A method for installing and debugging a structured light illumination system

Abstract

The application provides a mounting and debugging method of a structured light illumination system, comprising the following steps: mounting an illumination module; mounting a mirror assembly; fixing the illumination module and the mirror assembly in a structural frame of the structured light illumination system and adjusting an optical path; mounting the illumination module on an optical machine; mounting a first lens, a second lens and a third lens according to an optical path design; mounting a grating and a diaphragm according to the optical path design; and fine-tuning a structured light stripe. The mounting and debugging method follows the principle of from top to bottom and from simple to complex. Since the grating can divide a light beam into multiple beams, the grating is mounted last in order to simplify the mounting and debugging difficulty.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of super-resolution microscopy, and more particularly to a method for installing and debugging a structured light illumination system. BACKGROUND

[0002] In the past ten years, super-resolution microscopy technology has developed rapidly in the past few years. Currently, common super-resolution microscopy technologies include stimulated emission depletion microscopy (STED), structured illumination microscopy (SIM), photo-activated localization microscopy (PALM), and stochastic optical reconstruction microscopy (STORM).

[0003] Among the above technologies, SIM has the advantages of fast imaging speed, low photobleaching, and large imaging field of view, and is very suitable for application in the gene sequencing industry.

[0004] However, most of the products on the market are based on high-magnification small-field-of-view objectives, generally 60x or higher, with a maximum field of view of only 0.4mm. However, most sequencing instruments on the market have a field of view of more than 1mm, and there is no related technology method. That is, the existing structured light illumination technology is designed based on high-magnification objectives, with a very small field of view, which does not meet the high-throughput requirements of sequencing instruments. Moreover, there is currently no corresponding installation and debugging technology for structured light illumination systems. SUMMARY

[0005] Therefore, in order to solve the above problems, the present application provides a method for installing and debugging a structured light illumination system, and the technical solution is as follows:

[0006] A method for installing and debugging a structured light illumination system, the structured light illumination system comprising: a fiber interface, a collimating lens, a grating, a first lens, an aperture, a second lens, a mirror, and a third lens.

[0007] The installation and debugging method comprises:

[0008] installing an illumination module, the illumination module comprising the fiber interface and the collimating lens.

[0009] mounting a mirror assembly, the mirror assembly comprising a mirror base and the mirror;

[0010] fixing the illumination module and the mirror assembly within a structural frame of the structured light illumination system, wherein placing a collimation mirror at a first surface, adjusting the illumination module so that reflected light coincides with the fiber interface, placing a concentric target on the first surface, translating the illumination module so that the illumination spot coincides with the center of the concentric target, the first surface being a surface of the mirror assembly facing the illumination module;

[0011] mounting the illumination module on an optical bench;

[0012] mounting the first lens, the second lens and the third lens according to an optical path design;

[0013] mounting the grating and the diaphragm according to the optical path design;

[0014] fine-tuning the structured light fringe.

[0015] Preferably, in the above mounting and adjusting method, the mounting the illumination module comprises:

[0016] connecting a light source into the fiber interface, then aligning the illumination module to a target plane, adjusting the front and back positions of the collimation lens so that the edges of the light spot are sharp.

[0017] Preferably, in the above mounting and adjusting method, the mounting the mirror assembly comprises:

[0018] mounting the mirror on the mirror base, adjusting the mirror using a collimation telescope so that reflected light coincides with incident light.

[0019] Preferably, in the above mounting and adjusting method, the mounting the illumination module on an optical bench comprises:

[0020] attaching a concentric target at an illumination entrance hole of the optical bench, adjusting the height of the mirror so that the illumination spot coincides with the center of the concentric target, then placing the collimation mirror at a second surface, ensuring that the angle of the mirror is unchanged, the second surface being a plane on which the illumination entrance hole is located.

[0021] Preferably, in the above mounting and adjusting method, the mounting the first lens, the second lens and the third lens according to an optical path design comprises:

[0022] mounting the first lens and the second lens in the same lens barrel according to an optical path design, then fixing the lens barrel on the first surface.

