How to adjust the illumination path of microscope

Abstract

The present disclosure describes a method for adjusting the illumination light path of a microscope. The method uses a reasonable illumination light path structure and an accurate adjustment confirmation window position to gradually complete the fine adjustment of the illumination light path in different zones. First, the light source is collimated to form a parallel light beam; a field stop and an aperture stop are set on the light path to present a light source image at the aperture stop; after the position of the tube lens and the turntable is determined, the light path assembly from the light source to the tube lens is completed by observing the field stop image on the turntable; finally, an observation window is set on the side of the tube lens close to the position where the microscope objective lens is installed, and the position of the objective lens is determined by adjusting the observation light source image. According to the present disclosure, a method for adjusting the light source image can be provided that is easy to operate, can improve the illumination accuracy of the light source in the microscope imaging system, and improve the imaging quality of the microscopic detection light path.

Description

Technical Field

[0001] The present disclosure generally relates to an intelligent manufacturing equipment industry, and specifically relates to a method for adjusting a microscope illumination light path. Background Art

[0002] At present, optical microscopy is widely used in various fields of scientific and technological research, but ordinary optical microscopy cannot achieve three-dimensional imaging for objects with a certain thickness. With the continuous development of microscopy technology in recent years, confocal microscopy has become one of the important technologies in the field of optical microscopy. It has the characteristics of high precision, high resolution, non-contact and unique axial tomography scanning imaging, which can easily realize three-dimensional image reconstruction and has been widely used in micro-nano detection, precision measurement and life science research. Traditional confocal microscopy detection technology is based on the principle that the light source, the illuminated object point and the detector are conjugated with each other to perform single-point mechanical scanning. Therefore, the scanning speed is relatively slow, the mechanical control system is complex, and the vibration caused by the scanning limits the measurement accuracy, making it difficult to achieve real-time and fast three-dimensional measurement. In order to solve the shortcomings of slow imaging speed and small field of view of confocal microscope, parallel scanning confocal microscopy technology has emerged. Parallel scanning confocal microscopy technology improves the measurement speed of the original single-point confocal measurement, and the parallel scanning confocal microscopy detection technology based on Nipkow turntable has the advantages of simple structure, easy implementation, low cost and high image quality.

[0003] In the Nipkow disk parallel scanning confocal microscope, the illumination light path and the detection light path need to share the small hole on the Nipkow disk. At this time, the reasonable setting and accurate adjustment of the illumination light path are particularly important. The illumination light path is one of the indispensable and important light paths in microscopy technology. Making the illumination light path have good and uniform illumination conditions is a necessary condition for achieving high-precision and high-resolution measurements.

[0004] Patent document [CN110764271A] discloses a method for adjusting the position accuracy between lenses in a conjugate optical system, which uses a high-precision air-floating rotating shaft to replace the focusing axis of an internal focusing telescope, so that the adjustment accuracy of the lens optical axis can reach the micron level. However, the solution disclosed in the patent document is costly and complicated to operate, and is not suitable for the adjustment of the illumination light path of a parallel scanning confocal microscope. Summary of the invention

[0005] The present disclosure is made in view of the above-mentioned prior art, and its purpose is to provide a method that is easy to operate and can accurately adjust the illumination light path of a turntable parallel scanning confocal microscope. By setting a reasonable illumination light path structure and accurately adjusting the observation window position, the illumination light path is finely adjusted step by step in different zones, the illumination accuracy of the light source in the microscope imaging system is improved, and the imaging quality of the microscopic detection light path is improved.

[0006] To this end, the present disclosure provides a method for adjusting a microscope illumination light path, wherein the microscope illumination light path comprises a sleeve assembly having a light source and a field diaphragm, a lens barrel assembly having a light collecting lens and an aperture diaphragm, a spectroscopic chamber having a spectroscope and a rotating disk having a small aperture surface, a tube lens, and a microscope objective lens, and the method for adjusting the illumination light path comprises: arranging the light source to form a first parallel light beam entering the lens barrel assembly via the field diaphragm; arranging the aperture diaphragm at the focal plane of the light collecting lens, adjusting the posture of the light source so that the light source forms a centrally symmetrical image in the aperture diaphragm; arranging the rotating disk and the spectroscope in the spectroscopic chamber; forming A second parallel light beam is emitted from the tube lens into the rotating disk of the spectroscopic chamber, and the relative positions of the tube lens and the spectroscopic chamber are adjusted so that the second parallel light beam forms a preset image on the rotating disk; the first parallel light beam is emitted into the spectroscopic chamber via the lens barrel assembly and forms a light spot on the rotating disk; the relative positions of the field diaphragm and the light collecting lens are adjusted so that the light spot covers the pinhole surface; the image of the light source formed by the light beam emitted from the tube lens is received at different positions, and the installation position of the microscope objective lens is determined based on the position of the image of the light source that meets the preset requirements, and the microscope objective lens is installed.

[0007] The adjustment method disclosed in the present invention is based on the requirements of Kohler illumination conditions and complies with the illumination conditions of "pupil to window, window to pupil". The effect of this illumination condition is to form an image of the light source on the image side rear focal plane of the microscope objective lens, and to achieve a conjugate relationship between the field stop and the object side focal plane of the microscope objective lens. As a result, the surface of the illuminated object sample is illuminated smoothly and evenly without any shadows.

[0008] The installation and adjustment method disclosed in the present invention firstly collimates the light beam of the light source to collect the edge light so that it is emitted in the form of parallel light, so that the light energy is evenly distributed in the cross section perpendicular to the optical axis. Then the light passes through the field diaphragm and the light collecting mirror to form a light source image at the aperture diaphragm, and the posture of the light source is adjusted so that the light source forms a centrally symmetrical image in the aperture diaphragm, so that the optical axis of the parallel light beam passes through the center of the aperture diaphragm, and the posture of the light source can be adjusted so that the illumination light energy is not lost, thereby reducing the aberration caused by the optical axis not passing through the center of the aperture diaphragm. Furthermore, by adjusting the relative positions of the field diaphragm, the aperture diaphragm, the turntable and the tube lens in sections and zones, the image of the light source formed by the light beam emitted from the tube lens is received, and the imaging position of the image of the light source that meets the preset requirements is the installation position of the image-side rear focal plane of the microscope objective lens. In this case, the effect of forming the light source image on the image-side rear focal plane of the microscope objective lens can be achieved. At the same time, the field diaphragm is formed on the small hole surface of the turntable so that the field diaphragm image covers the small hole surface of the turntable, which can achieve the effect of the field diaphragm being conjugated with the object-side focal plane of the microscope objective lens.

