Illumination device, scanning imaging method and total internal reflection microscopy system

Abstract

The application relates to an illumination device, a scanning imaging method and a total internal reflection microscopic imaging system. The illumination device comprises a light source for providing a plurality of light rays; a collimating light path for receiving the plurality of light rays and collimating the plurality of light rays to be parallel to each other to reach a waveguide; and the waveguide containing a plurality of coupling incidence areas for coupling the plurality of light rays into the interior of the waveguide, the plurality of light rays being totally internally reflected in the waveguide and reflected to a target sample to enable a microscopic imaging device to image the target sample. The device can provide a uniform light source for the sample, illuminate the field of view of the imaging, and enable the minimum magnification of the total internal reflection microscopic imaging system to be not limited, thereby improving the scanning efficiency.

Description

Technical Field

[0001] This application relates to the field of optical technology, and in particular to an illumination device, a scanning imaging method, and a total internal reflection microscopy imaging system. Background Technology

[0002] With the development of cell detection technology, there is an increasing demand for pathological slide scanning imaging. Pathological slide scanners can be used for histological cell imaging and fluorescence analysis, assisting researchers in cell detection.

[0003] In traditional pathological slide scanning imaging systems, lasers are used as the light source for the illumination system, and the laser is reflected onto the carrier slide through a high-magnification total internal reflection objective to provide a light source for the carrier slide.

[0004] However, in current scanning imaging systems, the use of high-magnification total internal reflection objectives results in an excessively high minimum magnification for imaging, which in turn limits the field of view and reduces scanning efficiency. Summary of the Invention

[0005] Therefore, it is necessary to provide an illumination device, a scanning imaging method, and a total internal reflection microscopy imaging system to address the aforementioned technical problems.

[0006] In a first aspect, this application provides a lighting device. The device includes:

[0007] A light source, used to provide multiple light rays;

[0008] A collimated optical path is used to receive the multiple light rays and collimate them so that the multiple light rays are parallel to each other and reach the waveguide.

[0009] The waveguide contains multiple coupling incident regions, which are used to couple multiple light rays into the interior of the waveguide. The multiple light rays are totally internally reflected in the waveguide and reflected to the target sample, so that the microscopic imaging device can image the target sample.

[0010] In one embodiment, the light source is a light-emitting diode, a laser diode, or a laser lamp.

[0011] In one embodiment, the light source is located directly above the collimated optical path, and the light source, the collimated optical path, and the coupled incident region are at a preset angle.

[0012] In one embodiment, each of the coupling regions within the waveguide contains a grating with different parameters, the grating being used to optimize the plurality of light rays.

[0013] In one embodiment, the grating is an embossed grating or a holographic grating.

[0014] Secondly, this application also provides a scanning imaging method. The scanning imaging method is applied to a lighting device, wherein the lighting device is the lighting device described in any of the above embodiments, and the scanning imaging method includes:

[0015] Provide multiple light rays;

[0016] The system receives the multiple light rays, collimates them through a collimating optical path, and makes the multiple light rays parallel to each other before reaching the waveguide.

[0017] Multiple light rays are coupled into the interior of the waveguide through multiple coupling incident regions on the waveguide, and the waveguide performs total internal reflection of the multiple light rays, reflecting them to the target sample so that the microscopic imaging device can image the target sample.

[0018] In one embodiment, the target sample is placed above the waveguide, and multiple light rays are coupled into the interior of the waveguide through multiple coupling incident regions on the waveguide. The waveguide performs total internal reflection of the multiple light rays, reflecting them to the target sample so that the microscopic imaging device can image the target sample. This includes:

[0019] By optimizing the corresponding light rays in each of the multiple coupling incident regions on the waveguide, and coupling the light rays into the interior of the waveguide, the multiple light rays are subjected to multiple total internal reflections to the surface of the waveguide through the waveguide, so as to provide illumination for the target sample located above the waveguide, so that the microscopic imaging device can image the target sample.