[0023] Preferably, in the above installation and debugging method, the installing the first lens, the second lens and the third lens according to the optical path design further comprises:

[0024] After the first lens and the second lens are installed, the illumination module is separated from the optical machine;

[0025] After the illumination module is separated from the optical machine, the third lens is installed according to the optical path design, and then the illumination module is installed on the optical machine.

[0026] Preferably, in the above installation and debugging method, the optical machine comprises an automatic focusing module and a dichroic mirror, and the installing the third lens according to the optical path design comprises:

[0027] The engineering chip is placed on the object plane, the automatic focusing module is started and the camera is turned on, and the dichroic mirror is adjusted so that the illumination spot is centered in the camera field of view.

[0028] Preferably, in the above installation and debugging method, the installing the grating and the diaphragm according to the optical path design comprises:

[0029] The grating is installed on the structural frame of the structured light illumination system while the light source is turned on, and the diaphragm is placed between the first lens and the second lens while the position of the diaphragm is fine-tuned up and down so that the diaphragm is placed at the focal plane position of the first lens.

[0030] Preferably, in the above installation and debugging method, the fine-tuning the structured light stripe comprises:

[0031] The green light is turned on, the position of the third lens is adjusted so that the structured light meets the target requirement, and the dichroic mirror is adjusted so that the structured light spot is centered in the camera field of view.

[0032] The red light is turned on, and the structured light illumination system is rotated until the structured light stripe appears.

[0033] The grating and the dichroic mirror in the optical machine are controlled so that the structured light in the green light state and the structured light in the red light state are consistent.

[0034] Preferably, in the above installation and debugging method, before the illumination module is installed, the installation and debugging method further comprises:

[0035] The elements in the structured light illumination system are subjected to tolerance analysis.

[0036] Compared with the prior art, the present application has the following beneficial effects:

[0037] The application provides a mounting and debugging method of a structured light illumination system, which comprises the following steps: installing an illumination module; installing a mirror assembly; fixing the illumination module and the mirror assembly in a structured frame of the structured light illumination system, wherein the mirror assembly is placed at a first surface, the illumination module is adjusted so that reflected light coincides with a fiber interface, a concentric target is placed on the first surface, and the illumination module is translated so that an illumination spot coincides with a center of the concentric target, the first surface is a surface of the mirror assembly facing the illumination module; installing the illumination module on an optical engine; installing a first lens, a second lens and a third lens according to an optical path design; installing a grating and a diaphragm according to the optical path design; and fine-tuning structured light stripes. The mounting and debugging method mainly follows the principle of from top to bottom and from simple to complex. Since the grating divides a light beam into multiple beams, the grating is installed last to simplify the mounting and debugging difficulty. BRIEF DESCRIPTION OF DRAWINGS

[0038] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or the prior art description. Obviously, the drawings in the following description only constitute the embodiments of the present application, and for those skilled in the art, other drawings can also be obtained without creative labor on the basis of the provided drawings.

[0039] Figure 1 A simulation schematic diagram of the structured light illumination system is provided for the embodiments of the present application.

[0040] Figure 2 A structural schematic diagram of the structured light illumination system is provided for the embodiments of the present application.

[0041] Figure 3 A tool mounting and debugging schematic diagram is provided for the embodiments of the present application.

[0042] Figure 4 Another tool mounting and debugging schematic diagram is provided for the embodiments of the present application.

[0043] Figure 5 An illumination module installation schematic diagram is provided for the embodiments of the present application.

[0044] Figure 6 A mirror assembly installation schematic diagram is provided for the embodiments of the present application.

[0045] Figure 7 An installation schematic diagram of the illumination module and the mirror assembly is provided for the embodiments of the present application.

[0046] Figure 8 An installation schematic diagram of the illumination module and the optical engine is provided for the embodiments of the present application.

[0047] Figure 9 A first lens and a mounting schematic diagram of the second lens are provided for an embodiment of the present application.