[0009] In addition, in the adjustment method of this embodiment, optionally, the field stop has a non-circular through hole, and the field stop is a square stop, so that it is easy to observe the central symmetrical image formed by the light source in the aperture stop and the outer shape image of the field stop on the small hole surface of the rotating disk.

[0010] In addition, in the adjustment method involved in this embodiment, optionally, the sleeve assembly includes a collimating lens, and the relative positions of the light source and the collimating lens are adjusted to form a first parallel light beam that is incident on the collimating lens and enters the lens barrel assembly. In this case, the collimating lens can effectively correct the edge aberration of the field of view, reduce spherical aberration, and improve the utilization rate of light energy.

[0011] In addition, in the adjustment method of this embodiment, optionally, an observation window is provided at the aperture stop to receive the image formed by the light source at the aperture stop. Since it is difficult to obtain the image of the light source through the through hole of the aperture stop, by using the observation window provided at the aperture stop, the light beam passing through the through hole of the aperture stop can be obtained to form the image of the light source, and then the relative position of the aperture stop and the light collecting mirror can be determined.

[0012] In addition, in the adjustment method involved in this embodiment, optionally, an observation window is used to receive the image of the light source formed by the light beam emitted from the tube lens, and the observation window is moved along the optical axis until the image of the light source that meets the preset requirements appears on the observation window. In this way, the position of a clear light source image can be determined, and then the installation position of the image-side rear focal plane of the microscope objective lens can be determined, so that the installation position of the microscope objective lens can be determined according to the numerical relationship between the image-side focal plane of the microscope objective lens and the parfocal plane.

[0013] In addition, in the adjustment method of this embodiment, optionally, the observation window is a translucent object, and the translucent object is a translucent frosted glass. In this case, the image of the light source formed by the aperture diaphragm and the image of the light source formed by the light beam emitted from the tube lens are imaged on the translucent object, thereby facilitating visual observation of the corresponding imaging conditions.

[0014] In addition, in the adjustment method involved in this embodiment, optionally, after adjusting the relative position of the light source and the collimating lens, the positional relationship between the light source and the collimating lens is fixed; after adjusting the relative position of the field diaphragm and the light collecting lens, the positional relationship between the field diaphragm and the light collecting lens is fixed; after adjusting the relative position of the tube lens and the spectroscopic chamber, the positional relationship between the tube lens and the spectroscopic chamber is fixed. In this case, partition adjustment can be achieved, and during the adjustment process, it is ensured that other adjustment steps will not affect the relative positional relationship between the adjusted modules.

[0015] In addition, in the adjustment method of this embodiment, optionally, the sleeve assembly includes an outer rotating cylinder, and the side wall of the outer rotating cylinder is provided with an adjustment device for adjusting the position of the light source in the vertical plane of the optical axis, and the adjustment device includes a top screw. Thus, by using the adjustment device, the posture of the light source can be adjusted so that the light source forms a centrally symmetrical image in the aperture stop.

[0016] In addition, in the adjustment method involved in this embodiment, optionally, the second parallel light beam is obtained by a collimator, and the preset image is a collimator graticule image. Since the graticule image is imaged at infinity through two parallel light beams, the collimator can simulate the reflected light beam of the sample under test passing through the microscope objective lens and entering the turntable, thereby being able to more accurately adjust the relative position of the tube lens and the spectroscopic chamber.

[0017] In addition, in the application device involved in this embodiment, optionally, the sleeve assembly is connected to the lens barrel assembly by threads, and the tube lens is connected to the spectroscopic chamber by threads. Thus, through the threaded connection, the sleeve assembly can be rotated along the thread, so that the sleeve assembly moves along the optical axis, and the relative position between the sleeve assembly and the lens barrel assembly is adjusted, thereby adjusting the relative position between the field stop and the light collecting lens; through the threaded connection, the tube lens and the spectroscopic chamber can be rotated along the thread, so that the tube lens moves along the optical axis, and the relative position between the tube lens and the spectroscopic chamber is adjusted, thereby adjusting the relative position between the tube lens and the rotating disk.

[0018] According to the present disclosure, a method can be provided that is easy to operate and can accurately adjust the illumination light path of a turntable parallel scanning confocal microscope. By setting a reasonable illumination light path structure and accurately adjusting the observation window position, the illumination light path adjustment is gradually completed according to the partition, which can improve the illumination accuracy of the light source in the microscope imaging system and improve the imaging quality of the microscopic detection light path. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 Schematic diagram showing the optical path of the microscope according to the present embodiment example.

[0020] Figure 2 Schematic diagram showing an illumination light path of a microscope according to this embodiment example.

[0021] Figure 3 1 is a schematic diagram showing the structure of the illumination light path of the microscope involved in this embodiment example.

[0022] Figure 4A It is a flowchart showing a first embodiment of the installation and adjustment method involved in this embodiment example.

[0023] Figure 4B FIG. 1 is a flow chart showing a second embodiment of the assembling and adjusting method according to the present embodiment.

[0024] Figure 5 is a schematic diagram showing a scenario of obtaining a first parallel light beam involved in this embodiment example.

[0025] Figure 6 1 is a schematic diagram of a scene showing the posture of adjusting the light source involved in this embodiment example.

[0026] Figure 7 1 is a schematic diagram showing the relative positions of the adjustment tube mirror and the spectroscopic chamber involved in this embodiment example.

