Raman confocal microscopic imaging system

Abstract

The utility model relates to a Raman confocal microscopic imaging system, and the system comprises a Raman excitation mechanism which comprises a laser assembly and a first light splitting assembly which are arranged in a first direction; the wide-field illumination mechanism comprises a wide-field light source assembly and a second light splitting assembly which are arranged in the first direction; the wide-field detection mechanism comprises a first imaging assembly and a third light splitting assembly which are arranged along a first direction; the Raman detection mechanism comprises a second imaging assembly and a fourth light splitting assembly which are arranged along a second direction; the objective lens, the second light splitting assembly, the first light splitting assembly, the third light splitting assembly and the fourth light splitting assembly are sequentially arranged in the second direction. The Raman confocal microscopic imaging system is easy to disassemble, easy to integrate and stackable, and provides multi-dimensional information including sample components, features and surface morphology.

Description

Technical Field

[0001] This application relates to the field of imaging technology, and in particular, to a Raman confocal microscopy system. Background Art

[0002] All components of a common Raman confocal microscope are concentrated in an optical microscope. The system is compact, but the drawback is that the optical path height is determined and it is difficult to make large adjustments. Once the detection parameters of a Raman microscopy system, such as laser wavelength, spectrometer resolution, sample observation area, etc., are set, it is difficult to make large changes to the Raman microscopy system and it is difficult to add new Raman imaging functions, such as adding new wavelength excitations, adding spectrometers with higher resolution or wider spectral acquisition ranges. In addition, a common Raman confocal microscope is generally equipped with a wide-field camera for sample positioning and tracking, which can only reflect the general shape of the sample and cannot perform fine three-dimensional surface topography imaging. Summary of the Utility Model

[0003] In view of the above analysis, this application aims to provide a Raman confocal microscopy system that can be adapted to different Raman excitation light sources and Raman spectrometers, has easy disassembly, easy integration, and stackability, and provides multi-dimensional information including sample composition, characteristics, and surface morphology.

[0004] The objectives of this application are mainly achieved through the following technical solutions:

[0005] An embodiment of this application provides a Raman confocal microscopy system, including: a Raman excitation mechanism, including a laser component and a first beam splitting component arranged along a first direction; a wide-field illumination mechanism, including a wide-field light source component and a second beam splitting component arranged along the first direction; a wide-field detection mechanism, including a first imaging component and a third beam splitting component arranged along the first direction; a Raman detection mechanism, including a second imaging component and a fourth beam splitting component arranged along a second direction; an objective lens, and the objective lens, the second beam splitting component, the first beam splitting component, the third beam splitting component, and the fourth beam splitting component are arranged in sequence along the second direction; the laser component emits a first light ray towards the first beam splitting component, the first beam splitting component reflects the first light ray towards the objective lens, the first light ray forms a second light ray after being reflected by the detection object, and the fourth beam splitting component reflects the second light ray towards the second imaging component; the wide-field light source component emits a third light ray towards the second beam splitting component, the third beam splitting component reflects the third light ray towards the objective lens, the third light ray forms a fourth light ray after being reflected by the detection object, and the third beam splitting component reflects the fourth light ray towards the first imaging component.

[0006] According to an embodiment of the present application, the laser assembly includes a laser, an optical fiber collimator, a first filter, and a first attenuation sheet arranged in sequence along a first direction; the laser emits laser light towards the optical fiber collimator along the first direction, the wavelength of the laser light is 532nm, 638nm, 785nm or 1064nm, the monochromaticity of the laser light is ±0.01nm; the focal length of the optical fiber collimator is 18mm; the optical density of the first filter is greater than 4; the optical density of the first attenuation sheet is equal to 2.

[0007] According to an embodiment of the present application, the first beam splitting assembly includes a long-pass coated dichroic mirror or a first non-polarizing beam splitter prism, which can reflect the first light ray and the second light ray, and can transmit the third light ray and the fourth light ray.

[0008] According to an embodiment of the present application, the wide-field light source assembly includes a wide-field white light source, a first lens, and a second lens arranged in sequence along a first direction; the power of the wide-field white light source is 20W, and the clear aperture diameter is 6mm; the first lens and the second lens form a Köhler illumination system.

[0009] According to an embodiment of the present application, the second beam splitting assembly includes a second non-polarizing beam splitter prism, which can reflect the third light ray and can transmit the fourth light ray.