[0048] Figure 10 A schematic diagram of a structured light stripe is provided for an embodiment of the present application. DETAILED DESCRIPTION

[0049] The technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the protection scope of the present application.

[0050] In order to make the above-mentioned purposes, features and advantages of the present application more obvious and easy to understand, the present application will be further described in detail below with reference to the drawings and specific embodiments.

[0051] Reference Figure 1 , Figure 1 A simulation schematic diagram of a structured light illumination system is provided for an embodiment of the present application, referring to Figure 2 , Figure 2 A structural schematic diagram of a structured light illumination system is provided for an embodiment of the present application, the structured light illumination system comprising: a fiber interface 1, a collimating lens (L1) in the embodiment, Figure 1 The collimating lens is not shown in the embodiment, and is labeled with L1, Figure 2 marked as 2), a grating 3, a first lens (L2) in the embodiment, Figure 1 marked as L2, Figure 2 marked as 4), an aperture 5, a second lens (L3) in the embodiment, Figure 1 marked as L3, Figure 2 marked as 6), a mirror 7 and a third lens (L4) in the embodiment, Figure 1 marked as L4, Figure 2 marked as 8).

[0052] It should be noted that the aperture 5 includes but is not limited to a pinhole aperture.

[0053] As shown in Figure 1 , the light collimated by the collimating lens L1 is incident on the grating 3, and after passing through the grating 3, a plurality of interference beams are generated and incident on the first lens L2, which is collimated and focused on the aperture 5 by the first lens L2, so that the target diffraction beam passes through to the second lens L3, which is received by the objective lens 9 through the third lens L4.

[0054] It should be noted that the structural light illumination system provided by the present application is a structural light illumination system based on a large field of view optical system, and realizes super-resolution imaging. First, the required structural light period and the corresponding illumination lens parameters are calculated according to the system parameters, and in combination with the corresponding software design optimization method in the structural light illumination design, in order to ensure that the two light beams interfere, we think that using the image height function to control the light is more simple and effective.

[0055] Further, since the structural light illumination system is relatively complex in structure, and the generation of the structural light has high requirements on the optical path, unlike other simple illumination systems, it is necessary to study the corresponding installation and debugging method to reduce the installation and debugging difficulty and provide corresponding technical support for the productization of subsequent super-resolution technology.

[0056] When the installation and debugging method of the structural light illumination system is executed, first, tolerance analysis is performed, that is, before installing the illumination module, tolerance analysis is performed on the elements in the structural light illumination system, as follows:

[0057] Before the optical system is designed, reasonable tolerance analysis needs to be performed to accurately understand the influence of each element in the optical system on the imaging effect of the system. According to the results of the tolerance analysis, appropriate mechanical design and installation methods can be selected. For example, the eccentricity requirement of a lens is 0.02mm, which is within the range that can be guaranteed by mechanical design, so mechanical processing accuracy can be used to meet the requirements; otherwise, adjustable devices need to be added during mechanical design, and professional installation tools such as an eccentricity instrument are used to complete the installation.

[0058] As shown in Figure 1 and Figure 2 In this structural light illumination system, there are a grating 3, four lenses L1, L2, L3, L4, and a diaphragm 5. We will analyze the tolerance of each element in turn, mainly in terms of the angle difference between the two light beams.

[0059] As shown in the following table, the tolerance analysis function is as follows:

[0060]

[0061] For the grating 3, the grating 3 is an element with a periodic structure, so translating the grating 3 has no effect on the structural light stripes. For the case of grating 3 inclination, the grating formula changes to:

[0062] d(sinθ-sini)=kλ

[0063] Where i is the grating inclination angle, θ is the grating diffraction angle, and k is the diffraction order.

[0064] When k = 0, sinθ = sini, that is, for zero-order diffraction, the exit angle is equal to the incident angle. For k = +-1, we use the angle function in the software to read the angle difference as 0.06°, which is a very small number, and it can be said that the angle of grating 3 is not sensitive to the interference fringes.

