A non-labeled far-field super-resolution microscopic spectral imaging system and implementation method

Abstract

The application relates to a non-labeled far-field super-resolution microscopic spectral imaging system and an implementation method, which utilizes the characteristics of a plane achromatic super-resolution lens chip to generate a far-field super-resolution focus point, combines a dynamic adjustable spectral filtering device, does not sacrifice the spatial resolution of spectral imaging, builds the non-labeled far-field super-resolution microscopic spectral imaging system, and overcomes the problem that the existing super-resolution microscopic imaging system cannot obtain spectral information of the target physical and chemical characteristics.

Description

Technical Field

[0001] This invention relates to the field of optoelectronic chips and imaging technology, and more specifically, to a label-free far-field super-resolution microscopic spectral imaging system and its implementation method. Background Technology

[0002] The spatial resolution of optical microscopes is limited by the diffraction limit, making it difficult to exceed 200 nm. Existing super-resolution microscopy techniques, such as stimulated emission depletion (STED), single-molecule localization microscopy (SMLM), and structured illumination microscopy (SIM), all label the samples and only provide morphological information in the spatial dimension, failing to meet the research needs of complex life science questions such as the physicochemical characteristics of samples.

[0003] Although spectral imaging can simultaneously acquire spatial and spectral information of a target, possessing the characteristic of "image and spectrum integration," traditional spectral imaging cannot break through the diffraction limit to acquire spectral information at the spatial scale of a single molecule or even an atom. Therefore, it is urgent to combine super-resolution imaging technology with spectral detection technology to achieve super-resolution analysis of in-depth information such as the physicochemical properties of samples at the molecular and even atomic scales. Summary of the Invention

[0004] The technical problem to be solved by the present invention is how to overcome the difficulty of existing super-resolution microscopic imaging systems in obtaining spectral information of the physicochemical properties of the target. In order to overcome the above-mentioned defects of the prior art, the present invention provides a label-free far-field super-resolution microscopic spectral imaging system and a method for implementing it, comprising a label-free far-field super-resolution microscopic spectral imaging system and a method for implementing label-free far-field super-resolution microscopic spectral imaging.

[0005] This invention provides a label-free far-field super-resolution microscopic spectral imaging system, which includes:

[0006] A planar achromatic super-resolution lens chip is used to provide a far-field super-resolution point light source;

[0007] The stage is equipped with a sample displacement stage for placing the sample to be imaged;

[0008] A beam splitter is used to split light in an optical path to obtain a first beam and a second beam.

[0009] A dynamically adjustable spectral filtering module is used to dynamically split the first beam.

[0010] A spectral signal acquisition module is used to acquire the spectral information output by the dynamically adjustable spectral filtering module;

[0011] An imaging objective lens is used to magnify the second beam in real time to form a magnified super-resolution image;

[0012] An image signal acquisition module is used to acquire the super-resolution image in real time;

[0013] in,

[0014] The planar achromatic super-resolution lens chip, the stage, and the beam splitter are arranged sequentially along the propagation direction of the light emitted by the point light source. The beam splitter, the dynamically adjustable spectral filtering module, and the spectral signal acquisition module are arranged sequentially along the propagation direction of the first beam. The beam splitter, the imaging objective, and the image signal acquisition module are arranged sequentially along the propagation direction of the second beam.

[0015] The label-free far-field super-resolution microscopic spectral imaging system disclosed in this invention employs a planar achromatic super-resolution lens chip to provide a far-field super-resolution point light source. This planar achromatic super-resolution lens chip can achieve focusing beyond the diffraction limit in the far field, demonstrating significant technical advantages in the field of super-resolution imaging. The planar achromatic super-resolution lens chip utilizes micro-nano structures to precisely control the light field behind the lens, achieving specific interference at specific locations and thus generating super-oscillation phenomena. Therefore, the planar achromatic super-resolution lens chip can achieve focusing beyond the diffraction limit in the far field. Furthermore, using a planar achromatic super-resolution lens chip as an imaging illumination source offers advantages such as the ability to overcome the diffraction limit, customization, and mass production using silicon microfabrication processes.

[0016] Meanwhile, this invention constructs a label-free super-resolution microscopic spectral imaging system using a far-field super-resolution focal point generated by a planar super-oscillating lens chip as the incident light source. Considering the very small spatial resolution, a dynamically adjustable spectral filtering module is selected as the spectral dispersive method. This enables online acquisition of spectral information for samples at the molecular and even atomic scales. Combined with real-time acquisition of super-resolution images by an image signal acquisition module, it is possible to obtain the physicochemical characteristics of microscale samples while breaking through the spatial diffraction limit. Furthermore, the use of dynamically adjustable spectral filtering devices is specifically chosen to address the system's high spatial spectral resolution without sacrificing it.

