Illumination module and microscopic imaging system comprising same

Abstract

The utility model provides an illumination module and a microscopic imaging system comprising the same. The illumination module is used for the microscopic imaging system, and comprises an illumination light source module, the optical axis of which is in the horizontal direction; a rotating shaft of the rolling type baffle and the optical axis are located in the same horizontal plane, and the rotating shaft is perpendicular to the optical axis, so that the rolling type baffle can rotate around the illumination light source module, and the rolling type baffle is provided with a window; when the rolling type baffle rotates around the illumination light source module, at least part of illumination light emitted by the illumination light source module can be shielded; the normal direction of the plane mirror and the optical axis form a 45-degree angle, and the plane mirror is used for vertically and upwards reflecting the illumination light out.

Description

Technical Field

[0001] This utility model relates to microscopic imaging technology, and in particular to an illumination module and a microscopic imaging system including the same. Background Technology

[0002] With the rapid development of modern science and technology, microscopic imaging technology, especially stereoscopic microscopy, has been increasingly widely and deeply applied in many key fields such as biomedicine, materials science, and industrial inspection, thanks to its unique advantages. In the biomedical field, it has become an important tool for exploring the mysteries of microscopic life. Researchers can use stereoscopic microscopy to observe the growth and differentiation processes of cells, as well as the microscopic structure of tissues and organs, providing crucial evidence for disease diagnosis and treatment. In materials science, this technology is used to analyze the microstructure and defect distribution of materials, playing an indispensable role in the research and development and performance optimization of new materials. In the field of industrial inspection, this technology can accurately detect minute defects on the surface of products and the integrity of their internal structures, effectively ensuring product quality and safety.

[0003] Tilt illumination, a relatively mature method in microscopy, plays a crucial role in enhancing the imaging of transparent samples. In a microscopic imaging system, the propagation path of light through the condenser lens is critical. Tilt illumination cleverly utilizes this principle, allowing a portion of the light to pass through the condenser lens at a specific angle, thus creating a unique relief effect when imaging the sample. This relief effect significantly improves the contrast of transparent samples, making previously indistinguishable details clearly visible. For example, when observing transparent biological cells or tissue sections, tilt illumination makes cell boundaries, nuclei, and other structures more prominent, facilitating analysis by researchers.

[0004] However, traditional tilt-illumination imaging methods have many limitations. In implementing tilt-illumination models, a simple method is typically used: shifting the condenser lens or adding an obstruction to the condenser lens. While this method is relatively simple in structure and allows for convenient real-time observation of the sample through an eyepiece or camera, it faces numerous problems in practical operation.

[0005] On the one hand, traditional oblique illumination imaging has serious shortcomings in quantifying the proportion of blocked light. Currently, operators can only rely on subjective visual feedback to adjust the proportion of blocked light in hopes of achieving the ideal imaging effect. This leads to inconvenience in the operation process, because everyone's visual judgment is different, and it is difficult to precisely control the proportion of blocked light. At the same time, due to the lack of quantitative standards, repeated adjustments are often required, which not only consumes a lot of time and energy but also reduces work efficiency.

[0006] On the other hand, traditional FOV images suffer from uneven brightness. Due to the shape and position of obstructions and the complexity of light propagation, the light intensity received by different areas in the field of view is inconsistent. This uneven brightness can severely affect the observation results, causing some areas to be too bright or too dark when observing the sample, and some important details may be obscured, thus affecting the comprehensive analysis and accurate judgment of the sample.

[0007] Therefore, although tilted illumination has important value in microscopic imaging technology, the shortcomings of traditional tilted illumination imaging methods urgently need to be improved and perfected through new technologies and methods to meet the needs of various fields for high-precision and high-quality microscopic imaging. Utility Model Content

[0008] To address at least some of the technical problems, this application provides an illumination module and a microscopic imaging system including the same, which can meet the needs of various fields for high-precision, high-quality microscopic imaging, especially for stereoscopic microscopic imaging.

[0009] The first aspect of this application provides an illumination module for a microscopic imaging system, comprising: an illumination source module, wherein the optical axis of the illumination source module is horizontal; a rolling baffle, wherein the rotation axis of the rolling baffle is located in the same horizontal plane as the optical axis and is perpendicular to the optical axis, such that the rolling baffle can rotate around the illumination source module, the rolling baffle having a window, wherein the rolling baffle can at least partially block the illumination light emitted by the illumination source module when rotating around the illumination source module; and a plane reflector, wherein the normal direction of the plane reflector is set at 45 degrees to the optical axis, for reflecting the illumination light vertically upward.