[0065] For the collimating lens L1, it can be known from the above description that the zero-order exit angle of grating 3 is the same as the incident angle, so it can be known that the illumination angle of collimating lens L1 will be more sensitive. The software simulation result shows that when collimating lens L1 is tilted by 0.1°, the beam angles of +-1 order are different by 0.84°.

[0066] For the first lens L2, the second lens L3 and the third lens L4, it can be noted from the optical path diagram that the parameters of the three lenses are the same, and the structures of the first lens L2, the second lens L3 and the third lens L4 in the optical system are also symmetrical, so they are discussed and analyzed together. The software simulation result shows that when the lenses are decentered by 0.1mm, the beam angles of +-1 order are different by 1.7°; and when the lenses are tilted by 0.1°, the angles are different by 0.33°.

[0067] By summarizing the above analysis results, it is found that the angle of collimating lens L1 is more sensitive, and the decentering of the first lens L2, the second lens L3 and the third lens L4 is more sensitive, which should be paid attention to in the subsequent mechanical design and adjustment process.

[0068] Based on this, the embodiment of the present application also provides a mounting and debugging method of the structured light illumination system, which is shown in Figure 3 , Figure 3 A mounting and debugging tool provided by the embodiment of the present application is shown in Figure 4 , Figure 4 Another mounting and debugging tool provided by the embodiment of the present application is shown in Figure 3 The mounting and debugging tool shown in is an internal focusing telescope, model NWJ-3, which is mainly used for adjusting the angle of an optical element, Figure 4 The mounting and debugging tool shown in is a concentric target, which can be printed on paper and used for adjusting the decentering displacement of an optical element. It should be noted that the mounting and debugging tool of a reflecting mirror will be used later.

[0069] The mounting and debugging method of the structured light illumination system comprises the following steps:

[0070] Step 1: install the illumination module, which comprises a fiber interface 1 and a collimating lens 2.

[0071] Reference Figure 5 , Figure 5The installation schematic diagram of the illumination module provided by the embodiment of the present application is as follows: the light source is connected to the optical fiber interface 1, then the illumination module is aligned to the target plane (for example, the ceiling), the front and back positions of the collimating lens 2 are adjusted so that the edge of the light spot is sharp.

[0072] The second step is to install the mirror assembly, which includes a mirror base and the mirror 7.

[0073] Reference Figure 6 , Figure 6 The installation schematic diagram of the mirror assembly provided by the embodiment of the present application is as follows: the mirror 7 is installed on the mirror base, and the mirror 7 is adjusted using the telescope adjustment so that the reflected light coincides with the incident light.

[0074] The third step is to refer to Figure 7 , Figure 7 The installation schematic diagram of the illumination module and the mirror assembly provided by the embodiment of the present application is as follows: the illumination module and the mirror assembly are fixed in the structural frame of the structured light illumination system, the telescope adjustment is placed at the first surface S1, the illumination module is adjusted so that the reflected light coincides with the optical fiber interface 1, the concentric target is placed on the first surface S1, and the illumination spot is translated so as to coincide with the center of the concentric target, and the first surface S1 is the surface of the mirror assembly facing the illumination module.

[0075] The fourth step is to refer to Figure 8 , Figure 8 The installation schematic diagram of the illumination module and the optical machine provided by the embodiment of the present application is as follows: the illumination module is installed on the optical machine, the concentric target is attached to the illumination entrance hole 21 of the optical machine, the height of the mirror 7 is adjusted so that the illumination spot coincides with the center of the concentric target, then the telescope adjustment is placed at the second surface S2, the angle of the mirror 7 is ensured unchanged, the second surface S2 is the plane where the illumination entrance hole 21 is located, and this step is mainly used to make the illumination optical axis coincide with the optical axis of the optical machine and parallel to the second surface S2.