[0017] In one possible implementation, the planar achromatic super-resolution lens chip is a planar super-oscillating lens chip; the use of a planar super-resolution lens chip has far-field super-resolution characteristics, thereby breaking through the working distance limitation of far-field unmarked super-resolution microscopic spectral imaging.

[0018] In one possible implementation, the dynamically adjustable spectral filtering module includes a liquid crystal tunable filter, an acousto-optic tunable filter, or a dynamically adjustable spectral filtering chip; the adjustable spectral filtering module containing one of the above devices can ensure smooth spectral information acquisition without sacrificing the spatial resolution of the system.

[0019] In one possible implementation, the imaging objective lens has a magnification and numerical aperture selected according to the pixel size of the image signal acquisition module, and the focus point size, after magnification, contains at least 3-5 pixels; thereby ensuring that the image signal acquisition module acquires clear information.

[0020] Another technical solution of the present invention is to provide a method for achieving label-free far-field super-resolution microscopic spectral imaging, the method comprising the following steps:

[0021] S1: Design and select planar achromatic super-resolution lens chips for different working distances, focal sizes and working bandwidths;

[0022] S2: Select dynamically adjustable spectral filtering modules with different numbers of channels and working bands as the system's spectroscopic elements;

[0023] S3: The super-resolution microscopic spectral imaging system is built based on the planar achromatic super-resolution lens chip and the dynamically adjustable spectral filtering module;

[0024] S4: The spectral signal acquisition module and the image signal acquisition module are used to acquire the spectral and image information of the sample to be imaged in real time, and to obtain the scanned image sequence and spectral images under different spectral channels;

[0025] S5: The spectral image obtained in step S4 is processed using image stitching algorithms and spectral image processing methods to obtain the final super-resolution microscopic spectral image of the sample.

[0026] The label-free far-field super-resolution microscopic spectral imaging method provided by this invention utilizes the characteristic of planar achromatic super-resolution lens chips to generate far-field super-resolution focal points, and combines the advantage of dynamically adjustable spectral filtering devices without sacrificing the spatial resolution of spectral imaging to build a label-free far-field super-resolution microscopic spectral imaging system. Subsequently, scanning image sequences and spectral images under different spectral channels are obtained, enabling online acquisition of spectral information of samples at the molecular and even atomic scales. This overcomes the problem that existing super-resolution microscopic imaging systems cannot simultaneously obtain spectral information of the physicochemical properties of the target, providing a reliable tool for fields such as biological monitoring, new drug development, and life sciences.

[0027] In one possible implementation, step S3 includes the following steps:

[0028] S31: A plane wave is incident perpendicularly onto the upper surface of the planar achromatic super-resolution lens chip to provide a far-field super-resolution point light source;

[0029] S32: Arrange the planar achromatic super-resolution lens chip, the stage, and the beam splitter sequentially along the propagation direction of the signal emitted by the point light source;

[0030] S33: Place the sample to be imaged on the sample displacement stage of the stage so that the point light source shines on the sample to be imaged;

[0031] S34: The beam passing through the stage is split by a beam splitter to obtain a first beam and a second beam;

[0032] S35: The beam splitter, the dynamically adjustable spectral filtering module, and the spectral signal acquisition module are arranged sequentially along the propagation direction of the first beam, and the beam splitter, the imaging objective, and the image signal acquisition module are arranged sequentially along the propagation direction of the second beam.

[0033] In one possible implementation, step S4, the process of acquiring a spectral image through the spectral signal acquisition module, includes the following steps:

[0034] S41: Place the sample to be imaged on the sample displacement stage of the stage, and set the initial position of the sample displacement stage;

[0035] S42: Use a high-precision timing control algorithm to control the dynamically adjustable spectral filtering module to change a 1-wavelength channel, so as to trigger the spectral signal acquisition module to acquire a spectral image under that channel;

[0036] S43: Determine whether the dynamically adjustable spectral filtering module has switched all channels. If not, return to step S42. If yes, trigger the sample displacement stage to move one unit length along the X-axis and proceed to the next step.

[0037] S44: Determine whether the displacement value of the sample displacement stage along the X-axis has reached the preset range limit. If not, then rotate back to execute step S42; if yes, then trigger the sample displacement stage to return to the initial position and move a second unit length along the Y-axis, and execute the next step.

