Illumination light path of turntable parallel scanning confocal microscope

Abstract

The present disclosure describes a turntable parallel scanning confocal microscope illumination light path, the illumination light path comprising a sleeve assembly having a light source and a field diaphragm, a lens barrel assembly having a collector lens, a beam splitting chamber having a turntable, a tube lens and a microscope objective, the light source being configured to form a first parallel beam entering the lens barrel assembly via the field diaphragm; the light source is imaged on the focal plane of the light collecting mirror; the turntable is configured to receive a second parallel light beam emitted from the tube lens, the second parallel light beam is configured to form a preset image on the turntable, the turntable is configured to receive a first parallel light beam emitted into the light splitting chamber through the lens barrel assembly, and the first parallel light beam is configured to form a light spot covering a small hole surface of the turntable on the turntable; the microscope objective is configured to be installed at the position of an image of the light source which is formed by the light beam emitted from the tube mirror and meets the preset requirement. According to the invention, the illumination light path of the turntable parallel scanning confocal microscope, which is simple and convenient to operate and can be conveniently and accurately assembled and adjusted, can be provided.

Description

[0001] This application is a divisional application of the patent application with the application date of August 26, 2022, application number 2022110299739, and invention title of "Alignment Method for Microscope Illumination Optical Path". Technical Field

[0002] The present disclosure generally relates to the field of intelligent manufacturing equipment industry, and specifically to an illumination optical path of a turntable parallel scanning confocal microscope. Background Art

[0003] Currently, optical microscopy technology is widely used in various fields of scientific and technological research. However, ordinary optical microscopy technology cannot achieve three-dimensional imaging of objects with a certain thickness. With the continuous development of microscopy technology in recent years, confocal microscopy technology 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, and can easily realize three-dimensional image reconstruction, and has been widely used in fields such as micro-nano detection, precision measurement, and life science research. Traditional confocal microscopy detection technologies are all based on the principle of conjugate of a light source, an illuminated object point, and a detector, and perform single-point mechanical scanning. Therefore, the scanning speed is relatively slow, the mechanical control system is complex, and the vibration caused by scanning limits the measurement accuracy, and it is not easy to achieve real-time and fast three-dimensional measurement. To solve the disadvantages of slow imaging speed and small field of view of confocal microscopes, parallel scanning confocal microscopy technology has emerged. Parallel scanning confocal microscopy technology has improved the measurement speed of the original single-point confocal measurement, and the parallel scanning confocal detection technology based on the Nipkow turntable has the advantages of simple structure, easy implementation, low cost, and high image quality.

[0004] In a Nipkow turntable parallel scanning confocal microscope, the illumination optical path and the detection optical path need to share the small holes on the Nipkow turntable. At this time, the reasonable setting and accurate alignment of the illumination optical path are particularly important. The illumination optical path is one of the indispensable important optical paths in microscopy technology. Making the illumination optical path have good and uniform illumination conditions is a necessary condition for achieving high-precision and high-resolution measurement.

[0005] Patent document

CN110764271A

[0006] The present disclosure is completed in view of the above-mentioned state of the prior art, and its purpose is to provide a method for easily operating and accurately aligning and adjusting the illumination optical path of a rotary scanning confocal microscope. By using a reasonably arranged illumination optical path structure and accurately aligning and observing the position of the window, the fine adjustment of the illumination optical path is gradually completed in zones, the illumination accuracy of the light source in the microscope imaging system is improved, and the imaging quality of the microscopic detection optical path is improved.

[0007] To this end, the present disclosure provides a method for aligning and adjusting the illumination optical path of a microscope. The illumination optical path of the microscope includes a sleeve assembly having a light source and a field stop, a lens barrel assembly having a condenser and an aperture stop, a beam splitter chamber having a beam splitter and a rotary disk with a small hole surface, a tube lens, and a microscope objective. The method for aligning and adjusting the illumination optical path includes: setting the light source to form a first parallel beam that enters the lens barrel assembly through the field stop; setting the aperture stop at the focal plane of the condenser, and adjusting the attitude of the light source so that the light source forms a centrally symmetric image in the aperture stop; setting the rotary disk and the beam splitter in the beam splitter chamber; forming a second parallel beam and making the second parallel beam enter the rotary disk in the beam splitter chamber from the tube lens, and adjusting the relative position between the tube lens and the beam splitter chamber so that the second parallel beam forms a preset image on the rotary disk; making the first parallel beam enter the beam splitter chamber through the lens barrel assembly and form a light spot on the rotary disk; adjusting the relative position between the field stop and the condenser so that the formed light spot covers the small hole surface; receiving the image of the light source formed by the beam emitted from the tube lens at different positions, and determining the installation position of the microscope objective based on the position of the image of the light source that meets the preset requirements, and installing the microscope objective.

[0008] The alignment and adjustment method involved in the present disclosure is based on the requirements of Köhler illumination conditions and follows the illumination condition of "pupil to window, window to pupil". The presentation effect of this illumination condition is to image the light source on the image-side rear focal plane of the microscope objective and realize the conjugate relationship between the field stop and the object-side focal plane of the microscope objective. Thus, the surface of the illuminated object sample is smoothly and evenly illuminated without shadow.