[0076] Then, the first lens 4, the second lens 6 and the third lens 8 are installed according to the optical path design, which is specifically shown in the fifth step and the sixth step:

[0077] The fifth step is to refer to Figure 9 , Figure 9 The installation schematic diagram of the first lens and the second lens provided by the embodiment of the present application is as follows: the first lens 4 and the second lens 6 are installed in the same lens barrel according to the optical path design, and then the lens barrel is fixed on the first surface S1.

[0078] It should be noted that after the first lens 4 and the second lens 6 are installed, in order to facilitate the installation of the third lens 8, the illumination module can be separated from the optical machine at this time.

[0079] Step 6: After the illumination module is separated from the optical machine, the third lens 8 is installed according to the optical path design, and then the illumination module is installed on the optical machine.

[0080] Specifically, the optical machine includes an automatic focusing module and a dichroic mirror 22, the engineering chip is placed on the object plane, the automatic focusing module is started and the camera is turned on, and the dichroic mirror 22 is adjusted so that the illumination spot is centered in the camera field of view.

[0081] Step 7: Install the grating 3 and the diaphragm 5 according to the optical path design.

[0082] The grating 3 is installed on the structural frame of the structured light illumination system, and the light source is turned on at the same time; the diaphragm 5 is placed between the first lens 4 and the second lens 6, and the position of the diaphragm 5 is adjusted up and down at the same time, so that the diaphragm 5 is placed at the focal plane position of the first lens 4.

[0083] Step 8: Fine-tune the structured light stripe.

[0084] In this step, the green light is turned on, the position of the third lens 8 is adjusted so that the structured light meets the target requirements, the dichroic mirror is adjusted so that the structured light spot is centered in the camera field of view; then the red light is turned on, the structured light illumination system is rotated until the structured light stripe appears; the above steps are repeated until the stripes of the two lights are in good condition. Then control the grating and the dichroic mirror in the optical machine so that the structured light in the green light state is consistent with the structured light in the red light state.

[0085] Specifically, because the incidence angle of red light is relatively large, it is easy to appear vignetting, which causes a beam of light to be blocked, so when adjusting the structured light, green light is generally used first. Adjust the front and back positions of the third lens 8, and you can see that the structured light becomes clearer, and adjust the dichroic mirror 22 so that the structured light spot is centered in the camera field of view. Then turn on the red light, if there is no structured light stripe, slightly rotate the entire illumination system until the structured light stripe appears. At the same time, repeat the above steps until the stripes of the two lights are in good condition.

[0086] Rotate the grating by 90°, if the structured light stripe becomes weak, adjust the dichroic mirror 22 to make it clear. At the same time, repeat the above steps until the structured light at two angles reaches the same effect. Figure 10 , Figure 10 A schematic diagram of a structured light stripe provided by an embodiment of the present application is shown in FIG. 1. Figure 10 The bright spot in the figure is an automatic focusing spot, which can be ignored.

[0087] The installation and debugging method mainly follows the principle of from top to bottom and from simple to complex. Since the grating divides the light beam into multiple beams, in order to simplify the installation and debugging difficulty, the grating is installed last.

[0088] The above describes in detail the installation and debugging method of the structured light illumination system provided by the present application. The principles and implementation manners of the present application are described by using specific examples. The above description of the embodiments is only used to help understand the method of the present application and its core idea. Meanwhile, for those skilled in the art, according to the idea of the present application, the specific implementation manners and application ranges can be changed. Therefore, the content of the present description should not be understood as a limitation of the present application.

[0089] It should be noted that each embodiment in the present description is described in a progressive manner, and each embodiment mainly describes the difference from other embodiments. The same and similar parts of each embodiment can be understood by referring to each other. For the device disclosed by the embodiments, since it corresponds to the method disclosed by the embodiments, the description is relatively simple, and the related parts can be understood by referring to the method part.

[0090] It should be further noted that, in the present text, the relationship terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between the entities or operations. Moreover, the terms "include", "contain" or any other variants thereof are intended to cover non-exclusive inclusion, so that the process, method, article or device inherently includes a series of elements, or further includes the elements inherent to the process, method, article or device. Without more limitations, the element defined by the statement "including a" does not exclude the presence of other identical elements in the process, method, article or device including the element.