[0038] S45: Determine whether the displacement value of the sample displacement stage along the Y-axis has reached the preset range limit. If not, proceed to the next step; if yes, stop image acquisition.

[0039] S46: Use a high-precision timing control algorithm to control the dynamically adjustable spectral filtering module to change a 1-wavelength channel, so as to trigger the spectral signal acquisition module to acquire a spectral image under that channel;

[0040] S47: Determine whether the dynamically adjustable spectral filtering module has switched all channels. If not, return to step S46. If yes, trigger the sample displacement stage to move one second unit length along the Y-axis, and then return to step S45.

[0041] This scheme can control the dynamically adjustable spectral filtering module to traverse the spectral images under all spectral channels until the scanning of all points of the target within the entire imaging range is completed, and then stop the acquisition to finally obtain the spectral image, ensuring that the image is accurate and clear.

[0042] In one possible implementation, both the spectral signal acquisition module and the image signal acquisition module are sCMOS sensors; this facilitates the real-time acquisition of the spectral and image information of the target under test, and the acquisition of scanned image sequences and spectral images under different spectral channels. Attached Figure Description

[0043] Figure 1 This is a schematic diagram of the structure of a label-free far-field super-resolution microscopic spectral imaging system disclosed in an embodiment of the present invention;

[0044] Figure 2 This is a flowchart of the method disclosed in an embodiment of the present invention;

[0045] Figure 3 This is a flowchart corresponding to step S4 in the method disclosed in the embodiment of the present invention.

[0046] Explanation of reference numerals in the attached figures:

[0047] 1. Planar achromatic super-resolution lens chip; 2. Sample to be imaged; 3. Stage; 4. Dynamically adjustable spectral filtering module; 5. Spectral signal acquisition module; 6. Beam splitter; 7. Imaging objective lens; 8. Image signal acquisition module. Detailed Implementation

[0048] First, those skilled in the art should understand that these embodiments are merely used to explain the technical principles of the embodiments of this application and are not intended to limit the scope of protection of the embodiments of this application. Those skilled in the art can make adjustments as needed to adapt to specific application scenarios.

[0049] In the embodiments of this application, unless otherwise expressly specified and limited, "above" or "below" the second feature can mean that the first feature is in direct contact with the second feature, or that the first feature is in indirect contact with the second feature through an intermediate medium. Furthermore, "above," "on top of," and "over" the second feature can mean that the first feature is directly above or diagonally above the second feature, or simply that the first feature is at a higher horizontal level than the second feature. "Below," "below," and "under" the second feature can mean that the first feature is directly below or diagonally below the second feature, or simply that the first feature is at a lower horizontal level than the second feature.

[0050] The present application will now be described in further detail with reference to the accompanying drawings and specific embodiments.

[0051] This application discloses a label-free far-field super-resolution microscopic spectral imaging system. (See also...) Figure 1 As shown, the system includes a planar achromatic super-resolution lens chip 1, a stage 3 on which a sample displacement stage for placing the sample to be imaged 2 is provided, a beam splitter 6, a dynamically adjustable spectral filtering module 4, a spectral signal acquisition module 5, an imaging objective lens 7, and an image signal acquisition module 8.

[0052] Please continue reading Figure 1 As shown, in this system, the planar achromatic super-resolution lens chip 1 is used to provide a far-field super-resolution point light source; the beam splitter 6 is used to split the light path to obtain a first beam and a second beam; the dynamically adjustable spectral filtering module 4 is used to dynamically split the first beam; the spectral signal acquisition module 5 is used to acquire the spectral information output by the dynamically adjustable spectral filtering module 4; the imaging objective 7 is used to magnify the second beam in real time to form a magnified super-resolution image; and the image signal acquisition module 8 is used to acquire the super-resolution image in real time. Specifically, the planar achromatic super-resolution lens chip 1, the stage 3, and the beam splitter 6 are arranged sequentially along the propagation direction of the light emitted by the point light source; the beam splitter 6, the dynamically adjustable spectral filtering module 4, and the spectral signal acquisition module 5 are arranged sequentially along the propagation direction of the first beam; and the beam splitter 6, the imaging objective 7, and the image signal acquisition module 8 are arranged sequentially along the propagation direction of the second beam.

