image due to the switching of the objective lens, and moreover, the switching of the objective lens can be followed without any time difference and the observation can be started in a short time. It is possible to provide a scanning optical microscope capable of performing the above.

[0016]

Embodiments of the present invention will be described below with reference to the drawings. (First Embodiment) FIG. 1 shows a block diagram of a scanning optical microscope of the present invention.

In FIG. 1, 1 is a light source, 2 is a light quantity adjusting element, 3 is a beam splitter (optical path branching element), 4a and 4a.
Reference numeral b is a mirror, 5 is a scanning optical system, 6 is an objective lens holding revolver (hereinafter referred to as a revolver), 7 is an objective lens, 8 is a sample, 9 is a pinhole member, 10 is a photodetector, and 11 is a data input device. , 12 are arithmetic units, and 13 is a memory.

Of these, the sample 8 is an object to be observed with a microscope, and the light source 1 is a light emitting source for illuminating the sample 8 and uses, for example, a laser light source. The light quantity adjusting element 2 is provided on the outgoing light path L1 of the light source 1 and is for adjusting the light quantity from the light source 1. The light quantity adjusting element 2 can be configured to variably adjust the light emitted from the light source 1, for example, using a liquid crystal shutter, a polarizing filter, a diaphragm device using diaphragm blades, or the like. Since the light quantity adjusting element 2 is the basic adjusting mechanism for adjusting the emitted light from the light source 1, it is fixed after the optimum emitted light quantity is adjusted.

The beam splitter 3 is an output optical path L of the light source 1.
1 is an element for transmitting the light of which the light amount is adjusted through the light amount adjusting element 2 and reflecting the light traveling backward in the optical path in a predetermined direction. The incident light is transmitted through the beam splitter 3, and the light incident on the beam splitter 3 in the opposite path is arranged so that the light is reflected by the beam splitter 3 in a predetermined direction.

The mirror 4a is a reflecting mirror which is fixedly arranged at a fixed position facing the light source 1 through the beam splitter 3 and reflects the light emitted from the light source 1 in a predetermined direction. Further, the mirror 4b is a reflecting mirror that faces the reflection optical path L2 of the beam splitter 3 and is fixedly provided at a fixed position, and that reflects the light reflected by the beam splitter 3 to guide it to the photodetector 10. .

The scanning optical system 5 is composed of, for example, a galvano mirror 5a for scanning in the X-axis direction and a galvano mirror 5b for scanning in the Y-axis direction, and the galvano mirror 5b for scanning in the Y-axis direction enters the mirror 4a. The Galvano mirror 5a for scanning in the X-axis direction is arranged on the reflected light path L so that the incident and reflected optical axes thereof coincide with each other, and the incident and reflected optical axes are reflected on the sample 8 through one of the objective lenses 7 held by the revolver 6. And the galvano-mirror 5a for scanning in the X-axis direction and the galvano-mirror 5b for scanning in the Y-axis direction have a positional relationship such that their incident / reflecting optical axes coincide with the incident / reflecting optical axes of the counterpart mirrors. The galvano mirrors 5 are opposed to each other so that the incident / reflected optical axis of the galvano mirror 5a with respect to the sample 8 can be scanned by XY scanning by swinging each galvano mirror.

The revolver 6 is a disk-shaped objective lens holding member holding a plurality of objective lenses 7, and is rotatably held. By manually rotating the revolver 6, a plurality of objective lenses can be operated. A desired one of 7 can be moved to a position (optical path position) facing the sample 8 so that the optical path of the scanning optical system 5 facing the sample 8 can be moved through the selected objective lens to the sample 8. 8 can be opposed.

The plurality of objective lenses 7 held by the revolver 6 are lenses having different magnifications. By selecting the objective lens 7 for observation, the sample 8 can be observed at a desired magnification. .

The photodetector 10 has detection means (photoelectric conversion element) for detecting the amount of incident light and converting it into an electric signal, and an amplifier circuit for amplifying and outputting the converted electric signal. The photodetector 10 is arranged so that its light incident side faces the reflection optical axis of the mirror 4b, and detects the amount of light (luminance) from the sample 8 incident via the mirror 4b by the detection means to detect the amount of light. Is converted into an electric signal corresponding to, and is amplified and output by an amplifier circuit incorporated therein.

Further, the photodetector 10 is constructed so that the sensitivity is controlled according to the sensitivity control information (sensitivity adjustment signal) from the arithmetic unit 12 and the sensitivity-controlled light amount (luminance) detection value is output. For the sensitivity adjustment, for example, the detection means itself is configured to have its sensitivity adjustable, or a variable amplifier circuit is used as an amplifier circuit incorporated in the photodetector 10, and its amplification factor is adjusted according to the sensitivity control information. It can be performed by using a system.

The pinhole member 9 is, for example, a plate-shaped member made of a light shielding material and has one minute through hole formed therein, and is arranged so as to be insertable and removable between the photodetector 10 and the mirror 4b. It When the pinhole member 9 is inserted between the photodetector 10 and the mirror 4b, the through hole of the pinhole member 9 naturally leads to the incident optical path from the mirror 4b to the photodetector 10. It should be placed in a position where it can be secured.
Further, when the pinhole member 9 is inserted between the photodetector 10 and the mirror 4b, the pinhole member 9 is moved from the mirror 4b moving by the optical path scanning by the scanning optical system 5 to the photodetector 10 by the scanning optical system 5. The drive movement is controlled in synchronization with the scanning optical system 5 so as to follow the movement of the incident optical path to the.

The data input device 11 detects the switching to the objective lens newly provided for sample observation in association with the rotation operation of the revolver 6, and the type information and the objective lens used before the switching. A device that provides information.

The memory 13 is a storage means for storing characteristic information of each objective lens 7 held by the revolver 6,
The arithmetic unit 12 determines the type of objective lens inserted into the optical path input from the data input unit 11 before the conversion of the objective lens by the rotation operation of the revolver 6 before the change, and the objective inserted into the optical path after the revolver 6 moves. The characteristics of each objective lens are read from the memory 13 from the lens type data (type data), and the calculation is performed based on the read characteristics, so that the sensitivity of the photodetector 10 is changed. This is an apparatus for obtaining control information for controlling the sensitivity of the photodetector 10 corresponding to the characteristics of the objective lens.

Next, the operation of the present apparatus having such a configuration will be described. In the present apparatus of FIG. 1, the light emitted from the light source 1 is adjusted in light quantity by the light quantity adjusting element 2 for adjusting the light quantity, passes through the beam splitter 3, and then is reflected by the mirror 4a to enter the scanning optical system 5. The scanning optical system 5 is operated so as to be operated in the XY axis directions to become spot-like illumination light that scans a two-dimensional space. Then, the illumination light emitted from the scanning optical system 5 enters the objective lens 7 held by the revolver 6 and forms an image on the sample 8 to be measured.

