CN116430569A - Illumination device, scanning imaging method and total internal reflection microscopic imaging system - Google Patents

Abstract

The application relates to an illumination device, a scanning imaging method and a total internal reflection microscopic imaging system. The lighting device includes: a light source for providing a plurality of light rays; the collimating light path is used for receiving the plurality of light rays, and collimating the plurality of light rays so that the plurality of light rays mutually parallel reach the waveguide; the waveguide comprises a plurality of coupling incidence areas, the coupling incidence areas are used for coupling the light rays into the waveguide, the light rays are totally internally reflected in the waveguide and reflected to a target sample, and the microscopic imaging device is used for imaging the target sample. The device can provide a uniform light source for the sample, lightens the imaging visual field, ensures that the imaging minimum multiple of the total internal reflection microscopic imaging system is not limited, and improves the scanning efficiency.

Description

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

Technical Field

The present application relates to the field of optical technology, and in particular, to an illumination device, a scanning imaging method, and a total internal reflection microscopic imaging system.

Background

With the development of cell detection technology, there is a growing need for pathological section scanning imaging. The pathological section scanner can be used for histological cell imaging and fluorescence analysis, and assists scientific researchers to carry out cell detection.

In a traditional pathological section scanning imaging system, laser is used as a light source of an illumination system, and is reflected onto a slide through a large-multiple total internal reflection objective lens to provide a light source for the slide.

However, in the current scanning imaging system, the adoption of the total internal reflection objective lens with a large multiple can lead to the imaging multiple to have an excessively high minimum multiple, so that the imaging field of view is limited, and the scanning efficiency is reduced.

Disclosure of Invention

Based on this, it is necessary to provide an illumination device, a scanning imaging method and a total internal reflection microscopic imaging system in view of the above technical problems.

In a first aspect, the present application provides a lighting device. The device comprises:

a light source for providing a plurality of light rays;

the collimating light path is used for receiving the plurality of light rays, and collimating the plurality of light rays so that the plurality of light rays mutually parallel reach the waveguide;

the waveguide comprises a plurality of coupling incidence areas, the coupling incidence areas are used for coupling the light rays into the waveguide, the light rays are totally internally reflected in the waveguide and reflected to a target sample, and the microscopic imaging device is used for imaging the target sample.

In one embodiment, the light source is a light emitting diode, a laser diode, or a laser lamp.

In one embodiment, the light source is located directly above the collimating light path, and the light source, the collimating light path and the coupling incidence area form a preset angle.

In one embodiment, each of the coupling regions within the waveguide contains a grating of different parameters for optimizing the plurality of light rays.

In one embodiment, the grating is a relief grating or a holographic grating.

In a second aspect, the present application also provides a scanning imaging method. The scanning imaging method is applied to an illumination device, and the illumination device is any one of the illumination devices described in the foregoing embodiments, and the scanning imaging method includes:

providing a plurality of light rays;

receiving the plurality of light rays, and collimating the plurality of light rays through a collimation light path to enable the plurality of light rays to reach the waveguide in parallel;

and coupling the plurality of light rays into the waveguide through a plurality of coupling incidence areas on the waveguide, and carrying out total internal reflection on the plurality of light rays through the waveguide to reflect the light rays to a target sample so as to enable a microscopic imaging device to image the target sample.

In one embodiment, the target sample is placed above the waveguide, the coupling of the plurality of light rays into the interior of the waveguide through a plurality of coupling-in regions on the waveguide, total internal reflection of the plurality of light rays through the waveguide, reflection to the target sample, to cause a microscopic imaging device to image the target sample, comprising:

optimizing corresponding light rays in the light rays through each coupling incidence area in the coupling incidence areas on the waveguide, coupling the light rays into the waveguide, and carrying out total internal reflection on the light rays to the surface of the waveguide for multiple times through the waveguide to provide illumination for a target sample above the waveguide so as to enable a microscopic imaging device to image the target sample.