[0027] Figure 8 is a schematic diagram showing a scenario of adjusting the position of a microscope objective lens involved in this embodiment example. DETAILED DESCRIPTION

[0028] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] It should be noted that the terms "first", "second", "third" and "fourth" in the specification and claims of the present invention and the above-mentioned drawings are used to distinguish different objects rather than to describe a specific order. In addition, the terms "include" and "have" and any variations thereof are intended to cover non-exclusive inclusions. For example, a process, method, system, product or device that includes a series of steps or units is not limited to the listed steps or units, but may optionally include steps or units that are not listed, or may optionally include other steps or units that are inherent to these processes, methods, products or devices. In the following description, the same symbols are given to the same components, and repeated descriptions are omitted. In addition, the drawings are only schematic diagrams, and the ratio of the sizes of the components to each other or the shapes of the components may be different from the actual ones.

[0030] The embodiments of the present disclosure relate to a method for adjusting the illumination light path of a turntable parallel scanning confocal microscope. Through the adjustment method of the present disclosure, the fine adjustment of the illumination light path can be completed step by step in a zoned manner without using special instruments and equipment. The overall operation process is simple and it is also easy to troubleshoot problems. The "method for adjusting the illumination light path of a turntable parallel scanning confocal microscope" can also be referred to as the "adjustment method".

[0031] Hereinafter, the installation and adjustment method involved in this embodiment will be described in detail with reference to the accompanying drawings.

[0032] Figure 1 is a schematic diagram showing the optical path of a microscope according to the present embodiment example, Figure 2 is a schematic diagram showing an illumination light path of a microscope according to the present embodiment. Figure 3 1 is a schematic diagram showing the structure of the illumination light path of the microscope involved in this embodiment example.

[0033] In some examples, the microscope involved in the present disclosure may be a parallel scanning confocal microscope. In some examples, the microscope involved in the present disclosure may be a Nipkow spinning disk parallel scanning confocal microscope.

[0034] For some examples, see Figure 1 ,The optical path of a microscope can include an illumination optical path and a detection optical path.

[0035] For some examples, see Figure 2 and Figure 3 The illumination light path may include: a sleeve assembly 1, a lens barrel assembly 2, a spectroscopic chamber 3, a tube lens 4 and a microscope objective lens 5.

[0036] In some examples, the microscope may include a first connection device, which may enable the sleeve assembly 1 to move along the optical axis. The first connection device may connect the sleeve assembly 1 with the lens barrel assembly 2, and the sleeve assembly 1 and the lens barrel assembly 2 constitute a first module 100.

[0037] In some examples, the microscope may include a second connecting device, which can enable the tube lens 4 to move along the optical axis. The second connecting device can connect the spectroscopic chamber 3 with the tube lens 4, and the spectroscopic chamber 3 and the tube lens 4 are connected to form a second module 200.

[0038] In some examples, the first connecting device and the second connecting device may have a thread structure, a slot structure, or a guide rail structure.

[0039] For some examples, see Figure 2 and Figure 3 The light can be emitted by the light source 11, enter the lens barrel assembly 2 through the sleeve assembly 1, and then enter the spectroscopic chamber 3; the light passes through the spectroscopic chamber 3 and enters the tube lens 4, then the light enters the microscope objective lens 5, and finally the light is irradiated on the surface of the sample to be measured.

[0040] In some examples, the sleeve assembly 1 may include a light source 11 , a collimating lens 12 , and a field stop 13 .

[0041] In some examples, there may be preset dark grid patterns on the light source 11, and whether the image of the light source 11 is clear can be determined by the pattern of the dark grid patterns, thereby facilitating imaging observation in subsequent steps.

[0042] In some examples, the collimating lens 12 can effectively correct the aberration at the edge of the field of view and reduce the spherical aberration, thereby improving the utilization rate of light energy.

[0043] In some examples, the field stop 13 may be disposed between the collimating lens 12 and the collecting mirror 21 (described later).

[0044] In some examples, the sleeve assembly 1 may include an outer rotating cylinder 14 , and the light source 11 may be located inside the outer rotating cylinder 14 of the sleeve assembly 1 .

[0045] In some examples, the side wall of the outer rotating cylinder 14 of the sleeve assembly 1 may be provided with an adjustment device. In some examples, the adjustment device may be used to adjust the position of the light source 11. In some examples, the adjustment device may be used to adjust the position of the light source 11 in the vertical plane of the optical axis, and in some examples, the adjustment device is a top screw.

[0046] In some examples, the sleeve assembly 1 can include an inner rotating barrel 15. In some examples, the collimating lens 12 and the field stop 13 can be located within the inner rotating barrel 15 of the sleeve assembly 1.

[0047] In some examples, the light collecting mirror 21 may be disposed in the first lens barrel 24 of the lens barrel assembly 2 .

[0048] In some examples, the reflector 22 may be disposed within the second barrel 25 of the barrel assembly 2 .

[0049] In some examples, the light collecting lens 21 is also called a condenser lens or a condenser.

[0050] In some examples, the aperture stop 23 may be a through-hole portion of the second lens barrel 25 .

[0051] In some examples, the light beam entering the lens barrel assembly 2 may be converged by the light collecting mirror 21 and reflected by the reflecting mirror 22 to the through hole of the aperture stop 23 .

[0052] In some examples, an observation window may be provided at the through hole of the aperture stop 23 to obtain a light source image (described later).

[0053] In some examples, the spectroscope 31 and the rotating disk 32 may be placed in the spectroscopic chamber 3 .

[0054] For some examples, see Figure 1 , the beam splitter 31 can be used to merge the measurement light path and the illumination light path.

[0055] In some examples, the turntable 32 may be a turntable with a small hole surface. Specifically, the turntable 32 may be provided with small holes distributed in a certain arrangement rule. In some examples, the turntable 32 may be a Nipkow turntable.

[0056] In some examples, a portion having a small hole may also be referred to as a small hole surface.

[0057] In some examples, in the illumination light path, the light beam entering the spectroscopic chamber 3 can reach the rotating disk 32 through the spectroscope 31 , and reach the tube lens 4 through the small hole of the rotating disk 32 .