From the sample 8 which receives this light, reflected light or fluorescence is generated by this light. And this sample 8
The reflected light or fluorescent light from the light source travels in the same optical path as the illumination light in the opposite direction, reaches the beam splitter 3, is deflected by the beam splitter 3 to the photometric optical path, and is placed in a position conjugate with the sample image plane. By means of 9, only reflected light or fluorescence from the image plane is guided to the light receiving element of the photodetector 10.

Here, if the pinhole member 9 is inserted in the optical path, a three-dimensional image can be obtained by the confocal effect, but it is also possible to obtain a non-confocal image without inserting the pinhole member 9. It is possible.

The light incident on the photodetector 10 is converted by the photodetector 10 into an electric signal corresponding to the amount of light (luminance), amplified, and then input to a data processing device (not shown). Is visualized by.

In this apparatus, the sensitivity of the photodetector 10 can be changed by an instruction from the arithmetic unit 12. That is, the arithmetic unit 12 includes the data input unit 11
Type information from is input. Then, this type information is associated with the switching of the objective lens 7 by the rotation operation of the revolver 6, for example, the data input device 11 recognizes the type of the objective lens set in the optical path from the rotational position detection information of the revolver 6. By the method, the type is known and this is output as type information, and the type information of the objective lens before switching is also output.

That is, the data input device 11 obtains the type information of the objective lens inserted in the optical path at that time before the objective lens is switched, and the revolver 6 is also provided.
When the rotation operation is performed, the type information of the objective lens newly moved to the optical path position is obtained by this rotation operation. Therefore, the type information of both objective lenses is generated at the time of lens switching by the rotation operation of the revolver 6, and the arithmetic unit Give to twelve.
(Note that if the arithmetic unit 12 is provided with a function of holding the type information of the objective lens before the revolver rotation operation, the data input unit 11 does not have to be a configuration that simultaneously gives the type information of both objective lenses. Then, the arithmetic unit 12 obtains the characteristic of each objective lens from the memory 13 based on these data (object type information of the objective lens), and the objective lens is based on this. The calculation for obtaining the correction amount to obtain the optimum detection sensitivity according to the change of the objective lens characteristics based on the change of
A control value that is the sensitivity for which the correction corresponding to the obtained correction amount has been obtained is given to the photodetector 10, and the sensitivity of the photodetector 10 is instructed and controlled.

As a result, the sensitivity of the photodetector 10 is automatically controlled, and the light incident from the sample 8 is not changed without changing the output light amount of the light source 1 and without further adjustment of the light amount adjusting element 2. The detection output of the photodetector 10 will be adjusted to the optimum level, and after replacement of the objective lens, an image having the same brightness as that before replacement can be automatically obtained. In addition, such adjustments are automatically made when the objective lens is replaced, so the operability is dramatically improved, and observation can be started immediately after focus adjustment, which helps improve efficiency. .

As described above, the point light is applied to the sample to be observed through the desired objective lens, and the light from the sample obtained by two-dimensionally scanning the point light is detected by the detecting means. The electric signal corresponding to the amount of light is detected and
In the scanning optical microscope configured to image the electric signal as a luminance signal corresponding to the two-dimensional scanning of the point light, the detection means can adjust the sensitivity corresponding to the sensitivity adjustment signal, and The detection means capable of detecting the detected light and outputting it as a signal is controllable, and when the objective lens is replaced, the type of the objective lens currently in the optical path and the objective lens after the replacement Means for inputting information of the objective lens, storage means for storing characteristic information of each objective lens, and objective lens before replacement based on the characteristic information of the objective lens obtained from the storage means based on the type information of the objective lens. The objective lens is composed of an arithmetic means for calculating the light amount difference of the objective lens after the exchange to obtain the sensitivity adjustment information and for giving the sensitivity adjustment information as the sensitivity adjustment signal to the detection means. Based by replacing, calculating means characteristic information of the objective lens after replacement before and exchange,
The light amount difference between the objective lens before replacement and the objective lens after replacement is calculated to obtain sensitivity adjustment information from this, and this sensitivity adjustment information is given to the detection means as a sensitivity adjustment signal, whereby the detection sensitivity of the detection means Was automatically adjusted to a level that compensates for the difference in light amount before and after the replacement of the objective lens.

Therefore, when the objective lens is replaced, the sensitivity of the detecting means is optimally adjusted according to the characteristics of the replaced objective lens, and an image with no change in brightness before and after replacement can be obtained. .

In this embodiment, an example in which the revolver 6 is rotationally driven by manual operation to switch the objective lens is shown, but there is also a microscope apparatus of an electrically driven revolver. An application example will be described below as a second embodiment.

(Second Embodiment) A second embodiment of the present invention will be described with reference to FIG. In FIG. 2, 1 is a light source, 2 is a light quantity adjusting element, 3 is a beam splitter, 4a,
4b is a mirror, 5 is a scanning optical system, 6a is an electric revolver for holding an objective lens (hereinafter referred to as an electric revolver), 7 is an objective lens, 8 is a sample, 9 is a pinhole member, 10 is a photodetector, and 11a is data. An input device, 12 is a computing device, and 13 is a memory. Reference numeral 20 is an operation unit.

In this apparatus, the revolver 6 holding the objective lens 7 in the first embodiment is an electric revolver 6a,
When the objective lens control signal is received, the motor M is driven and controlled so that the objective lens corresponding to the signal comes to the optical path position, the revolver is rotationally driven, and the objective lens 7 can be automatically switched. is there.

The operation unit 20 is a device for giving a command for switching the objective lens, etc., and is an input for issuing a command (objective lens switching command) for switching to an objective lens having a desired magnification when operated by an operator (observer). The operation can be performed.

The data input device 11a receives an objective lens switching command issuing operation input from the operating unit 20 by the input operation of the operating unit 20, and the objective lens selected and designated by the observer based on the command is brought to the optical path position. It has a function of outputting a proper drive control signal to the electric revolver 6a and outputting the type information of the objective lens selected and designated by the observer to the arithmetic unit 12.

The sample 8 is an object to be observed with a microscope, and the light source 1 is a light emitting source for illuminating the sample 8 and uses, for example, a laser light source. The light quantity adjusting element 2 is provided on the outgoing light path L1 of the light source 1 and is for adjusting the light quantity from the light source 1. The light quantity adjusting element 2 can be configured to variably adjust the light emitted from the light source 1, for example, using a liquid crystal shutter, a polarizing filter, a diaphragm device using diaphragm blades, or the like. Since the light quantity adjusting element 2 is the basic adjusting mechanism for adjusting the light emitted from the light source 1, the light quantity adjusting element 2 is also fixed in this embodiment after being adjusted to the optimum light emitting quantity.