[0009] The alignment method involved in the present disclosure first collimates the light beam of the light source, which can converge the marginal light rays to emit them in the form of parallel light, so as to make the distribution of light energy uniform within the cross-section perpendicular to the optical axis. Then, the light rays pass through the field stop and the condenser lens, and form an image of the light source at the aperture stop. By adjusting the posture of the light source so that the light source forms a centrosymmetric image in the aperture stop, it is possible to make the optical axis of the parallel light beam pass through the center of the aperture stop, and adjust the posture of the light source so that there is no loss of illumination light energy, thereby reducing the aberration caused by the optical axis not passing through the center of the aperture stop. Further, by segmentally and regionally adjusting the relative positions among the field stop, the aperture stop, the turntable and the tube lens, and receiving the image of the light source formed by the light beam emitted from the tube lens, 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. In this case, the effect of forming the image of the light source on the image-side rear focal plane of the microscope objective can be achieved. At the same time, by making the field stop form an image of the field stop covering the small hole surface of the turntable at the small hole surface of the turntable, the effect of conjugating the field stop with the object-side focal plane of the microscope objective can be achieved.

[0010] In addition, in the alignment method involved in the present embodiment, optionally, the field stop has a non-circular through hole, and the field stop is a square stop. Thus, it is easy to observe the centrosymmetric image of the light source formed in the aperture stop and the external shape image of the field stop on the small hole surface of the turntable.

[0011] In addition, in the alignment method involved in the present embodiment, optionally, the sleeve assembly includes a collimating lens. By adjusting the relative position between the light source and the collimating lens to form a first parallel light beam entering the lens barrel assembly in the collimating lens. In this case, the collimating lens can effectively correct the field edge aberration, reduce the spherical aberration, and improve the light energy utilization rate.

[0012] In addition, in the alignment method involved in the present embodiment, optionally, an observation window is arranged at the aperture stop to receive the image of the light source formed at the aperture stop. Since it is difficult to obtain the image of the light source as the light beam passes through the through hole of the aperture stop, by using the observation window arranged at the aperture stop, it is possible to obtain the light beam passing through the through hole of the aperture stop to form an image of the light source, and further determine the relative position between the aperture stop and the condenser lens.

[0013] In addition, in the alignment method according to 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 direction until the image of the light source that meets the preset requirements appears on the observation window. Thus, the position of the clear image of the light source can be determined, and then the installation position of the image-side rear focal plane of the microscope objective can be determined. Therefore, according to the numerical relationship between the image-side focal plane and the parfocal plane of the microscope objective, the installation position of the microscope objective can be determined.

[0014] In addition, in the alignment method according to this embodiment, optionally, the observation window is a translucent object, and the translucent object is a translucent ground glass. In this case, the image of the light source formed by the image of the light source formed by the aperture stop and the light beam emitted from the tube lens is imaged on the translucent object, thereby facilitating visual observation of the corresponding imaging situation.

[0015] In addition, in the alignment method according to this embodiment, optionally, after adjusting the relative position between 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 between the field stop and the condenser, the positional relationship between the field stop and the condenser is fixed; after adjusting the relative position between the tube lens and the spectroscopic chamber, the positional relationship between the tube lens and the spectroscopic chamber is fixed. In this case, zonal alignment can be achieved, and during the alignment process, it is ensured that other alignment steps will not affect the relative positional relationship between the aligned modules.

[0016] In addition, in the alignment method according to this embodiment, optionally, the sleeve assembly includes an outer rotating cylinder, and an adjusting device for adjusting the position of the light source in the plane perpendicular to the optical axis is provided on the side wall of the outer rotating cylinder, and the adjusting device includes a setscrew. Thus, by using this adjusting device, the attitude of the light source can be adjusted so that the light source forms a centrosymmetric image in the aperture stop.

[0017] In addition, in the alignment method according to this embodiment, optionally, the second parallel light beam is obtained through a collimator, and the preset image is a collimator reticle image. Since the reticle image is imaged at infinity through the two parallel light beams, the collimator can simulate the reflected light beam of the measured sample passing through the microscope objective and entering the turntable, and thus the relative position between the tube lens and the spectroscopic chamber can be adjusted more accurately.

[0018] In addition, in the application device according to this embodiment, optionally, the sleeve assembly and the objective lens barrel assembly are connected by threads, and the tube lens and the spectroscopic chamber are connected by threads. Thus, through the threaded connection, the sleeve assembly can be rotated along the thread, causing the sleeve assembly to move in the optical axis direction, adjusting the relative position between the sleeve assembly and the lens barrel assembly, and thereby adjusting the relative position between the field stop and the condenser; through the threaded connection, the tube lens and the spectroscopic chamber can be rotated along the thread, causing the tube lens to move in the optical axis direction, adjusting the relative position between the tube lens and the spectroscopic chamber, and thereby adjusting the relative position between the tube lens and the turntable.

[0019] According to the present disclosure, a method that is easy to operate and can accurately align and adjust the turntable and parallel scan the confocal microscope illumination optical path can be provided. By setting a reasonable illumination optical path structure and accurately aligning and adjusting the position of the observation window, and completing the illumination optical path calibration step by step according to the partition, the illumination accuracy of the light source in the microscope imaging system can be improved, and the imaging quality of the microscopic detection optical path can be improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 is a schematic diagram showing the optical path of the microscope according to the example of this embodiment.

[0021] Figure 2 is a schematic diagram showing the illumination optical path of the microscope according to the example of this embodiment.

[0022] Figure 3 is a schematic structural diagram showing the illumination optical path of the microscope according to the example of this embodiment.

[0023] Figure 4A is a schematic flow diagram showing the first embodiment of the alignment and adjustment method according to the example of this embodiment.

[0024] Figure 4B is a schematic flow diagram showing the second embodiment of the alignment and adjustment method according to the example of this embodiment.

[0025] Figure 5 is a schematic diagram showing the scene of obtaining the first parallel light beam according to the example of this embodiment.

[0026] Figure 6 is a schematic diagram showing the scene of adjusting the attitude of the light source according to the example of this embodiment.