[0058] In some examples, as described above, the image-side back focal plane and the parfocal plane of the microscope objective 5 may be a uniform fixed value.

[0059] In some examples, the spectroscopic chamber 3 may have a window or an openable side. In this case, the imaging on the rotating disk 32 can be observed visually or determined by an instrument.

[0060] In some examples, after the light source 11 emits light and evenly illuminates the surface of the sample to be tested, the surface of the sample to be tested reflects the illumination light, and the reflected light enters the microscope objective 5, enters the tube lens 4, the turntable 32, and the spectroscope 31, is reflected 90° at the spectroscope 31, and enters the detection light path 6 after reflection, and finally the information of the sample to be tested is imaged on the imaging device of the detection light path 6.

[0061] In some examples, the adjustment method may adjust the illumination light path in sections and partitions. Figure 4A It is a flowchart showing a first embodiment of the installation and adjustment method involved in this embodiment example. Figure 4B FIG. 1 is a flow chart showing a second embodiment of the assembling and adjusting method according to the present embodiment.

[0062] For some examples, see Figure 4A , the installation and adjustment method may include: forming a first parallel light beam (step S100); adjusting the imaging of the light source 11 image on the aperture diaphragm 23 (step S200); setting the turntable 32 and the spectroscope 31 in the spectroscope chamber 3 (step S300); adjusting the tube lens 4 so that the tube lens 4 focuses the image (step S400); allowing the first parallel light beam to enter the tube lens 4 (step S500); adjusting the imaging of the field diaphragm 13 on the turntable 32 (step S600); determining the position of the microscope objective 5, and installing the microscope objective 5 (step S700). Based on the requirements of the Kohler illumination conditions, the "pupil to window, window to pupil" illumination conditions are followed. The presentation effect of this illumination condition is to form the image of the light source 11 on the image-side rear focal plane of the microscope objective 5, and to achieve a conjugate relationship between the field diaphragm 13 and the object-side focal plane of the microscope objective 5. As a result, the illumination of the surface of the illuminated object sample is smooth and uniform without any shadows. By setting a reasonable illumination light path structure and accurately adjusting the observation window position, the illumination light path is finely adjusted step by step in different zones, the illumination accuracy of the light source 11 in the microscope imaging system is improved, and the imaging quality of the microscopic detection light path is improved.

[0063] For some examples, see Figure 2 and Figure 8 The imaging process of the light source 11 in the illumination light path can be as follows: Figure 2 and Figure 8As shown by the solid line, the light source 11 forms a light source image for the first time at the aperture stop 23, and the second light source image position is placed on the image side rear focal plane of the microscope objective 5. In this case, the light source image passes through the microscope objective 5 and forms an image at infinity on the side of the sample to be tested. Since the sample to be tested is at a finite distance, the illumination on the surface of the sample to be tested will not be affected by the shadow of the light source filament, so the illumination light on the surface of the sample to be tested is uniform and there is no shadow. The imaging process of the field stop 13 in the illumination light path can be seen in Figure 2 and Figure 8 As shown by the middle dotted line, the field diaphragm 13 forms an image of the field diaphragm 13 for the first time at the turntable 32, and the image is a conjugate image of the field diaphragm 13. Meanwhile, the field diaphragm 13 and the object focal plane of the microscope objective 5 are in a conjugate relationship, and thus the field diaphragm 13 determines the range of the illuminated object surface.

[0064] In some examples, the order between step S100 and step S200 and between step S300 and step S400 can be changed. Specifically, step S100 and step S200 can be placed before step S300 and step S400; step S100 and step S200 can be placed after step S300 and step S400. Figure 4B , while executing step S100 and step S200 in sequence, step S300 and step S400 can be executed synchronously.

[0065] Figure 5 is a schematic diagram showing a scenario of obtaining a first parallel light beam involved in this embodiment example.

[0066] In some examples, in step S100, a first parallel light beam may be formed. In this case, the light beam of the light source 11 may be collimated to collect the edge light beams so that the light beams are emitted in the form of parallel light beams, so that the light energy is evenly distributed in the cross section perpendicular to the optical axis.

[0067] In some examples, the position of the light source 11 can be adjusted to change the relative position between the light source 11 and the focus of the collimating lens 12, and the light source 11 is approximately located near the focus of the collimating lens 12. In this case, the light source 11 passes through the collimating lens 12 to form a first parallel light beam, and the first parallel light beam passes through the field aperture 13 and enters the lens barrel kit 2. In this case, the collimating lens 12 can effectively correct the aberration at the edge of the field of view, reduce spherical aberration, and improve the utilization rate of light energy. At the same time, the light is emitted in the form of a parallel light beam, so that the light energy is evenly distributed in the vertical cross section of the optical axis, and the shape of the light source 11 is imaged at infinity.

[0068] In some examples, the initial position of the light source 11 is located near the theoretical focal length value of the collimating lens 12. Figure 5, an observation window is set on the side of the outer side of the inner rotating cylinder 15 away from the light source 11, and the observation window is moved along the optical axis direction, and the shape and size of the light spot on the observation window are observed at the same time. If the outer diameter of the light spot changes with the movement of the observation window, it is necessary to further adjust the outer rotating cylinder 14, and then adjust the focal position of the light source 11 in the collimating lens 12, until the observation window is at different selected positions, and it is observed that the shape and size of the light spot remain consistent, and the outer diameter of the light spot does not change with the movement of the observation window, and remains consistent. In some examples, 3-5 positions can be selected to place the observation window to observe the shape and size of the light spot. Then, the outer rotating cylinder 14 is fixed, the light source collimation adjustment is completed, and step S100 is completed. It should be noted that "the shape and size of the light spot remain consistent" can be understood in the following way: the shape and size of the light spot is determined by visual inspection, and the shape and size of the light spot is determined to remain consistent based on the visual inspection; or the shape and size of the light spot is obtained by instruments and equipment, and the shape and size of the light spot is determined to remain consistent based on preset rules.

