JP4034759B2 - Microscope illumination adjustment method and microscope - Google Patents

Description

  The present invention relates to an illumination adjustment method for a microscope and a microscope.

  A Kohler illumination optical system is known as an illumination optical system used for illumination on a specimen surface on which an observation target is placed in a microscope. The Koehler illumination optical system includes at least a light source such as a gas discharge lamp and a gas-filled lamp, a collector lens, a field stop, an aperture stop, and an objective lens. The Koehler illumination optical system illuminates the specimen surface by projecting the lamp arc or lamp filament of the light source to the position of the aperture stop by the collector lens, and projecting this light source image to the pupil of the objective lens as a secondary light source. There is little illumination unevenness. In addition, since the field stop image is projected onto the sample surface by the objective lens, the irradiation field is clearly determined.

  In such an illumination optical system, the illumination state is adjusted, for example, when the microscope is assembled or maintained, or when the light source is replaced. Although this adjustment can be performed by visual operation, in this case, the adjustment is performed based on the adjustment standard of the adjuster, so that the lighting state may change each time the adjustment is made. Therefore, it has been required to automatically adjust the illumination optical system so that the illumination state is always constant regardless of the adjuster. As such an automatic adjustment method, for example, there is a technique described in Patent Document 1 described later.

The technique described in Patent Document 1 is based on the technical idea that the object field is illuminated uniformly when the light output is maximized behind the pupil plane of the microscope objective lens or behind the pupil plane of the illumination optical path. The integrated light output of the light beam applied to the objective field is measured by a detector, and the lamp is automatically adjusted relative to the illumination light path so that the light output detected by the detector is maximized. .
JP 2002-277748 A (FIG. 1 etc.)

  However, although the technical idea of Patent Document 1 is established when the illumination optical system is in an ideal state, the actual illumination optical system has light emission unevenness of the light source, lens aberration, and setting error. (Eccentricity of the optical axis, etc.) and the like, and therefore, even if the technique described in Patent Document 1 is applied to an actual microscope, illumination unevenness cannot always be sufficiently reduced.

  The present invention has been made in view of the above-described circumstances, and an object thereof is to provide an illumination adjustment method for a microscope and a microscope that can reduce illumination unevenness for an actual microscope.

In order to achieve the above object, the present invention provides the following means.
The present invention uses a light source with uneven light emission , projects a light source image onto a position of an aperture stop by a collector lens, and projects the light source image as a secondary light source onto a pupil of an objective lens to illuminate a specimen surface. a microscope illumination adjusting method used in a focusing step for focusing the light source image is moved in the optical axis direction the collector lens to the sample surface, the light source, the optical axis of the collector lens A light source position adjusting step for moving the sample surface to a position where the amount of light applied to the sample surface is maximized on a plane perpendicular to the sample surface ; a central portion located at the center of the sample surface ; and a periphery located outside the central portion the amount of light irradiated respectively for the area measured by the light amount measuring device, a position difference between the irradiation light amount in the peripheral region and the irradiation light amount in the center portion is within a reference range, the Korekutare And a illumination unevenness suppression step of moving's in the optical axis direction.

In this microscope illumination adjustment method, first, the position of the collector lens and the light source is adjusted so that the light source image is focused on the sample surface and the amount of light applied to the sample surface is maximized. Is in a state of being uniformly illuminated.
Then, in this state, the collector lens is moved on the optical axis while actually measuring the amount of light emitted for the central portion and the peripheral region on the specimen surface by the light amount measuring device, and the actual irradiation is performed in the central portion and the peripheral region . Keep the difference in light intensity within the reference range.
Thus, in this illumination adjustment method for a microscope, since the amount of light irradiated to each part on the sample surface is made uniform based on the actual measurement value of the light amount measuring device, the light emission unevenness of the light source of the illumination optical system and the lens aberration, For an actual microscope in which a setting deviation or the like occurs, illumination can be adjusted appropriately according to the state.

Further, in this microscope illumination adjustment method, the amount of light applied to each measurement region is measured using a light amount measuring device, and therefore high-precision adjustment can be stably performed without depending on the adjuster.
Furthermore, since the light quantity is measured only for a part of the sample surface, the light quantity can be measured and analyzed in a short time compared to the case where the light quantity is measured for the entire sample surface, and the adjustment work is performed. Can be done quickly.