[0027] Figure 7 is a schematic diagram showing the scene of adjusting the relative position between the tube lens and the spectroscopic chamber according to the example of this embodiment.

[0028] Figure 8 is a schematic diagram showing the scene of adjusting the position of the microscopic objective lens according to the example of this embodiment. Detailed Embodiments

[0029] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0030] It should be noted that the terms "first", "second", "third", "fourth", etc. in the specification and claims of the present invention and the above-mentioned accompanying drawings are used to distinguish different objects, rather than to describe a specific order. In addition, the terms "comprising" and "having" and any variations thereof are intended to cover non-exclusive inclusion. 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 further include steps or units not listed, or may optionally further include other steps or units inherent to these processes, methods, products, or devices. In the following description, the same reference numerals are given to the same components, and repeated descriptions are omitted. In addition, the drawings are only schematic diagrams, and the proportional relationship of the sizes between components or the shapes of components, etc. may be different from the actual ones.

[0031] The embodiments of the present disclosure relate to a method for adjusting and aligning the illumination optical path of a spinning disk parallel scanning confocal microscope. Through the adjustment and alignment method involved in the present disclosure, the fine adjustment work of the illumination optical path can be gradually completed in zones without using special instrument equipment. The overall operation process is simple and it is also easy to troubleshoot problems. The "method for adjusting and aligning the illumination optical path of a spinning disk parallel scanning confocal microscope" can also be referred to as the "adjustment and alignment method".

[0032] Hereinafter, the adjustment and alignment method involved in the present embodiment will be described in detail in conjunction with the accompanying drawings.

[0033] Figure 1 is a schematic diagram showing the optical path of the microscope involved in the example of the present embodiment, Figure 2 is a schematic diagram showing the illumination optical path of the microscope involved in the example of the present embodiment, Figure 3 is a structural schematic diagram showing the illumination optical path of the microscope involved in the example of the present embodiment.

[0034] 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.

[0035] In some examples, referring to Figure 1 , the optical path of the microscope may include an illumination optical path and a detection optical path.

[0036] In some examples, referring to Figure 2 and Figure 3 , the illumination optical path may include: a sleeve assembly 1, a lens barrel assembly 2, a beam splitting chamber 3, a tube lens 4, and a microscope objective 5.

[0037] In some examples, the microscope may include a first connecting device, and the first connecting device may enable the sleeve assembly 1 to move along the optical axis. The first connecting device may connect the sleeve assembly 1 to the lens barrel assembly 2, and the sleeve assembly 1 and the lens barrel assembly 2 form a first module 100.

[0038] In some examples, the microscope may include a second connecting device, and the second connecting device may enable the tube lens 4 to move along the optical axis. The second connecting device may connect the beam splitting chamber 3 to the tube lens 4, and the connection between the beam splitting chamber 3 and the tube lens 4 forms a second module 200.

[0039] In some examples, the first connecting device and the second connecting device may have a threaded structure, a card slot structure, or a guide rail structure.

[0040] In some examples, referring to Figure 2 and Figure 3 , light may be emitted from a light source 11, enter the lens barrel assembly 2 through the sleeve assembly 1, and then enter the beam splitting chamber 3; the light passes through the beam splitting chamber 3, enters the tube lens 4, and then the light enters the microscope objective 5, and finally the light irradiates on the surface of the sample to be measured.

[0041] In some examples, the sleeve assembly 1 may include a light source 11, a collimating lens 12, and a field stop 13.

[0042] In some examples, there may be a preset dark grid pattern on the light source 11, and the clarity of the image of the light source 11 can be determined through the pattern of the dark grid pattern, thereby facilitating imaging observation in subsequent steps.

[0043] In some examples, the collimating lens 12 can effectively correct the off-axis aberration and reduce the spherical aberration. Thus, the light energy utilization rate can be improved.

[0044] In some examples, the field stop 13 may be disposed between the collimating lens 12 and a condenser lens 21 (described later).

[0045] 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.

[0046] In some examples, an adjustment device may be provided on the side wall of the outer rotating cylinder 14 of the sleeve assembly 1. 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 plane perpendicular to the optical axis. In some examples, the adjustment device is a setscrew.

[0047] In some examples, the sleeve assembly 1 may include an inner rotating cylinder 15. In some examples, the collimating lens 12 and the field stop 13 may be located inside the inner rotating cylinder 15 of the sleeve assembly 1.

[0048] In some examples, the condenser lens 21 may be placed in the first lens barrel 24 of the lens barrel assembly 2.

[0049] In some examples, the reflecting mirror 22 may be placed inside the second lens barrel 25 of the lens barrel assembly 2.

[0050] In some examples, the condenser lens 21 is also referred to as a condenser or a concentrator.

[0051] In some examples, the aperture stop 23 may be the through-hole portion of the second lens barrel 25.

[0052] In some examples, the light beam entering the lens barrel assembly 2 may be converged by the condenser lens 21 and reflected by the reflecting mirror 22 to the through-hole of the aperture stop 23.

[0053] In some examples, an observation window may be provided at the through-hole of the aperture stop 23 to obtain the image of the light source (described later).

[0054] In some examples, the beam splitter 31 and the turntable 32 may be placed in the beam splitting chamber 3.

[0055] In some examples, referring to Figure 1 , the beam splitter 31 may be used to combine the measurement optical path and the illumination optical path.

[0056] In some examples, the turntable 32 may be a turntable with a small hole surface. Specifically, a certain number of small holes may be provided on the turntable 32 in a regular arrangement. In some examples, the turntable 32 may be a Nipkow turntable.

[0057] In some examples, the part with small holes may also be referred to as the small hole surface.