In a third aspect, the present application also provides a total internal reflection microscopy imaging system comprising:

a lighting device comprising a light source for providing a plurality of light rays; the collimating light path is used for receiving the plurality of light rays, and collimating the plurality of light rays so that the plurality of light rays mutually parallel reach the waveguide; the waveguide comprises a plurality of coupling incidence areas, a plurality of coupling incidence areas are used for coupling the plurality of light rays into the waveguide, the light rays are totally internally reflected in the waveguide and reflected to a target sample, and the microscopic imaging device is used for imaging the target sample;

the microscopic imaging device is used for microscopic imaging of the target sample.

In one embodiment, the total internal reflection microscopy imaging system further comprises:

a sample holder for holding the target sample;

and the scanning platform is positioned below the microscopic imaging device and is used for controlling the microscopic imaging device to move so as to scan and image the target sample.

In one embodiment, the microimaging device comprises:

an objective lens for magnified imaging of the target sample;

the filter box is used for collecting signals of fluorescence with specific wavelengths;

and the detector is used for collecting target information.

The illumination device, the scanning imaging method and the total internal reflection microscopic imaging system, and the illumination device is characterized in that the device comprises: a light source for providing a plurality of light rays; the collimating light path is used for receiving the plurality of light rays, and collimating the plurality of light rays so that the plurality of light rays mutually parallel reach the waveguide; the waveguide comprises a plurality of coupling incidence areas, the coupling incidence areas are used for coupling the light rays into the waveguide, the light rays are totally internally reflected in the waveguide and reflected to a target sample, and the microscopic imaging device is used for imaging the target sample. By adopting the illumination device, a plurality of light rays are optimized through a plurality of coupling incidence areas in the waveguide, the light rays are coupled into the waveguide, the waveguide totally internally reflects the light rays, a uniform light source is provided for a sample, the imaging view field is illuminated, the imaging minimum multiple of the total internal reflection microscopic imaging system is not limited, and the scanning efficiency is improved.

Drawings

FIG. 1 is a schematic diagram of a total internal reflection microscopy imaging system in accordance with one embodiment;

FIG. 2 is a schematic diagram of the structure of an illumination system in one embodiment;

FIG. 3 is a schematic diagram of a waveguide design for one embodiment with multiple coupling-in regions;

FIG. 4 (a) is a schematic side view of a waveguide in one embodiment;

FIG. 4 (b) is a schematic diagram of the front structure of a waveguide in one embodiment;

FIG. 5 is a flow chart of a scanning imaging method in one embodiment.

Detailed Description

In order to make the objects, technical solutions and advantages of the present application more apparent, the present application will be further described in detail with reference to the accompanying drawings and examples. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the present application.

It should be understood that the terms "first," "second," "third," and "fourth," etc. in the claims, specification, and drawings of this disclosure are used for distinguishing between different objects and not for describing a particular sequential order. The terms "comprises" and "comprising" when used in the specification and claims of the present disclosure, specify the presence of stated features, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof.

It is also to be understood that the terminology used in the description of the present disclosure is for the purpose of describing particular embodiments only, and is not intended to be limiting of the disclosure. As used in the specification and claims of this disclosure, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It should be further understood that the term "and/or" as used in the present disclosure and claims refers to any and all possible combinations of one or more of the associated listed items, and includes such combinations.

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

The illumination device provided in the embodiment of the present application may be applied to a total internal reflection microscopic imaging system 100 as shown in fig. 1. In total internal reflection microscopy imaging system 100, illumination device 110 and microscopy imaging device 120 are included. Optionally, the total internal reflection microscopy imaging system 100 further comprises a scanning platform 130 and a sample holder 140. Wherein, the illumination device 110 is used for providing uniform illumination for the target sample on the sample holder 140, so as to facilitate the microscopic imaging device 120 to perform microscopic imaging on the target sample. The sample holder 140 is used to hold a target sample and the scanning platform 130 is used to control the movement of the microscopic imaging device 120.