  Here, in this illumination adjustment method for a microscope, the measurement of the amount of light irradiated on the sample surface may be performed by a light amount measuring device installed on the sample surface, and a flat reflector or fluorescent plate is installed on the sample surface. However, the measurement may be performed by a light quantity measuring device installed at a position conjugate with the sample surface by an optical system including the reflecting plate (or fluorescent plate). When a fluorescent plate is installed on the sample surface, the amount of light applied to the fluorescent plate is obtained based on the amount of fluorescent light emitted from the fluorescent plate.

In this microscope illumination adjustment method, after the light source position adjustment step, the focusing step and the light source position adjustment step may be performed again and then the illumination unevenness suppression step may be performed.

In this microscope illumination adjustment method, after the initial focusing step and the initial light source position adjustment step are performed to bring the light source position closer to the appropriate position, the focusing step is performed again to further improve the position of the collector lens. Since the process shifts to the illumination unevenness suppressing process in an appropriate state, the illumination can be adjusted more precisely.
Here, the second light source position adjustment step may be performed after the second focusing step. In this case, since the process proceeds to the illumination unevenness suppressing step with the position of the light source being more appropriate, the illumination can be adjusted more precisely.
Further, after the initial focusing step and the light source position adjusting step, the focusing step and the light source position adjusting step may be repeated a plurality of times. In this case, each time the focusing process and the light source position adjustment process are repeated, the position can be adjusted more finely by reducing the unit movement amount when moving the collector lens and the light source. The lighting can be adjusted more precisely.

  In this microscope illumination adjustment method, a central region located at the center of the specimen surface and a peripheral region made of a region different from the central region may be used as the measurement regions.

  In this microscope illumination adjustment method, the amount of light irradiated to the central region located at the center of the sample surface and the amount of light irradiated to the peripheral region are measured, and adjustment is performed so as to suppress the difference between these amounts of light. Illumination unevenness between the center area and the peripheral area of the sample surface can be effectively suppressed.

  In this microscope illumination adjustment method, a plurality of the peripheral regions may be provided.

  In this microscope illumination adjustment method, it is possible to reduce illumination unevenness between the peripheral regions by performing adjustment so as to suppress the difference in the amount of light between the peripheral regions of the specimen surface. In particular, when the peripheral regions are arranged at equal intervals with point symmetry with respect to the center of the sample surface (for example, when four peripheral regions are arranged about 90 ° around the center of the sample surface), The deviation of the light amount with respect to the center can be reduced.

  Moreover, in this illumination adjustment method for a microscope, the microscope has a two-dimensional imaging sensor that images the specimen on the specimen surface, and the measurement region on the specimen surface is output from the output of the two-dimensional imaging sensor. You may obtain | require the light quantity irradiated.

  In this microscope illumination adjustment method, a two-dimensional imaging sensor (for example, a CCD camera or a CMOS camera) provided in the microscope is used as a light quantity measuring device. For this reason, since it is not necessary to prepare a light quantity measuring apparatus newly, the illumination of a microscope can be adjusted at low cost. Here, the amount of light applied to the measurement area may be obtained directly from the magnitude of the output signal of the two-dimensional image sensor, etc., or by analyzing image information captured by the two-dimensional image sensor. Also good.

  In this microscope illumination adjustment method, an area captured by a plurality of adjacent light receiving elements among the light receiving elements constituting the two-dimensional imaging sensor may be used as the measurement area.

  In this microscope illumination adjustment method, an area captured by a plurality of adjacent light receiving elements (a set of areas imaged by each adjacent light receiving element) is used as a measurement area, thereby reducing the influence of noise on each light receiving element. Thus, it is possible to accurately measure the amount of light applied to the measurement region.

  Here, if the number of light receiving elements corresponding to one measurement region is too large, the detection sensitivity deteriorates, and it becomes difficult to accurately measure the amount of light irradiated to the measurement region. The time required will increase. Therefore, the number of light receiving elements corresponding to one measurement region is preferably 4 to 64 elements.