[0010] According to an embodiment of the present application, the first imaging assembly includes a second attenuation sheet, a tube lens, and a color camera arranged in sequence along a first direction; the optical density of the second attenuation sheet is 2; the tube lens is a positive lens with a focal length of 200mm; the color camera is a complementary metal oxide semiconductor sensor camera, and the distance between the color camera and the tube lens is 200mm.

[0011] According to an embodiment of the present application, the third beam splitting assembly includes a third non-polarizing beam splitter prism, which can reflect part of the fourth light ray and can transmit the third light ray and part of the fourth light ray; the transmittance of the third non-polarizing beam splitter prism for the fourth light ray is 10:90.

[0012] According to an embodiment of the present application, the second imaging assembly includes a third attenuation sheet, a second filter, a third lens, a pinhole, a fourth lens, an optical fiber coupler, and a Raman spectrometer arranged in sequence along a first direction; the optical density of the third attenuation sheet is 2; the second filter is a long-pass filter with an optical density greater than 6; the third lens is a doublet achromatic convex lens with a focal length of 50mm; the pinhole is a stepless variable pinhole with a diameter ranging from 0 - 500μm and a distance of 50mm from the third lens; the fourth lens is a doublet achromatic convex lens with a focal length of 50mm, and the third lens and the fourth lens form a four-focal length system; the optical fiber coupler has a focal length of 18.4mm, a numerical aperture of 0.15, and a clear aperture diameter of 5mm.

[0013] According to an embodiment of the present application, the fourth light splitting component includes a reflector, and the light transmission size of the reflector is 25 mm.

[0014] According to an embodiment of the present application, it also includes an electric stage, which is arranged on a side of the objective lens away from the second beam splitting component along the second direction.

[0015] The Raman confocal microscopy system provided in the embodiments of the present application has at least the following advantages:

[0016] 1. In the Raman confocal microscopy system of the embodiment of the present application, the Raman excitation mechanism, the wide-field illumination mechanism, the wide-field detection mechanism and the Raman detection mechanism are independent of each other, and the relative positions and parameters can be adjusted according to needs, thereby broadening the scope of use of the Raman confocal microscopy system of the embodiment of the present application.

[0017] 2. In the Raman confocal microscopy system of the embodiment of the present application, the working states of the Raman excitation mechanism, the wide-field illumination mechanism, the wide-field detection mechanism and the Raman detection mechanism can be adjusted, so that the Raman confocal microscopy system of the embodiment of the present application can switch between the three modes of wide-field imaging, reflective confocal imaging and Raman confocal imaging, which can further broaden the scope of use of the Raman confocal microscopy system of the embodiment of the present application and provide multi-dimensional information of the sample.

[0018] In the present application, the above-mentioned technical solutions can also be combined with each other to achieve more preferred combination solutions. Other features and advantages of the present application will be described in the subsequent description, and some advantages can become obvious from the description, or can be understood by practicing the present application. The purpose and other advantages of the present application can be realized and obtained through the contents particularly pointed out in the description and the drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The accompanying drawings are only for the purpose of illustrating specific embodiments and are not to be considered as limiting the present application. The same reference symbols denote the same components throughout the accompanying drawings.

[0020] Figure 1 A schematic diagram of an optical path of a Raman confocal microscopy imaging system according to an embodiment of the present application.

[0021] Figure 2 Another optical path schematic diagram of the Raman confocal microscopy imaging system according to an embodiment of the present application.

[0022] Figure 3 A schematic diagram of a Raman confocal microscopy system according to an embodiment of the present application.

[0023] Figure 4 Another schematic diagram of a Raman confocal microscopic imaging system according to an embodiment of the present application.

[0024] Figure 5 Another optical path schematic diagram of the Raman confocal microscopy imaging system according to the embodiment of the present application.

[0025] Figure 6 Another optical path schematic diagram of the Raman confocal microscopy imaging system according to the embodiment of the present application.

[0026] Figure 7 Another optical path schematic diagram of the Raman confocal microscopy imaging system according to the embodiment of the present application.