The beam splitter 3 is an output optical path L of the light source 1.
1 is an element for transmitting the light of which the light amount is adjusted through the light amount adjusting element 2 and reflecting the light traveling backward in the optical path in a predetermined direction. The incident light is transmitted through the beam splitter 3, and the light incident on the beam splitter 3 in the opposite path is arranged so that the light is reflected by the beam splitter 3 in a predetermined direction.

The mirror 4a is a reflecting mirror that is fixedly arranged at a fixed position facing the light source 1 through the beam splitter 3 and reflects the light emitted from the light source 1 in a predetermined direction. Further, the mirror 4b is a reflecting mirror that faces the reflection optical path L2 of the beam splitter 3 and is fixedly provided at a fixed position, and that reflects the light reflected by the beam splitter 3 to guide it to the photodetector 10. .

The scanning optical system 5 comprises, for example, a galvano-mirror 5a for X-axis scanning and a galvano-mirror 5b for Y-axis scanning, and the galvano-mirror 5b for Y-axis scanning is mounted on the mirror 4a. The galvano-mirror 5a for scanning in the X-axis direction is arranged on the reflected light path L so that the incident and reflected light axes thereof coincide with each other, and the objective lens 7 held by the electric revolver 6a.
Of the galvano mirror 5a for scanning in the X-axis direction and the galvano mirror 5b for scanning in the Y-axis direction are arranged so that their incident / reflecting optical axes are aligned with each other. They are made to face each other with a positional relationship that coincides with the incident / reflected optical axis of the other side mirror, and by swinging each galvanomirror, the incident / reflected optical axis of the galvanomirror 5a with respect to the sample 8 is XY scanned. I am able to do it.

The electric revolver 6a includes a plurality of objective lenses 7
Is a disk-shaped objective lens holding member that holds a plurality of objective lenses by rotatably holding the electric revolver 6a by a motor in response to a signal from the data input device 11a. A desired one of 7 can be moved to a position facing the sample 8 so that the optical path on the sample 8 facing side of the scanning optical system 5 faces the sample 8 via the selected objective lens. be able to.

The plurality of objective lenses 7 held by the electric revolver 6a are lenses having different magnifications. By selecting the objective lens 7 for observation, the sample 8 can be observed at a desired magnification. is there.

The photodetector 10 has a detection means (photoelectric conversion element) for detecting the amount of incident light and converting it into an electric signal, and an amplifier circuit for amplifying and outputting the converted electric signal. The photodetector 10 is arranged so that its light incident side faces the reflection optical axis of the mirror 4b, and detects the amount of light (luminance) from the sample 8 incident via the mirror 4b by the detection means to detect the amount of light. Is converted into an electric signal corresponding to, and is amplified and output by an amplifier circuit incorporated therein.

Further, the photodetector 10 is constructed so that the sensitivity is controlled according to the sensitivity control information from the arithmetic unit 12 and the detected light amount (luminance) detection value is output. Sensitivity adjustment is performed by, for example, a method in which the detection means itself has a structure capable of adjusting its sensitivity, or the photodetector 10
This can be performed by using a variable amplifier circuit as the built-in amplifier circuit and adjusting the amplification factor according to the sensitivity control information.

The pinhole member 9 is, for example, a plate-shaped member made of a light shielding material and has one minute through hole formed therein. The pinhole member 9 is arranged so that it can be inserted and removed between the photodetector 10 and the mirror 4b. It When the pinhole member 9 is inserted between the photodetector 10 and the mirror 4b, the through hole of the pinhole member 9 naturally leads to the incident optical path from the mirror 4b to the photodetector 10. It should be placed in a position where it can be secured.
Further, when the pinhole member 9 is inserted between the photodetector 10 and the mirror 4b, the pinhole member 9 is moved from the mirror 4b moving by the optical path scanning by the scanning optical system 5 to the photodetector 10 by the scanning optical system 5. The drive movement is controlled in synchronization with the scanning optical system 5 so as to follow the movement of the incident optical path to the.

The data input device 11a is an electric revolver 6a.
It is an apparatus that detects the objective lens currently used for sample observation in conjunction with the rotation operation of, and provides the type information and the information of the objective lens used at the previous time point.

The memory 13 is a storage means for storing the characteristic information of each objective lens 7 held by the electric revolver 6a, and the arithmetic unit 12 is a data input device prior to the conversion of the objective lens by the rotating operation of the electric revolver 6a. The characteristics of each objective lens are stored in the memory 13 based on the data (type data) of the type of the objective lens inserted into the optical path input from 11a before the change and the type of the objective lens inserted into the optical path after the revolver is moved. Further, the control information is read out and an operation is performed based on this to obtain control information for controlling the sensitivity of the photodetector 10 in accordance with the changed characteristic of the objective lens so that the sensitivity of the photodetector 10 is optimized. It is a device.

The basic operation of this apparatus having such a configuration is the same as that of the first embodiment. That is, the amount of light emitted from the light source 1 is adjusted by the light amount adjusting element 2 for adjusting the light amount,
After passing through the beam splitter 3, it is reflected by the mirror 4a and enters the scanning optical system 5, and is operated by the scanning optical system 5 so as to be operated in the XY axis directions to scan a two-dimensional space. It becomes the illumination light. Then, the illumination light emitted from the scanning optical system 5 is incident on the objective lens 7 held by the electric revolver 6a, and the sample 8 to be measured is measured.
Image on top.

From the sample 8 which receives this light, reflected light or fluorescence is generated by this light. And this sample 8
The reflected light or fluorescent light from the light source travels in the same optical path as the illumination light in the opposite direction, reaches the beam splitter 3, is deflected by the beam splitter 3 to the photometric optical path, and is placed in a position conjugate with the sample image plane. By means of 9, only reflected light or fluorescence from the image plane is guided to the light receiving element of the photodetector 10.

Here, if the pinhole member 9 is inserted in the optical path, a three-dimensional image can be obtained by the confocal effect, but it is also possible to obtain a non-confocal image without inserting the pinhole member 9. It is possible.

The light incident on the photodetector 10 is converted by the photodetector 10 into an electric signal corresponding to the amount of light (luminance), amplified, and then input to a data processing device (not shown). Is visualized by.

In this apparatus, when the objective lens 7 is switched, the observer operates the operation section 20 to set a desired magnification, and the data input apparatus 11a causes the electric revolver 6 to operate.
A control signal is given to a, and thereby the electric revolver 6a is rotationally driven so that the objective lens having the designated magnification comes into the optical path.