The microscope according to the present invention uses a light source with uneven light emission , projects a light source image onto a position of an aperture stop using a collector lens, and projects the light source image onto a pupil of an objective lens as a secondary light source, thereby illuminating the specimen surface. a microscope according to a collector lens position adjusting device for moving the collector lens in the optical axis direction, and the light source position adjustment device for moving on a plane perpendicular to the light source with respect to the optical axis of the collector lens, the A light quantity measuring device that measures the amount of light emitted for each of a central portion located at the center of the sample surface and a peripheral region located outside the central portion, and the collector lens position adjustment based on the measurement value of the light quantity measuring device device and a control device for controlling the operation of the light source position adjustment device, the difference is the reference range of light quantity in the peripheral region and the irradiation amount of light at the central portion To fit, it includes moving the collector lens in the optical axis direction to the control.

  Since the microscope configured as described above has a collector lens position adjustment device, a light source position adjustment device, and a control device that controls these operations, illumination using the microscope illumination adjustment method according to the present invention is performed. Adjustments can be made automatically.

  According to the illumination adjustment method for a microscope according to the present invention, since the amount of light irradiated on the sample surface is made uniform based on the actual measurement value, an actual microscope in which there is aberration of the lens of the illumination optical system, a setting deviation, or the like. In addition, the illumination can be appropriately adjusted according to the state.

  Further, according to the microscope according to the present invention, the illumination adjustment method can be applied appropriately and automatically by applying the microscope illumination method according to the present invention.

Hereinafter, embodiments according to the present invention will be described with reference to the drawings.
[First embodiment]
Hereinafter, a first embodiment of the present invention will be described with reference to FIGS. 1 to 6.
As shown in FIG. 1, a microscope 1 according to the present embodiment includes a microscope main body 2, a stage 3 on which a measurement target is installed, an observation optical system 4 used for observing a specimen surface on the stage 3, and a specimen surface. And an illumination optical system 5 for illuminating.
The stage 3 is provided to be movable relative to the microscope body 2.

The observation optical system 4 has an objective lens 11 and an imaging lens 12 as in the observation optical system used for a general microscope. The objective lens 11 and the imaging lens 12 are arranged in a state in which their optical axes coincide with each other and are separated by the same distance as the sum of their focal lengths.
In the present embodiment, a two-dimensional imaging sensor 13 that captures an image of the specimen surface obtained through the objective lens 11 and the imaging lens 12 is provided at the focal position behind the imaging lens 12. Yes. As the two-dimensional imaging sensor 13, for example, a CCD camera, a CMOS camera, or the like that has a plurality of light receiving elements arranged two-dimensionally to form an imaging region is used.

  The illumination optical system 5 has a configuration similar to that of an illumination optical system generally used for Kohler illumination. Specifically, the illumination optical system 5 includes a light source 21 such as a gas discharge lamp or a gas-filled lamp (specifically, a halogen lamp, a xenon lamp, a mercury lamp, etc.) and a collector that collects light rays emitted from the light source 21. The lens 22 and a reflecting mirror 23 that reflects the light beam emitted from the light source 21 in the direction other than the collector lens 22 and enters the collector lens 22 in a feedback manner.

  A field stop 24 that restricts the irradiation range of the sample surface is provided behind the collector lens 22 in the optical axis direction at a position conjugate with the focal plane of the objective lens 11. Includes a relay lens 25 that forms an image of the field stop 24 at infinity, and an aperture stop that adjusts the diameter of the exit pupil of the objective lens 11 on the focal plane behind the relay lens 25 in the optical axis direction. 26 is provided. Here, the field stop 24 and the aperture stop 26 can be independently adjusted in aperture diameter.

  Of the illumination optical system 5, the portion constituted by each of the above-described constituent members is disposed with its optical axis inclined with respect to the optical axis of the observation optical system 4. At a position where the optical axis of the optical system 4 intersects, a flat plate mirror 27 that reflects the light beam that has passed through the aperture stop 26 toward the objective lens 11 and transmits the light beam that travels from the objective lens 11 toward the imaging lens 12. Is provided. That is, the illumination optical system 5 of the present embodiment is a coaxial incident illumination optical system that projects the light emitted from the light source 21 onto the sample surface via the observation optical system 4. Here, as the mirror 27, for example, a half mirror or a dichroic mirror is used.

The illumination optical system 5 includes a collector lens position adjusting device 31 that moves the collector lens 22 in the optical axis direction, a light source position adjusting device 32 that moves the light source 21 relative to the collector lens 22, and two A control device 33 that controls the operation of the collector lens position adjusting device 31 and the light source position adjusting device 32 based on the output of the two-dimensional imaging sensor 13 is provided.
In the present embodiment, the light source position adjusting device 32 is configured to move the light source 21 on a plane orthogonal to the optical axis of the collector lens 22.