[0027] Sequence numbers of reference numerals:

[0028] 1. Raman excitation mechanism; 11. Laser assembly; 111. Laser; 112. Fiber collimator; 113. First filter; 114. First attenuation sheet; 12. First beam splitting assembly; 121. Long-pass coated dichroic mirror; 122. First non-polarizing beam splitting prism;

[0029] 2. Wide-field illumination mechanism; 21. Wide-field light source assembly; 211. Wide-field white light source; 212. First lens; 213. Second lens; 22. Second beam splitting assembly; 221. Second non-polarizing beam splitting prism;

[0030] 3. Wide-field detection mechanism; 31. First imaging assembly; 311. Second attenuation sheet; 312. Tube lens; 313. Color camera; 32. Third beam splitting assembly; 321. Third non-polarizing beam splitting prism;

[0031] 4. Raman detection mechanism; 41. Second imaging assembly; 411. Third attenuation sheet; 412. Second filter; 413. Third lens; 414. Pinhole; 415. Fourth lens; 416. Fiber coupler; 417. Raman spectrometer; 42. Fourth beam splitting assembly; 421. Mirror;

[0032] 5. Objective lens;

[0033] 6. Electronic stage;

[0034] 71. Upright microscope stand; 72. Inverted microscope;

[0035] X. First direction; Y. Second direction. Detailed implementation manners

[0036] The features and exemplary embodiments of various aspects of the present application will be described in detail below. In the following detailed description, many specific details are set forth in order to provide a comprehensive understanding of the present application. However, it will be apparent to those skilled in the art that the present application may be practiced without some of these specific details. The following description of the embodiments is only intended to provide a better understanding of the present application by showing examples of the present application.

[0037] It should be noted that, without conflict, the embodiments in the present application and the features in the embodiments may be combined with each other. The embodiments will be described in detail below with reference to the accompanying drawings.

[0038] Relative terms such as first and second are used solely to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the term "comprising", "including" or any other variant thereof is intended to cover non-exclusive inclusion, such that a process, method, article or device comprising a series of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article or device. Without further limitation, an element defined by the statement "comprising..." does not exclude the presence of additional identical elements in the process, method, article or device comprising the said element.

[0039] It should be understood that when describing the structure of a component, when a layer or a region is referred to as being "above" or "over" another layer or another region, it may mean directly above the other layer or another region, or there may be other layers or regions between it and the other layer or another region. And if the component is turned over, this layer or this region will be "below" or "under" the other layer or another region.

[0040] In addition, the term "and / or" in this document is merely a description of the association relationship of associated objects, indicating that three relationships may exist. For example, A and / or B may represent: A exists alone, A and B exist simultaneously, and B exists alone. In addition, the character " / " in this document generally represents an "or" relationship between the associated objects before and after.

[0041] It should be understood that in the embodiments of the present application, "B corresponding to A" means that B is associated with A, and B can be determined according to A. However, it should also be understood that determining B according to A does not mean determining B solely according to A, and B can also be determined according to A and / or other information.

[0042] The applicant has found that all components of a Raman confocal microscope are concentrated in an optical microscope, making the system compact. However, the drawback is that it is difficult to significantly adjust the positions of the various components, and the optical path height is determined. After determining the detection parameters of the Raman detection system, such as the laser wavelength, spectrometer resolution, sample observation area, etc., it is difficult to make major changes to the optical path of the Raman confocal microscope, and it is also difficult to add other imaging functions, such as adding new wavelength excitation, adding a spectrometer with higher resolution or a wider spectral acquisition range. Considering that a Raman confocal microscope is generally equipped with a wide-field camera for sample positioning and tracking, it can only reflect the general shape of the sample and cannot perform fine three-dimensional surface topography imaging.

[0043] In view of the above analysis, the applicant has proposed a Raman confocal microscopy imaging system, including a Raman excitation mechanism, a wide-field illumination mechanism, a wide-field detection mechanism, and a Raman detection mechanism. The Raman excitation mechanism, the wide-field illumination mechanism, the wide-field detection mechanism, and the Raman detection mechanism are independent of each other, and their relative positions and parameters can be adjusted according to requirements. They can be adapted to different Raman excitation light sources and Raman spectrometers, and have the characteristics of easy disassembly, easy integration, and stackability. The working states of the Raman excitation mechanism, the wide-field illumination mechanism, the wide-field detection mechanism, and the Raman detection mechanism can also be adjusted, so that the Raman confocal microscopy imaging system of the embodiments of the present application can switch among three modes: wide-field imaging, reflection confocal imaging, and Raman confocal imaging, thereby providing multi-dimensional information including sample composition, characteristics, and surface morphology, helping to understand characteristics such as the surface roughness and particle size of the sample, and further providing chemical and physical information of the sample, which is applicable to multiple fields, such as materials science, biology, medicine, and environmental science, etc.

[0044] Figure 1 It is a schematic diagram of an optical path of the Raman confocal microscopy imaging system of the embodiments of the present application.