[0010] In a first aspect of this application, the lighting module is disposed within a base, the base having a light outlet from which the lighting light is emitted.

[0011] According to the design of the lighting module in this application, the rolling baffle can rotate around the lighting source module and partially block the lighting light. Compared with the traditional method, it can more flexibly control the angle and intensity of light illumination, effectively improving the contrast when imaging transparent samples and forming a clearer relief effect. The optical axis of the lighting source module is horizontal, and combined with the plane mirror to reflect the light vertically upward, so that the light path extends mainly in the horizontal direction. This makes full use of the horizontal space of the base, making the base more compact in size. Moreover, each module and component can be easily and firmly fixed to the bottom surface of the base, ensuring the stability and efficiency of the light propagation path.

[0012] In a first aspect of this application, the lighting source module includes a white LED component, a heat sink, and a beam shaping lens component. The white LED component is disposed on the heat sink, and the beam shaping lens component is disposed on the light-emitting side of the white LED component and is fixedly connected to the heat sink by fasteners.

[0013] The illumination source module employs a combination of white LED components, a heat sink, and a beam-shaping lens assembly. The white LED components provide a stable illumination source, while the heat sink effectively addresses LED heat generation, extending its lifespan and ensuring illumination stability. The beam-shaping lens assembly shapes the illumination light, and after subsequent optical processing, it further enhances image clarity and uniformity, optimizing the observation of transparent samples.

[0014] In the first aspect of this application, the white LED assembly includes at least two horizontally arranged white LED light-emitting chips, and the beam shaping lens assembly also includes at least two horizontally arranged beam shaping lenses, each corresponding to one of the white LED light-emitting chips, to shape the illumination light.

[0015] Multiple horizontally arranged white LED chips and beam-shaping lenses increase the intensity and coverage of the illumination light, ensuring that all parts of the sample are adequately illuminated. Each beam-shaping lens works in conjunction with a corresponding LED chip to shape the LED beam (i.e., the illumination light), improving the uniformity of illumination. When observing the sample, this effectively avoids areas that are too dark or too bright, enhancing the relief effect while improving the overall image quality.

[0016] In the first aspect of this application, the rolling baffle includes two bearings and a semi-cylindrical light-shielding plate having the window. The two bearings are respectively fixedly disposed on the left and right sides of the lighting source module. The light-shielding plate is rotatably connected to the two bearings through two shafts, thereby enabling it to rotate around the lighting source module.

[0017] The structural design of the rolling baffle in this application makes its rotation more stable and flexible, ensuring smooth rotation. By placing the bearings on the left and right sides of the lighting source module, the rotation axis of the rolling baffle and the optical axis are ensured to be perpendicular to each other in the same horizontal plane. This allows the rotation of the rolling baffle to evenly and gradually block the lighting light, ensuring a linear relationship between the rotation angle of the rolling baffle and the blocking ratio of the lighting light. This facilitates precise control of the baffle's position, thereby more accurately adjusting the blocking ratio of the lighting light.

[0018] In a first aspect of this application, one of the two shafts extends out of the side of the base, and a knob is provided at the end of the shaft extending out of the side of the base for manually rotating the rolling baffle.

[0019] A knob is provided at the end of the shaft to facilitate manual rotation of the rolling baffle. Users can directly and conveniently control the rotation angle of the baffle and adjust the light blocking situation in real time while observing directly with the naked eye.

[0020] In a first aspect of this application, the lighting module further includes a collimating lens for collimating the lighting light.

[0021] In the first aspect of this application, the collimating lens is a Fresnel lens, and the surface of the Fresnel lens without microstructure is further fitted with a sheet-like light-diffusing device for uniformly distributing the illumination light.

[0022] Collimating lenses converge the illumination light to form a collimated beam, improving the efficiency of illumination. Using Fresnel lenses as collimating lenses effectively reduces lens thickness and weight, lowering costs while maintaining collimation performance. The uniform illumination effect of sheet-like light-diffusing devices ensures more even illumination of the sample, preventing uneven light distribution in the field of view (FOV) image and further improving image uniformity and clarity, resulting in a more natural and realistic relief effect.

[0023] In a first aspect of this application, the window is a rectangular window, and at least one of the two borders of the rectangular window used to block the illumination light is provided with a chamfer, thereby forming the border into a sharp edge.

[0024] The rectangular window border features a chamfered edge, creating a sharp, razor-sharp design that produces a clearer and more pronounced transition between light and shadow when the light source is blocked, thus optimizing the contrast of the embossed image. Compared to a standard rectangular border, this design results in a more realistic embossed effect, highlighting the sample's detailed features and helping users to more accurately observe and analyze the sample's microstructure.