[0058] In some examples, in the illumination optical path, the light beam entering the beam splitting chamber 3 may reach the turntable 32 through the beam splitter 31 and reach the tube lens 4 through the small holes of the turntable 32.

[0059] In some examples, as described above, the image-side rear focal plane of the microscope objective 5 and the parfocal plane may be a unified fixed value.

[0060] In some examples, the beam splitting chamber 3 may have a window or an openable side. In this case, the imaging on the turntable 32 can be observed visually or determined by an instrument.

[0061] In some examples, after the light source 11 emits light rays that uniformly illuminate the surface of the sample to be measured, the surface of the sample to be measured reflects the illuminating light rays. The reflected light enters the microscopic objective lens 5, passes through the tube lens 4, the turntable 32, and the beam splitter 31. A 90° reflection occurs at the beam splitter 31, and after reflection, it enters the detection optical path 6. Finally, the information of the sample to be measured is imaged on the imaging device of the detection optical path 6.

[0062] In some examples, the alignment method can align the illumination optical path section by section and region by region. Figure 4A FIG. is a schematic flow chart of the first embodiment of the alignment method involved in the example of this embodiment. Figure 4B FIG. is a schematic flow chart of the second embodiment of the alignment method involved in the example of this embodiment.

[0063] In some examples, referring to Figure 4A , the alignment method may include: forming a first parallel light beam (step S100); adjusting the imaging of the image of the light source 11 on the aperture stop 23 (step S200); arranging the turntable 32 and the beam splitter 31 in the beam splitting chamber 3 (step S300); adjusting the tube lens 4 to make the tube lens 4 focus and form an image (step S400); making the first parallel light beam enter the tube lens 4 (step S500); adjusting the imaging of the field stop 13 on the turntable 32 (step S600); determining the position of the microscopic objective lens 5 and installing the microscopic objective lens 5 (step S700). Based on the requirements of Köhler illumination conditions, following the illumination condition of "pupil to window, window to pupil", 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 microscopic objective lens 5, and to conjugate the field stop 13 with the object-side focal plane of the microscopic objective lens 5. Thus, the surface of the illuminated object sample is illuminated smoothly and uniformly without shadows. By using a reasonable illumination optical path structure and an accurate alignment observation window position, the fine-tuning of the illumination optical path is gradually completed in sections, improving the illumination accuracy of the light source 11 in the microscope imaging system and improving the imaging quality of the microscopic detection optical path.

[0064] In some examples, referring to Figure 2 and Figure 8 , the imaging process of the light source 11 in the illumination optical path can be as Figure 2 and Figure 8As shown by the solid line, the light source 11 forms an image of the light source for the first time at the aperture stop 23, and the position for forming the second image of the light source is placed on the image-side rear focal plane of the microscope objective 5. In this case, after passing through the microscope objective 5, the image of the light source is imaged at infinity on the side of the measured sample. Since the measured sample is at a finite distance, the illumination on the surface of the measured sample will not be affected by the shadow of the light source filament. Therefore, the illumination light on the surface of the measured sample is uniformly illuminated and there is no shadow. The imaging process of the field stop 13 in the illumination optical path can be seen in Figure 2 and Figure 8 As shown by the dashed line, the field stop 13 forms an image of the field stop 13 for the first time at the turntable 32. This image is the conjugate image of the field stop 13. At the same time, the field stop 13 and the object-side focal plane of the microscope objective 5 are in a conjugate relationship. Thus, the field stop 13 determines the range of the illuminated object surface.

[0065] 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 located before step S300 and step S400; step S100 and step S200 can be located after step S300 and step S400. In some examples, see Figure 4B , while step S100 and step S200 are being executed in sequence, step S300 and step S400 can be executed synchronously.

[0066] Figure 5 is a schematic diagram of the scene for obtaining the first parallel light beam involved in the example of this embodiment.

[0067] In some examples, in step S100, a first parallel light beam can be formed. In this case, by collimating the light beam of the light source 11, the marginal rays can be focused and emitted in the form of parallel light, so as to make the distribution of light energy uniform in the cross-section perpendicular to the optical axis.

[0068] 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 forms a first parallel light beam through the collimating lens 12, and the first parallel light beam passes through the field stop 13 and enters the lens barrel assembly 2. In this case, the collimating lens 12 can effectively correct the off-axis aberration, reduce the spherical aberration, and improve the light energy utilization rate. At the same time, the light rays are emitted in the form of parallel light beams, so as to make the distribution of light energy uniform in the cross-section perpendicular to the optical axis, and image the outer shape of the light source 11 at infinity.

[0069] In some examples, the initial position of the light source 11 is near the theoretical focal length value of the collimating lens 12. Refer to Figure 5, an observation window is provided on the side of the outer rotating cylinder 15 away from the light source 11. By moving the observation window along the optical axis direction and observing the shape and size of the light spot on the observation window at the same time, if the outer diameter of the light spot changes as the observation window moves, it is necessary to further adjust the outer rotating cylinder 14, and then adjust the focal position of the light source 11 on the collimating lens 12 until the shape and size of the light spot are observed to be consistent at different selected positions of the observation window, and the outer diameter of the light spot does not change as the observation window moves, maintaining a consistent state. In some examples, the observation window can be placed at 3 - 5 positions to observe the shape and size of the light spot. Then, fix the outer rotating cylinder 14, and the collimation adjustment of the light source is completed, and step S100 is completed. It should be noted that "the shape and size of the light spot are consistent" can be understood in the following ways: determine the shape and size of the light spot by visual inspection, and determine that the shape and size of the light spot are consistent based on the visually determined shape and size of the light spot; or obtain the shape and size of the light spot through instrument equipment, and determine that the shape and size of the light spot are consistent based on a preset rule.