[0020] Thirdly, this application also provides a total internal reflection microscopy imaging system, the total internal reflection microscopy imaging system comprising:

[0021] An illumination device includes a light source for providing multiple light rays; a collimating optical path for receiving the multiple light rays and collimating them so that they are parallel to each other and reach a waveguide; the waveguide contains multiple coupling incident regions for coupling the multiple light rays into the interior of the waveguide, whereby the multiple light rays undergo total internal reflection within the waveguide and are reflected to a target sample, thereby enabling a microscopic imaging device to image the target sample;

[0022] The microscopic imaging device is used to perform microscopic imaging on the target sample.

[0023] In one embodiment, the total internal reflection microscopy system further includes:

[0024] Sample holder, the sample holder being used to fix the target sample;

[0025] A scanning platform is located below the microscopic imaging device and is used to control the movement of the microscopic imaging device to scan and image the target sample.

[0026] In one embodiment, the microscopic imaging apparatus includes:

[0027] Objective lens, used for magnified imaging of the target sample;

[0028] A filter cartridge used to collect fluorescence signals at specific wavelengths;

[0029] A detector used to collect target information.

[0030] The aforementioned illumination device, scanning imaging method, and total internal reflection microscopy imaging system include an illumination device characterized in that the device comprises: a light source for providing multiple light rays; a collimating optical path for receiving the multiple light rays and collimating them so that the multiple light rays are parallel to each other and reach a waveguide; the waveguide contains multiple coupling incident regions for coupling the multiple light rays into the interior of the waveguide, and the multiple light rays undergo total internal reflection in the waveguide, reflecting them to a target sample so that the microscopy imaging device can image the target sample. Using this illumination device, multiple light rays are optimized through multiple coupling incident regions within the waveguide, and multiple light rays are coupled into the interior of the waveguide. The waveguide performs total internal reflection of the multiple light rays, providing a uniform light source for the sample, illuminating the imaging field of view, and allowing the minimum magnification of the total internal reflection microscopy imaging system to be unrestricted, thus improving scanning efficiency. Attached Figure Description

[0031] Figure 1 This is a schematic diagram of the total internal reflection microscopy imaging system in one embodiment;

[0032] Figure 2 This is a schematic diagram of the lighting system in one embodiment;

[0033] Figure 3 A schematic diagram of a waveguide design with multiple coupled incident regions in one embodiment;

[0034] Figure 4(a) is a schematic diagram of the side structure of the waveguide in one embodiment;

[0035] Figure 4(b) is a schematic diagram of the front structure of the waveguide in one embodiment;

[0036] Figure 5 This is a schematic flowchart of a scanning imaging method in one embodiment. Detailed Implementation

[0037] To make the objectives, technical solutions, and advantages of this application clearer, the following detailed description is provided in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the scope of this application.

[0038] It should be understood that the terms "first," "second," "third," and "fourth," etc., in the claims, specification, and drawings of this disclosure are used to distinguish different objects, not to describe a specific order. The terms "comprising" and "including" as used in the specification and claims of this disclosure indicate the presence of the described features, integrals, steps, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, integrals, steps, operations, elements, components, and / or collections thereof.

[0039] It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of this disclosure. As used in this disclosure and claims, the singular forms “a,” “an,” and “the” are intended to include the plural forms unless the context clearly indicates otherwise. It should also be understood that the term “and / or” as used in this disclosure and claims refers to any combination and all possible combinations of one or more of the associated listed items, and includes such combinations.

[0040] As used in this specification and claims, the term "if" may be interpreted, depending on the context, as "when," "once," "in response to determination," or "in response to detection." Similarly, the phrase "if determined" or "if [described condition or event] is detected" may be interpreted, depending on the context, as "once determined," "in response to determination," "once [described condition or event] is detected," or "in response to detection of [described condition or event]."

[0041] The lighting device provided in this application embodiment can be applied to, for example... Figure 1 The total internal reflection microscopy (TIV) imaging system 100 shown includes an illumination device 110 and a microscopic imaging device 120. Optionally, the TIV imaging system 100 further includes a scanning platform 130 and a sample holder 140. The illumination device 110 provides uniform illumination to the target sample on the sample holder 140, facilitating microscopic imaging of the target sample by the microscopic imaging device 120. The sample holder 140 is used to fix the target sample, and the scanning platform 130 is used to control the movement of the microscopic imaging device 120.