Hereinafter, the illumination adjustment work of the microscope 1 will be described. This illumination adjustment work is performed at the time of assembly and maintenance of the microscope 1 and at the time of replacement of the light source 21, and is performed according to the procedure shown in the flowchart of FIG.
Here, among the steps of the illumination adjustment work described below, the steps after the focusing step are performed by the control device 33 based on the output of the two-dimensional imaging sensor 13 of the collector lens position adjusting device 31 and the light source position adjusting device 32. This is done automatically by controlling the operation. These steps may be performed manually by the adjuster based on the output of the two-dimensional imaging sensor 13.

[Initialization process]
First, the setting of the microscope 1 is initialized (step S1). Specifically, as shown in FIG. 1, the adjuster installs a plate-shaped illumination adjustment plate 36 on the specimen surface on the stage 3, and in this state, the stage 3 is moved so that the illumination adjustment plate 36 is moved. Move to the focal position of the objective lens 11. As the illumination adjustment plate 36, for example, a reflecting plate that reflects a light beam irradiated by the illumination optical system 5, a fluorescent plate that emits a light having a light amount corresponding to the irradiation light amount by being irradiated with the light beam by the illumination optical system 5, and the like Is used.

  Here, a stage position adjusting device that moves the stage 3 to the microscope 1 and an automatic focusing device that controls the operation of the stage position adjusting device so that the sample surface on the stage 3 is positioned at the focal position of the objective lens 11 are provided. In the case where it is provided, the operation after the installation of the illumination adjustment plate 36 in the initialization process can be automated as in the other processes.

  The automatic focusing device, for example, based on the output of the two-dimensional imaging sensor 13, the contrast ratio between the area illuminated by the illumination optical system 4 and the other area of the illumination adjustment plate 36 on the specimen surface. And the position of the stage 3 is adjusted so that this contrast ratio is maximized. In other words, among the light receiving elements constituting the two-dimensional imaging sensor 13, the output ratio of the light receiving elements contributing to the shooting of the illuminated area of the illumination adjustment plate 36 to the light receiving elements contributing to the shooting of other areas is calculated. In other words, the position of the stage 3 is adjusted so that this output ratio is maximized.

  Here, during the illumination adjustment work, when the light incident on the light receiving element of the two-dimensional image sensor 13 is too bright and overexposed, the exposure time of the two-dimensional image sensor 13 is shortened to receive light. The amount of light incident on the element is kept within an appropriate range. On the other hand, when the light ray incident on the light receiving element is too dark, the exposure time of the two-dimensional imaging sensor 13 is lengthened to sufficiently secure the amount of light incident on the light receiving element. This operation can be performed using, for example, an automatic exposure time control function mounted on the two-dimensional imaging sensor 13 or the illumination optical system 4.

[Focus process]
Next, the position of the collector lens 22 in the optical axis direction (hereinafter referred to as the Z-axis direction) is adjusted to focus the light source image on the sample surface (step S2). In this focusing step, the position of the collector lens 22 is adjusted by utilizing the fact that when the light source image is focused on the sample surface, the amount of light emitted from the light source 21 to the sample surface is maximized. In this focusing step, the illumination of the sample surface by the illumination optical system 5 is not Koehler illumination but critical illumination.

In the present embodiment, the control device 33 sets the amount of light of the imaging area A _all (see FIG. 3) captured by the two-dimensional imaging sensor 13 on the sample surface, that is, each of the entire imaging areas of the two-dimensional imaging sensor 13. The operation of the collector lens position adjusting device 31 is controlled based on the average output value I _all of the light receiving elements. Here, the average output value _Iall is a value obtained by dividing the integrated value of the outputs of all the light receiving elements of the two-dimensional imaging sensor 13 by the total number of light receiving elements.

As shown in FIG. 4, the average output value I _all, the collector lens 22 as the vertex a point in the focus position Z _F, the position of the collector lens 22 gradually decreases as out of the focus position Z _F. Controller 33, By utilizing this, by moving the collector lens 22 to a position where the average output value I _all is maximized, moving the collector lens 22 to the in-focus position Z _F. Through this focusing step, a circular illumination spot S by the illumination optical system 5 is formed on the sample surface.