Alternatively, the microimaging device 120 may be an integrated imaging device incorporating an objective lens, detector and filter, and various hardware devices for imaging. The embodiment of the present invention does not limit the types and the number of hardware devices that the microscopic imaging device 120 can integrate. Specifically, the internal connection structure and the achievable application functions of the microimaging device 120 are described in detail in the subsequent embodiments of the present invention.

In one embodiment, as shown in fig. 2, an illumination device 110 is provided, such that the illumination device 110 is used in the total internal reflection microscopy imaging system 100 of fig. 1, the illumination device 110 comprising: a light source for providing a plurality of light rays; the collimating light path is used for receiving a plurality of light rays, and collimating the light rays so that the light rays are mutually parallel to reach the waveguide; the light source comprises a waveguide, a light source and a microscopic imaging device, wherein the waveguide comprises a plurality of coupling incidence areas, the coupling incidence areas are used for coupling a plurality of light rays into the interior of the waveguide, and the light rays are totally internally reflected in the waveguide and reflected to a target sample so as to enable the microscopic imaging device to image the target sample.

In practice, the illumination device 110 includes a light source 200, a collimated light path 300, and a waveguide 400. The light source 200 is used for providing a plurality of light rays with different directions. The collimated light path 300 is located below the light source 200. The light rays propagate through the air and a portion of the light rays directed downward reach the collimated light path 300. The parameters of the light rays are different, so that the light rays are divergent. The collimating light path 300 is configured to receive a plurality of light rays, collimate the plurality of light rays, and correct parameters of light in the plurality of light rays to enable the plurality of light rays to be parallel to each other. The light rays reach the waveguide 400 in parallel with each other. The target sample is positioned above the waveguide 400 and the waveguide 400 includes a plurality of coupling-in regions 410. After the multiple light rays reach the different coupling-in regions 410, the different coupling-in regions optimize the multiple light rays and couple the multiple light rays into the interior of the waveguide. The waveguide provides illumination for the target sample by multiple total internal reflections of the plurality of light rays to the target sample to cause the microimaging device 120 to microimage the target sample.

Alternatively, the collimating optical path 300 may be a collimating mirror, a laser collimator, or the like, and the implementation form of the collimating optical path 300 in the embodiment of the present application is not limited.

The lighting device is used for providing a plurality of light rays; the collimating light path is used for receiving a plurality of light rays, and collimating the light rays so that the light rays are mutually parallel to reach the waveguide; the light source comprises a waveguide, a light source and a microscopic imaging device, wherein the waveguide comprises a plurality of coupling incidence areas, the coupling incidence areas are used for coupling a plurality of light rays into the interior of the waveguide, and the light rays are totally internally reflected in the waveguide and reflected to a target sample so as to enable the microscopic imaging device to image the target sample. By adopting the device, the plurality of light rays are optimized through the plurality of coupling incidence areas in the waveguide, the plurality of light rays are coupled into the waveguide, the waveguide totally internally reflects the plurality of light rays, a uniform light source is provided for a sample, the imaging field of view is illuminated, the imaging minimum multiple of the total internal reflection microscopic imaging system is not limited, and the scanning efficiency is improved.

In one embodiment, the light source is a light emitting diode, a laser diode, or a laser lamp.

In implementations, the light source 200 in the lighting device 110 may be a light emitting diode (Light Emitting Diode, LED), a Laser Diode (LD), or a laser lamp.

For example, as shown in fig. 2, the light source 200 is an LED lamp that provides a plurality of divergent light rays in different directions. Wherein the divergent light rays are not parallel to each other.

Optionally, the light source 200 in the lighting device 110 is determined to be a light emitting diode or a laser light according to the requirement. The light emitting diode is convenient to obtain. The laser lamp can provide a light beam with higher quality and higher collimation. The embodiments of the present application are not limited herein with respect to the type of light source.

In this embodiment, the light source provides a plurality of light rays for the lighting device, so that the plurality of light rays can be conveniently processed subsequently. In addition, if the light source is an LED, the convenience of establishing the lighting device can be improved.