[0069] In some examples, after adjusting the relative position of the light source 11 and the collimating lens 12, the positional relationship between the light source 11 and the collimating lens 12 is fixed. In this case, partition adjustment can be achieved, and during the adjustment process, it is ensured that other adjustment steps will not affect the relative positional relationship between the adjusted modules.

[0070] In some examples, the adjustment can be performed manually. In some examples, as the observation window moves, the change in the outer diameter of the light spot can be determined by visually observing the observation window. In this case, the adjustment can be performed conveniently.

[0071] In some examples, digital imaging observation technology can be used to implement automatic software calibration, and the digital imaging observation system includes a scanning / control unit, an image acquisition / processing unit, and a computer-aided measurement / control unit.

[0072] In some examples, multiple observation windows may be set at the same time, and it is determined whether the outer diameters, shapes and sizes of the light spots in the multiple observation windows are all consistent, so as to determine whether the light beam passing through the collimating lens 12 is collimated.

[0073] In some examples, the relative positions of the light source 11 and the collimating lens 12 can be adjusted to form a first parallel light beam incident on the collimating lens 12 and entering the lens barrel assembly 2. In some examples, the collimating lens 12 uses a spherical lens group to collimate the light source light.

[0074] In some examples, the light beam of the light source 11 in the illumination light path is collimated by the collimating lens 12 and then emitted as parallel light. At a position close to the collimating lens 12, the light in the cross section of the optical axis is evenly distributed, and it is suitable to set the field stop 13 here. Setting the field stop 13 here can effectively limit the light beam, thereby effectively limiting the imaging range of the light source 11.

[0075] In some examples, the field stop 13 is a limit to the field of view of the optical path system, which determines the range of the illuminated object surface. The aperture value of the field stop 13 is a preset value of the optical design of the microscope illumination light path.

[0076] In some examples, as described above, the field stop 13 may be disposed between the collimating lens 12 and the collecting lens 21 .

[0077] In some examples, the field diaphragm 13 can be set outside one focal length and within two focal lengths on the light source side of the light collecting mirror 21. In this case, the conjugate image of the field diaphragm 13 through the light collecting mirror 21 is presented on the turntable 32, and its conjugate image is an enlarged real image. The light in the image plane is evenly distributed, which meets the lighting requirements on the turntable 32. At the same time, the field diaphragm 13 and the object focal plane of the microscope objective 5 are a pair of conjugate planes, thereby ensuring that the illumination light illuminates the surface of the illuminated object sample smoothly and evenly without any shadows.

[0078] In some examples, the field stop 13 has a non-circular through hole. In some examples, the field stop 13 is a square stop in shape. In this case, during the adjustment process, it is easy to observe the shape image of the field stop 13 on the small hole surface of the turntable 32, but the through hole of the field stop 13 is not limited to a square. For example, the through hole of the field stop 13 can be a star or irregular shape. In some cases, since the frame of the housing or other components of the lens barrel assembly 2 (such as the outer frame of the lens) may also serve as the actual effective field stop, and in the optical path system, the shape of the frame of the housing or other components of the first lens barrel 24 is generally circular, rectangular, polygonal or elliptical, therefore, by using the field stop 13 with a non-circular through hole, it can be determined that the field stop 13 provided on the sleeve assembly 1 has a limiting effect on the light beam. In other words, compared to the method of using the outer frame of a lens as a field stop or omitting the field stop, the present invention uses a field stop 13 with a non-circular through hole during optical path adjustment, which makes it easier for the adjuster to distinguish the image of the field stop 13 and achieve accurate adjustment.

[0079] In some examples, the field diaphragm 13 is a variable aperture diaphragm, which can adapt to different optical design requirements. According to the optical index requirements, the microscope can be replaced with a variable aperture of different apertures.

[0080] Figure 6 1 is a schematic diagram of a scene showing the posture of adjusting the light source involved in this embodiment example.

[0081] In some examples, after step S100 , step S200 may be performed.

[0082] In some examples, in step S200, the imaging of the light source 11 in the aperture stop 23 may be adjusted. In this case, the posture of the light source 11 is adjusted so that the light source 11 forms a centrally symmetrical image in the aperture stop 23, so that the optical axis of the parallel light beam passes through the center of the aperture stop 23, and the posture of the light source 11 can be adjusted so that the illumination light energy is not lost, thereby reducing the aberration caused by the optical axis not passing through the center of the aperture stop 23.

[0083] In some examples, reference Figure 3 and Figure 6 The lens barrel assembly 2 may include a first lens barrel 24 and a second lens barrel 25 .

[0084] In some examples, the light collecting mirror 21 can be placed in the first lens barrel 24, and the reflector 22 can be placed in the second lens barrel 25; the aperture stop 23 is a through hole portion of the second lens barrel 25. The first connecting device can connect the sleeve assembly 1 with the lens barrel assembly 2, and the first lens barrel 24 can be moved along the optical axis through the first connecting device. The light from the sleeve assembly 1 passes through the light collecting mirror 21, irradiates the reflector 22, is reflected by the reflector 22 to the aperture stop 23, and then passes through the aperture stop 23.

[0085] In some examples, in a microscope illumination system, a light collecting mirror 21 can be set after the collimated light path of the light source (i.e., the light path of the first parallel light beam formed in step S100). In this case, the light collecting mirror 21 increases the energy density of the illumination beam, so that the microscope has high resolution and contrast characteristics and increases the illumination light energy.

[0086] In some examples, the sleeve assembly 1 can be connected to the first lens barrel 24 of the lens barrel assembly 2. In some examples, the sleeve assembly 1 is connected to the lens barrel assembly 2 by a thread. In this case, the sleeve assembly 1 can move along the optical axis direction, thereby adjusting the relative position of the sleeve assembly 1 and the lens barrel assembly 2.

[0087] In some examples, an observation window may be provided at the through hole of the aperture stop 23. In this case, the imaging of the light source 11 at the aperture stop 23 can be observed through the observation window. In other words, the observation window may be provided at the aperture stop 23 to receive the image of the light source 11 formed at the aperture stop 23. Since the light beam passes through the through hole of the aperture stop 23, it is difficult to obtain the image of the light source. By using the observation window provided at the aperture stop 23, the light beam passing through the through hole of the aperture stop 23 can be obtained to form the image of the light source, and then the relative position of the aperture stop 23 and the light collecting mirror 21 can be determined.