[Light source position adjustment process]
Next, the position of the light source 21 on a plane substantially perpendicular to the optical axis of the collector lens 22 is adjusted, and the light source 21 is moved onto the optical axis of the collector lens 22 (step S3).
When the light source 21 is positioned on the optical axis of the collector lens 22, the illumination spot S is positioned at the central portion A ₀ (see FIG. 3) of the imaging area A _{all on} the sample surface. amount of light applied to the a ₀ is maximized. In this light source position adjusting step, the position of the light source 21 is adjusted using this fact. The central portion A ₀ is a circular area smaller than the illumination spot S. When the light source 21 is located on the optical axis of the collector lens 22, the entire central portion A ₀ is illuminated. It fits within the spot S.

In the present embodiment, the control device 33 has an average output value of a light receiving element that contributes to photographing of the central portion A _{0 in} the imaging region of the two-dimensional imaging sensor 13 (that is, a light receiving element constituting the central portion A ₀ of the imaging region). Based on I ₀ , the operation of the light source position adjusting device 32 is controlled.
Here, the two-dimensional image sensor 13, the imaging of the central portion A _0, there is a configuration in which a plurality of light receiving elements are used. This reduces the influence of the noise light receiving elements have, it is possible to the amount of light radiated on the central portion A ₀ to accurately measure. Incidentally, the number of light receiving elements corresponding to the central portion A ₀ is too large, the detection sensitivity is deteriorated, on to accurately measure the amount of light radiated on the central portion A ₀ becomes difficult, the measurement and analysis of the quantity of light The time required for this will increase. Therefore, the number of contributing light receiving element to the imaging of the central portion A ₀ is preferably set to 4 to 64 elements.

  For example, a contrast method is used as a method of finding the position of the light source 21 that can obtain the maximum light output. This method uses the current position of the light source 21 as a reference to determine the moving direction of the light source 21 and the amount of change in the output value of the two-dimensional imaging sensor 13 after the movement, and the amount of change in the output value per unit movement amount is obtained. This is a method of moving the light source 21 to the position where the output value is maximized by moving the light source 21 in the direction in which it becomes the largest (however the output value increases), and thereafter repeating this operation.

Here, for example, the control device 33 is configured to control the position of the light source 21 on an orthogonal coordinate system including an X axis and a Y axis on a plane substantially orthogonal to the optical axis (Z axis).
In addition, the size of the central portion A ₀ and the illumination spot S is sufficiently secured, and in the illumination position adjustment process, at least a part of the illumination spot S usually covers the central portion A ₀ . Although the it, rarely illumination spot S is sometimes not applied to the central portion a _0. In this case, even if the light source 21 is moved, the average output value I ₀ does not change.
Therefore, in the present embodiment, when the average output value I ₀ does not change even when the light source 21 is moved in this way, the control device 33 turns the light source 21 so that the spot S covers the central portion A _0. It is configured to move.

Specifically, when the average output value I ₀ does not change even when the light source 21 is moved in this way, the control device 33 does not change the average output value I ₀ until the average output value I ₀ changes. The light source 21 is moved along one of the axes. If the average output value I ₀ does not change even after performing this operation, the control device 33 moves the light source 21 along the other axis by a predetermined amount, and then again moves along the one axis. This operation is repeated until the average output value I ₀ changes.

Here, when the light source 21 is greatly deviated from the optical axis of the collector lens 22 immediately after the initialization process, the light source 21 is placed on the specimen surface only by performing the focusing process and the light source position adjusting process once each. It is also conceivable that the image is not completely formed. In this case, the focusing process and the light source position adjusting process are repeated a plurality of times in this order, so that the light source 21 is more reliably imaged on the sample surface. Specifically, the control device 33 is configured to repeat these steps a predetermined number of times, or the average output value I ₀ at the center when the light source position adjustment step is finished and the previous light source position adjustment step. In the configuration, the focusing process and the light source position adjusting process are repeated until the difference from the average output value I ₀ at the center of the light source is within a reference range (for example, within 5% of the average output value I ₀ at the current center) To do. In addition, each time the focusing process and the light source position adjustment process are repeated, the adjustment of illumination can be performed more precisely by decreasing the unit movement amount when moving the collector lens and the light source.