In one embodiment, as shown in fig. 2, the light source is located directly above the collimated light path, with the light source, the collimated light path, and the coupling incidence zone being at a predetermined angle.

In implementations, the light source 200 in the illumination device 110 may be located directly above the collimated light path 300. The light source 200, the collimated light path 300 and the coupling incidence area 410 in the illumination device 110 are at a preset angle.

Specifically, as shown in fig. 2, fig. 2 is a schematic structural diagram of the lighting device 110. The light source 200 in the illumination device 110 may employ LEDs and the collimation light path 300 may employ LED collimation light paths. The LED collimation light path is directly below the LEDs. The waveguide 400 in the illumination device 110 is located below the collimated light path of the LED. A coupling-in region 410 within the waveguide 400 is located below the LED collimation path. The LED, LED collimation path, and coupling-in region 410 are on the same vertical line. Wherein, the preset angle may be 0 degrees.

Alternatively, if the light source 200 is a laser lamp, the emission direction of the laser lamp is adjusted to be directly below. Since the plurality of light rays emitted by the laser lamp have high collimation, most of the plurality of light rays provided by the laser lamp can reach the collimation light path 300.

Optionally, the preset angle is determined according to a micro-nano structure design of the grating in the coupling incidence area, and different micro-nano structures have different incidence angles.

In this embodiment, the collimating light path is disposed directly below the light source, so that more light can reach the collimating light path, the collimating light path collimates a plurality of light, and through a preset incident angle, more light can reach the waveguide and reach the target sample through total internal reflection, so as to provide illumination with better quality for the target sample.

In one embodiment, as shown in FIG. 3, each coupling region within the waveguide contains a grating of different parameters for optimizing the plurality of light rays.

In implementation, fig. 3 is a waveguide structure diagram of multiple coupling-in regions. The waveguide 400 in the illumination device 110 contains a plurality of coupling-in regions 410 as seen from the front of the waveguide. x is the length of the waveguide 400 and y is the width of the waveguide 400. 1. 2, 3 are the coupling incidence area 1, the coupling incidence area 2 and the coupling incidence area 3, respectively.

Wherein the waveguide 400 in the illumination device 110 may be a glass sheet of variable thickness. The glass sheet has a plurality of coupling-in regions 410. Each coupling-in region 410 contains a corresponding grating 420. The multiple gratings 420 have different micro-nano structure designs. Wherein micro-nano structure is a nano-scale technology that can be used to design, develop and manufacture micro-structures and mechanical devices. By micro-nanostructure is meant that the micro-structures and mechanical systems are designed, fabricated and perfected on an atomic scale using the correct atomic scale components (typically from microelectronics and biological systems) in combination with precisely controlled mechanical techniques. It uses structures built on the micrometer scale to achieve many physical phenomena such as energy conversion, energy storage, radiation detection, etc. The upper side or surface of the glass sheet is engraved to provide a plurality of coupling-in regions 410, depending on the micro-nano structure design. Because the parameters of the plurality of light rays provided by the light source 200 in the lighting device 110 are different. For example, the plurality of parameters such as wavelength, frequency, etc. of the light are different. Each ray needs to be optimized according to the different parameters of the ray. Thus, different micro-nano structures are designed. The micro-nano structure of each coupling region corresponds to an illumination wavelength.