[0088] In some examples, the initial position of the aperture stop 23 can be set near the focal plane of the light collecting mirror 21 to control the aperture angle of the illumination system. The light beam of the light source 11 passes through the collimating lens 12 to form a first parallel light beam, and the first parallel light beam passes through the light collecting mirror 21 to be imaged at the aperture stop 23.

[0089] In some examples, the imaging of the light source 11 on the observation window can be observed by moving the first lens barrel 24 along the optical axis, and the observation window is set at the through hole of the aperture stop 23. In this embodiment, the light source 11 can have a preset dark grid pattern, and when a preset clear dark pattern image of the light source wick appears in the observation window, the first lens barrel 24 can be fixed. Thus, it can be determined that the aperture stop 23 is located at the focal plane position of the light collecting lens 21, thereby ensuring that the aperture stop 23, the light collecting lens 21 and the collimating lens 12 are located on the same optical axis and are placed perpendicular to the optical axis without tilt, ensuring that the light source image will not have aberrations.

[0090] In some examples, after adjusting the relative position of the field stop 23 and the light collecting mirror 21, the positional relationship between the field stop 23 and the light collecting mirror 21 is fixed. In this case, partition adjustment can be achieved, and during the adjustment process, it is ensured that other adjustment steps will not affect the relative positional relationship between the adjusted modules.

[0091] In some examples, after determining that the aperture stop 23 is located at the focal plane position of the light collecting mirror 21, the posture of the light source 11 can be adjusted so that the light source 11 forms a centrally symmetrical image in the aperture stop 23. In some examples, the side wall of the outer rotating cylinder 14 of the sleeve assembly 1 is provided with an adjustment device, which can be used to adjust the position of the light source 11 up, down, left, and right in the vertical plane of the optical axis. In some examples, the adjustment device is a top screw. Thus, by using the adjustment device, the posture of the light source 11 can be adjusted so that the light source 11 forms a centrally symmetrical image in the aperture stop. Furthermore, by adjusting the posture of the light source 11, the position of the light source 11 is adjusted up, down, left, and right in the plane vertical to the optical axis, and the observation window is observed at the same time, so that the image of the light source 11 is completely symmetrical in the aperture stop 23, and the edge of the light source image is completely within the aperture range of the aperture stop 23, thereby ensuring that the illumination light energy is not lost.

[0092] In some examples, the observation window in step S200 may be a translucent object, and in some examples, the translucent object may be a translucent frosted glass, in which case it is convenient for visual observation. At the same time, the image formed by the light source 11 at the aperture stop 23 is formed on the translucent object, thereby facilitating visual observation of the imaging situation.

[0093] In some examples, after step S200 , step S300 may be performed.

[0094] In some examples, in step S300 , the rotating disk 32 and the spectroscope 31 may be disposed in the spectroscopic chamber 3 , and their position distribution may be preset values ​​for optical design.

[0095] In some examples, as described above, many small holes may be distributed on the turntable 32, and the light beam of the light source 11 covers the range of all the small holes (i.e., the scanning area). When the turntable 32 rotates at high speed, a small hole scans a corresponding area on the sample to be tested, thereby achieving a complete scan of the sample to be tested.

[0096] In some examples, after step S300 , step S400 may be performed.

[0097] In some examples, in step S400 , the tube lens 4 may be adjusted to enable the tube lens 4 to focus the image. Figure 7 A schematic diagram of a scene showing the relative positions of the adjustment tube lens 4 and the spectroscopic chamber 3 according to the embodiment example is shown.

[0098] For some examples, see Figure 7 , the tube lens 4 can be connected to the spectroscopic chamber 3.

[0099] In some examples, after the tube lens 4 is connected to the spectroscopic chamber 3 , a second parallel light beam may be formed and emitted from the tube lens 4 into the spectroscopic chamber 3 .

[0100] In some examples, the second parallel light beam can be generated by a parallel light tube 7 and have a preset image, and the second parallel light beam is emitted from the tube lens 4 into the spectroscopic chamber 3, and the relative positions of the tube lens 4 and the spectroscopic chamber 3 are adjusted so that the parallel light beam forms a preset image on the turntable 32.

[0101] In some examples, the preset image may be a collimator graticule image that matches the collimator 7. Since the graticule image is imaged at infinity through the second parallel light beam, the collimator 7 can simulate the reflected light beam of the sample under test passing through the microscope objective 5 and entering the turntable 32, thereby being able to more accurately adjust the relative position of the tube lens 4 and the spectroscopic chamber 3.

[0102] In some examples, the tube lens 4 is connected to the spectroscopic chamber 3 by threads. In this case, the tube lens 4 and the spectroscopic chamber 3 can be rotated along the threads to move the tube lens 4 along the optical axis, thereby adjusting the relative position between the tube lens 4 and the spectroscopic chamber 3, and thus adjusting the relative position between the tube lens 4 and the rotating disk 32.

[0103] In some examples, the tube lens 4 and the spectroscopic chamber 3 are connected as a whole and placed under the collimator 7. The collimator 7 forms a second parallel light beam that is emitted from the tube lens 4 into the spectroscopic chamber 3. The relative positions of the tube lens 4 and the spectroscopic chamber 3 are adjusted so that the second parallel light beam appears as a clear image of the graticule of the collimator 7 on the turntable 32. In this case, by simulating the scene in which the surface of the sample under test reflects the illumination light, the reflected light enters the microscope objective 5, enters the tube lens 4 as a parallel light beam, and forms an image on the turntable 32, the relative position of the tube lens 4 and the turntable 32 is determined to ensure the integrity of the optical path.

[0104] In some examples, after adjusting the relative position of the tube lens 4 and the spectroscopic chamber 3, the positional relationship between the tube lens 4 and the spectroscopic chamber 3 is fixed. In this case, partition adjustment can be achieved, and during the adjustment process, it is ensured that other adjustment steps will not affect the relative positional relationship between the adjusted modules.