[0042] Optionally, the microscopic imaging device 120 can be an integrated imaging device, which integrates objectives, detectors, filters, and other hardware devices for imaging. This embodiment of the invention does not limit the types and number of hardware devices that can be integrated into the microscopic imaging device 120. Specifically, the internal connection structure and achievable application functions of the microscopic imaging device 120 will be described in detail in subsequent embodiments of the invention.

[0043] In one embodiment, such as Figure 2 As shown, a lighting device 110 is provided for use in... Figure 1 In the total internal reflection microscopy imaging system 100, the illumination device 110 includes: a light source for providing multiple light rays; a collimating optical path for receiving multiple light rays and collimating the multiple light rays so that the multiple light rays are parallel to each other and reach the waveguide; and a waveguide containing multiple coupling incident regions for coupling multiple light rays into the interior of the waveguide. The multiple light rays undergo total internal reflection in the waveguide and are reflected to the target sample so that the microscopy imaging device can image the target sample.

[0044] In implementation, the illumination device 110 includes a light source 200, a collimating optical path 300, and a waveguide 400. The light source 200 provides multiple rays with different directions. The collimating optical path 300 is located below the light source 200. Multiple rays propagate through the air, with some rays pointing downwards reaching the collimating optical path 300. The parameters of these rays are different, making them divergent. The collimating optical path 300 receives and collimates these rays, correcting their parameters to make them parallel to each other. These parallel rays then reach the waveguide 400. The target sample is located above the waveguide 400, which contains multiple coupling incident regions 410. After reaching different coupling incident regions 410, the rays are optimized and coupled into the waveguide. The waveguide performs multiple total internal reflections of multiple light rays to reach the target sample, providing illumination to the target sample so that the microscopic imaging device 120 can perform microscopic imaging on the target sample.

[0045] Optionally, the collimating optical path 300 can be a collimating lens, a laser collimator, etc. The implementation of the collimating optical path 300 in this application embodiment is not limited.

[0046] The aforementioned illumination device includes a light source for providing multiple light rays; a collimating optical path for receiving and collimating these rays to ensure they are parallel to each other before reaching the waveguide; and a waveguide containing multiple coupling incident regions for coupling these rays into the waveguide's interior. These rays undergo total internal reflection within the waveguide, reflecting onto the target sample to enable the microscopic imaging device to image the sample. This device optimizes the multiple light rays through multiple coupling incident regions within the waveguide and couples them into the waveguide's interior. The waveguide's total internal reflection provides a uniform light source for the sample, illuminating the imaging field of view and eliminating limitations on the minimum magnification of the total internal reflection microscopic imaging system, thus improving scanning efficiency.

[0047] In one embodiment, the light source is a light-emitting diode, a laser diode, or a laser lamp.

[0048] In practice, the light source 200 in the lighting device 110 can be a light-emitting diode (LED), a laser diode (LD), or a laser lamp.

[0049] For example, such as Figure 2 As shown, the light source 200 is an LED light, which provides multiple divergent light rays in different directions. These multiple divergent light rays are not parallel to each other.

[0050] Optionally, the light source 200 in the lighting device 110 may be a light-emitting diode (LED) or a laser lamp, depending on the requirements. LEDs are more readily available. Laser lamps can provide a high-quality beam with high collimation. This application embodiment does not limit the type of light source.

[0051] In this embodiment, the light source provides multiple light rays to the lighting device, facilitating subsequent processing of these multiple light rays. Furthermore, if the light source is an LED, it can improve the ease of setting up the lighting device.

[0052] In one embodiment, such as Figure 2 As shown, the light source is located directly above the collimated optical path, and the light source, collimated optical path, and coupled incident area form a preset angle.

[0053] In practice, the light source 200 in the lighting device 110 can be located directly above the collimated light path 300. The light source 200, the collimated light path 300, and the coupled incident area 410 in the lighting device 110 are at a preset angle.

[0054] Specifically, such as Figure 2 As shown, Figure 2This is a schematic diagram of the lighting device 110. The light source 200 in the lighting device 110 can be an LED, and the collimating light path 300 can also be an LED collimating light path. The LED collimating light path is directly below the LED. The waveguide 400 in the lighting device 110 is located below the LED collimating light path. The coupling incident region 410 within the waveguide 400 is located below the LED collimating light path. The LED, the LED collimating light path, and the coupling incident region 410 are all located on the same vertical line. The preset angle can be 0 degrees.