[0070] In some examples, after adjusting the relative positions 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, zonal alignment can be achieved, and during the alignment process, it is ensured that other alignment steps will not affect the relative positional relationship between the aligned modules.

[0071] In some examples, the adjustment can be carried out manually. In some examples, as the observation window moves, the observation window can be visually observed to judge the change in the outer diameter of the light spot. In this case, the adjustment can be carried out conveniently.

[0072] In some examples, software automatic adjustment can be achieved using digital imaging observation technology. The digital imaging observation system includes a scanning / control unit, an image acquisition / processing unit, and a computer-aided measurement / control unit.

[0073] In some examples, multiple observation windows can be set simultaneously, and it is judged whether the shape and size of the outer diameter of the light spots on the multiple observation windows are all consistent, so as to judge whether the light beam passing through the collimating lens 12 is collimated.

[0074] 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 entering the lens barrel assembly 2 on the collimating lens 12. In some examples, the collimating lens 12 uses a spherical mirror group to collimate the light rays of the light source.

[0075] In some examples, after the light beam of the light source 11 in the illumination optical path is collimated by the collimating lens 12, it exits in the form of parallel light. At a position close to the collimating lens 12, the light rays on the cross-section of the optical axis are evenly distributed. It is suitable to set the field stop 13 here. Setting the field stop 13 here can effectively limit the light beam, and thus can effectively limit the imaging range of the light source 11.

[0076] In some examples, the field stop 13 is a limitation on the field of view of the optical path system, which determines the size of the range of the illuminated object surface. The aperture value of the field stop 13 is a preset value in the optical design of the microscope illumination optical path.

[0077] In some examples, as described above, the field stop 13 can be disposed between the collimating lens 12 and the condenser lens 21.

[0078] In some examples, the field stop 13 can be disposed beyond the focal length and within twice the focal length on the light source side of the condenser lens 21. In this case, the conjugate image of the field stop 13 through the condenser lens 21 is presented on the turntable 32, and its conjugate image is an enlarged real image with uniform light distribution in the image plane, meeting the illumination requirements on the turntable 32. At the same time, the field stop 13 and the object-side focal plane of the microscope objective 5 are a pair of conjugate planes. Thus, it is ensured that the illumination light is smooth and uniform on the surface of the illuminated object sample without shadow.

[0079] In some examples, the field stop 13 has a non-circular through hole. In some examples, the outer shape of the field stop 13 is a square aperture. In this case, during the alignment process, it is easy to observe the outer shape image of the field stop 13 on the small hole surface of the turntable 32. However, 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 star-shaped or irregular. In some cases, since the outer shell of the lens barrel assembly 2 or the frame of other components (such as the outer frame of the lens) may also act as the actual effective field stop, and in the optical path system, the shape of the outer shell of the first lens barrel 24 or the frame of other components 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 disposed in the sleeve assembly 1 restricts the light beam. In other words, compared with the method of using the outer frame of a certain lens as the field stop or omitting the field stop, in the optical path adjustment of the present disclosure, by using the field stop 13 with a non-circular through hole, it is easier for the adjuster to distinguish the image of the field stop 13 and achieve accurate adjustment.

[0080] In some examples, the field stop 13 is an aperture variable stop. Thus, it can adapt to different optical design requirements. According to the optical index requirements, the microscope can be replaced with variable stops of different apertures.

[0081] Figure 6 It is a schematic diagram showing the scene of adjusting the attitude of the light source involved in the example of the present embodiment.

[0082] In some examples, after step S100, step S200 can be executed.

[0083] In some examples, in step S200, the imaging of the light source 11 in the aperture stop 23 can be calibrated. In this case, by adjusting the attitude of the light source 11 so that the light source 11 forms a centrosymmetric image in the aperture stop 23, it is possible to make the optical axis of the parallel light beam pass through the center of the aperture stop 23, and the attitude of the light source 11 can be adjusted so that the illumination light energy is lossless, thereby reducing the aberration caused by the optical axis not passing through the center of the aperture stop 23.

[0084] In some examples, referring to Figure 3 and Figure 6 , the lens barrel assembly 2 may include a first lens barrel 24 and a second lens barrel 25.

[0085] In some examples, the condenser lens 21 may be placed inside the first lens barrel 24, and the reflector 22 may be placed inside the second lens barrel 25; the aperture stop 23 is a through-hole part of the second lens barrel 25. The first connecting device can connect the sleeve assembly 1 and the lens barrel assembly 2, and the first lens barrel 24 can be moved along the optical axis direction through the first connecting device. The light rays from the sleeve assembly 1 are irradiated on the reflector 22 through the condenser lens 21, reflected by the reflector 22 to the aperture stop 23, and then pass through the aperture stop 23.

[0086] In some examples, in the microscopic illumination system, a condenser lens 21 can be provided after the light source collimation optical path (i.e., the optical path for forming the first parallel light beam in step S100). In this case, the condenser lens 21 increases the energy density of the illumination light beam, enables the microscope to have high resolution and contrast characteristics, and improves the illumination light energy.

[0087] In some examples, the sleeve assembly 1 and the first lens barrel 24 of the lens barrel assembly 2 can be connected. In some examples, the sleeve assembly 1 and the lens barrel assembly 2 are connected by threads. In this case, the sleeve assembly 1 can move along the optical axis direction, thereby adjusting the relative position between the sleeve assembly 1 and the lens barrel assembly 2.