[Lighting unevenness suppression process]
Next, the amount of light irradiated on each of the plurality of measurement areas on the sample surface is measured based on the output of the two-dimensional imaging sensor 13, and the collector lens 22 is measured based on the difference in the amount of irradiation light on each measurement area on the optical axis. Move to a position that falls within the range (step S4). Thereby, the illumination of the sample surface by the illumination optical system 5 becomes Koehler illumination.
In the present embodiment, as shown in FIG. 3, on the sample surface, the inner circumferential edge sets a peripheral region A ₁ of the annular contact with the outer peripheral edge of the central portion A ₀ surrounds the periphery of the central portion A _0, and the peripheral region a ₁ and the central portion a ₀ is used as a measurement region.

Specifically, the control device 33 detects the average output value I _{0 of the} light receiving element that contributes to the photographing of the central portion A _{0 in} the imaging region of the two-dimensional imaging sensor 13 and the light receiving element that contributes to the photographing of the peripheral region A _1. the difference between the average output value I ₁ of is configured for controlling the operation of the collector lens position adjusting device 31 so as to be within the reference range (eg, within 5% of the average output value I _0).

By moving the collector lens 22 on the optical axis, the ratio (I ₁ / I ₀ ) between the average output value I ₀ and the average output value I ₁ changes as shown in the graph of FIG.
Here, for each state of points A, B, and C on the graph of FIG. 5, the distance and position from the center on the sample surface and the output value of the light receiving element of the two-dimensional imaging sensor 13 corresponding to the position. The relationship with I is shown in the graph of FIG. 6A shows the state at point A in FIG. 5, FIG. 6B shows the state at point B in FIG. 5, and FIG. 6C shows the state at point C in FIG.
When the state after the light source position adjustment process is different from the state of the point B (for example, the state of the point A or the point C in FIG. 5), the control device 33 has the value of I ₁ / I ₀ . The position of the collector lens 22 is adjusted so that the point B in FIG.

Thus, in this microscope 1, when adjusting the illumination, the amount of light irradiated onto the sample surface is made uniform based on the actual output of each light receiving element of the two-dimensional imaging sensor 13. Even if the light emission unevenness 21, lens aberration, setting deviation, or the like occurs, the illumination of the microscope 1 can be appropriately adjusted according to the state.
In the present embodiment, the average output value I ₀ of the light receiving element that contributes to the photographing of the central portion A _{0 in} the imaging region of the two-dimensional imaging sensor 13 and the average output value I of the light receiving element that contributes to the photographing of the peripheral region A _1. the difference between the ₁ performs become so adjusted (within 5% for example, the average output value I ₀₎ reference range, effectively suppressing uneven illumination between the central area and the peripheral area of the sample surface be able to.

Moreover, since the light quantity irradiated to each measurement area | region is measured using the two-dimensional imaging sensor 13, it can perform highly accurate adjustment stably irrespective of an adjuster. Furthermore, when readjustment is performed for the microscope 1 that has been adjusted once by the microscope illumination adjustment method, the adjustment value can be made reproducible by setting the measured value at the time of the previous adjustment as a target value. .
Furthermore, since the light quantity is measured only for a part of the sample surface, the light quantity can be measured and analyzed in a shorter time than the case where the light quantity is measured for the entire specimen surface, and the illumination is adjusted. Can be done quickly.

In this microscope illumination adjustment method, since the two-dimensional imaging sensor 13 provided in the microscope 1 is used as a light amount measuring device, it is not necessary to prepare a new light amount measuring device.
For this reason, the illumination of the microscope 1 can be adjusted at low cost.

In the present embodiment, the peripheral region A ₁ on the sample surface, but the inner peripheral edge has a shape in contact with the outer peripheral edge of the central portion A _0, without being limited thereto, the shape and arrangement of the peripheral regions A ₁ Can be set arbitrarily. For example, the peripheral area A ₁ may be arranged such that the inner peripheral edge is separated from the outer peripheral edge of the central portion A ₀ as shown in FIG.

[Second Embodiment]
Next, a second embodiment of the present invention will be described with reference to FIG.
The microscope according to the present embodiment is mainly characterized in that a plurality of peripheral regions on the specimen surface are provided in the microscope 1 shown in the first embodiment.