[0105] In some examples, after step S400 , step S500 may be performed.

[0106] In some examples, in step S500, the first parallel light beam can be injected into the tube lens 4. Specifically, the first module 100 can be placed on the second module 200, and the second module 200 can be placed on the side of the light collecting mirror 21 away from the light source 11, at a position greater than 2 times the focal length of the light collecting mirror 21, so that the first parallel light beam is injected into the spectroscopic chamber 3 through the sleeve assembly 1 and the lens barrel assembly 2, and enters the tube lens 4 through the spectroscopic chamber 3.

[0107] In some examples, after step S500 , step S600 may be performed.

[0108] In some examples, in step S600, the imaging of the field stop 13 on the turntable 32 can be adjusted. In this case, by adjusting the relative positions of the field stop 13, the turntable 32 and the tube lens 4 in sections and regions, and making the field stop 13 form the image of the field stop 13 on the small hole surface of the turntable 32, the image of the field stop 13 covers the small hole surface of the turntable, so that the field stop 13 and the object side focal plane of the microscope objective 5 can be conjugated.

[0109] In some examples, an infinite conjugate image microscope objective lens 5 and a tube lens 4 can be selected to cooperate to achieve the purpose of microscopic imaging. In a turntable parallel scanning confocal microscope, the pinhole on the turntable 32 is shared by the illumination light path and the detection imaging light path. The microscopic imaging image plane, i.e., the conjugate image plane of the object plane, is on the pinhole surface of the turntable 32. Therefore, the image of the field diaphragm 13 needs to be presented on the pinhole surface of the turntable 32, and the image spot of the field diaphragm 13 completely covers the pinhole surface within the imaging range of the turntable 32. It should be noted that the imaging range can be understood in the following way: during the operation of the parallel scanning confocal microscope, the turntable 32 will rotate along the central axis of the turntable 32, and at the same time, a part of the turntable 32 will be located in the illumination light path and the measurement light path. On the turntable 32, the area located in the illumination light path and the measurement light path can be called the imaging range.

[0110] In some examples, the position of the sleeve assembly 1 can be adjusted by moving along the optical axis. In this case, the light source 11, the collimating lens group 12, and the field stop 13 are moved along the optical axis relative to the lens barrel assembly 2 as an integral module, and the relative position of the field stop 13 and the light collecting lens 21 is adjusted so that a clear image of the field stop 13 is formed on the rotating disk 32, and the image spot of the field stop 13 completely covers the small hole surface within the imaging range of the rotating disk 32, see Figure 2 and Figure 8 As shown, the dotted line in the figure is the imaging process of the field stop 13. In some examples, as described above, the spectroscopic chamber 3 may have an openable side, in which case the image spot of the field stop 13 on the rotating disk 32 can be observed.

[0111] In some examples, the actual image spot size of the field stop 13 is the design value of the illumination light path. The design value of the illumination light path can be calculated using the Gaussian formula.

[0112] In some examples, after step S600, step S700 may be performed. In some examples, in step S700, the position of the microscope objective lens 5 may be determined, and the microscope objective lens 5 may be installed. In this case, by placing an observation window on the side of the tube lens 4 away from the spectroscopic chamber 3, the observation window is observed until it receives an image of the light source 11 that meets the preset requirements. At this time, the position of the observation window is the image-side rear focal plane of the microscope objective lens 5. Through the design parameters of the microscope objective lens 5, the numerical relationship between the image-side rear focal plane of the microscope objective lens 5 and the parfocal plane can be determined, and then the installation position of the microscope objective lens 5 can be determined, so that the light source can be imaged on the image-side rear focal plane of the microscope objective lens 5.

[0113] Figure 8 is a schematic diagram showing a scenario of adjusting the position of a microscope objective lens involved in this embodiment example.

[0114] In some examples, before assembling the microscope objective lens 5 in the overall optical path, it is necessary to first find a clear light source image formed by the light beam of the light source 11 after passing through the sleeve assembly 1, the lens barrel assembly 2, the spectroscopic chamber 3 and the tube lens 4, and mark the position of the clear light source image, and set the image-side rear focal plane of the microscope objective lens 5 at this position. In this case, the light source image is imaged at infinity on one side of the sample to be measured after passing through the microscope objective lens 5. Since the sample to be measured is at a finite distance, the illumination of the sample surface to be measured will not be affected by the shadow of the light source filament, so the illumination light on the surface of the sample to be measured is uniformly illuminated and there is no shadow. In other words, see Figure 2 and Figure 8 As shown by the solid line in the middle, the light source 11 presents a first real image at the aperture stop 23 through the collimating lens 12 and the collecting lens 21. The first image then passes through the tube lens 4 and forms a second real image between the tube lens 4 and the microscope objective lens 5. Therefore, the microscope objective lens 5 can be assembled based on the positions of the first real image and the second real image.

[0115] In some examples, reference Figure 8 An observation window can be placed on the side of the tube lens 4 away from the spectroscopic chamber 3, and the observation window can be moved along the optical axis to receive the image of the light source 11 formed by the light beam emitted from the tube lens 4 at different positions until a clear image of the light source 11 that meets the preset requirements appears on the observation window, and the position of the observation window at this time is marked.

[0116] In some examples, the observation window can be used to receive the image of the light source 11 formed by the light beam emitted from the tube lens 4, and the observation window can be moved along the optical axis until the image of the light source 11 that meets the preset requirements appears on the observation window. In this way, the position of the clear image of the light source 11 can be determined, and then the installation position of the image-side rear focal plane of the microscope objective lens 5 can be determined, so that the installation position of the microscope objective lens 5 can be determined according to the numerical relationship between the image-side rear focal plane of the microscope objective lens 5 and the parfocal plane.

[0117] In some examples, the preset requirement may be that the clarity and shape of the image of the light source 11 meet the requirements. In some examples, the preset requirement may be that the operator can clearly distinguish the shape of the image of the light source 11. In some examples, it may be determined by image processing whether the image of the light source 11 meets the preset requirements, for example, the preset requirement may be that the resolution (or gradient) of the image of the light source 11 meets the requirements.