For example, as shown in fig. 2, a plurality of light rays reach the interior of the waveguide after passing through the coupling incidence region 410 on the waveguide, and the plurality of light rays undergo multiple total internal reflections in the waveguide, resulting in a larger total internal reflection illumination region. The total internal reflection illumination area is shown in fig. 4, and fig. 4 is a schematic view of a waveguide 400. Fig. 4 (a) is a schematic view of a side structure of a waveguide, and fig. 4 (b) is a schematic view of a front structure of a waveguide. According to the waveguide side structure schematic of fig. 4 (a), the coupling-in region 410 is located above or on the surface of the waveguide 400. According to the schematic diagram of the front structure of the waveguide in fig. 4 (b), x and y are the length and width of the waveguide 400, and x1 and y1 are the length and width of the coupling region on the waveguide 400, respectively. The area of the coupling region determines the illumination width of the light source 200 within the waveguide 400. In particular, the length of the total internal reflection may cover the length x of the entire waveguide and the width of the total internal reflection may cover the width y of the entire waveguide. Thus, a large total internal reflection illumination area can be obtained by passing a plurality of light rays through the waveguide 400 and the coupling-in area 410 in the illumination device 110. Thus, the length and width of coupling region 410, the parameters of grating 420 within coupling region 410, and the light source 200 and collimated light path 300 need to be co-designed. By the cooperative design of the light source 200, the collimated light path 300 and the waveguide 400, a large field of view total internal reflection illumination system can be obtained.

Optionally, parameters such as size, depth, and inclination of the grating 420 in the illumination device 110 are designed according to the wavelength range of the plurality of light rays, and the incident angle range is determined. The parameters of grating 420 are not limited in the embodiments of the present application.

Alternatively, the thickness of the waveguide 400 in the lighting device 110 may be 1mm (micrometer) or 2mm, and the specific thickness of the waveguide 400 is not limited in the embodiment of the present application.

In this embodiment, by setting a plurality of coupling regions and gratings, a plurality of light rays incident into the waveguide are optimized for multiple times, and the waveguide performs multiple total internal reflection on the light rays, so that the illumination region of total internal reflection can be enlarged. And according to application requirements, the waveguide can be flexibly designed, so that the total internal reflection illumination range can be designed and optimized, and the light energy utilization rate is improved.

In one embodiment, the grating is a relief grating or a holographic grating.

In implementations, the grating 410 in the illumination device 110 may be a relief grating or a holographic grating. The waveguide 400 or the grating 410 may be subjected to a coating process to improve the light energy utilization of the waveguide.

Alternatively, a surface relief grating may be used as the relief grating. The holographic grating may be a volume holographic grating. A surface relief grating is a grating (similar to a surface engraving) formed after performing photolithography, etching, imprinting, etc. operations on the surface of a substrate. The volume holographic grating is formed by forming interference fringes inside the photosensitive material, and a diffraction grating (similar to internal development) is generated after exposure. Whether the grating is a relief grating or a holographic grating is determined according to requirements. The surface relief grating has a uniform refractive index profile, but the surface microstructure has a period. The volume hologram grating has a periodically varying refractive index profile, but a smooth surface. The embodiment of the application does not limit the form of the grating.

In this embodiment, multiple light rays can be coupled through the relief grating or the holographic grating, so that the light energy utilization rate is improved.

In one embodiment, there is provided a scanning imaging method that can be applied to the illumination device in the above embodiment, as shown in fig. 5, the scanning imaging method including:

step 502 provides a plurality of rays.

In practice, in a lighting device, a plurality of light rays are provided by emitting light to the surroundings by a light source in the lighting device. The propagation paths of the different lights are different, and part of the lights propagate downwards to reach the collimated light rays. Wherein the parameters of the plurality of light rays are different.

Alternatively, the plurality of light rays may be provided by an LED, a laser diode, or a laser lamp, and the embodiment of the present application is not limited herein to the device for providing light rays.

In step 504, a plurality of light rays are received, and the plurality of light rays are collimated by the collimating light path, so that the plurality of light rays reach the waveguide in parallel with each other.

The propagation paths of the light rays are different, so that the light rays diverge, i.e. two adjacent light rays are separated more and more after being propagated.

In implementations, the collimated light path receives a plurality of light rays. The collimating light path collimates the plurality of light rays so that the plurality of light rays reach the waveguide in parallel with each other. Collimation is the process of changing divergent light into parallel light.

In step 506, a plurality of light rays are coupled into the waveguide through a plurality of coupling incidence areas on the waveguide, and are totally internally reflected by the waveguide to the target sample, so that the microscopic imaging device images the target sample.