[0045] Figure 2 It is another schematic diagram of an optical path of the Raman confocal microscopy imaging system of the embodiments of the present application.

[0046] Figure 3 It is a schematic diagram of a device of the Raman confocal microscopy imaging system of the embodiments of the present application. Figure 4 It is another schematic diagram of a device of the Raman confocal microscopy imaging system of the embodiments of the present application.

[0047] Please refer to Figures 1 to 4, the embodiment of the present application provides a Raman confocal microscopy imaging system, including: a Raman excitation mechanism 1, including a laser component 11 and a first beam splitting component 12 arranged along the first direction X; a wide-field illumination mechanism 2, including a wide-field light source component 21 and a second beam splitting component 22 arranged along the first direction X; a wide-field detection mechanism 3, including a first imaging component 31 and a third beam splitting component 32 arranged along the first direction X; a Raman detection mechanism 4, including a second imaging component 41 and a fourth beam splitting component 42 arranged along the second direction Y; an objective lens 5, and the objective lens 5, the second beam splitting component 22, the first beam splitting component 12, the third beam splitting component 32, and the fourth beam splitting component 42 are arranged in sequence along the second direction Y; the laser component 11 of the Raman excitation mechanism 1 emits a first light ray towards the first beam splitting component 12, the first beam splitting component 12 reflects the first light ray towards the objective lens 5, the first light ray forms a second light ray after being reflected by the detection object, and the fourth beam splitting component 42 reflects the second light ray towards the second imaging component 41; the wide-field light source component 21 of the wide-field illumination mechanism 2 emits a third light ray towards the second beam splitting component 22, the third beam splitting component 32 reflects the third light ray towards the objective lens 5, the third light ray forms a fourth light ray after being reflected by the detection object, and the third beam splitting component 32 reflects the fourth light ray towards the first imaging component 31.

[0048] In the embodiment of the present application, the first direction X and the second direction Y are two intersecting directions. For the convenience of arranging each component, the second direction Y can be perpendicular to the first direction X. Exemplarily, when the Raman confocal microscopy imaging system of the embodiment of the present application is located in the horizontal plane, the second direction Y can be the vertical direction, and the first direction X can be the horizontal direction.

[0049] In the embodiment of the present application, the laser component 11 of the Raman excitation mechanism 1 can emit a first light ray along the first direction X, and the first light ray is the laser used in the Raman confocal imaging process. The first light ray is incident on the first beam splitting component 12 and is reflected towards the objective lens 5 along the second direction Y. After the first light ray irradiates the sample, it is reflected to form a second light ray. After the second light ray passes through the objective lens 5 along the second direction Y, it also passes through the first beam splitting component 12 and is incident on the fourth beam splitting component 42 of the Raman detection mechanism 4, and is reflected by the fourth beam splitting component 42 and is incident on the second imaging component 41 along the first direction X for Raman confocal imaging.

[0050] In the embodiment of the present application, the wide-field light source component 21 of the wide-field illumination mechanism 2 can emit a third light ray along the first direction X, and the third light ray is the white light source used in the wide-field imaging process. The third light ray is incident on the second beam splitting component 22 and is reflected towards the objective lens 5 along the second direction Y. After the third light ray irradiates the sample, it is reflected to form a third light ray. After the third light ray passes through the objective lens 5 along the second direction Y, it also passes through the second beam splitting component 22 and is incident on the third beam splitting component 32 of the wide-field detection mechanism 3, and is reflected by the third beam splitting component 32 and is incident on the first imaging component 31 along the first direction X for wide-field imaging.

[0051] The objective lens 5, the second beam splitting component 22, the first beam splitting component 12, the third beam splitting component 32 and the fourth beam splitting component 42 are arranged in sequence along the second direction Y, so that the light before and after the light is reflected at the sample can be transmitted along the second direction Y, and the light path from the second beam splitting component 22 to the sample is consistent. The spacing between any two adjacent ones can be determined according to the parameters of each mechanism. In order to facilitate the adjustment of the spacing, the objective lens 5, the wide-field illumination mechanism 2, the Raman excitation mechanism 1, the wide-field detection mechanism 3 and the Raman detection mechanism 4 can be arranged on a guide rail arranged along the second direction Y.