[0118] In some examples, the image-side rear focal plane of the microscope objective lens 5 may be arranged near or at the position. This position is the position where the image-side rear focal plane of the microscope objective lens 5 is placed.

[0119] In some examples, as described above, the image-side back focal plane of the microscope objective lens 5 and the parfocal plane of the microscope objective lens 5 are uniformly fixed values. In this case, the installation position of the microscope objective lens 5 can be determined based on the uniformly fixed value of the image-side back focal plane of the microscope objective lens 5 and the parfocal plane of the microscope objective lens 5.

[0120] In some examples, as described above, the image-side rear focal plane and the parfocal plane of the microscope objective lens 5 may be uniform fixed values, thereby making it possible to determine the installation position of the microscope objective lens 5 and install the microscope objective lens 5 .

[0121] In some examples, the observation window in the above step S700 is a translucent object, and in some examples, the translucent object is a translucent frosted glass. In this case, it is convenient for visual observation, and at the same time, the image of the light source 11 formed by the light beam emitted from the tube lens 4 is formed on the translucent object, thereby facilitating visual observation of the corresponding imaging situation.

[0122] Various embodiments of the present invention are described above in the detailed description. Although these descriptions directly describe the above embodiments, it should be understood that modifications and / or variations of the specific embodiments shown and described herein may be conceived by those skilled in the art. Any such modifications or variations that fall within the scope of this specification are also intended to be included therein. Unless otherwise specified, it is the intention of the inventor that the words and phrases in the specification and claims are given the ordinary and customary meanings of ordinary technicians.

[0123] The above description of various embodiments of the present invention known to the applicant at the time of filing this application has been presented and is intended for the purpose of illustration and description. This description is not intended to be exhaustive of the present invention, nor to limit the present invention to the exact form disclosed, and many modifications and variations may be made in accordance with the above teachings. The described embodiments serve to explain the principles of the present invention and its practical application, and to enable others skilled in the art to utilize the present invention in various embodiments and various modifications suitable for the intended specific use. Therefore, it is intended that the present invention is not limited to the specific embodiments disclosed for implementing the present invention.

[0124] Although specific embodiments of the present invention have been shown and described, it is apparent to those skilled in the art that variations and modifications may be made based on the teachings of the present invention without departing from the present invention and its broader aspects, and the appended claims will cover within their scope all such changes and modifications that are within the true spirit and scope of the present invention. It will be understood by those skilled in the art that, in general, the terms used in the present invention are generally intended to be "open" terms (e.g., the term "including" should be interpreted as "including but not limited to", the term "having" should be interpreted as "at least having", the term "including" should be interpreted as "including but not limited to", etc.).

Claims

1. A method for adjusting a microscope illumination light path, wherein the microscope illumination light path comprises a sleeve assembly having a light source and a field diaphragm, a lens barrel assembly having a light collecting lens and an aperture diaphragm, a spectroscopic chamber having a spectroscope and a rotating disk having a small aperture surface, a tube lens, and a microscope objective lens, wherein: The method for adjusting the illumination light path comprises: The light source is arranged to form a first parallel light beam entering the lens barrel assembly through the field aperture; The aperture stop is set at the focal plane of the light collecting mirror, and the posture of the light source is adjusted so that the light source forms a centrally symmetrical image in the aperture stop; The rotating disk and the spectroscope are arranged in the spectroscopic chamber; Forming a second parallel light beam and allowing the second parallel light beam to enter the rotating disk of the spectroscopic chamber from the tube lens, and adjusting the relative position of the tube lens and the spectroscopic chamber so that the second parallel light beam forms a preset image on the rotating disk; Allow the first parallel light beam to enter the spectroscopic chamber through the lens barrel assembly and form a light spot on the rotating disk; Adjusting the relative positions of the field aperture and the light collecting mirror to form the light spot covering the pinhole surface; The image of the light source formed by the light beam emitted from the tube lens is received at different positions, and the installation position of the microscope objective lens is determined based on the position of the image of the light source that meets the preset requirements, and the microscope objective lens is installed.

2. The adjustment method according to claim 1, characterized in that: The field diaphragm has a non-circular through hole, and the field diaphragm is a square diaphragm.

3. The adjustment method according to claim 1, characterized in that: The sleeve assembly includes a collimating lens, and the relative positions of the light source and the collimating lens are adjusted to form a first parallel light beam incident on the collimating lens and entering the lens barrel assembly.

4. The adjustment method according to claim 1, characterized in that: An observation window is arranged at the aperture stop to receive an image formed by the light source at the aperture stop.

5. The adjustment method according to claim 1, characterized in that: The image of the light source formed by the light beam emitted from the tube lens is received by using an observation window, and the observation window is moved along the optical axis until the image of the light source that meets the preset requirements appears on the observation window.

6. The adjustment method according to claim 4 or 5, characterized in that: The observation window is a translucent object, and the translucent object is a translucent frosted glass.

7. The adjustment method according to claim 3, characterized in that: After adjusting the relative position of the light source and the collimating lens, the positional relationship between the light source and the collimating lens is fixed; after adjusting the relative position of the field diaphragm and the light collecting lens, the positional relationship between the field diaphragm and the light collecting lens is fixed; after adjusting the relative position of the tube lens and the spectroscopic chamber, the positional relationship between the tube lens and the spectroscopic chamber is fixed.

8. The adjustment method according to claim 1, characterized in that: The sleeve assembly comprises an outer rotating cylinder, and a side wall of the outer rotating cylinder is provided with an adjusting device for adjusting the position of the light source in a plane vertical to the optical axis, and the adjusting device comprises a top screw.

9. The adjustment method according to claim 1, characterized in that: The second parallel light beam is obtained through a collimator, and the preset image is a collimator graticule image.

10. The adjustment method according to claim 1, characterized in that: The sleeve assembly is connected to the lens barrel assembly via threads, and the tube lens is connected to the spectroscopic chamber via threads.

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