[0055] Optionally, if the light source 200 is a laser lamp, the emission direction of the laser lamp is adjusted to directly downwards. Because the multiple rays emitted by the laser lamp have high collimation, most of the rays provided by the laser lamp can reach the collimated optical path 300.

[0056] Optionally, the preset angle is determined based on the micro / nano structure design of the grating within the coupling incident region. Different micro / nano structures have different incident angles. This application embodiment does not limit the incident angle.

[0057] In this embodiment, the collimating optical path is placed directly below the light source, allowing more light to reach the collimating optical path. This collimates multiple light rays, and through a preset incident angle, more light can reach the waveguide and undergo total internal reflection to reach the target sample, providing better illumination for the target sample.

[0058] In one embodiment, such as Figure 3 As shown, each coupling region within the waveguide contains gratings with different parameters, which are used to optimize multiple rays.

[0059] In implementation, Figure 3 This is a waveguide structure diagram showing multiple coupled incident regions. Viewed from the front, the waveguide 400 in the illumination device 110 contains multiple coupled incident regions 410. x represents the length of the waveguide 400, and y represents the width of the waveguide 400. 1, 2, and 3 represent coupled incident region 1, coupled incident region 2, and coupled incident region 3, respectively.

[0060] In the illumination device 110, the waveguide 400 can be a glass sheet with variable thickness. This glass sheet contains multiple coupling incident regions 410. Each coupling incident region 410 contains a corresponding grating 420. The multiple gratings 420 have different micro / nanostructure designs. Micro / nanostructures are a nanoscale technology used to design, develop, and manufacture microstructures and mechanical devices. Micro / nanostructures refer to the design, fabrication, and refinement of microstructures and mechanical systems at the atomic scale using precise atomic-level components (typically from microelectronic devices and biological systems) combined with precisely controlled mechanical techniques. It utilizes structures built at the micrometer scale to achieve many physical phenomena, such as energy conversion, energy storage, and radiation detection. Multiple coupling incident regions 410 are obtained by engraving the top or surface of the glass sheet according to different micro / nanostructure designs. Because the light source 200 in the illumination device 110 provides multiple rays with different parameters—for example, different wavelengths and frequencies—each ray needs to be optimized according to its parameters. Therefore, different micro / nanostructures are designed. Each micro / nano structure in the coupling region corresponds to an illumination wavelength.

[0061] For example, such as Figure 2 As shown, multiple light rays pass through the coupling incident region 410 on the waveguide and reach the interior of the waveguide. These rays undergo multiple total internal reflections within the waveguide, resulting in a large total internal reflection illumination area. The total internal reflection illumination area is shown in Figure 4, which is a schematic diagram of waveguide 400. Figure 4(a) shows a schematic diagram of the waveguide side structure, and Figure 4(b) shows a schematic diagram of the waveguide front structure. According to Figure 4(a), the coupling incident region 410 is located above or on the surface of waveguide 400. According to Figure 4(b), x and y are the length and width of waveguide 400, respectively, and x1 and y1 are the length and width of the coupling region on waveguide 400. The area of ​​the coupling region determines the illumination width of the light source 200 within waveguide 400. Specifically, the length of the total internal reflection can cover the entire length x of the waveguide, and the width of the total internal reflection can cover the entire width y of the waveguide. Therefore, multiple light rays passing through the waveguide 400 and the coupling incident region 410 in the illumination device 110 can achieve a large total internal reflection illumination area. Thus, the length and width of the coupling region 410, the parameters of the grating 420 within the coupling region 410, and the light source 200 and collimating optical path 300 need to be designed collaboratively. By utilizing the collaborative design of the light source 200, the collimating optical path 300, and the waveguide 400, a large field-of-view total internal reflection illumination system can be obtained.

[0062] Optionally, the design of parameters such as size, depth, and slope of the grating 420 in the lighting device 110 needs to be determined based on the wavelength range and incident angle range of multiple light rays. This application embodiment does not limit the parameters of the grating 420.