[0053] Please continue reading Figure 1 As shown, in this system, the planar achromatic super-resolution lens chip 1 is a planar super-oscillating lens chip. The dynamically adjustable spectral filtering module 4 includes a liquid crystal tunable filter, an acousto-optic tunable filter, or a dynamically adjustable spectral filtering chip. In the imaging objective lens 7: the magnification of the objective lens is set to 100x, and the numerical aperture of the air lens is set to 0.9.

[0054] Please continue reading Figure 1As shown, after the incident plane wave is perpendicularly incident on the upper surface of the planar achromatic super-resolution lens chip 1, it is focused in the far field by the lens, and the resulting point light source hits the sample 2 to be imaged. The sample 2 is placed on the hollow stage 3. The light passing through the stage is split by the beam splitter 6. One beam of light is dynamically split by the dynamically adjustable spectral filtering module 4. The spectral information after splitting is acquired and captured by the spectral signal acquisition module 5. The other beam of light after splitting is magnified by the imaging objective lens 7, and the resulting image is collected and captured by the image signal acquisition module 8. Finally, the two modules simultaneously acquire the image and spectral signal of the same super-resolution point focused on the incident sample.

[0055] The following discloses one spectral imaging implementation method corresponding to this system. See [link to relevant documentation]. Figure 2 and Figure 3 As shown, the method includes the following steps:

[0056] S1: A planar achromatic super-resolution lens chip with a wide band design operating wavelength of 400nm-1000nm, a focal size of 195nm, an operating distance of 1mm, and a diameter of 12mm.

[0057] S2: Select a MEMS-FP dynamically adjustable spectral filter chip as the dynamically adjustable spectral filter module 4. This type of dynamically adjustable spectral filter chip is a spectroscopic element with an operating wavelength of 400nm-650nm and a minimum spectral bandwidth of 20nm.

[0058] S3: See also Figure 1 As shown, a system is built based on a planar achromatic super-resolution lens chip 1 and a dynamically adjustable spectral filtering module 4 as follows: Figure 1 The super-resolution microscopic spectral imaging system is shown. The specific process is as follows:

[0059] S31: A plane wave is incident perpendicularly onto the upper surface of the planar achromatic super-resolution lens chip 1 to provide a far-field super-resolution point light source.

[0060] S32: Arrange the planar achromatic super-resolution lens chip 1, the stage 3, and the beam splitter 6 sequentially along the direction of the signal propagation emitted by the point light source.

[0061] S33: Place the sample 2 to be imaged on the sample displacement stage of the stage 3 so that the point light source shines on the sample 2 to be imaged. The sample 2 to be imaged is a glass slide spin-coated with multiple colored microspheres.

[0062] S34: The beam passing through the stage 3 is split by the beam splitter 6 to obtain the first beam and the second beam.

[0063] S35: See also Figure 1As shown, the beam splitter 6, MEMS-FP dynamically adjustable spectral filtering module 4, and spectral signal acquisition module 5 are arranged sequentially along the propagation direction of the first beam, and the beam splitter 6, imaging objective lens 7, and image signal acquisition module 8 are arranged sequentially along the propagation direction of the second beam.

[0064] S4: The spectral signal acquisition module 5 and the image signal acquisition module 8 are used to acquire the spectral and image information of the sample 2 to be imaged in real time, thereby obtaining the scanned image sequence and spectral images under different spectral channels. In this embodiment, both the spectral signal acquisition module 5 and the image signal acquisition module 8 are high quantum efficiency sCMOS sensors, which is beneficial for the real-time acquisition of the spectral and image information of the target to be measured, and the acquisition of the scanned image sequence and spectral images under different spectral channels.

[0065] See Figure 3 As shown, in step S4, the process of acquiring the spectral image through the spectral signal acquisition module 5 is as follows:

[0066] S41: Place the sample to be imaged 2 on the sample displacement stage of the stage 3, and set the initial position of the sample displacement stage.

[0067] S42: Using a high-precision timing control algorithm, the MEMS-FP dynamically adjustable spectral filtering module 4 changes a wavelength channel to trigger the spectral signal acquisition module 5 to acquire a spectral image under that channel.

[0068] S43: Determine whether the MEMS-FP dynamic adjustable spectral filter module 4 has switched all channels. If not, return to step S42. If yes, trigger the sample displacement stage to move one unit length along the X-axis and proceed to the next step.

[0069] S44: Determine whether the displacement value of the sample displacement stage along the X-axis has reached the preset range limit. If not, then execute step S42. If yes, then trigger the sample displacement stage to return to the initial position and move one second unit length along the Y-axis, and execute the next step.

[0070] S45: Determine whether the displacement value of the sample displacement stage along the Y-axis has reached the preset range limit. If not, proceed to the next step; if yes, stop image acquisition.