In the present embodiment, as shown in FIG. 8, instead of setting the annular peripheral area A ₁ , the four peripheral areas A ₂ , A ₃ , A ₄ , the a _5, are arranged at approximately 90 ° intervals around the center of the sample surface. Here, in the present embodiment, the area A _{all to} be imaged is rectangular, and the peripheral areas A _{2 to} A ₅ are arranged near the corners of the area A _{all to} be imaged.

Such intends line illumination adjustment of the arranged peripheral region. Of the illumination adjustment at the light source position adjusting step, after the average output value I ₀ of contributing light receiving element to the imaging of the central portion A _0, and adjusted the position of the light source 21 so as to maximize, further, all for the peripheral area _a 2 to a _5, the average output value _I 2 ~I ₅ corresponding light receiving elements to adjust the position of the light source 21 to fit within the reference range.

For example, as the light source position adjustment step, first, the light source 21 is moved so that the value of (I ₂ + I ₃ ) − (I ₄ + I ₅ ) is minimized (approximately 0), and then (I ₂ + I ₄ ) − ( The light source 21 is moved so that the value of I ₃ + I ₅ ) is minimized (approximately 0). Thus, for all of the peripheral areas A ₂ to A _5, becomes the average output value I ₂ ~I ₅ corresponding light receiving element is minimal, since the deviation of the light amount is reduced with respect to the center of the sample surface, uniform near the sample surface Illumination suitable for observation of the specimen surface can be obtained.

Here, the shape and arrangement of the measurement region can be arbitrary.
For example, in the example shown in FIG. 9, the central portion A _0, has a rectangular similar and orientation as the imaging area A _all be the same. Also, the central portion A ₀ and similar and orientation greater rectangular region than further central A ₀ be the same, the central portion A ₀ and concentrically to set the peripheral region A of each rectangular area in the quarters and ₆ to a _9, are configured to respectively cover the four-quadrant around the center of the sample surface by these peripheral areas a ₆ to a _9.

Further, for example, in the example shown in FIG. 10, the central portion A ₀ is a circular region, and an annular region that is concentric with the central portion A ₀ and larger in diameter than the central portion A ₀ is defined by a straight line that passes through the center of the sample surface. The area is divided into four equal parts, which are the peripheral areas A _{10 to} A _13, and the four quadrants around the center of the specimen surface are covered by these peripheral areas A _{10 to} A ₁₃ .

  Here, in each of the above-described embodiments, the example in which the measurement of the amount of light applied to the sample surface is performed using the two-dimensional imaging sensor 13 provided in the observation optical system 4 is shown, but the present invention is not limited to this. The dimensional imaging sensor 13 may be installed on the sample surface.

  In each of the above embodiments, the example in which the amount of light irradiated to each measurement area on the sample surface is measured using the two-dimensional imaging sensor 13 is shown, but the measurement area is not limited to this and is irradiated to each measurement area. The measurement of the amount of light may be performed using, for example, a photodetector provided corresponding to each measurement region.

It is a figure showing roughly the composition of the microscope concerning a first embodiment of the present invention. It is a flowchart which shows the adjustment procedure of the illumination optical system of the microscope which concerns on 1st embodiment of this invention. It is a figure which shows the adjustment method of the illumination optical system of the microscope which concerns on 1st embodiment of this invention. It is a figure which shows the adjustment method of the illumination optical system of the microscope which concerns on 1st embodiment of this invention. It is a figure which shows the adjustment method of the illumination optical system of the microscope which concerns on 1st embodiment of this invention. It is a figure which shows the example of the illumination state of the sample surface by an illumination optical system. It is a figure which shows the other example of the adjustment method of the illumination optical system of the microscope which concerns on 1st embodiment of this invention. It is a figure which shows the adjustment method of the illumination optical system of the microscope which concerns on 2nd embodiment of this invention. It is a figure which shows the other example of the adjustment method of the illumination optical system of the microscope which concerns on 2nd embodiment of this invention. It is a figure which shows the other example of the adjustment method of the illumination optical system of the microscope which concerns on 2nd embodiment of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Microscope 11 Objective lens 13 Two-dimensional imaging sensor (light quantity measuring device)
22 collector lens 26 aperture stop 31 collector lens position adjusting device 32 light source position adjusting device 33 control device A ₀ central region (measurement region)
A ₁ to A ₁₃ peripheral region (measurement region)

Claims (12)