[0025] In a first aspect of this application, the lighting module further includes a focusing lens disposed at the light outlet.

[0026] A condenser lens is placed at the light outlet to converge the light, thus forming an oblique illumination path. This improves light utilization and provides the sample with more concentrated and brighter illumination. This not only enhances image clarity but also improves the three-dimensionality of the relief effect to some extent, allowing users to observe sample details and features more clearly and meeting the demand for high-resolution imaging.

[0027] In a first aspect of this application, the lighting module further includes a stepper motor for driving the rolling baffle to rotate stepwise around the lighting source module.

[0028] A stepper motor drives the rolling baffle to rotate in steps, enabling precise control of its rotation. Compared to traditional manual adjustments relying on subjective visual feedback, the stepper motor can precisely adjust the occlusion ratio of the illumination light according to a preset step distance, eliminating the need for repeated adjustments and resulting in accurate and efficient operation. Furthermore, precise control ensures the stability and consistency of each image capture, which is beneficial for subsequent image processing and analysis.

[0029] A second aspect of this application provides a microscopic imaging system, including the illumination module of the first aspect.

[0030] The microscopic imaging system of this application adopts the above-mentioned illumination module, which integrates the advantages of each part of the illumination module structure. It can provide the system with stable, uniform and precisely adjustable illumination light, significantly improve the imaging quality of the imaging system for transparent samples, form clear and realistic relief images, and meet the high requirements of microscopic imaging in many fields such as biomedicine, materials science, and industrial testing. Attached Figure Description

[0031] Figure 1 This is a three-dimensional structural diagram of the lighting module according to this application;

[0032] Figure 2 This is a cross-sectional schematic diagram of the lighting module according to this application along the optical axis.

[0033] Figure 3 Exploded view of the lighting source module and the rolling baffle in the lighting module according to this application;

[0034] Figure 4 This is a comparative schematic diagram showing the rolling baffle according to this application in different rotational positions;

[0035] Figure 5 A flowchart illustrating automatic image capture using the microscopic imaging system of this application;

[0036] Figure 6 This is a flowchart for image correction processing of FOV images;

[0037] Figure 7 A flowchart for image processing using the differential phase contrast method;

[0038] Figure 8 This is a flowchart of a relief image generation method according to an embodiment of this application. Detailed Implementation

[0039] The present application will be further described below with reference to specific embodiments and accompanying drawings. It is to be understood that the illustrative embodiments of this disclosure are merely for explaining the present application and not for limiting it. Furthermore, for ease of description, the accompanying drawings show only the parts relevant to the present application, and not all of the structures or processes.

[0040] The following specific embodiments illustrate the implementation of this application. Those skilled in the art can easily understand other advantages and effects of this application from the content disclosed in this specification. Although the description of this application is presented in conjunction with preferred embodiments, this does not mean that the features of this utility model are limited to this embodiment. On the contrary, the purpose of describing the utility model in conjunction with embodiments is to cover other options or modifications that may be derived based on the claims of this application. To provide a thorough understanding of this application, many specific details will be included in the following description. This application may also be implemented without using these details. Furthermore, to avoid confusion or obscuring the focus of this application, some specific details will be omitted in the description. It should be noted that, unless otherwise specified, the embodiments and features in the embodiments of this application can be combined with each other.

[0041] Unless the context otherwise specifies, the terms “contains,” “has,” and “includes” are synonyms. The phrase “A / B” means “A or B.” The phrase “A and / or B” means “(A and B) or (A or B).”

[0042] It should be understood that although terms such as "first," "second," etc., may be used herein to describe various components, units, or data, these components, units, or data should not be limited by these terms. These terms are used merely to distinguish one feature from another. For example, without departing from the scope of the exemplary embodiments, a first feature may be referred to as a second feature, and similarly, a second feature may be referred to as a first feature.

[0043] It should be understood that although directional terms such as "up," "down," "left," "right," "front," and "back" may be used here to describe the positional relationship between the various components, these directional terms are only for the convenience of understanding and are not intended to limit the scope of protection of this application.

[0044] It should be noted that in this specification, similar reference numerals and letters in the accompanying drawings indicate similar items. Therefore, once an item is defined in one drawing, it does not need to be further defined and explained in subsequent drawings.

[0045] To make the objectives, technical solutions, and advantages of this application clearer, the embodiments of this application will be described in further detail below with reference to the accompanying drawings.