[0063] Optionally, the thickness of the waveguide 400 in the lighting device 110 can be 1 mm (millimeter) or 2 mm. In this embodiment, the specific thickness of the waveguide 400 is not limited.

[0064] In this embodiment, by setting multiple coupling regions and gratings, multiple light rays incident on the waveguide are optimized multiple times. The waveguide performs multiple total internal reflections on these light rays, which can expand the total internal reflection illumination area. Furthermore, according to application requirements, the waveguide can be flexibly designed to optimize the illumination wavelength and the total internal reflection illumination range, thereby improving light energy utilization.

[0065] In one embodiment, the grating is an embossed grating or a holographic grating.

[0066] In practice, the grating 410 in the lighting device 110 can be an embossed grating or a holographic grating. The waveguide 400 or the grating 410 can be coated to improve the light energy utilization of the waveguide.

[0067] Optionally, the relief grating can be a surface relief grating. The holographic grating can be a volume holographic grating. A surface relief grating is formed by performing photolithography, etching, imprinting, or other operations on the surface of a substrate (similar to surface engraving). A volume holographic grating forms interference fringes within the photosensitive material, which then generate a diffraction grating after exposure (similar to internal development). The choice between a relief grating and a holographic grating is determined based on requirements. A surface relief grating has a uniform refractive index distribution, but its surface microstructure is periodic. A volume holographic grating has a periodically varying refractive index distribution, but its surface is smooth. This application does not limit the form of the grating in its embodiments.

[0068] In this embodiment, multiple light rays can be coupled using an embossed grating or a holographic grating, thereby improving the utilization rate of light energy.

[0069] In one embodiment, a scanning imaging method is provided, which can be applied to the illumination device in the above embodiments, such as... Figure 5 As shown, the scanning imaging method includes:

[0070] Step 502: Provide multiple light rays.

[0071] In practice, within a lighting device, multiple light rays are provided by the light source emitting light in all directions. These different light rays travel along different paths, with some rays propagating downwards to reach collimated rays. Furthermore, these multiple light rays have different parameters.

[0072] Optionally, multiple light rays can be provided by LEDs, laser diodes, or laser lights. This application does not limit the device for providing light rays.

[0073] Step 504: Receive multiple light rays, collimate the multiple light rays through a collimating optical path, and make the multiple light rays parallel to each other as they reach the waveguide.

[0074] In this process, multiple light rays travel along different paths, causing them to diverge. That is, two initially adjacent light rays will gradually drift further apart as they propagate.

[0075] In practice, the collimating optical path receives multiple light rays. The collimating optical path collimates these multiple light rays, making them parallel to each other as they reach the waveguide. Collimation, in essence, transforms diverging light rays into parallel ones.

[0076] Step 506: Multiple light rays are coupled into the interior of the waveguide through multiple coupling incident regions on the waveguide, and the multiple light rays are reflected to the target sample through total internal reflection by the waveguide, so that the microscopic imaging device can image the target sample.

[0077] There are multiple coupling incident regions on the waveguide.

[0078] In practice, multiple light rays arrive at the waveguide in parallel. Multiple coupling incident regions on the waveguide optimize the multiple light rays and couple them into the interior of the waveguide. The waveguide then reflects all the light rays internally to the target sample, providing a uniform light source for the target sample and facilitating imaging of the target sample by the microscopic imaging device.

[0079] Specifically, multiple light rays arrive at the waveguide in parallel. Each coupling incident region contains a corresponding grating. Different gratings have different grating parameters. Based on the grating parameters, each coupling incident region has a corresponding illumination wavelength. Therefore, each coupling incident region within the waveguide optimizes the corresponding ray among the multiple rays, coupling the ray into the waveguide interior. The waveguide then performs total internal reflection of the multiple rays to the target sample. Total internal reflection refers to the phenomenon where, when light travels from a medium with a higher refractive index to a medium with a lower refractive index, if the angle of incidence is greater than the critical angle (the ray is away from the normal), the refracted ray will disappear, and all incident rays will be reflected and not enter the lower refractive index medium.