Wherein there are multiple coupling-in regions on the waveguide.

In practice, multiple rays arrive parallel to each other at the waveguide. The multiple coupling incidence areas on the waveguide optimize the multiple light rays, the multiple light rays are coupled into the waveguide, the multiple light rays are totally internally reflected to the target sample through the waveguide, a uniform light source is provided for the target sample, and the microscopic imaging device is convenient for imaging the target sample.

Specifically, a plurality of light rays reach the waveguide in parallel with each other. Corresponding gratings are present in each coupling-in region. Different gratings have different grating parameters. Depending on the grating parameters, there is a corresponding illumination wavelength for each coupling-in region. Thus, each coupling-in region within the waveguide optimizes a corresponding light ray of the plurality of light rays, coupling the light rays into the interior of the waveguide. The waveguide totally internally reflects a plurality of light rays to the target sample. Total internal reflection, where light enters a medium of lower refractive index from a medium of higher refractive index, if the incident angle is greater than the critical angle (the light is far from normal), the refracted light will disappear and all incident light will be reflected without entering a medium of lower refractive index.

In this embodiment, a plurality of light rays are optimized through a plurality of coupling incidence areas on the waveguide, and are coupled into the waveguide, the waveguide totally internally reflects the plurality of light rays, provides a uniform light source for a sample, illuminates the imaging field of view, and enables the imaging minimum multiple of the total internal reflection microscopic imaging system not to be limited, thereby improving the scanning efficiency.

In one embodiment, the target sample is placed over the waveguide, and the specific process of step 506 includes:

and optimizing corresponding light rays in the light rays through each coupling incidence area in the coupling incidence areas on the waveguide, coupling the light rays into the waveguide, and carrying out multiple total internal reflection on the light rays to the surface of the waveguide through the waveguide to provide illumination for a target sample above the waveguide so as to enable the microscopic imaging device to image the target sample.

In practice, the target sample is placed over the waveguide. Different coupling areas are designed according to different parameters of a plurality of light rays. Each coupling-in region has a corresponding light ray. Each of a plurality of coupling-in regions within the waveguide optimizes a corresponding light ray of the plurality of light rays and couples the light rays into an interior of the waveguide. The waveguide totally internally reflects a plurality of light rays. Each of the plurality of light rays reaches the surface of the waveguide through multiple total internal reflections, providing sufficient illumination for a target sample on the waveguide. Wherein the area illuminated by the plurality of light rays is the entire waveguide in the illumination device. When a focal layer is present in the target sample, light in the light rays totally internally reflected by the waveguide in-coupling entrance area can illuminate the focal layer.

Alternatively, the target sample may be a pathological section or a plant section, etc. The examples of the present application are not limited herein to the target sample.

In this embodiment, placing the target sample over the waveguide can illuminate the target sample, providing sufficient illumination for the target sample. And when the focal plane layer is illuminated by the illumination, fluorescent substances of the non-focal plane layer are prevented from being excited to become a background, and the imaging signal-to-noise ratio is improved.

In one embodiment, as shown in fig. 1, a total internal reflection microscopic imaging system 100 is provided, and the scanning imaging method of the above embodiment can be applied to the total internal reflection microscopic imaging system 100, as shown in fig. 1, where the total internal reflection microscopic imaging system 100 includes:

a lighting device 110 comprising a light source 200 for providing a plurality of light rays; a collimating light path 300, configured to receive a plurality of light rays, and collimate the plurality of light rays so that the plurality of light rays reach the waveguide in parallel with each other; a waveguide 400 having a plurality of coupling-in regions 410, the plurality of coupling-in regions 410 for coupling a plurality of light rays into an interior of the waveguide, the plurality of light rays being totally internally reflected in the waveguide 400 to a target sample for imaging the target sample by the microimaging device 120;

microscopic imaging device 120 is used to microscopic image the target sample.

In practice, 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 lighting device 110 are described in the above embodiments, and the embodiments of the present application are not repeated here. The microscopic imaging device 120 is used for microscopic imaging of the target sample.