At the same time, the data input device 11a outputs the type information of the old and new objective lenses set in the optical path when the objective lens 7 is switched. That is, the data input device 11a displays the type information of the objective lens inserted in the optical path before switching the objective lens and the type information of the objective lens newly selected and moved to the optical path position by the rotational driving operation of the electric revolver 6a. Generating and computing device 1
Give to 2. (Note that if the arithmetic unit 12 is provided with a function of holding the type information of the objective lens before the revolver rotation driving operation, the data input device 11a does not have to be a configuration that simultaneously gives the type information of both objective lenses. Then, the arithmetic unit 12 obtains the characteristic of each objective lens from the memory 13 based on these data (objective lens type information), and the objective is calculated based on this. The calculation is performed to obtain the correction amount that provides the optimum detection sensitivity according to the change in the objective lens characteristics based on the lens change, and the control value that is the sensitivity with the correction corresponding to the obtained correction amount is obtained and is calculated. It is given to the photodetector 10 to instruct and control the sensitivity of the photodetector 10.

Since the sensitivity of the photodetector 10 is changed according to the control value in accordance with an instruction from the arithmetic unit 12, it is possible to observe an image having the same brightness as before even if the objective lens is switched.

In the above description, the revolver is an electric revolver for electrically switching the lens, and the detecting means is one capable of sensitivity adjustment control. When the lens type information is input from the operation unit, the electric revolver is activated. On the other hand, a control signal is output so that the position of the conversion destination objective lens becomes the optical path position, and the electric revolver itself automatically switches the lens to the optical path position. , A function of generating a sensitivity adjustment signal for obtaining sensitivity such that the brightness is maintained at a predetermined value according to the characteristics of the lens and giving the sensitivity adjustment signal to the detection means. When the information is given, the electric revolver automatically performs conversion to the designated objective lens by this, and the calculating means changes the lens according to the switching of the lens. The sensitivity adjusting signal is generated to obtain a sensitivity such that the brightness is maintained at a predetermined value according to the brightness, and the detecting means outputs the brightness having a predetermined value (that is, the brightness corresponding to the characteristic of the lens switched by the sensitivity adjusting signal). , The same brightness as before switching) is adjusted so that the sensitivity is maintained.

Therefore, the objective lens can be replaced automatically, and the brightness necessary for the replacement of the objective lens is automatically adjusted, so that the operability is dramatically improved. If you adjust the focus, you can immediately start observing, which helps improve efficiency.

The first and second embodiments have been described above with reference to the embodiments of the scanning optical microscope for automatically adjusting the brightness of the image changed by switching the objective lens by the revolver. The function of automatically adjusting the brightness of the image to a constant level is a function incorporated in the photodetector 10 adopted in the present invention.

Therefore, the embodiments of the present invention will be described as the third and fourth embodiments with a focus on a specific configuration example of the photodetector 10. First, the third embodiment will be described.

(Third Embodiment) FIG. 3 shows a block diagram of the third embodiment. As shown in the figure, in the third embodiment, the photodetector 10 includes a photodiode, a phototransistor, a P
Photoelectric conversion element 14 such as SD and this photoelectric conversion element 14
It is composed of a variable gain amplifier 15 which is a variable amplification factor amplifier for converting the current output from the device into a voltage signal and further amplifying its amplitude.

In the case of this configuration, the variable gain amplifier 15 corresponds to the sensitivity control information (sensitivity adjustment signal) from the arithmetic unit 12.
The sensitivity is adjusted by changing the amplification factor of.
Therefore, in this embodiment, in the photodetector 10, the light reflected by the mirror 4b and incident is converted by the photoelectric conversion element 14 into an electric signal (current signal) corresponding to the amount of incident light, and is output from the photoelectric conversion element 14. By supplying an electric signal (current signal) to the variable gain amplifier 15,
Reference numeral 5 converts the current signal into a voltage signal, further amplifies the amplitude of the voltage signal with a gain corresponding to the sensitivity control information (sensitivity adjustment signal) from the arithmetic unit 12, and outputs the amplified signal.

In this way, according to an instruction from the arithmetic unit 12, the signal can be amplified at a required amplification factor, and the sensitivity can be automatically adjusted so that an image having a predetermined brightness is obtained. .

(Fourth Embodiment) Next, a fourth embodiment will be described with reference to FIG.
Will be explained. In the example of FIG. 4, the photodetector 10 outputs an electric signal according to the amount of incident photons, and the output level of the electric signal can be adjusted by the level value of the applied high voltage, such as a photomultiplier. The photoelectric conversion element 14a is used, and a variable high voltage circuit 16 is provided as a circuit for controlling the detection sensitivity of the photoelectric conversion element 14a. Then, the sensitivity control signal from the arithmetic unit 12 is input to the variable high voltage circuit 16. Further, the output signal from the photoelectric conversion element 14a is output to a data processing device (not shown) via the amplifier 17 having a constant amplification factor.

Even with such a configuration, the sensitivity can be automatically adjusted by an instruction from the arithmetic unit 12 so that an image having a predetermined brightness is obtained. In any of the above examples, the change in the brightness of the image caused by the switching of the objective lens is corrected by the photodetector 10 so as not to change. However, instead of such correction by the photodetector 10, the amount of light emitted from the light source 1 may be adjusted so that the change in the brightness of the image does not occur due to the switching of the objective lens. . Next, such an embodiment will be described as a fifth embodiment.

(Fifth Embodiment) A fifth embodiment will be described with reference to FIGS. FIG. 5 is a block diagram showing the overall construction of the fifth embodiment, and FIG. 6 is a perspective view showing the concrete construction of the light quantity adjusting device used in this embodiment.

In FIG. 5, 1 is a light source, 2A is a light quantity adjusting element according to the present invention, 3 is a beam splitter, 4a and 4a.
b is a mirror, 5 is a scanning optical system, 6 is a revolver, 7 is an objective lens, 8 is a sample, 9 is a pinhole member, 10A is a photodetector, 11 is a data input device, 12A is a computing device, and 13
Is a memory.

Of these, the sample 8 is an object to be observed with a microscope, the light source 1 is a light emitting source for illuminating the sample 8, and the light quantity adjusting element 2A is provided on the emission optical path L1 of the light source 1. It is for adjusting the amount of light from the light source 1. The light amount adjusting element 2A is configured by using an ND filter capable of continuously variably adjusting the emitted light of the light source 1, and
The D filter is arranged such that a part of the D filter is inserted in the outgoing optical path from the light source 1, and the ND filter is configured to be rotatable and movable with respect to the outgoing optical path. A drive device is provided for this rotational movement, and this drive device is driven to rotate by the control output of the arithmetic unit 12A.

In the previous embodiment, the light amount adjusting element 2 is fixed after setting the transmitted light amount to the optimum value, but in this embodiment, the light amount adjusting element 2A can be variably adjusted at any time by the control output of the arithmetic unit 12A. It is a composition.