[0052] The Raman confocal microscopy imaging system of the embodiment of the present application has two working modes. When only the wide-field illumination mechanism 2 and the wide-field detection mechanism 3 are working, the Raman confocal microscopy imaging system of the embodiment of the present application is in the wide-field imaging mode, and the sample can be located and tracked; when the Raman excitation mechanism 1 and the Raman detection mechanism 4 are working, the Raman confocal microscopy imaging system of the embodiment of the present application is in the confocal imaging mode, and the physical and chemical properties of the sample can be obtained. Therefore, the Raman confocal microscopy imaging system of the embodiment of the present application can provide multi-dimensional information of the sample.

[0053] The Raman confocal microscopy system of the embodiment of the present application also includes an electric stage. The electric stage is arranged on the side of the objective lens 5 away from the second light splitting component 22 along the second direction Y. When the Raman confocal microscopy system of the embodiment of the present application is placed on a horizontal plane, the electric stage can move in two orthogonal directions in the horizontal plane, with an effective stroke of 300mm*300mm and a resolution of 100nm. The electric stage can also be raised and lowered, that is, moved along the second direction Y, with a stroke of 15mm.

[0054] The objective lens 5, the wide-field illumination mechanism 2, the Raman excitation mechanism 1, the wide-field detection mechanism 3 and the Raman detection mechanism 4 can be arranged on an upright microscope stand or on an inverted microscope. The upright microscope architecture is based on a gantry design and can be compatible with electric stages and sample operation tables of different sizes and models, such as material stretching tables, high and low temperature hot and cold tables, in-situ tables, etc. The various mechanisms of the Raman confocal microscopy imaging system of the embodiment of the present application can also be compatible with the commercial inverted microscope 72 structure, which is realized through an adapter module.

[0055] Figure 5 Another optical path schematic diagram of the Raman confocal microscopy imaging system according to an embodiment of the present application. Figure 6 Another optical path schematic diagram of the Raman confocal microscopy imaging system according to an embodiment of the present application. Figure 7 Another optical path schematic diagram of the Raman confocal microscopy imaging system according to an embodiment of the present application.

[0056] Further, please refer to Figures 5 to 7 , the laser assembly 11 includes a laser 111, an optical fiber collimator 112, a first filter 113, and a first attenuation sheet 114 arranged in sequence along the first direction X; the laser 111 emits laser light towards the optical fiber collimator 112 along the first direction X, the wavelength of the laser light is 532 nm, and the monochromaticity of the laser light is ±0.01 nm; the focal length of the optical fiber collimator 112 is 18 mm; the optical density of the first filter 113 is greater than 4; the optical density of the first attenuation sheet 114 is equal to 2.

[0057] The laser 111 can adopt a semiconductor single-mode fiber-coupled laser 111, and the wavelength of the emitted monochromatic laser light can be 532 nm, 638 nm, 785 nm, or 1064 nm. In this embodiment of the present application, the case where the laser 111 emits 532 nm laser light is taken as an example for illustration. The monochromaticity of the laser light is ±0.01 nm, the power stability is less than or equal to 1%, the output power is 0 - 200 mW, and the adjustment accuracy is 0.1%. Therefore, the laser 111 can emit stable and uniform laser light. The optical fiber collimator 112 can make the monochromatic laser light form parallel light, with a focal length of 18 mm, and the spot size after the optical fiber is input and collimated is 10 mm. The first filter 113 is used to filter out stray light, and a 532 nm narrow-band filter can be adopted, and the optical density (optical density, OD, dimensionless number) is greater than 4. The first attenuation sheet 114 is used to attenuate the laser light intensity, and the optical density of the first attenuation sheet 114 is equal to 2. After collimation, green light filtering, and attenuation, the laser assembly 11 can emit uniform and moderately intense laser light rays, that is, the first light rays.

[0058] Further, continue to refer to Figures 5 to 7 , the first beam splitting component 12 includes a long-pass coated dichroic mirror 121 or a first non-polarizing beam splitting prism 122, which can reflect the first light rays and the second light rays, and can transmit the third light rays and the fourth light rays.

[0059] It should be noted that the confocal imaging mode of the Raman confocal microscopy imaging system in this embodiment of the present application further includes two sub-modes: a reflection confocal imaging mode and a Raman confocal imaging mode.

[0060] The first beam splitting component 12 can use a long-pass coated dichroic mirror 121 or a first non-polarizing beam splitting prism 122, which can be replaced according to different requirements. The long-pass coated dichroic mirror 121 is used for Raman confocal imaging, reflecting the first light rays and transmitting the second light rays, and the optical density is greater than 6. The non-polarizing beam splitting prism is used for reflection confocal imaging, which can reflect part of the first light rays and transmit part of the second light rays.