Optionally, a plurality of devices, such as an objective lens, a filter cartridge, a detector, etc., are integrated into the microscopic imaging device 120. The number and type of devices integrated with the microscopic imaging device are not limited in the embodiments of the present application.

Alternatively, the current pathological diagnosis is mainly image analysis, and the total internal reflection system 100 based on the waveguide 400 can realize single molecule positioning of biological macromolecules such as proteins, nucleic acids and the like by combining algorithms. Realizing the integration of molecular pathology and histopathology and cytopathology.

In this embodiment, the illumination device and the microscopic imaging device are separated, and the light beams are totally internally reflected by the waveguides having different coupling incidence areas, so that a large-multiple total internal reflection objective lens is not required, the limitation of the large-multiple objective lens on the imaging field of view is avoided, and a large-field total internal reflection illumination system is obtained.

In one embodiment, the total internal reflection system 100 further comprises:

a sample holder 130, the sample holder 130 for holding a target sample;

and the scanning platform 140 is positioned below the microscopic imaging device, and the scanning platform 140 is used for controlling the microscopic imaging device to move so as to scan and image the target sample.

In practice, the total internal reflection system 100 further includes a sample holder 130 and a scanning platform 140. As shown in fig. 1, a sample holder 130 is located below the illumination device 110, and the sample holder 130 is used to hold a target sample. The microscopic imaging device 120 is positioned above a scanning platform 140, which scanning platform 140 is used to control the microscopic imaging device 120 to move left and right so that the illuminated portion of the target sample can be scanned for imaging.

Specifically, the scan stage 140 includes a scan stage controller 141 and a workstation 142. The scanning stage controller 141 is used to control the left and right movement of the microscopic imaging device. The workstation 142 contains a display screen for immediate display of microscopic imaging. A first communication link between the scanning platform controller 141 and the workstation 142. Wherein the scanning platform controller 141 is coupled to a second communication link of the microimaging device for transmitting movement information. Workstation 142 is in third communication with a detector in microscopic imaging device 120. The third communication connection is for transmitting microscopic imaging information.

Wherein the relative positions of the illumination device 110, the target sample and the microimaging device 120 are fixed by the designed structure. The embodiment of the application does not limit the structural member.

In this embodiment, the target sample can be fixed by the sample holder, and the scanning observation position can be flexibly controlled by the scanning platform, so that the observation of the target sample is facilitated.

In one embodiment, a microimaging apparatus includes:

an objective lens 121 for magnified imaging of a target sample;

the filter box 122 is used for collecting signals of fluorescence with specific wavelength;

and a detector 123 for collecting target information.

In practice, microscopic imaging device 120 includes an objective lens 121, a filter cassette 122, and a detector 123. The objective lens 121 is a lens group formed by combining a plurality of lenses. As shown in fig. 1, OBJ in fig. 1 is an objective lens 121. The objective lens 121 is located below the target sample. The object sample can be magnified and imaged by the objective lens 121. The objective lens 121 can be driven to move left and right by moving the microscopic imaging device 120, thereby changing the observation place. By adjusting the objective lens, the magnification of the objective lens can be changed, thereby obtaining a larger observation field of view. The filter cassette 122 is positioned under the objective lens 121, and the filter cassette 122 forms a predetermined angle with the objective lens 121. The filter cartridge 122 is used for signals of fluorescence of a specific wavelength. The detector 123 is at the same level as the cassette 122. The detector 123 is used to collect microscopic imaging information of the target.

In this embodiment, the target sample is scanned and imaged by each integrated device in the microscopic imaging device, so as to facilitate observation of the target sample.

It should be understood that, although the steps in the flowcharts related to the embodiments described above are sequentially shown as indicated by arrows, these steps are not necessarily sequentially performed in the order indicated by the arrows. The steps are not strictly limited to the order of execution unless explicitly recited herein, and the steps may be executed in other orders. Moreover, at least some of the steps in the flowcharts described in the above embodiments may include a plurality of steps or a plurality of stages, which are not necessarily performed at the same time, but may be performed at different times, and the order of the steps or stages is not necessarily performed sequentially, but may be performed alternately or alternately with at least some of the other steps or stages.