[0046] Figure 1 This is a three-dimensional structural diagram of the lighting module according to this application. Figure 2 This is a schematic cross-sectional view of the lighting module according to this application along the optical axis. Figure 1 and Figure 2 As shown, the lighting module 1 includes: a lighting source module 2, a rolling baffle 3, a collimating lens 4, and a plane reflector 5.

[0047] Figure 3 This is an exploded mechanical view of the lighting source module and the rolling baffle in the lighting module according to this application. Figure 3 As shown, the optical axis of the lighting source module 2 is horizontal, meaning the illumination light emitted by the lighting source module propagates horizontally. The lighting source module 2 includes: a white LED component 21, a heat sink base 22, and a beam shaping lens component 23. The white LED component 21 is an integrated component including a white LED chip and an LED driving circuit, for example, the white LED chip and LED driving circuit are integrated on an integrated circuit board. The white LED component 21 is disposed on the heat sink base 22, meaning the non-emitting side of the white LED component 21 is tightly attached to a groove in the heat sink base 22. It can be fixed by fasteners (e.g., screws), or by clips provided on the heat sink base 22, or by using thermally conductive adhesive (e.g., thermally conductive silicone). The heat sink base 22 can be made of metal materials, such as copper or aluminum alloy, or ceramic materials, such as alumina ceramic or aluminum nitride ceramic. The back of the heat sink base has multiple protruding heat sink fins to facilitate heat dissipation. A beam-shaping lens assembly 23 is disposed on the emitting surface side of the white LED assembly 21 and is fixed to the heat dissipation base 22 by fasteners. The beam-shaping lens assembly 23 includes a cover plate 231, a beam-shaping lens 232, and a lens holder 233. The beam-shaping lens 232 is fixed to a groove in the lens holder 233 by the cover plate 231. The white LED assembly may include one or more white LED chips, for example, two horizontally arranged white LED chips, thereby increasing the coverage of the illumination field. The position of the beam-shaping lens 232 corresponds one-to-one with the white LED chip. The divergence angle of the LED emitted beam may vary in different directions; the beam-shaping lens can be used to shape the LED emitted beam to obtain the desired illumination field range. The beam-shaping lens 232 can be an aspherical lens, and its aspherical parameters can be designed according to the divergence angle of the LED emitted beam and the illumination field range.

[0048] The rolling baffle 3 is a generally semi-cylindrical light-blocking baffle capable of rotating around an axis. This axis is located in the same horizontal plane as the optical axis of the lighting source module and is perpendicular to it, allowing the rolling baffle 3 to rotate around the lighting source module 2. The rolling baffle 3 has a rectangular window 31, which at least partially blocks the illumination light as it rotates around the lighting source module 2. Specifically, when the rectangular window 31 is aligned with the front of the lighting source module 2 (i.e., in the direction of the optical axis), the illumination light can pass through the rectangular window 31 without any obstruction. As the rolling baffle 3 rotates, the rectangular window 31 gradually deviates from the direction of the optical axis, thus gradually blocking the illumination light until it is completely blocked. It should be noted that the window can also be other shapes besides rectangles, such as circles.

[0049] Figure 4 This is a comparative schematic diagram showing the rolling baffle according to this application in different rotational positions. Figure 4 The left image shows a rectangular window of a rolling baffle facing the optical axis, allowing the illumination light to pass through the rectangular window without any obstruction. Figure 4 The middle image shows a situation where a rolling baffle partially blocks the lighting light; Figure 4 The right image shows a situation where a rolling baffle completely blocks the light.

[0050] It should be noted that the illumination module involved in this application can be set in a base, such as the base of a microscopic imaging system. The base has a light outlet, and the illumination module can be a transmissive illumination module. The illumination light is emitted upward from the light outlet, thereby illuminating the sample.