[0061] Further, continue to refer to Figures 5 to 7, the second imaging component 41 includes a third attenuation sheet 411, a second filter 412, a third lens 413, a pinhole 414, a fourth lens 415, an optical fiber coupling mirror 416, and a Raman spectrometer 417 arranged in sequence along the first direction X; the optical density of the third attenuation sheet 411 is 2; the second filter 412 is a long-pass filter with an optical density greater than 6; the third lens 413 is a doublet achromatic convex lens with a focal length of 50 mm; the pinhole 414 is a continuously variable pinhole 414 with a diameter varying range of 0 - 500 μm and a distance of 50 mm from the third lens 413; the fourth lens 415 is a doublet achromatic convex lens with a focal length of 50 mm, and the third lens 413 and the fourth lens 415 form a four-focal length system; the optical fiber coupling mirror 416 has a focal length of 18.4 mm, a numerical aperture of 0.15, and a clear aperture diameter of 5 mm.

[0062] The third attenuation sheet 411 is used to attenuate the fourth light ray with an optical density of 2. The second filter 412 is used to filter out stray light and can be a 537 nm long-pass filter with an optical density greater than 6. The third lens 413 and the fourth lens 415 form a four-focal length system, which can improve the imaging resolution, reduce chromatic aberration and crosstalk. The pinhole 414 is used for confocal spatial filtering and can adopt a continuously variable pinhole 414 with a diameter varying range of 0 - 500 μm. The optical fiber coupling mirror 416 is used to collect Raman signals, with a focal length of 18.4 mm, a numerical aperture of 0.15, a clear aperture diameter of 5 mm, and the optical fiber interface type can be SMA905. The Raman spectrometer 417 has 1024 x 58 pixels, can adopt a back-illuminated thin-type FFT-CCD, is configured with a 1800 / 850 mm grating, and the spectral detection range is 100 - 2000 cm -1 , and the spectral resolution is 4 cm -1 , with a clear aperture diameter of 5 mm, for the second light ray.

[0063] Further, continue to refer to Figures 5 to 7 , the fourth beam splitting component 42 includes a reflecting mirror 421, and the clear aperture size of the reflecting mirror 421 is 25 mm.

[0064] The fourth beam splitting component 42 is in the form of a reflecting mirror 421, which can be a silver-plated reflecting mirror 421, and the clear aperture size of the reflecting mirror 421 is 25 mm.

[0065] When the Raman confocal microscopy imaging system of the embodiment of the present application is in the reflection confocal imaging mode, the first beam splitting component 12 uses a non-polarizing beam splitting prism, moves the third attenuation sheet 411 into the optical path, moves the second filter 412 out of the optical path, and records the value of each pixel point by using the single-point intensity integration mode of the spectrometer, so as to obtain the surface topography map of the sample. When the Raman confocal microscopy imaging system of the embodiment of the present application is in the Raman confocal imaging mode, the first beam splitting component 12 uses a long-pass coated dichroic mirror 121, moves the third attenuation sheet 411 out of the optical path, moves the second filter 412 into the optical path, uses the spectral detection mode of the spectrometer to collect the Raman spectral data of the sample, and selects the Raman spectrum acquisition of the characteristic peak implementation area. Therefore, the Raman confocal microscopy imaging system of the embodiment of the present application can realize dual-mode confocal imaging, can not only observe three-dimensional surface structure information, but also observe Raman component information. This provides more comprehensive information for the observation and analysis of material samples.

[0066] The selection of the diameter of the pinhole 414 can be controlled by a stepless motor, and the optimal pinhole 414 can be matched according to the selection of the objective lens 5 and the laser 111 to match the signal intensity of reflection or Raman confocal; the focal lengths of all doublet lens groups have been verified by optical design, which is beneficial to the correction of aberration, improves the uniformity of confocal scanning, and reduces the distortion of the entire field of view, so that the resolution, transmittance, and image uniformity of the confocal imaging system are significantly improved. The positions of the third lens 413 and the fourth lens 415 relative to the pinhole 414 can be adjusted in the first direction X and the second direction Y. Through the adjustment in the second direction Y, each component in the second imaging component 41 has a common optical axis; through the adjustment in the first direction X, precise focusing on the pinhole 414 can be achieved. Since there may be an error of several micrometers in the installation of the pinhole 414. Therefore, the pinhole 414 assembly should be installed in a precision translation stage so that the pinhole 414 can be aligned with the focal points of the third lens 413 and the fourth lens 415.