[0080] In this embodiment, multiple light rays are optimized through multiple coupling incident regions on the waveguide and coupled into the waveguide. The waveguide performs total internal reflection of the multiple light rays, providing a uniform light source for the sample, illuminating the imaging field of view, and making the minimum magnification of the total internal reflection microscopy imaging system unrestricted, thus improving the scanning efficiency.

[0081] In one embodiment, the target sample is placed above the waveguide, and the specific processing steps of step 506 include:

[0082] By optimizing the corresponding light rays in each of the multiple coupled incident regions on the waveguide, the light rays are coupled into the interior of the waveguide. The multiple light rays are then subjected to multiple total internal reflections to the surface of the waveguide, providing illumination to the target sample located above the waveguide, so that the microscopic imaging device can image the target sample.

[0083] In practice, the target sample is placed above the waveguide. Different coupling regions are designed based on the varying parameters of multiple light rays. Each coupling incident region corresponds to a light ray. Within the waveguide, each coupling incident region optimizes the corresponding light ray from the multiple light rays and couples it into the waveguide's interior. The waveguide performs total internal reflection on the multiple light rays. Each light ray, after multiple total internal reflections, reaches the waveguide surface, providing sufficient illumination for the target sample on the waveguide. The area illuminated by the multiple light rays constitutes the entire waveguide in the illumination device. When a focal plane layer exists in the target sample, the light from the total internal reflections of the light rays within the coupling incident regions of the waveguide can illuminate this focal plane layer.

[0084] Optionally, the target sample can be a pathological section or a plant section, etc. This application does not limit the target sample in its embodiments.

[0085] In this embodiment, placing the target sample above the waveguide illuminates it, providing sufficient light. Furthermore, this illumination prevents the fluorescent material in the non-focal layers from being excited and becoming background, thus improving the imaging signal-to-noise ratio.

[0086] In one embodiment, such as Figure 1 As shown, a total internal reflection microscopy imaging system 100 is provided. The scanning imaging method of the above embodiment can be applied to the total internal reflection microscopy imaging system 100, such as... Figure 1 As shown, the total internal reflection microscopy imaging system 100 includes:

[0087] The illumination device 110 includes a light source 200 for providing multiple light rays; a collimating optical path 300 for receiving and collimating the multiple light rays so that they are parallel to each other and reach the waveguide; and a waveguide 400 containing multiple coupling incident regions 410 for coupling multiple light rays into the interior of the waveguide. The multiple light rays undergo total internal reflection in the waveguide 400 and are reflected to the target sample so that the microscopic imaging device 120 can image the target sample.

[0088] Microscopic imaging device 120 is used to perform microscopic imaging on target samples.

[0089] In implementation, the total internal reflection microscopy imaging system 100 includes an illumination device 110 and a microscopy imaging device 120. The specific structure and function of the illumination device 110 have been described in the above embodiments and will not be repeated here. The microscopy imaging device 120 is used to perform microscopic imaging on the target sample.

[0090] Optionally, the microscopic imaging apparatus 120 integrates multiple devices, such as objectives, filter boxes, and detectors. This application embodiment does not limit the number or type of devices integrated into the microscopic imaging apparatus.

[0091] Optionally, current pathological diagnosis still relies primarily on image analysis. However, a total internal reflection system based on waveguide 400 combined with algorithms can achieve single-molecule localization of biological macromolecules such as proteins and nucleic acids. This enables the integration of molecular pathology with histopathology and cellular pathology.

[0092] In this embodiment, by separating the illumination device and the microscopic imaging device, and by using waveguides with different coupling incident regions to perform total internal reflection of multiple light rays, a large field of view total internal reflection illumination system is obtained without the need for a high-magnification total internal reflection objective lens, thus avoiding the limitation of the imaging field of view by a high-magnification objective lens.

[0093] In one embodiment, the total internal reflection system 100 further includes:

[0094] Sample holder 130, sample holder 130 is used to fix the target sample;

[0095] The scanning platform 140 is located below the microscopic imaging device and is used to control the movement of the microscopic imaging device so as to scan and image the target sample.