The beam splitter 3 is an optical path L emitted from the light source 1.
1, which is an element for transmitting the light of which the light amount is adjusted through the light amount adjusting element 2A and for reflecting the light traveling backward in the optical path in a predetermined direction from the light source 1 to the beam splitter 3. The incident light is the beam splitter 3
The light that has passed through the beam splitter 3 and has entered the beam splitter 3 in the opposite path is arranged so as to be reflected by the beam splitter 3 in a predetermined direction.

The mirror 4a is a reflecting mirror that is fixedly arranged at a fixed position facing the light source 1 via the beam splitter 3 and reflects the light emitted from the light source 1 in a predetermined direction. The mirror 4b is a reflecting mirror that faces the reflected light path L2 of the beam splitter 3, is fixedly provided at a fixed position, and reflects the light reflected by the beam splitter 3 to guide it to the photodetector 10A. .

The scanning optical system 5 is composed of, for example, a galvano mirror 5a for scanning in the X-axis direction and a galvano mirror 5b for scanning in the Y-axis direction, and the galvano mirror 5b for scanning in the Y-axis direction enters the mirror 4a. The Galvano mirror 5a for scanning in the X-axis direction is arranged on the reflected light path L so that the incident and reflected optical axes thereof coincide with each other, and the incident and reflected optical axes are reflected on the sample 8 through one of the objective lenses 7 held by the revolver 6. And the galvano-mirror 5a for scanning in the X-axis direction and the galvano-mirror 5b for scanning in the Y-axis direction have a positional relationship such that their incident / reflecting optical axes coincide with the incident / reflecting optical axes of the counterpart mirrors. The galvano mirrors 5 are opposed to each other so that the incident / reflected optical axis of the galvano mirror 5a with respect to the sample 8 can be scanned by XY scanning by swinging each galvano mirror.

The revolver 6 is an objective lens holding member that holds a plurality of objective lenses 7, and is rotatably held. Of the plurality of objective lenses 7, the revolver 6 is manually rotated. Of the scanning optical system 5 can be moved to a position (optical path position) facing the sample 8 so that the optical path on the side facing the sample 8 of the scanning optical system 5 faces the sample 8 via the selected objective lens. Can be made.

The plurality of objective lenses 7 held by the revolver 6 are lenses having different magnifications. By selecting the objective lens 7 for observation, the sample 8 can be observed at a desired magnification. .

The photodetector 10A has a detection means (photoelectric conversion element) for detecting the amount of incident light and converting it into an electric signal, and an amplifier circuit for amplifying and outputting the converted electric signal. The photodetector 10A is arranged so that its light-incident side faces the reflection optical axis of the mirror 4b, and the amount of light (luminance) from the sample 8 incident via the mirror 4b is detected by the detecting means to detect the amount of light. Is converted into an electric signal corresponding to, and is amplified and output by an amplifier circuit incorporated therein.

Also, unlike the previous embodiment, the photodetector 10A is used with fixed sensitivity (fixed amplification factor). The pinhole member 9 is, for example, a plate-shaped member made of a light shielding material and has one minute through hole formed therein, and is arranged so as to be insertable and removable between the photodetector 10A and the mirror 4b.
When the pinhole member 9 is inserted between the photodetector 10A and the mirror 4b, the through hole of the pinhole member 9 naturally goes from the mirror 4b to the photodetector 10.
It should be placed at a position where the incident optical path to A can be secured. Further, when the pinhole member 9 is inserted between the photodetector 10A and the mirror 4b, the pinhole member 9 moves from the mirror 4b which moves by the optical path scanning by the scanning optical system 5 to the photodetector 10A. The drive movement is controlled in synchronization with the scanning optical system 5 so as to follow the movement of the incident optical path to the.

The data input device 11 detects the switching to the objective lens newly provided for sample observation in association with the rotation operation of the revolver 6, and the type information and the objective lens used before the switching. A device that provides information.

The memory 13 is a storage means for storing characteristic information of each objective lens 7 held by the revolver 6,
The arithmetic unit 12A determines the type of the objective lens inserted into the optical path input from the data input device 11 before the conversion of the objective lens by the rotation operation of the revolver 6 before the change, and the objective inserted into the optical path after the revolver 6 moves. The characteristic of each objective lens is read from the memory 13 from the data of the lens type (type data), and the calculation is performed based on this, so that the output signal level of the photodetector 10A with fixed sensitivity is optimized. In addition, a control output for adjusting the light transmission amount of the light amount adjustment element 2A is generated.

Therefore, the light transmission position of the ND filter of the light quantity adjusting element 2A is selected in accordance with the changed characteristic of the objective lens (the position of the ND filter which comes to the position of the emission light path from the light source 1 is adjusted. By doing so, the light quantity can be controlled so that the quantity of light emitted from the light source 1 becomes the quantity of incident light such that the brightness of the image due to the signal of the detection output from the photodetector 10A does not change from that before the switching of the objective lens. it can.

In this embodiment, as a light quantity adjusting element 2A which is provided on the light emitting side of the light source 1 and adjusts the quantity of light emitted from the light source 1, a concrete configuration example thereof is shown in FIG.

As shown in the figure, the light amount adjusting device 2A has an ND filter (transmission amount continuously changing ND filter) 22 created so that the light transmission amount is continuously changed as if a gradation is applied. To use. The transmission amount continuously changing ND filter 22 is a disc-shaped member, and is configured to be rotatable around the center thereof as a rotation center. The continuous transmission amount change ND filter 22 exhibits a change in transmitted light amount such that an area outside the center of the disc is a light transmission amount of 100% and gradually decreases to a predetermined percentage.
The prepared optical filter is arranged with its central axis decentered with respect to the emission optical path (emission optical axis) of the light source 1.

Due to this eccentricity, the transmission amount continuously changing ND filter 22 has a structure capable of continuously changing the transmission amount in the emission optical path L1 of the light source 1. The permeation amount continuous change ND filter 22 has a gear shape at the periphery thereof, and by meshing with the gear of a motor 19 having a gear on its rotation shaft, the rotation of the motor 19 is transmitted by a gear transmission mechanism and transmitted. The amount of light can be adjusted steplessly.

Further, the light amount adjusting device 2A is provided with a control device 18 for controlling the rotational drive of the motor 19, and further, a position detecting sensor 21 for detecting the rotational position of the ND filter 22 for continuously changing the transmission amount. Is provided. The position detection sensor 21 is composed of, for example, a photoelectric detection element having a structure in which a light emitting element and a light receiving element are arranged to face each other through an ND filter 22 for continuously changing the transmission amount.
A slit for optical position detection is formed around the D filter 22.

The controller 18 uses the position detecting sensor 21.
In addition to knowing which position of the transmission amount of the continuously changing transmission amount ND filter 22 is on the emission optical path of the light source 1, the desired transmission amount of light based on the sensitivity control information from the arithmetic unit 12 is received. Has a function of controlling so that the position of is on the outgoing optical path of the light source 1.