[0088] In some examples, an observation window can 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 can be set on the aperture stop 23 to receive the image formed by the light source 11 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 light source image. By using the observation window provided on the aperture stop 23, the light beam passing through the through-hole of the aperture stop 23 can be obtained to form the light source image, and then the relative position between the aperture stop 23 and the condenser lens 21 can be determined.

[0089] In some examples, the initial position of the aperture stop 23 can be set near the focal plane of the condenser lens 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 then passes through the condenser lens 21 and is imaged at the aperture stop 23.

[0090] In some examples, by moving the first lens barrel 24 along the optical axis direction, the imaging of the light source 11 on the observation window can be observed. The observation window is arranged at the through hole of the aperture stop 23. In this embodiment, the light source 11 can have a preset dark grid pattern. When a preset clear dark pattern image of the light source wick appears on 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 condenser lens 21, so as to ensure that the aperture stop 23, the condenser lens 21 and the collimating lens 12 are on the same optical axis and are placed perpendicular to the optical axis without inclination, ensuring that there is no aberration in the light source image.

[0091] In some examples, after adjusting the relative position between the field stop 23 and the condenser lens 21, the positional relationship between the field stop 23 and the condenser lens 21 is fixed. In this case, zonal alignment can be achieved, and during the alignment process, it is ensured that other alignment steps will not affect the relative positional relationship between the aligned modules.

[0092] In some examples, after determining that the aperture stop 23 is located at the focal plane position of the condenser lens 21, the attitude of the light source 11 can be adjusted so that the light source 11 forms a centrally symmetric image in the aperture stop 23. In some examples, an adjusting device is provided on the side wall of the outer rotating cylinder 14 of the sleeve assembly 1, and this adjusting device can be used to adjust the position of the light source 11 in the up, down, left, and right directions in the plane perpendicular to the optical axis. In some examples, this adjusting device is a setscrew. Thus, by using this adjusting device, the attitude of the light source 11 can be adjusted so that the light source 11 forms a centrally symmetric image in the aperture stop. Further, by adjusting the attitude of the light source 11 and adjusting the position of the light source 11 in the up, down, left, and right directions in the plane perpendicular to the optical axis, and at the same time observing the observation window, so that the image of the light source 11 is complete and symmetric within the aperture stop 23, and the edge of the light source image is completely within the aperture range of the aperture stop 23. Thus, it is ensured that there is no loss of illumination light energy.

[0093] In some examples, the observation window in step S200 can be a translucent object. In some examples, the translucent object can be a translucent ground glass, in which case it is convenient for visual observation. At the same time, the image formed by the light source 11 in the aperture stop 23 is imaged on the translucent object, so it is convenient to visually observe the imaging situation.

[0094] In some examples, after step S200, step S300 can be executed.

[0095] In some examples, in step S300, the turntable 32 and the beam splitter 31 can be arranged in the beam splitting chamber 3, and their positional distribution can be a preset value for optical design.

[0096] In some examples, as described above, a number of small holes can 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 a high speed, one small hole scans a corresponding area on the sample to be measured, so as to achieve a complete scan of the sample to be measured.

[0097] In some examples, after step S300, step S400 can be executed.

[0098] In some examples, in step S400, the tube lens 4 can be adjusted so that the tube lens 4 focuses to form an image. Figure 7 A schematic diagram of the scenario for adjusting the relative position between the tube lens 4 and the beam splitting chamber 3 involved in the example of this embodiment is shown.

[0099] In some examples, referring to Figure 7 , the tube lens 4 and the beam splitting chamber 3 can be connected.

[0100] In some examples, after the tube lens 4 and the beam splitting chamber 3 are connected, a second parallel beam can be formed and the second parallel beam can be made to enter the beam splitting chamber 3 from the tube lens 4.

[0101] In some examples, the second parallel beam can be generated by the collimator 7, have a preset image, and the second parallel beam is made to enter the beam splitting chamber 3 from the tube lens 4, and the relative position between the tube lens 4 and the beam splitting chamber 3 is adjusted so that the parallel beam forms a preset image on the turntable 32.

[0102] In some examples, the preset image can be a collimator reticle image matching the collimator 7. Since the reticle image is imaged at infinity through the second parallel beam, the collimator 7 can simulate the reflected beam of the sample to be measured passing through the microscope objective 5 and entering the turntable 32, and thus the relative position between the tube lens 4 and the beam splitting chamber 3 can be adjusted more accurately.

[0103] In some examples, the tube lens 4 and the beam splitting chamber 3 are connected by a thread. In this case, the tube lens 4 and the beam splitting chamber 3 can be rotated along the thread, and the tube lens 4 can be moved along the optical axis direction. Thus, the relative position between the tube lens 4 and the beam splitting chamber 3 can be adjusted, and thereby the relative position between the tube lens 4 and the turntable 32 can be adjusted.

[0104] In some examples, after the endoscope 4 is connected to the spectroscopic chamber 3, the whole is placed under the collimator 7. The collimator 7 forms a second parallel light beam that enters the spectroscopic chamber 3 through the endoscope 4. The relative positions of the endoscope 4 and the spectroscopic chamber 3 are adjusted so that a clear image of the reticle of the collimator 7 appears on the turntable 32 for the second parallel light beam. In this case, by simulating the reflection of the illumination light by the surface of the sample to be measured, the reflected light enters the microscope objective 5, enters the endoscope 4 as a parallel light beam, and forms an image on the turntable 32. Thus, the relative positions of the endoscope 4 and the turntable 32 are determined to ensure the integrity of the optical path.

[0105] In some examples, after adjusting the relative positions of the endoscope 4 and the spectroscopic chamber 3, the positional relationship between the endoscope 4 and the spectroscopic chamber 3 is fixed. In this case, sectional alignment can be achieved, and during the alignment process, it is ensured that other alignment steps will not affect the relative positional relationship between the aligned modules.