[0071] S46: Use a high-precision timing control algorithm to control the MEMS-FP dynamically adjustable spectral filtering module 4 to change a wavelength channel, so as to trigger the spectral signal acquisition module 5 to acquire a spectral image under that channel.

[0072] S47: Determine whether the MEMS-FP dynamic adjustable spectral filter module 4 has switched all channels. If not, return to step S46. If yes, trigger the sample displacement stage to move a second unit length along the Y-axis, and then return to step S45.

[0073] For details, see Figure 3 As shown, firstly, the sample 2 to be imaged is placed on the sample displacement stage of the stage 3, and the initial position 0 of the sample displacement stage is set; the high-precision timing control algorithm controls the dynamically adjustable spectral filtering module to change one wavelength channel, i.e., switch from 400nm to 420nm; the spectral signal acquisition module 5 is triggered to acquire a spectral image under this channel; it is determined whether all channels have been switched and spectral image acquisition has been completed. If not, the timing control of the dynamically adjustable spectral filtering module 4 continues to change one wavelength channel, with a channel switching interval of 20nm; if all spectral channels have been acquired, the sample displacement stage is triggered to move 10nm along the X-axis, and then the image information at the corresponding position is acquired. After acquisition, it is determined whether the displacement value in the X direction has reached the preset value. Set a range limit, with a preset range of 0-20μm. If the limit is not reached, continue timing control of the MEMS-FP dynamically adjustable spectral filtering module 4 to change one wavelength channel, traverse all filtering channels at that position and acquire the corresponding spectral image. Acquire one spectral image every 20nm in the 400nm-650nm wavelength range. Until the X-direction moves to the preset range limit, the displacement stage returns to the initial position in the X-direction and moves one unit length (20nm) in the Y-direction. In sequence, the displacement stage acquires the image information at the corresponding position for each step and adjusts the MEMS-FP dynamically adjustable spectral filtering module 4 to traverse the spectral images under all spectral channels until the scanning of all points of the target within the entire imaging range is completed, at which point the acquisition stops.

[0074] S5: The acquired spectral image is processed using a point-by-point image stitching algorithm and spectral image processing methods to obtain the final super-resolution microscopic spectral image of the sample.

[0075] The system disclosed in this embodiment can acquire the physicochemical characteristics of microscale samples while breaking through the spatial diffraction limit. It employs a planar super-resolution lens chip with far-field super-resolution characteristics, thus overcoming the working distance limitation of far-field label-free super-resolution microscopic spectral imaging. Simultaneously, the spectroscopic element involved in this embodiment is a MEMS-FP dynamically tunable spectral filter device, selected to leverage the system's high spatial spectral resolution without sacrificing its spatial resolution.

[0076] In the description of the embodiments of this application, it should be noted that the terms "inner" and "outer" and other terms indicating direction or positional relationship are based on the direction or positional relationship shown in the drawings. This is only for the convenience of description and does not indicate or imply that the device or component must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, it should not be construed as a limitation of this application.

[0077] In the description of this application, the references to terms such as "an embodiment," "some embodiments," "in this embodiment," "specific example," or "some examples," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in a suitable manner in any one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0078] The above description is merely a specific embodiment of this application, but the scope of protection of this application is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in this application should be included within the scope of protection of this application. Therefore, the scope of protection of this application should be determined by the scope of the claims.

Claims

1. A label-free far-field super-resolution microscopic spectral imaging system, characterized in that, include: A planar achromatic super-resolution lens chip (1) is used to provide a far-field super-resolution point light source; The stage (3) is provided with a sample displacement stage for placing the sample (2) to be imaged; Beam splitter (6) is used to split the optical path to obtain the first beam and the second beam; The dynamically adjustable spectral filtering module (4) is used to dynamically split the first beam. The spectral signal acquisition module (5) is used to acquire the spectral information output by the dynamically adjustable spectral filtering module (4); Imaging objective (7) is used to magnify the second beam in real time to form a magnified super-resolution image; Image signal acquisition module (8) is used to acquire the super-resolution image in real time; in, The planar achromatic super-resolution lens chip (1), the stage (3), and the beam splitter (6) are arranged sequentially along the propagation direction of the light emitted by the point light source. The beam splitter (6), the dynamically adjustable spectral filtering module (4), and the spectral signal acquisition module (5) are arranged sequentially along the propagation direction of the first beam. The beam splitter (6), the imaging objective (7), and the image signal acquisition module (8) are arranged sequentially along the propagation direction of the second beam.