The technical features of the above embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.

The above examples only represent a few embodiments of the present application, which are described in more detail and are not to be construed as limiting the scope of the present application. It should be noted that it would be apparent to those skilled in the art that various modifications and improvements could be made without departing from the spirit of the present application, which would be within the scope of the present application. Accordingly, the scope of protection of the present application shall be subject to the appended claims.

Claims (10)

1. A lighting device, the device comprising:

a light source for providing a plurality of light rays;

the collimating light path is used for receiving the plurality of light rays, and collimating the plurality of light rays so that the plurality of light rays mutually parallel reach the waveguide;

the waveguide comprises a plurality of coupling incidence areas, the coupling incidence areas are used for coupling the light rays into the waveguide, the light rays are totally internally reflected in the waveguide and reflected to a target sample, and the microscopic imaging device is used for imaging the target sample.

2. A lighting device as recited in claim 1, wherein said light source is a light emitting diode, a laser diode, or a laser lamp.

3. A lighting device as recited in claim 1, wherein said light source is located directly above said collimated light path, said light source, said collimated light path being at a predetermined angle to said coupling-in region.

4. A lighting device as recited in claim 1, wherein each of said coupling regions within said waveguide comprises a grating of different parameters, said grating being configured to optimize said plurality of light rays.

5. A lighting device as recited in claim 4, wherein said grating is a relief grating or a holographic grating.

6. A scanning imaging method, characterized in that the scanning imaging method is applied to an illumination device as claimed in any one of claims 1 to 5, the scanning imaging method comprising:

providing a plurality of light rays;

receiving the plurality of light rays, and collimating the plurality of light rays through a collimation light path to enable the plurality of light rays to reach the waveguide in parallel;

and coupling the plurality of light rays into the waveguide through a plurality of coupling incidence areas on the waveguide, and carrying out total internal reflection on the plurality of light rays through the waveguide to reflect the light rays to a target sample so as to enable a microscopic imaging device to image the target sample.

7. The scanning imaging method of claim 6, wherein said target sample is placed above said waveguide, said coupling said plurality of light rays into the interior of said waveguide through a plurality of coupling-in regions on said waveguide, said plurality of light rays being totally internally reflected by said waveguide and reflected to the target sample for imaging said target sample by a microscopic imaging device, comprising:

optimizing corresponding light rays in the light rays through each coupling incidence area in the coupling incidence areas on the waveguide, coupling the light rays into the waveguide, and carrying out total internal reflection on the light rays to the surface of the waveguide for multiple times through the waveguide to provide illumination for a target sample above the waveguide so as to enable a microscopic imaging device to image the target sample.

8. A total internal reflection microscopy imaging system, the total internal reflection microscopy imaging system comprising:

a lighting device comprising a light source for providing a plurality of light rays; the collimating light path is used for receiving the plurality of light rays, and collimating the plurality of light rays so that the plurality of light rays mutually parallel reach the waveguide; the waveguide comprises a plurality of coupling incidence areas, a plurality of coupling incidence areas are used for coupling the plurality of light rays into the waveguide, the light rays are totally internally reflected in the waveguide and reflected to a target sample, and the microscopic imaging device is used for imaging the target sample;

the microscopic imaging device is used for microscopic imaging of the target sample.

9. The total internal reflection microscopy imaging system of claim 8, further comprising:

a sample holder for holding the target sample;

and the scanning platform is positioned below the microscopic imaging device and is used for controlling the microscopic imaging device to move so as to scan and image the target sample.

10. The total internal reflection microscopy imaging system of claim 8, wherein the microscopy imaging means comprises:

an objective lens for magnified imaging of the target sample;

the filter box is used for collecting signals of fluorescence with specific wavelengths;

and the detector is used for collecting target information.

Images