[0096] In implementation, the total internal reflection system 100 also includes a sample holder 130 and a scanning platform 140. For example... Figure 1 As shown, the sample holder 130 is located below the illumination device 110 and is used to fix the target sample. The microscopic imaging device 120 is located above the scanning platform 140, which is used to control the left and right movement of the microscopic imaging device 120 so that the illuminated part of the target sample can be scanned and imaged.

[0097] Specifically, the scanning platform 140 includes a scanning platform controller 141 and a workstation 142. The scanning platform controller 141 controls the left and right movement of the microscopic imaging device. The workstation 142 includes a display screen for real-time display of microscopic images. A first communication connection exists between the scanning platform controller 141 and the workstation 142. A second communication connection exists between the scanning platform controller 141 and the microscopic imaging device, used for transmitting movement information. A third communication connection exists between the workstation 142 and the detector in the microscopic imaging device 120, used for transmitting microscopic imaging information.

[0098] The relative positions of the illumination device 110, the target sample, and the microscopic imaging device 120 are fixed by a designed structural component. This embodiment of the application does not limit the scope of this structural component.

[0099] In this embodiment, the target sample can be fixed by the sample holder, and the scanning platform can flexibly control the scanning observation point, which facilitates the observation of the target sample.

[0100] In one embodiment, the microscopic imaging apparatus includes:

[0101] Objective lens 121 is used for magnified imaging of the target sample;

[0102] Filter box 122 is used to collect fluorescence signals at a specific wavelength;

[0103] Detector 123 is used to collect target information.

[0104] In implementation, the microscopic imaging apparatus 120 includes an objective lens 121, a filter box 122, and a detector 123. The objective lens 121 refers to a lens group composed of several lenses. Figure 1 As shown, Figure 1 In this diagram, OBJ represents objective lens 121. Objective lens 121 is located below the target sample. Magnified imaging of the target sample is achieved through objective lens 121. By moving the microscopic imaging device 120, objective lens 121 can be moved left and right, thereby changing the observation point. By adjusting the objective lens, the magnification can be changed, thus obtaining a larger field of view. Filter box 122 is located below objective lens 121, and the filter box 122 forms a preset angle with objective lens 121. Filter box 122 is used for fluorescence signals of a specific wavelength. Detector 123 is at the same height as filter box 122. Detector 123 is used to collect target microscopic imaging information.

[0105] In this embodiment, the target sample is scanned and imaged by the integrated devices in the microscopic imaging device, which facilitates the observation of the target sample.

[0106] It should be understood that although the steps in the flowcharts of the embodiments described above are shown sequentially according to the arrows, these steps are not necessarily executed in the order indicated by the arrows. Unless explicitly stated herein, there is no strict order restriction on the execution of these steps, and they can be executed in other orders. Moreover, at least some steps in the flowcharts of the embodiments described above may include multiple steps or multiple stages. These steps or stages are not necessarily completed at the same time, but can be executed at different times. The execution order of these steps or stages is not necessarily sequential, but can be performed alternately or in turn with other steps or at least some of the steps or stages of other steps.

[0107] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0108] The embodiments described above are merely illustrative of several implementation methods of this application, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of this patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this application should be determined by the appended claims.

Claims

1. A lighting device, characterized in that, The device includes: A light source, used to provide multiple light rays; A collimated optical path is used to receive the multiple light rays and collimate them so that the multiple light rays are parallel to each other and reach the waveguide. The waveguide contains multiple coupling incident regions, which are used to couple multiple light rays into the interior of the waveguide. The multiple light rays undergo total internal reflection within the waveguide and are reflected to the target sample, enabling the microscopic imaging device to image the target sample. Each coupling incident region has a corresponding illumination wavelength. The area illuminated by the multiple light rays is the entire waveguide in the illumination device. When a focal plane layer exists in the target sample, the light rays undergoing total internal reflection through the coupling incident regions within the waveguide illuminate the focal plane layer, improving the imaging signal-to-noise ratio. The light source is a laser lamp, and the laser lamp emits light directly downwards; the light source is located directly above the collimating optical path; the light source, the collimating optical path, and the coupling incident region are located on the same vertical line; the length of the total internal reflection covers the length of the waveguide, and the width of the total internal reflection covers the width of the waveguide; the length and width of the coupling incident region, the parameters of the grating within the coupling incident region, and the light source and the collimating optical path are designed in a coordinated manner to obtain a total internal reflection illumination system with a large field of view, and to ensure that the minimum magnification of the microscopic imaging system is not limited; Each of the coupled incident regions within the waveguide contains a grating with different parameters, which is used to optimize the corresponding rays among the multiple rays.