[0091] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present application. Various modifications to the embodiments will be apparent to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present application. Therefore, the present application will not be limited to the embodiments shown herein, but will conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method of installation and commissioning of a structured light illumination system, characterized in that, The structured light illumination system comprises a fiber interface, a collimating lens, a grating, a first lens, a diaphragm, a second lens, a mirror and a third lens; The installation and adjustment method comprises: installing an illumination module, the illumination module comprising the fiber interface and the collimating lens; installing a mirror assembly, the mirror assembly comprising a mirror base and the mirror; fixing the illumination module and the mirror assembly in a structural frame of the structured light illumination system, wherein the installation and adjustment of the mirror is placed at a first surface, the illumination module is adjusted so that reflected light coincides with the fiber interface, a concentric target is placed on the first surface, the illumination module is translated so that an illumination spot coincides with the center of the concentric target, and the first surface is a surface of the mirror assembly facing the illumination module; installing the illumination module on an optical machine; installing the first lens, the second lens and the third lens according to an optical path design; installing the grating and the diaphragm according to the optical path design; fine-tuning structured light stripes.

2. The method of installing and commissioning of claim 1, wherein, The installation of the illumination module comprises: connecting a light source into the fiber interface, and then aligning the illumination module to a target plane, and adjusting the front and back positions of the collimating lens so that the edges of the light spot are sharp.

3. The method of installing and commissioning of claim 1, wherein, The installation of the mirror assembly comprises: installing the mirror on the mirror base, and adjusting the mirror using an installation and adjustment telescope so that reflected light coincides with incident light.

4. The method of installing and commissioning of claim 1, wherein, The installation of the illumination module on the optical machine comprises: attaching a concentric target to an illumination entrance hole of the optical machine, adjusting the height of the mirror so that an illumination spot coincides with the center of the concentric target, and then placing the installation and adjustment of the mirror at a second surface, ensuring that the angle of the mirror is unchanged, and the second surface is a plane where the illumination entrance hole is located.

5. The method of installing and commissioning of claim 1, wherein, The installation of the first lens, the second lens and the third lens according to the optical path design comprises: installing the first lens and the second lens in the same lens barrel according to the optical path design, and then fixing the lens barrel on the first surface.

6. The method of installing and commissioning of claim 5, wherein, The installation of the first lens, the second lens and the third lens according to the optical path design further comprises: after the installation of the first lens and the second lens is completed, separating the illumination module from the optical machine; after the separation of the illumination module from the optical machine, installing the third lens according to the optical path design, and then installing the illumination module on the optical machine.

7. The method of installing and commissioning of claim 6, wherein, The optical machine comprises an automatic focusing module and a dichroic mirror, and the installation of the third lens according to the optical path design comprises: placing an engineering chip on an object plane, starting the automatic focusing module and turning on a camera, and adjusting the dichroic mirror so that an illumination spot is centered in a field of view of the camera.

8. The method of installing and commissioning of claim 1, wherein, The installation of the grating and the diaphragm according to the optical path design comprises: installing the grating on a structural frame of the structured light illumination system while turning on a light source, and placing the diaphragm between the first lens and the second lens while fine-tuning the position of the diaphragm up and down so that the diaphragm is placed at a focal plane position of the first lens.

9. The method of installing and commissioning of claim 7, wherein, The fine-tuning of the structured light stripes comprises: Turning on green light, adjusting the position of the third lens to make the structured light meet the target requirements, and adjusting the dichroic mirror to make the structured light spot centered in the camera field of view; Turning on red light, rotating the structured light illumination system until the structured light stripes appear; Controlling the grating and the dichroic mirror in the light machine to make the structured light in the green light state consistent with the structured light in the red light state.

10. The method of installing and commissioning of claim 1, wherein, Before installing the lighting module, the installation and debugging method further comprises: Performing tolerance analysis on the elements in the structured light illumination system.

Images