[0051] According to a specific embodiment of this application, such as Figure 3As shown, the rolling baffle 3 includes two bearings 32 and 33, two fan-shaped connectors 34 and 35, and a light-shielding plate 36 with the rectangular window 31. Bearings 32 and 33 are fixedly disposed on the left and right sides of the lighting source module 2, respectively. Preferably, bearings 32 and 33 are directly fixed to the bottom surface of the base by fasteners, thereby supporting the rotation of the rolling baffle 3. However, this invention is not limited to this; bearings 32 and 33 can also be fixed to the side walls of the left and right sides of the lighting source module 2. The fan-shaped connectors 34 and 35 have through holes on their central sides, which can respectively cooperate with one end of the shafts 37 and 38, and be fixed together by snaps or pins to rotate. The other ends of the shafts 37 and 38 are respectively connected to the bearings 32 and 33 for rotational connection. The circumferential sides of the fan-shaped connectors 34 and 35 are fixedly connected to the light-shielding plate 36 by fasteners. The mechanical design of the rolling baffle described above facilitates its assembly and allows for convenient replacement of the light-shielding plate. However, this application is not limited to this mechanical design. For example, the connecting part can be rod-shaped instead of fan-shaped, or the light-shielding plate can be an integrally formed semi-cylindrical structure, i.e., the connecting part and the light-shielding plate are an integral structure. By placing the bearings on the left and right sides of the lighting source module, it can be ensured that the rotation axis of the rolling baffle and the optical axis are perpendicular to each other in the same horizontal plane. This allows the rotation of the rolling baffle to evenly and gradually block the lighting light, ensuring a linear relationship between the rotation angle of the rolling baffle and the blocking ratio of the lighting light. This facilitates precise control of the baffle's position, thereby more accurately adjusting the blocking ratio of the lighting light.

[0052] Further as Figure 3 As shown, after one end of the shaft 37 is inserted into the bearing 32, it extends further outward to the side of the base (not shown), and a knob is provided at its end so that the rolling baffle can be rotated manually.

[0053] like Figure 1 and Figure 2 As shown, the collimating lens 4 is located downstream of the rolling baffle 3 in the optical path and is vertically mounted on the optical path by a fixing device. It is used to collimate the illumination light, making it approximately parallel after passing through the collimating lens 4. Preferably, the collimating lens 4 is a Fresnel lens, which can effectively reduce the thickness and weight of the lens, lowering costs. Furthermore, the planar shape of the Fresnel lens facilitates its use in conjunction with other sheet-like optical devices. More preferably, a sheet-like light-diffusing device is attached to the structureless surface of the Fresnel lens for homogenizing the illumination light. The sheet-like light-diffusing device is preferably frosted glass, but it can also be a light-diffusing film with a microprism or microlens array structure. It should be noted that in some embodiments, the collimating lens can be omitted, for example, in cases where a beam-shaping lens has already been used to substantially collimate the illumination light.

[0054] like Figure 1 and Figure 2 As shown, the plane mirror 5 is located downstream of the collimating lens 4 in the optical path. It is tilted and positioned on the optical path by a fixing device. The normal direction of the plane mirror 5 is at a 45-degree angle to the optical axis, thus reflecting the illumination light vertically upwards. Further as... Figure 3 As shown, during the rotation of the light-shielding plate 36, the two side borders 311 and 312 of the rectangular window 31 cut and block the illumination light. According to one embodiment of this application, the side borders 311 and 312 are provided with chamfers, thus forming sharp blade edges. The sharp blade edges approach ideal geometric straight lines. When the blade edges slowly move to block the light, the sharp blade edges can bring obvious and clear shadow changes, and the transition between the blocked and unblocked parts of the illumination light is clear, which can produce a clearer and more obvious light and dark transition effect and optimize the contrast of the relief image. When the light-shielding plate 36 is rotated downward to cut and block the illumination light, the predetermined technical effect can be achieved by chamfering only the upper side border 311. Alternatively, when the light-shielding plate 36 is rotated upward to cut and block the illumination light, the predetermined technical effect can be achieved by chamfering only the lower side border 312. Compared to conventional microscopic imaging techniques that use ordinary light-shielding devices to achieve oblique illumination, this application sets the border of the rectangular window to a sharp blade, which makes the resulting relief effect more realistic, highlights the detailed features of the sample, and helps users to observe and analyze the microstructure of the sample more accurately.

[0055] In this application, the lighting module also includes a stepper motor for driving a rolling baffle to rotate stepwise around the lighting source module. In microscopic imaging technology, this design of a stepper motor driving the rolling baffle to rotate stepwise plays a crucial role. Traditional lighting adjustment methods often rely on subjective visual feedback for manual adjustment, which is not only cumbersome but also makes it difficult to accurately control the occlusion ratio of the lighting light. Repeated adjustments are often required to achieve a relatively satisfactory effect, which undoubtedly reduces work efficiency and makes it difficult to ensure consistency in each adjustment.

[0056] In contrast, the application of stepper motors has greatly improved this situation. They enable precise control of the rotation of the rolling baffle, accurately adjusting the occlusion ratio of the illumination light according to a preset step distance. This means that operators no longer need to rely on subjective judgment for repeated adjustments, significantly improving operational accuracy and efficiency. By presetting different step distances, stepper motors can flexibly control the rotation angle of the rolling baffle, thereby precisely adjusting the incident angle and intensity of the illumination light to provide ideal lighting conditions for the sample.