[0106] In some examples, after step S400, step S500 can be executed.

[0107] In some examples, in step S500, the first parallel light beam can be made to enter the endoscope 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 condenser lens 21 away from the light source 11, at a position more than twice the focal length of the condenser lens 21, so that the first parallel light beam enters the spectroscopic chamber 3 through the sleeve assembly 1 and the lens barrel assembly 2 and enters the endoscope 4 through the spectroscopic chamber 3.

[0108] In some examples, after step S500, step S600 can be executed.

[0109] In some examples, in step S600, the imaging of the field stop 13 on the turntable 32 can be calibrated. In this case, by adjusting the relative positions of the field stop 13, the turntable 32, and the endoscope 4 section by section and region by region, and making the field stop 13 form an image of the field stop 13 on the small hole surface of the turntable 32, and the image of the field stop 13 covers the small hole surface of the turntable, the field stop 13 can be conjugated with the object focal plane of the microscope objective 5.

[0110] In some examples, an infinite conjugate image microscope objective 5 and a tube lens 4 can be selected to cooperate to achieve the purpose of microscopic imaging. In a spinning disk parallel scanning confocal microscope, the small holes on the spinning disk 32 are shared by the illumination optical path and the detection imaging optical path. The microscopic imaging image plane, which is the conjugate image plane of the object plane, is on the small hole plane of the spinning disk 32. Therefore, the image of the field stop 13 needs to be presented on the small hole plane of the spinning disk 32, and the spot of the image plane of the field stop 13 completely covers the small hole plane within the imaging range of the spinning disk 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 spinning disk 32 rotates along the central axis of the spinning disk 32. At the same time, a part of the area of the spinning disk 32 is located in the illumination optical path and the measurement optical path. On the spinning disk 32, the area located in the illumination optical path and the measurement optical path can be called the imaging range.

[0111] In some examples, the position of the adjustment sleeve assembly 1 can be adjusted by moving it along the optical axis direction. In this case, the light source 11, the collimating lens group 12, and the field stop 13 move along the optical axis as a whole module relative to the lens barrel assembly 2, so as to adjust the relative position between the field stop 13 and the condenser lens 21, so that a clear image of the field stop 13 is formed on the spinning disk 32, and the spot of the image plane of the field stop 13 completely covers the small hole plane within the imaging range of the spinning 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 can have an openable side. In this case, the spot of the image plane of the field stop 13 on the spinning disk 32 can be observed.

[0112] In some examples, the actual size of the spot of the image plane of the field stop 13 is the design value of the illumination optical path. The design value of the illumination optical path can be obtained by calculating with the Gaussian formula.

[0113] In some examples, after step S600, step S700 can be executed. In some examples, in step S700, the position of the microscope objective 5 can be determined and the microscope objective 5 can be installed. In this case, by placing an observation window on the side of the tube lens 4 away from the spectroscopic chamber 3 and observing the observation window until the image of the light source 11 that meets the preset requirements is received, the position of this observation window at this time is the image-side rear focal plane of the microscope objective 5. Through the design parameters of the microscope objective 5, the numerical relationship between the image-side rear focal plane of the microscope objective 5 and the parfocal plane can be determined, and then the installation position of the microscope objective 5 can be determined, so that the light source can be imaged on the image-side rear focal plane of the microscope objective 5.

[0114] Figure 8 is a schematic diagram showing the scenario of adjusting the position of the microscope objective involved in the example of this embodiment.

[0115] In some examples, before assembling the microscope objective 5 in the overall optical path, it is first necessary to find the clear image of the light source 11 formed after the light beam of the light source 11 passes through the sleeve assembly 1, the lens barrel assembly 2, the beam splitting chamber 3 and the tube lens 4, and mark the position of the clear image of the light source. The image-side rear focal plane of the microscope objective 5 is set at this position. In this case, the image of the light source passes through the microscope objective 5 and forms an image at infinity on the side of the sample to be measured. Since the sample to be measured is at a finite distance, the illumination on the surface of the sample to be measured will not be affected by the shadow of the light source filament. Therefore, the illumination light on the surface of the sample to be measured is uniformly illuminated and there is no shadow. In other words, referring to Figure 2 and Figure 8 As shown by the solid line in, the light source 11 presents a first real image at the aperture stop 23 through the collimating lens 12 and the condenser lens 21. The first image forms a second real image between the tube lens 4 and the microscope objective 5 after passing through the tube lens 4. Thus, the microscope objective 5 can be assembled based on the positions of the first real image and the second real image.

[0116] In some examples, referring to Figure 8 , an observation window can be placed on the side of the tube lens 4 away from the beam splitting chamber 3, and the observation window is moved along the optical axis direction 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 mark the position of the observation window at this time.

[0117] 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 is moved along the optical axis direction until a clear image of the light source 11 that meets the preset requirements appears on the observation window. Thus, the position of the clear image of the light source 11 can be determined, and further the installation position of the image-side rear focal plane of the microscope objective 5 can be determined. Thereby, according to the numerical relationship between the image-side rear focal plane and the parfocal plane of the microscope objective 5, the installation position of the microscope objective 5 can be determined.

[0118] In some examples, the preset requirements can be that the clarity and shape of the image of the light source 11 meet the requirements. In some examples, the preset requirements can be that the operator can clearly distinguish the shape of the image of the light source 11. In some examples, it can be determined whether the image of the light source 11 meets the preset requirements through an image processing method. For example, the preset requirements can be that the resolution (or gradient) of the image of the light source 11 meets the requirements.