In such a configuration, the arithmetic unit 12A
The sensitivity adjustment amount calculated and calculated by is input to the controller 18 of the transmission amount continuous change ND filter 22 for adjusting the light amount of the light source 1, and the motor 19 for rotationally driving the transmission amount continuous change ND filter 22 is driven. The rotation is controlled to rotate the transmission amount continuously changing ND filter 22 to a position where it gives a desired light amount.

The position of the transmission amount continuous change ND filter 22 is detected by the position detecting sensor 21, and the detection output is input to the control device 18, and the rotation of the motor 19 is controlled while using this as feedback information of the position information. Then, the position of the ND filter 22 for continuously changing the transmission amount is controlled so as to exhibit the required transmission amount.

The operation will be described in more detail. In the present apparatus shown in FIGS. 5 and 6, the light emitted from the light source 1 is adjusted in the light amount by the light amount adjusting element 2A for adjusting the light amount, passes through the beam splitter 3, and then is reflected by the mirror 4a to be scanned by the scanning optical system 5. Incident on the X-axis by this scanning optical system 5.
It becomes the spot-like illumination light that is operated to be operated in the Y-axis direction and scans the two-dimensional space. Then, the scanning optical system 5
The emitted illumination light enters the objective lens 7 held by the revolver 6 and forms an image on the sample 8 to be measured.

From the sample 8 which receives this light, reflected light or fluorescence is generated by this light. And this sample 8
The reflected light or fluorescent light from the light source travels in the same optical path as the illumination light in the opposite direction, reaches the beam splitter 3, is deflected by the beam splitter 3 to the photometric optical path, and is placed in a position conjugate with the sample image plane. By means of 9, only reflected light or fluorescence from the image plane is guided to the light receiving element of the photodetector 10A.

Here, if the pinhole member 9 is inserted in the optical path, a three-dimensional image can be obtained due to the confocal effect, but it is also possible to obtain a non-confocal image without inserting the pinhole member 9. It is possible.

The light incident on the photodetector 10A is converted by the photodetector 10 into an electric signal corresponding to the amount of light (luminance), amplified, and then input to a data processing device (not shown). Is visualized by.

In this apparatus, the sensitivity of the photodetector 10A is fixed, and instead, the transmitted light amount of the light amount adjusting element 2A is changed so as to eliminate the fluctuation of the brightness of the image due to the switching of the objective lens. . That is, the light quantity adjusting element 2A
Can change the amount of transmitted light according to an instruction from the arithmetic unit 12A.

The type information from the data input device 11 is input to the arithmetic unit 12A. Then, this type information is associated with the switching of the objective lens 7 by the rotation operation of the revolver 6, for example, the data input device 11 recognizes the type of the objective lens set in the optical path from the rotational position detection information of the revolver 6. By the method, the type is known and this is output as type information, and the type information of the objective lens before switching is also output.

That is, the data input device 11 obtains the type information of the objective lens inserted in the optical path at that time before the objective lens is switched, and the revolver 6 is also provided.
When the rotation operation is performed, the type information of the objective lens newly moved to the optical path position is obtained by this rotation operation. Therefore, the type information of both objective lenses is generated at the time of lens switching by the rotation operation of the revolver 6, and the arithmetic unit Give to 12A.
(Note that if the arithmetic unit 12 is provided with a function of holding the type information of the objective lens before the revolver rotation operation, the data input unit 11 does not have to be a configuration that simultaneously gives the type information of both objective lenses. The arithmetic unit 12A obtains the characteristics of each objective lens from the memory 13 based on these data (objective lens type information), and the objective lens is based on this data. The calculation for obtaining the correction amount that gives the optimum detection sensitivity according to the change in the objective lens characteristics based on the change of The amount of light emitted from the light source 1 is applied to the adjusting element 2A to control the amount of light so that the detection output of the photodetector 10A reaches an optimum level.

As a result, the light quantity of the spot light with which the sample 8 is irradiated is optimally automatically controlled according to the characteristics of the objective lens, and the level of the detection signal of the photodetector 10A with fixed sensitivity is also optimized.

Therefore, the sensitivity of the photodetector 10A is fixed and the amount of light emitted from the light source 1 is adjusted according to the characteristics of the objective lens, and the detection output of the photodetector 10A for the light incident from the sample 8 is optimum. Since the level is adjusted, it is possible to automatically obtain an image having the same brightness as before the replacement even after the replacement of the objective lens. In addition, such adjustments are automatically made when the objective lens is replaced, so the operability is dramatically improved, and observation can be started immediately after focus adjustment, which helps improve efficiency. .

In this embodiment, the configuration using the ND filter 22 for continuously changing the transmission amount is shown. However, the present invention is not limited to this, and the transmission light amount is changed by changing the polarization axis directions of two polarizing elements by rotation. The configuration may be adjusted. Further, regarding the position detecting sensor 21, the light amount of the light source light that has passed through the light amount adjusting mechanism may be detected by the light receiving element using the beam splitter 3.

The concept of this embodiment can be applied to the structure of the second embodiment as it is, as shown in FIG. In that case, the light quantity adjusting element 2 in the second embodiment is replaced with the light quantity adjusting element 2A, and the photodetector 10 is replaced with the photodetector 10 described above.
A, the arithmetic unit 12 is replaced with the arithmetic unit 12A, and the arithmetic unit 12A uses the light amount adjusting element 2A.
Is controlled.

In such a structure, the light emitted from the light source 1 is adjusted in the light amount by the light amount adjusting element 2A for adjusting the light amount, passes through the beam splitter 3, and then is reflected by the mirror 4a to be reflected by the scanning optical system 5. It is incident and is operated by the scanning optical system 5 so as to be operated in the XY axis directions.
The spot-like illumination light scans the dimensional space. Then, the illumination light emitted from the scanning optical system 5 is transmitted to the electric revolver 6
The light enters the objective lens 7 held by a and forms an image on the sample 8 to be measured.

From the sample 8 that receives this light, reflected light or fluorescence is generated by this light. And this sample 8
The reflected light or fluorescent light from the light source travels in the same optical path as the illumination light in the opposite direction, reaches the beam splitter 3, is deflected by the beam splitter 3 to the photometric optical path, and is placed in a position conjugate with the sample image plane. By means of 9, only reflected light or fluorescence from the image plane is guided to the light receiving element of the photodetector 10A.

Here, if the pinhole member 9 is inserted in the optical path, a three-dimensional image can be obtained by the confocal effect, but it is also possible to obtain a non-confocal image without inserting the pinhole member 9. It is possible.

The light incident on the photodetector 10A is converted by the photodetector 10A into an electric signal corresponding to the amount of light (luminance), amplified, and then input to a data processing device (not shown). Is visualized by.