[0057] More importantly, this precise control ensures the stability and consistency of each imaging process. In microscopic imaging, stable and consistent illumination conditions are fundamental to acquiring high-quality images. Stable illumination avoids variations in image brightness caused by light fluctuations, ensuring the consistent and reliable quality of each image. Consistent illumination conditions also make images captured at different times comparable, which is crucial for subsequent image analysis and processing.

[0058] In this application, as Figure 1 and Figure 2 As shown, the lighting module also includes a focusing lens 6 disposed at the light outlet of the base. The focusing lens 6 is used to converge the lighting light, thereby forming an inclined lighting light path.

[0059] This application also provides a microscopic imaging system including the illumination module described above. In addition to the illumination module, the microscopic imaging system also includes an objective lens, an eyepiece, a camera, and a computer device. The microscopic imaging system according to this application has both manual and automatic modes. In manual mode, the user can manually adjust a knob to rotate a rolling baffle to adjust the shading ratio and observe the relief image of the sample through the eyepiece. In automatic mode, the computer device controls a stepper motor and camera via software. By setting an appropriate step distance and shooting interval, the stepper motor drives the rolling baffle to stop at different positions, and the camera captures an image at each position. After capturing the images, the software stitches these images together.

[0060] This application also provides a microscopic imaging method applied to the microscopic imaging system described above. The illumination module of the microscopic imaging system includes a stepper motor, which drives a rolling baffle to rotate stepwise around the illumination source module. The microscopic imaging method is described below with reference to the accompanying drawings.

[0061] Figure 5 A flowchart illustrating the automatic image capture using the microscopic imaging system described in this application. Figure 5 As shown, the first step is to perform hardware initialization on the microscopic imaging system. This includes driving the rolling baffle back to its initial position via a stepper motor, i.e., the rolling baffle is facing the optical axis and does not block the illumination light at all. Hardware initialization may also include calibrating the position reference of the stepper motor to ensure that all components of the illumination module, the camera, and the computer device are in a ready state.

[0062] Next, the number of FOV (Field of View) images, n, to be captured is determined, where n is an integer greater than or equal to 1, and the step angle of the rolling baffle is calculated by the computer device. The number of FOV images, n, can be determined by the user based on personal experience, or recommended by the computer device based on the optimal settings of the microscope imaging system. The step angle of the rolling baffle is calculated by dividing the angle from its initial position to its position when it completely blocks the illumination light by n.

[0063] Then, control the rolling baffle to rotate to the position corresponding to step i, where 1≤i≤n, and the camera takes a picture to obtain FOV images. Repeat this step until a set of FOV images including n FOV images is obtained.

[0064] Figure 6 This is a flowchart for image correction processing of FOV images. (Example:) Figure 6 As shown, first, an FOV image is loaded, then shadow correction is performed on the FOV image, and then image brightness normalization processing is performed on the shadow-corrected FOV image. FOV = (FOV - Min(FOV)) / (Max(FOV) - Min(FOV)) to obtain the corrected FOV image. Image shadow correction is a key technology in the fields of computer vision and image processing, aiming to eliminate or reduce the shadow effects in images caused by uneven lighting, object occlusion, or environmental factors, such as uneven lighting around the edges and center of the image, and to restore the true brightness, color, and detail information of the image. According to one embodiment of this application, the method for shadow correction of an FOV image is to control a rolling baffle to stepwise rotate and capture n reference background images without a sample, and then subtract the n FOV images captured with a sample from the reference background images respectively to achieve the purpose of shadow correction. However, this utility model is not limited to this, and those skilled in the art can conceive of using other conventional shadow correction methods, such as algorithms based on Retinex theory, algorithms based on deep learning, and regularization algorithms based on energy minimization.

[0065] The formula FOV = (FOV - Min(FOV)) / (Max(FOV) - Min(FOV)) is a commonly used method for image brightness normalization. Its core principle is to linearly map image pixel values ​​to the 0-1 range. Specifically, for each pixel in an FOV image, its original pixel value is FOV, Max(FOV) represents the maximum value of all pixel values ​​in the image, and Min(FOV) represents the minimum value of all pixel values. By scaling the value of each pixel proportionally to the range of 0 to 1, the image brightness is normalized. By linearly mapping pixel values ​​to a uniform range, brightness offsets between different FOV images can be eliminated, ensuring a consistent brightness baseline when stitching multiple FOV images and avoiding artifacts or blurred edges during stitching. Moreover, normalizing low-contrast relief images can stretch the dynamic range of the original image, making dark details clearer and bright details richer, thus making it easier to identify minute height differences on the sample surface and improving the relief effect.