[0067] Further, continue to refer to Figures 5 to 7 , the wide-field light source assembly 21 includes a wide-field white light source 211, a first lens 212, and a second lens 213 arranged in sequence along the first direction X; the power of the wide-field white light source 211 is 20W, and the clear aperture diameter is 6mm; the first lens 212 and the second lens 213 form a Köhler illumination system.

[0068] The wide-field white light source 211 can emit white light, that is, the third light ray, and a high-power LED white light cold light source can be used, with a power of 20W and a clear aperture diameter of 6mm. The focal lengths of the first lens 212 and the second lens 213 are both 50mm, and the two form a Köhler illumination system, which can provide wide-field white light illumination for the sample.

[0069] Further, continue to refer to Figures 5 to 7, the second beam splitting component 22 includes a second non-polarizing beam splitting prism 221, which can reflect the third light ray and transmit the fourth light ray.

[0070] The second beam splitting component 22 is in the form of a non-polarizing beam splitting prism, which can reflect the third light ray and transmit the fourth light ray. When the Raman confocal microscopy imaging system of the embodiment of the present application is in the wide-field imaging mode, the second non-polarizing beam splitting prism 221 is moved into the optical path; when the Raman confocal microscopy imaging system of the embodiment of the present application is in the confocal imaging mode, the second non-polarizing beam splitting prism 221 is moved out of the optical path.

[0071] Further, continue to refer to Figures 5 to 7 , the first imaging component 31 includes a second attenuation sheet 311, a tube lens 312, and a color camera 313 arranged in sequence along the first direction X; the optical density of the second attenuation sheet 311 is 2; the tube lens 312 is a positive lens with a focal length of 200 mm; the color camera 313 is a complementary metal oxide semiconductor sensor camera, and the distance between the color camera 313 and the tube lens 312 is 200 mm.

[0072] The second attenuation sheet 311 is used to attenuate the wide-field received optical signal, that is, attenuate the fourth light ray, and the optical density of the second attenuation sheet 311 is 2. The tube lens 312 is in the form of a positive lens with a focal length of 200 mm. The color camera 313 is used to detect the wide-field image, and a complementary metal oxide semiconductor (CMOS) sensor camera is adopted. The distance between the color camera 313 and the tube lens 312 is 200 mm, so that the color camera 313 is located at the intersection point of the tube lens 312.

[0073] Further, continue to refer to Figures 5 to 7 , the third beam splitting component 32 includes a third non-polarizing beam splitting prism 321, which can reflect part of the fourth light ray and transmit the third light ray and part of the fourth light ray; the transmittance of the third non-polarizing beam splitting prism 321 to the fourth light ray is 10:90.

[0074] The third beam splitting component 32 is in the form of a non-polarizing beam splitting prism, which can reflect part of the fourth light ray and transmit the third light ray and part of the fourth light ray. The transmittance of the third non-polarizing beam splitting prism 321 to the fourth light ray is 10:90. When the Raman confocal microscopy imaging system of the embodiment of the present application is in the wide-field imaging mode, the third non-polarizing beam splitting prism 321 is moved into the optical path; when the Raman confocal microscopy imaging system of the embodiment of the present application is in the confocal imaging mode, the third non-polarizing beam splitting prism 321 is moved out of the optical path.

[0075] In summary, the embodiment of the present application provides a Raman confocal microscopy imaging system. The Raman excitation mechanism, wide-field illumination mechanism, wide-field detection mechanism, and Raman detection mechanism are independent of each other. The relative positions and parameters can be adjusted according to requirements, and different Raman excitation light sources and Raman spectrometers can be adapted. It has the characteristics of easy disassembly, easy integration, and stackability. The working states of the Raman excitation mechanism, wide-field illumination mechanism, wide-field detection mechanism, and Raman detection mechanism can also be adjusted, so that the Raman confocal microscopy imaging system of the embodiment of the present application can switch among three modes: wide-field imaging, reflection confocal imaging, and Raman confocal imaging, thereby providing multi-dimensional information including sample composition, features, and surface morphology, helping to understand features such as the roughness and particle size of the sample surface, and further providing chemical and physical information of the sample. It is applicable to multiple fields, such as materials science, biology, medicine, and environmental science, etc.