A microscope used for a microscope that uses a light source with uneven light emission , projects a light source image to the position of an aperture stop by a collector lens, and projects the light source image as a secondary light source onto the pupil of the objective lens to illuminate the specimen surface. Lighting adjustment method,
A focusing step of moving the collector lens in the optical axis direction and focusing the light source image on the sample surface;
The light source, on a plane perpendicular to the optical axis of the collector lens, the light source position adjustment step of moving to a position where the amount of light to be irradiated on the sample surface is maximum,
The amount of light irradiated respectively for the peripheral region outside the central portion and the central portion is located in the center of the sample surface was measured by the light amount measuring apparatus, the irradiation light amount in the peripheral region and the irradiation amount of light at the central portion A microscope illumination adjustment method comprising: an illumination unevenness suppressing step of moving the collector lens in the optical axis direction at a position where the difference falls within a reference range. The microscope illumination adjustment method according to claim 1 , wherein after the light source position adjustment step, the focusing step and the light source position adjustment step are performed again , and then the illumination unevenness suppression step is performed. The peripheral region is composed of a first region to a fourth region,
The focusing step has a reference range different from the reference range in the illumination unevenness suppressing step,
The microscope according to claim 1 or 2 , wherein the light source is moved in the focusing step so that each of the irradiation light amounts in the first to fourth regions falls within the different reference range. Lighting adjustment method. When the measurement values of the light quantity in the first region to the fourth region are I _{2 to} I ₅ respectively ,
The light source is moved so that the value of (I ₂ + I ₃ ) − (I ₄ + I ₅ ) is minimized, and then the value of (I ₂ + I ₄ ) − (I ₃ + I ₅ ) is minimized. 4. The microscope illumination adjustment method according to claim 3, wherein the light source is moved . Instead of the difference between the irradiation light amount in the central portion and the irradiation light amount in the peripheral region, the collector lens is used in the illumination unevenness suppressing step so that the ratio of the irradiation light amount in the central portion and the peripheral region becomes a predetermined value. The illumination adjustment method for a microscope according to any one of claims 1 to 4, wherein the illumination is adjusted in the optical axis direction . The central region is smaller than the illumination spot on the specimen surface,
The focusing step is performed using the sum of the measured values in the central portion and the peripheral region,
The light source position adjustment step is performed using only the measured value in the central portion,
6. The microscope illumination adjustment method according to claim 1 , wherein the illumination unevenness suppressing step is performed using a difference or a ratio of measured values in the central portion and the peripheral region . A microscope that uses a light source with uneven light emission , projects a light source image to a position of an aperture stop by a collector lens, and projects the light source image as a secondary light source onto a pupil of an objective lens to illuminate a specimen surface,
A collector lens position adjusting device for moving the collector lens in the optical axis direction;
A light source position adjustment device for moving in a plane perpendicular to the optical axis of the collector lens the light source,
A light amount measuring device that measures the amount of light irradiated on each of a central portion located at the center of the sample surface and a peripheral region located outside the central portion ;
A control device for controlling the operation of the collector lens position adjusting device and the light source position adjusting device based on the measurement value of the light quantity measuring device ;
The microscope including the control in which the collector lens is moved in an optical axis direction so that a difference between an irradiation light amount in the central portion and an irradiation light amount in the peripheral region falls within a reference range .   The central portion is smaller than the illumination spot on the specimen surface,
  The microscope according to claim 7, wherein the peripheral region has an inner peripheral edge larger than an outer peripheral edge of the central portion and is disposed at a position surrounding the central portion.   The microscope according to claim 7 or 8, wherein the peripheral region includes four regions divided into four equal parts by a straight line passing through a center of the central portion.   The central portion is smaller than the illumination spot on the specimen surface,
  The microscope according to claim 7 or 8, wherein the peripheral region is composed of four regions that are point-symmetric with respect to the center of the specimen surface and that are arranged at intervals of 90 degrees around the center.   The microscope has a two-dimensional imaging sensor for photographing a specimen on the specimen surface,
  The microscope according to any one of claims 7 to 10, wherein an amount of light irradiated to the measurement region on the specimen surface is obtained from an output of the two-dimensional imaging sensor.   The microscope according to claim 11, wherein an area captured by a plurality of adjacent light receiving elements among the light receiving elements constituting the two-dimensional imaging sensor is the measurement area.

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