[0066] Figure 7 This is a flowchart illustrating image processing using the Differential Phase Contrast (DPC) method.

[0067] Among them, INV (Inverse) image refers to the image that is inverse with ... Figure 5 The automatic shooting process shown corresponds to the image obtained by the reverse rotation of the rolling baffle, and the images are captured step by step to obtain the INV image set. Specifically, the forward rotation of the rolling baffle, i.e. Figure 4 After taking n FOV images in a counter-clockwise direction when viewed from the knob side, the rolling baffle is reset, and then rotated in the opposite direction with the same step angle. Figure 4 Take n INV images in a clockwise direction when viewed from the knob side. For example... Figure 7 As shown, firstly, the FOV image and INV image are loaded separately, and then shadow correction is performed on the FOV image and INV image respectively. The shadow correction steps are the same as those described above. Figure 6 The same applies as described above, and will not be repeated here. Furthermore, the shadow correction step here is optional; shadow correction can be omitted from the FOV and INV images, and subsequent image processing can proceed directly.

[0068] Next, difference and sum operations are performed on the FOV and INV images, respectively. The difference operation, IMG1 = FOV - INV, highlights pixel grayscale differences caused by changes in illumination direction. For example, for a raised structure on a sample surface, the left edge is bright and the right edge is shadowed under forward illumination, while the right edge is bright and the left edge is shadowed under reverse illumination. The difference operation amplifies this edge feature of brightness reversal. For flat background areas (without relief structures), the grayscale levels of the FOV and INV images are similar, and IMG1 approaches 0 (noise level). For relief edge areas, the grayscale differences between the FOV and INV images are significant, and the absolute value of IMG1 is larger, forming high-contrast edges. The sum operation, IMG2 = FOV + INV, averages the noise, suppressing global illumination fluctuations and fixed-pattern noise.

[0069] Then, the image undergoes DPC processing, where the ratio operation IMG0 = IMG1 / IMG2 normalizes the contrast of the relief structure and eliminates the influence of illumination intensity on absolute grayscale values. For the same relief structure, the ratio of the grayscale difference between forward and reverse illumination (IMG1) to the background brightness (IMG2) more robustly reflects the change in the angle between the surface normal vector and the illumination direction (i.e., the relief height gradient). For raised structures, the grayscale difference between the FOV and INV images varies with the surface slope; the ratio operation amplifies this difference, making the contrast of the relief edges independent of the background brightness.

[0070] The minimum value shift operation IMG0 = IMG0 - Min(IMG0) shifts the grayscale range of IMG0 from [-1,1] to [0,2] to avoid negative values ​​affecting subsequent image processing (such as display, normalization, etc.) and to highlight the relative differences of the relief structure. The minimum value (usually corresponding to the concave area or background) is set to 0, the maximum value (corresponding to the steep edge) is retained as the peak value, and the intermediate value is scaled proportionally.

[0071] Finally, image brightness normalization is performed: IMG0 = (IMG0 - Min(IMG0) / (Max(IMG0) - Min(IMG0))), thus obtaining the DPC image. This image brightness normalization process is related to... Figure 6 Similar to those in the text, I will not elaborate further here.

[0072] Figure 8 This is a flowchart of a relief image generation method according to an embodiment of this application. Figure 8 As shown, firstly, an image set is acquired using an automatic shooting process. In this step, one can choose to acquire the FOV image set only by driving the scrolling baffle in the forward direction, or one can drive the scrolling baffle in the forward direction and then drive it in reverse direction to acquire both the FOV and INV image sets.

[0073] Next, it is determined whether an INV image set is available. If an INV image set is not available, then... Figure 6 The image correction method shown is used for image processing to perform subsequent image stitching; if an INV image set is available, then... Figure 7 The method shown performs DPC processing on the image to obtain a DPC image for subsequent image stitching.