In this apparatus, when the objective lens 7 is switched, the observer operates the operation section 20 to set a desired magnification, so that the data input apparatus 11a causes the electric revolver 6 to operate.
A control signal is given to a, and thereby the electric revolver 6a is rotationally driven so that the objective lens having the designated magnification comes into the optical path.

At the same time, the data input device 11a outputs the type information of the old and new objective lenses set in the optical path when the objective lens 7 is switched. That is, the data input device 11a displays the type information of the objective lens inserted in the optical path before switching the objective lens and the type information of the objective lens newly selected and moved to the optical path position by the rotational driving operation of the electric revolver 6a. Generating and computing device 1
Give to 2A. (Note that if the arithmetic device 12A has a function of holding the type information of the objective lens before the revolver rotation driving operation, the data input device 11a does not have to be a configuration that simultaneously gives the type information of both objective lenses. Then, the arithmetic unit 12A can obtain these data (objective lens type information).
Based on the above, the characteristic of each objective lens is obtained from the memory 13, and based on this, the calculation for obtaining the correction amount to obtain the optimum detection sensitivity according to the change of the objective lens characteristic based on the change of the objective lens is performed. The obtained control value which is the sensitivity with which the correction corresponding to the correction amount is obtained is given to the light amount adjusting element 2A, and the light amount emitted from the light source 1 is set to the optimum level for the detection output of the photodetector 10A. The light amount is controlled so that

As a result, the light quantity of the spot light with which the sample 8 is irradiated is automatically controlled optimally in accordance with the characteristics of the objective lens, whereby the level of the detection signal of the photodetector 10A with fixed sensitivity is also optimized.

Therefore, the sensitivity of the photodetector 10A is fixed and the amount of light emitted from the light source 1 is adjusted according to the characteristics of the objective lens, and the detection output of the photodetector 10A with respect to the light incident from the sample 8 is optimum. Since the level is adjusted, it is possible to automatically obtain an image having the same brightness as before the replacement even after the replacement of the objective lens. In addition, such adjustments are automatically made when the objective lens is replaced, so the operability is dramatically improved, and observation can be started immediately after focus adjustment, which helps improve efficiency. .

(Sixth Embodiment) Next, a method of calculating the light amount difference which is essential for control in the arithmetic units 12 and 12A will be described as a sixth embodiment.

In this embodiment, as the data to be stored in the memory 13 used in each of the embodiments including FIGS. 1 to 3, the type of the objective lens 7 attached to the revolver 6 or the electric revolver 6a is used. In response to the,
The aperture angle of the lens, the transmittance of the objective lens at the wavelength of the light used, and the focal length of the objective lens are prepared. Then, these data are stored in the memory 13 so that these data can be read corresponding to the type information of the objective lens.

Here, the aperture angle of the lens is θ, the transmittance of the objective lens at the wavelength of the light used is T, and
The focal length of the objective lens is f. Arithmetic unit 12,
In the arithmetic unit 12A, these are read from the memory 13 in correspondence with the objective lens type information, and the light amount difference is calculated by the following procedure.

When the type information of the objective lens after conversion (after switching) is input from the data input device 11 (or 11a), the arithmetic unit 12 or 12A stores it as data corresponding to the type information of the objective lens 7. Opening angle θ
₂ , the transmittance T ₂ and the focal length f ₂ are read from the memory 13 and the data input device 11 (or 11a) is also used.
Based on the type information of the current objective lens (objective lens before switching) input by the above, the aperture angle θ ₁ , the transmittance T _1, and the focal length f ₁ of the objective lens are stored in the memory 13 as data corresponding to the type information. Read more. And
From these data obtained from the memory 13, the following formula
Calculate the brightness ratio based on (1).

That is, assuming that the brightness of the objective lens after switching is I ₂ and the brightness of the objective lens before switching (current objective lens) is I ₁ , I ₁ is T ₁ (f ₁ tan θ
_{Since 1} ) ² and I ₂ are expressed as T ₂ (f ₂ tan θ ₂ ) ² , the brightness ratio I ₂ / I ₁ before and after switching the objective lens is I ₂ / I ₁ = T ₂ ( It can be obtained by calculating f ₂ tan θ ₂ ) ² / T ₁ (f ₁ tan θ ₁ ) ² (1). Sensitivity control information that makes the brightness of the image the same before and after switching the objective lens according to the brightness ratio I ₂ / I ₁ before and after switching the objective lens obtained by this It is generated and given to the photodetector 10 (in the case of the first to third embodiments) or the light quantity adjusting element 2A (in the case of the fifth embodiment). This makes it possible to observe a change in brightness before and after switching the objective lens by the photodetector 10 or the light amount adjusting element 2A to observe an image of constant brightness.

(Seventh Embodiment) Next, another example of the calculation method of the light amount difference will be described as a seventh embodiment. The calculation of the light amount difference as the seventh embodiment can be selected by rotating the revolver by devising the data stored in the memory 13 used in each of the embodiments including FIGS. 1 to 3. This makes it easy to obtain the brightness ratio between the objective lenses.

For example, the data stored in the memory 13 is the average value of the brightness of the image of the standard sample 8S measured by each objective lens or the brightness of the arbitrarily designated part. Then, from these data, the brightness ratio between different objective lenses can be obtained, and the arithmetic processing can be simplified.

The arithmetic methods in the sixth and seventh embodiments described above may be used alone or in combination. Further, in each of the above-described embodiments, the description has been made of measuring (detecting) the amount of light such as reflected light and fluorescence from the sample with the photodetector, but in addition, the transmitted light from the sample is measured (detected) with the photodetector. It is also possible to carry out various modifications.

Although various embodiments have been described above, in short, the present invention controls the detection sensitivity of the photodetector in the scanning optical microscope so as to correspond to the objective lens used for observation, or is used for scanning the sample. The light quantity of the point light source is adjusted, and the adjustment of the change in the brightness of the image due to the switching of the objective lens can be automated, and the switching of the objective lens can be followed without any time difference. It is a thing. The present invention can be summarized as follows. [1] The present invention relates to the case of replacing an objective lens in a scanning optical microscope which is attached to a microscope and measures the amount of light such as reflected light, transmitted light, and fluorescence from a sample. , Means for inputting the type of objective lens after replacement with the objective lens currently in the optical path, memory for storing characteristics of each objective lens, objective lens before replacement and objective after replacement from information in the memory A configuration is provided that includes a calculation unit that calculates the difference in the light amount of the lens and a unit that adjusts the sensitivity based on the output of the calculation unit.

According to the scanning optical microscope having such a configuration, when the objective lens is exchanged, the means for inputting the type of the objective lens is the objective lens currently in the optical path and the type information of the objective lens after the exchange. Is given to the calculation means,
The calculation means obtains characteristic information corresponding to the type information of the objective lens from the storage means for storing the characteristic of each objective lens, and calculates the light amount difference between the objective lens before the exchange and the objective lens after the exchange based on the characteristic information. Then, the sensitivity of the scanning optical microscope is adjusted by the sensitivity adjusting means in accordance with the light amount difference output obtained by the calculating means.