[0119] In some examples, the image-side rear focal plane of the microscope objective 5 can be set near this position or at this position. This position is the position where the image-side rear focal plane of the microscope objective 5 is placed.

[0120] In some examples, as described above, the image-side rear focal plane of the microscope objective lens 5 and the parfocal plane of the microscope objective lens 5 are unified fixed values. In this case, the installation position of the microscope objective lens 5 can be determined based on the unified fixed values of the image-side rear focal plane of the microscope objective lens 5 and the parfocal plane of the microscope objective lens 5.

[0121] In some examples, as described above, the image-side rear focal plane of the microscope objective lens 5 and the parfocal plane can be unified fixed values. Thus, the installation position of the microscope objective lens 5 can be determined and the microscope objective lens 5 can be installed.

[0122] In some examples, the observation window in the above step S700 is a translucent object. In some examples, the translucent object is a translucent ground glass. In this case, it is convenient for visual observation. At the same time, the image of the light source 11 formed by the light beam emitted from the tube lens 4 is imaged on the translucent object. Thus, it is convenient to visually observe the corresponding imaging situation.

[0123] The above has described various embodiments of the present invention in the specific implementation manners. Although these descriptions directly describe the above embodiments, it should be understood that those skilled in the art can think of modifications and / or deformations of the specific embodiments shown and described herein. Any such modifications or deformations falling within the scope of this specification are also intended to be included therein. Unless otherwise specified, the intention of the inventor is that the words and phrases in the specification and claims be given the ordinary and customary meaning of those of ordinary skill in the art.

[0124] 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 purposes 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 forms disclosed, and many modifications and variations are possible in light of the above teachings. The described embodiments are used to explain the principles of the present invention and its practical applications, and enable other technicians in the art to utilize the present invention in various embodiments and various modifications suitable for the specific purposes contemplated. Therefore, it is intended that the present invention not be limited to the specific embodiments disclosed for carrying out the present invention.

[0125] Although specific embodiments of the present invention have been shown and described, it will be apparent to those skilled in the art that, based on the teachings of the present invention, variations and modifications can be made without departing from the present invention and its broader aspects. Therefore, the appended claims will cover all such changes and modifications within the true spirit and scope of the present invention. Those skilled in the art will understand that, generally speaking, the terms used in the present invention are generally intended to be "open" terms (e.g., the term "comprising" should be interpreted as "comprising but not limited to", the term "having" should be interpreted as "having at least", the term "including" should be interpreted as "including but not limited to", etc.).

Claims

1. A lighting optical path of a turntable parallel scanning confocal microscope, characterized in that, The illumination optical path includes a sleeve assembly having a light source and a field stop, a lens barrel assembly having a condenser lens, a beam splitter chamber having the turntable, a tube lens, and a microscope objective. The light source is configured to form a first parallel light beam that enters the lens barrel assembly via the field stop; the light source is imaged on the focal plane of the condenser lens; the turntable is configured to receive a second parallel light beam that is incident from the tube lens, the second parallel light beam is configured to form a preset image on the turntable, the turntable is configured to receive the first parallel light beam that is incident into the beam splitter chamber via the lens barrel assembly, and the first parallel light beam is configured to form a light spot covering the small hole surface of the turntable on the turntable; the microscope objective is configured to be mounted at the position of the image of the light source that meets the preset requirements formed by the light beam emitted from the tube lens.

2. The illumination optical path of the turntable parallel scanning confocal microscope according to claim 1, wherein The lens barrel assembly further includes an aperture stop. The aperture stop of the lens barrel assembly is arranged on the focal plane of the condenser lens, and the posture of the light source is adjusted so that the light source forms a centrosymmetric image in the aperture stop.

3. The illumination optical path of the turntable parallel scanning confocal microscope according to claim 2, characterized in that, Adjust the posture of the light source so that the light source forms a centrosymmetric image in the aperture stop.

4. The illumination optical path of the turntable parallel scanning confocal microscope according to claim 1, characterized in that, Adjust the relative position between the tube lens and the beam splitter chamber so that the second parallel light beam forms a preset image on the turntable.

5. The illumination optical path of the turntable parallel scanning confocal microscope according to claim 4, characterized in that Let the first parallel light beam enter the beam splitter chamber via the lens barrel assembly and form a light spot on the turntable, and adjust the relative position between the field stop and the condenser lens to form a light spot covering the small hole surface of the turntable.

6. The illumination optical path of the turntable parallel scanning confocal microscope according to claim 5, characterized in that, The sleeve assembly includes a collimating lens, and the relative position between the light source and the collimating lens is adjusted to form the first parallel light beam on the collimating lens.

7. The illumination optical path of the turntable parallel scanning confocal microscope according to claim 6, characterized in that, After adjusting the relative position between the light source and the collimating lens, fix the positional relationship between the light source and the collimating lens; after adjusting the relative position between the field stop and the condenser lens, fix the positional relationship between the field stop and the condenser lens; after adjusting the relative position between the tube lens and the beam splitter chamber, fix the positional relationship between the tube lens and the beam splitter chamber.

8. The illumination optical path of the turntable parallel scanning confocal microscope according to claim 1, characterized in that Receive the image of the light source formed by the light beam emitted from the tube lens at different positions to obtain the position of the image of the light source that meets the preset requirements.

9. The illumination optical path of the turntable parallel scanning confocal microscope according to claim 1 or 8, characterized in that Use the observation window to receive the image of the light source formed by the light beam emitted from the tube lens, and move the observation window along the optical axis direction until the image of the light source that meets the preset requirements appears on the observation window.

10. The illumination optical path of the turntable parallel scanning confocal microscope according to claim 1, wherein The field stop has a non-circular through hole.

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