2. The label-free far-field super-resolution microscopic spectral imaging system according to claim 1, characterized in that, The planar achromatic super-resolution lens chip (1) is a planar super-oscillating lens chip.

3. The label-free far-field super-resolution microscopic spectral imaging system according to claim 2, characterized in that, The dynamically adjustable spectral filtering module (4) includes a liquid crystal tunable filter, an acousto-optic tunable filter, or a dynamically adjustable spectral filtering chip.

4. The label-free far-field super-resolution microscopic spectral imaging system according to any one of claims 1-3, characterized in that, In the imaging objective (7): the magnification and numerical aperture of the objective are selected according to the pixel size of the image signal acquisition module (8), and the focus size contains 3-5 pixels after magnification.

5. A method for achieving label-free far-field super-resolution microscopic spectral imaging, characterized in that, The label-free far-field super-resolution microscopic spectral imaging system according to any one of claims 1-4 includes the following steps: S1: Design and select planar achromatic super-resolution lens chips for different working distances, focal sizes and working bandwidths (1). S2: Select dynamic adjustable spectral filtering modules (4) with different numbers of channels and working bands as the system's spectroscopic elements; S3: The super-resolution microscopic spectral imaging system is built based on the planar achromatic super-resolution lens chip (1) and the dynamically adjustable spectral filtering module (4); S4: The spectral signal acquisition module (5) and the image signal acquisition module (8) are used to acquire the spectral and image information of the sample to be imaged (2) in real time, and to obtain the scanned image sequence and spectral images under different spectral channels; S5: The spectral image obtained in step S4 is processed using image stitching algorithms and spectral image processing methods to obtain the final super-resolution microscopic spectral image of the sample.

6. The method for achieving label-free far-field super-resolution microscopic spectral imaging according to claim 5, characterized in that, Step S3 includes the following steps: S31: A plane wave is incident perpendicularly onto the upper surface of the planar achromatic super-resolution lens chip (1) to provide a far-field super-resolution point light source; S32: Arrange the planar achromatic super-resolution lens chip (1), the stage (3), and the beam splitter (6) sequentially along the direction of the light emitted by the point light source; S33: Place the sample to be imaged (2) on the sample displacement stage of the stage (3) so that the point light source hits the sample to be imaged (2); S34: The beam passing through the stage (3) is split by the beam splitter (6) to obtain the first beam and the second beam; S35: The beam splitter (6), the dynamically adjustable spectral filtering module (4), and the spectral signal acquisition module (5) are arranged sequentially along the propagation direction of the first beam, and the beam splitter (6), the imaging objective (7), and the image signal acquisition module (8) are arranged sequentially along the propagation direction of the second beam.

7. The method for achieving label-free far-field super-resolution microscopic spectral imaging according to claim 6, characterized in that, In step S4, the process of acquiring spectral images through the spectral signal acquisition module (5) includes the following steps: S41: Place the sample to be imaged (2) on the sample displacement stage of the stage (3) and set the initial position of the sample displacement stage; S42: Using a high-precision timing control algorithm, the dynamically adjustable spectral filtering module (4) changes a 1-wavelength channel to trigger the spectral signal acquisition module (5) to acquire a spectral image under that channel; S43: Determine whether the dynamically adjustable spectral filtering module (4) has switched all channels. If not, then repeat step S42. If yes, then trigger the sample displacement stage to move one unit length along the X-axis and proceed to the next step. S44: Determine whether the displacement value of the sample displacement stage along the X-axis has reached the preset range limit. If not, then rotate back to execute step S42; if yes, then trigger the sample displacement stage to return to the initial position and move a second unit length along the Y-axis, and execute the next step. S45: Determine whether the displacement value of the sample displacement stage along the Y-axis has reached the preset range limit. If not, proceed to the next step; if yes, stop image acquisition. S46: Using a high-precision timing control algorithm, the dynamically adjustable spectral filtering module (4) changes a 1-wavelength channel to trigger the spectral signal acquisition module (5) to acquire a spectral image under that channel; S47: Determine whether the dynamically adjustable spectral filtering module (4) has switched all channels. If not, then execute step S46 in reverse order. If yes, trigger the sample displacement stage to move one second unit length along the Y-axis, and then execute step S45 in reverse order.

8. The method for achieving label-free far-field super-resolution microscopic spectral imaging according to claim 7, characterized in that, Both the spectral signal acquisition module (5) and the image signal acquisition module (8) are sCMOS sensors.

Images