[0076] As described above, the above is only a preferred specific embodiment of the present application, but the protection scope of the present application is not limited thereto. Any changes or substitutions that can be easily thought of by those skilled in the art within the technical scope disclosed in the present application should be covered within the protection scope of the present application.

Claims

1. A Raman confocal microscopy imaging system, characterized in that: include: A Raman excitation mechanism includes a laser component and a first light splitting component arranged along a first direction; A wide-field lighting mechanism, comprising a wide-field light source assembly and a second light splitting assembly arranged along the first direction; A wide-field detection mechanism comprises a first imaging component and a third light splitting component arranged along a first direction; A Raman detection mechanism, comprising a second imaging component and a fourth light splitting component arranged along a second direction; An objective lens, wherein the objective lens, the second beam splitting component, the first beam splitting component, the third beam splitting component and the fourth beam splitting component are arranged in sequence along a second direction; The laser component emits a first light ray toward the first beam splitter component, the first beam splitter component reflects the first light ray toward the objective lens, the first light ray forms a second light ray after being reflected by a detection object, and the fourth beam splitter component reflects the second light ray toward the second imaging component; the wide-field light source component emits a third light ray toward the second beam splitter component, the third beam splitter component reflects the third light ray toward the objective lens, the third light ray forms a fourth light ray after being reflected by a detection object, and the third beam splitter component reflects the fourth light ray toward the first imaging component.

2. The Raman confocal microscopy system according to claim 1, characterized in that: The laser assembly includes a laser, a fiber optic collimator, a first filter and a first attenuation plate arranged in sequence along the first direction; the laser emits laser light toward the fiber optic collimator along the first direction, the wavelength of the laser light is 532nm, 638nm, 785nm or 1064nm, and the monochromaticity of the laser light is ±0.01nm; the focal length of the fiber optic collimator is 18mm; the optical density of the first filter is greater than 4; and the optical density of the first attenuation plate is equal to 2.

3. The Raman confocal microscopy system according to claim 1, characterized in that: The first beam splitter component includes a long-pass coated dichroic mirror or a first non-polarized beam splitter prism, which can reflect the first light and the second light and transmit the third light and the fourth light.

4. The Raman confocal microscopy system according to claim 1, characterized in that: The wide-field light source assembly includes a wide-field white light source, a first lens, and a second lens arranged in sequence along the first direction; the power of the wide-field white light source is 20W, and the light transmission diameter is 6mm; the first lens and the second lens form a Kohler illumination system.

5. The Raman confocal microscopy imaging system according to claim 1, characterized in that: The second light splitting component includes a second non-polarizing light splitting prism, and the second non-polarizing light splitting prism can reflect the third light and transmit the fourth light.

6. The Raman confocal microscopy imaging system according to claim 1, characterized in that: The first imaging component includes a second attenuation plate, a tube lens and a color camera arranged in sequence along the first direction; the optical density of the second attenuation plate is 2; the tube lens is a positive lens with a focal length of 200 mm; the color camera is a complementary metal oxide semiconductor sensor camera, and the distance between the color camera and the tube lens is 200 mm.

7. The Raman confocal microscopy imaging system according to claim 1, characterized in that: The third beam splitter component includes a third non-polarizing beam splitter prism, which can reflect part of the fourth light and transmit part of the third light and the fourth light; the transmittance ratio of the third non-polarizing beam splitter prism to the fourth light is 10:

90.

8. The Raman confocal microscopy imaging system according to claim 1, characterized in that: The second imaging component includes a third attenuation plate, a second filter, a third lens, a pinhole, a fourth lens, a fiber coupling mirror and a Raman spectrometer arranged in sequence along the first direction; the optical density of the third attenuation plate is 2; the second filter is a long-pass filter with an optical density greater than 6; the third lens is a double-cemented achromatic convex lens with a focal length of 50 mm; the pinhole is a continuously variable pinhole with a diameter variation range of 0-500 μm and a distance from the third lens of 50 mm; the fourth lens is a double-cemented achromatic convex lens with a focal length of 50 mm, and the third lens and the fourth lens form a four-focal length system; the focal length of the fiber coupling mirror is 18.4 mm, the numerical aperture is 0.15, and the light transmission diameter is 5 mm.

9. The Raman confocal microscopy imaging system according to claim 1, characterized in that: The fourth light splitting component comprises a reflector, and the light transmission size of the reflector is 25 mm.

10. The Raman confocal microscopy imaging system according to claim 1, characterized in that: It also includes an electric stage, which is arranged on a side of the objective lens away from the second light splitting component along the second direction.

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