[0074] Then, the processed images are stitched together to obtain an embossed image. First, the optimal embossed area for each image is determined. Using the sharp border of a rectangular window with chamfered edges as a cutting edge, as the edge slowly moves to block light, it creates a clear and distinct shadow change. The transition between the blocked and unblocked parts of the illumination is clear, producing a sharper and more obvious light-dark transition effect, optimizing the contrast of the embossed image. Therefore, the area near the cutting edge is the optimal embossed area. The computer device can determine the optimal embossed area for each image based on the position of the cutting edge's stepping rotation and the distance of each step. The first image is obtained as the base image. Then, the optimal embossed area of ​​the second image is cropped. The cropped optimal embossed area and the corresponding area of ​​the base image are feathered and stitched together. This stitching process is repeated until a complete embossed image is obtained. Feathering is a technique that achieves smooth blending of image areas through gradual changes in transparency, and it is widely used in image stitching, compositing, and restoration. This method creates a transition zone in the overlapping area of ​​images, allowing pixel values ​​to gradually transition from one image to the other, eliminating stitching artifacts and improving visual continuity. Specifically, a transparency mask is defined in the overlapping area of ​​the two images, making edge pixels completely transparent (100% transparency) and gradually transitioning to complete opacity (0% transparency) towards the center. It should be further noted that the first image is chosen as the Base image for ease of understanding and to facilitate sequential operation by the computer device, stitching the optimal relief areas of subsequent images (second, third, and so on up to the nth image) with the Base image. However, this application is not limited to this; any image can be selected as the Base image for image stitching. Furthermore, Figure 8 Some of the steps shown are optional; for example, the image correction step can be omitted.

[0075] In summary, this application provides an illumination module, a microscopic imaging system, and a microscopic imaging method for a microscopic imaging system. The technical solution of this application, in particular, meets the demand for high-precision, high-quality stereoscopic microscopic imaging. It should be further noted that the above are merely specific embodiments of this application, but the scope of protection of this application is not limited thereto. Any variations or substitutions conceived by those skilled in the art within the scope of the technology disclosed in this application should be included within the scope of protection of this application. Where there is no conflict, the embodiments and features in the embodiments of this application can be combined with each other. Therefore, the scope of protection of this application should be determined by the scope of the claims.

Claims

1. An illumination module for a microscopic imaging system, characterized in that, include: The lighting source module has its optical axis in the horizontal direction. A rolling baffle, wherein the axis of rotation of the rolling baffle is located in the same horizontal plane as the optical axis and is perpendicular to the optical axis, so that the rolling baffle can rotate around the lighting source module. The rolling baffle has a window that can at least partially block the lighting light emitted by the lighting source module when the rolling baffle rotates around the lighting source module. A plane mirror, wherein the normal direction of the plane mirror is set at 45 degrees to the optical axis, is used to reflect the illumination light vertically upward.

2. The lighting module as described in claim 1, characterized in that, The lighting module is disposed inside the base, and the base has a light outlet, from which the lighting light is emitted.

3. The lighting module as described in claim 1, characterized in that, The lighting source module includes a white LED component, a heat sink base, and a beam shaping lens assembly. The white LED component is disposed on the heat sink base, and the beam shaping lens assembly is disposed on the light-emitting side of the white LED component and is fixedly connected to the heat sink base by fasteners.

4. The lighting module as described in claim 3, characterized in that, The white LED assembly includes at least two horizontally arranged white LED light-emitting chips, and the beam shaping lens assembly also includes at least two horizontally arranged beam shaping lenses. The beam shaping lenses correspond one-to-one with the white LED light-emitting chips to shape the illumination light.

5. The lighting module as described in claim 2, characterized in that, The rolling baffle includes two bearings and a semi-cylindrical light-shielding plate with the window. The two bearings are fixedly installed on the left and right sides of the lighting source module, respectively. The light-shielding plate is rotatably connected to the two bearings through two shafts, so that it can rotate around the lighting source module.

6. The lighting module as described in claim 5, characterized in that, One of the two shafts extends out of the side of the base, and a knob is provided at the end of the shaft extending out of the side of the base for manually rotating the rolling baffle.

7. The lighting module as described in claim 1, characterized in that, It also includes a collimating lens, which is used to collimate the illumination light.

8. The lighting module as described in claim 7, characterized in that, The collimating lens is a Fresnel lens, and the surface of the Fresnel lens without microstructure is also attached with a sheet-like light-diffusing device for uniformly distributing the illumination light.

9. The lighting module as described in claim 1, characterized in that, The window is a rectangular window.

10. The lighting module as described in claim 9, characterized in that, At least one of the two borders of the rectangular window used to block the illumination light is chamfered, thereby forming a sharp edge.

11. The lighting module as described in claim 2, characterized in that, It also includes a focusing lens disposed at the light outlet.

12. The lighting module as described in any one of claims 1-11, characterized in that, It also includes a stepper motor for driving the rolling baffle to rotate stepwise around the lighting source module.

13. A microscopic imaging system, characterized in that, Includes the lighting module as described in any one of claims 1-12.

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