In this way, the sensitivity of the scanning optical microscope is adjusted as the objective lens is replaced, and an image with the same brightness can be obtained before and after replacement. Therefore, according to the present invention, it becomes possible to automatically adjust the change in the brightness of the image due to the switching of the objective lens, and moreover, the switching of the objective lens can be followed without any time difference and the observation can be started in a short time. It is possible to provide a scanning optical microscope capable of performing the above.

[2] In addition, in the configuration of [1], the objective lens exchanging means for automatically exchanging the input type of objective lens is adopted. As a result, it is possible to automatically switch to the designated objective lens only by designating the type of the objective lens, and it becomes possible to automate the adjustment for the change in the brightness of the image due to the switching of the objective lens.

[3] Further, in the configuration of [2], the sensitivity adjusting means is constituted by means for adjusting the gain of the amplifier for amplifying the signal from the light receiving element. [4] Also, in the configuration of [2] or [3],
The sensitivity adjusting means is configured by means for adjusting the sensitivity of the light receiving element.

[5] Further, in the configuration of [2], the sensitivity adjusting means is configured by means for adjusting the light amount of the illumination light. By adopting the configuration as described in [3] to [5], it becomes possible to automate the adjustment with respect to the change in the brightness of the image due to the switching of the objective lens, and moreover, to follow the switching of the objective lens without any time difference. Become.

[6] In addition, in the configurations of [1] to [5], the information of the objective lens stored in the memory is
The basic characteristics of the objective lens such as aperture angle, transmittance, and focal length are used.

Then, from these data, the ratio of brightness between the objective lens currently used and the objective lens to be switched next to be used is obtained and used for the sensitivity adjustment control to obtain the objective ratio. It is possible to obtain the adjustment amount for the change in the brightness of the image due to the switching of the lens.

[7] In the configurations [1] to [5], the characteristics of the objective lens stored in the memory are
The value was based on the photometric value obtained by measuring the standard sample in advance. Then, based on these data, the ratio of brightness between the objective lens currently used and the objective lens to be switched next to be used is obtained and used for sensitivity adjustment control to switch the objective lens. It is possible to obtain an adjustment amount with respect to a change in the brightness of the image due to, and to simplify the arithmetic processing.

[0127]

As described above, according to the present invention,
The change in brightness of the image before and after the objective lens replacement can be minimized, and the sensitivity can be adjusted based on the already acquired data, so that it can be adjusted without any time lag with the objective lens replacement. Thus, it is possible to provide a scanning optical microscope that enables rapid observation.

[Brief description of drawings]

FIG. 1 is a diagram for explaining an embodiment of the present invention,
FIG. 1 is a block diagram showing a configuration example of a scanning optical microscope according to a first embodiment of the present invention.

FIG. 2 is a diagram for explaining an embodiment of the present invention,
The block diagram which shows the structural example of the scanning optical microscope concerning the 2nd Example of this invention.

FIG. 3 is a diagram for explaining an embodiment of the present invention,
The block diagram which shows the structural example of the scanning optical microscope concerning the 3rd Example of this invention.

FIG. 4 is a diagram for explaining an embodiment of the present invention,
The block diagram which shows the structural example of the scanning optical microscope concerning the 4th Example of this invention.

FIG. 5 is a diagram for explaining an embodiment of the present invention,
The block diagram which shows the structural example of the scanning optical microscope concerning the 5th Example of this invention.

FIG. 6 is a diagram for explaining an embodiment of the present invention,
The figure which shows the structural example of 2 A of light quantity adjustment elements used for the 5th Example of this invention.

FIG. 7 is a diagram for explaining an embodiment of the present invention,
The block diagram which shows the structural example of the scanning optical microscope which applied the 5th Example of this invention to the structure which used the electric revolver.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 ... Light source (laser light source) 2, 2A ... Light quantity adjusting element 3 ... Beam splitter (optical path branching element) 5 ... Scanning optical system 6 ... Objective lens holding revolver (revolver) 6A ... Objective lens holding electric revolver (electric revolver) 7 ... Objective lens 8 ... Sample 10, 10A ... Photodetector 11, 11a ... Data input device 12, 12A ... Arithmetic device 13 ... Memory 20 ... Operation part.

Claims (3)

[Claims] 1. A scanning optical microscope which obtains a microscope image of a sample by irradiating the sample with spot light that is two-dimensionally scanned through an objective lens of the optical microscope and measuring the amount of light from the sample obtained thereby. When exchanging the objective lens, means for giving the type information of the objective lens after the exchange with the current objective lens in the optical path of the spot light, and the optical characteristic information of each objective lens corresponding to the type information of the objective lens. The storage means for storing, and the objective lens before the exchange and the objective after the exchange from the optical characteristic information corresponding to the type information of the objective lens after the exchange with the current objective lens in the optical path of the spot light obtained from the storage means. A scanning optical microscope comprising: a calculating means for calculating a light amount difference of a lens; and a means for adjusting sensitivity by an output of the calculating means. 2. A scanning optical microscope which obtains a microscope image of a sample by irradiating the sample with spot light which is two-dimensionally scanned through an objective lens of the optical microscope and measuring the amount of light from the sample obtained thereby. When exchanging the objective lens, means for giving the type information of the objective lens after the exchange with the current objective lens in the optical path of the spot light, and the optical characteristic information of each objective lens corresponding to the type information of the objective lens. The storage means for storing, and the objective lens before the exchange and the objective after the exchange from the optical characteristic information corresponding to the type information of the objective lens after the exchange with the current objective lens in the optical path of the spot light obtained from the storage means. In addition to having a calculating means for calculating the light amount difference of the lens, a light receiving element for converting incident light into an electric signal and an amplifying means for amplifying a signal from the light receiving element, A detecting means for measuring the amount of light from the sample, said detecting means sensitivity scanning optical microscope characterized by comprising a means for adjusting the output of the calculation means. 3. A scanning optical microscope which obtains a microscope image of a sample by irradiating the sample with spot light which is two-dimensionally scanned through an objective lens of the optical microscope and measuring the amount of light from the sample obtained thereby. When exchanging the objective lens, means for giving the type information of the objective lens after the exchange with the current objective lens in the optical path of the spot light, and the optical characteristic information of each objective lens corresponding to the type information of the objective lens. The storage means for storing, and the objective lens before the exchange and the objective after the exchange from the optical characteristic information corresponding to the type information of the objective lens after the exchange with the current objective lens in the optical path of the spot light obtained from the storage means. A scanning type comprising: a calculation unit that calculates a light amount difference of a lens; and a light amount adjustment unit that adjusts the light amount of the spot light according to the output of the calculation unit. Manabu microscope.