2. The lighting device according to claim 1, characterized in that, The multiple light rays pass through the coupling incident area on the waveguide and reach the inside of the waveguide. The multiple light rays undergo multiple total internal reflections inside the waveguide to obtain a total internal reflection illumination area.

3. The lighting device according to claim 1, characterized in that, The grating is an embossed grating or a holographic grating.

4. The lighting device according to claim 3, characterized in that, The waveguide or the grating is coated.

5. The lighting device according to claim 4, characterized in that, The relief grating is a surface relief grating; the holographic grating is a volume holographic grating.

6. A scanning imaging method, characterized in that, The scanning imaging method is applied to an illumination device, wherein the illumination device is the illumination device described in any one of claims 1 to 5, and the scanning imaging method comprises: Provide multiple light rays; The system receives the multiple light rays, collimates them through a collimating optical path, and makes the multiple light rays parallel to each other before reaching the waveguide. Multiple light rays are coupled into the interior of the waveguide through multiple coupling incident regions on the waveguide, and the waveguide performs total internal reflection of the multiple light rays, reflecting them to the target sample so that the microscopic imaging device can image the target sample.

7. The scanning imaging method according to claim 6, characterized in that, The target sample is placed above the waveguide. Multiple light rays are coupled into the interior of the waveguide through multiple coupling incident regions on the waveguide. The waveguide performs total internal reflection of the multiple light rays, reflecting them to the target sample so that the microscopic imaging device can image the target sample. This includes: By optimizing the corresponding light rays in each of the multiple coupling incident regions on the waveguide, and coupling the light rays into the interior of the waveguide, the multiple light rays are subjected to multiple total internal reflections to the surface of the waveguide through the waveguide, so as to provide illumination for the target sample located above the waveguide, so that the microscopic imaging device can image the target sample.

8. A total internal reflection microscopy imaging system, characterized in that, The total internal reflection microscopy imaging system includes: An illumination device includes a light source for providing multiple light rays; a collimating optical path for receiving and collimating the multiple light rays so that they are parallel to each other and reach a waveguide; the waveguide contains multiple coupling incident regions for coupling the multiple light rays into the interior of the waveguide, where the multiple light rays undergo total internal reflection within the waveguide and are reflected to a target sample, enabling a microscopic imaging device to image the target sample; each coupling incident region has a corresponding illumination wavelength; the area illuminated by the multiple light rays is the entire waveguide in the illumination device; when a focal plane layer exists in the target sample, the light rays undergoing total internal reflection through the coupling incident regions within the waveguide illuminate the focal plane layer, improving the imaging signal-to-noise ratio; The light source is a laser lamp, and the laser lamp emits light directly downwards; the light source is located directly above the collimating optical path; the light source, the collimating optical path, and the coupling incident region are located on the same vertical line; the length of the total internal reflection covers the length of the waveguide, and the width of the total internal reflection covers the width of the waveguide; the length and width of the coupling incident region, the parameters of the grating within the coupling incident region, and the light source and the collimating optical path are designed in a coordinated manner to obtain a total internal reflection illumination system with a large field of view, and to ensure that the minimum magnification of the microscopic imaging system is not limited; The microscopic imaging device is used to perform microscopic imaging on the target sample.

9. The total internal reflection microscopy imaging system according to claim 8, characterized in that, The total internal reflection microscopy imaging system also includes: Sample holder, the sample holder being used to fix the target sample; A scanning platform is located below the microscopic imaging device and is used to control the movement of the microscopic imaging device to scan and image the target sample.

10. The total internal reflection microscopy imaging system according to claim 8, characterized in that, The microscopic imaging device includes: Objective lens, used for magnified imaging of the target sample; A filter cartridge used to collect fluorescence signals at specific wavelengths; A detector used to collect target information.

Images