Unmarked far-field super-resolution microscopic spectrum imaging system and implementation method

Abstract

The invention relates to an unmarked far-field super-resolution microscopic spectrum imaging system and an implementation method, and the system is constructed by using the characteristic that a planar achromatic super-resolution lens chip generates a far-field super-resolution focus and combining the advantage that a dynamic adjustable spectrum filter does not sacrifice the spectral imaging spatial resolution. The problem that a super-resolution microscopic imaging system in the prior art cannot obtain spectral information of target physicochemical properties at the same time is solved.

Description

Technical Field

[0001] The present invention relates to the field of optoelectronic chips and imaging technology, and in particular to a label-free far-field super-resolution micro-spectral imaging system and an implementation method thereof. Background Art

[0002] The spatial resolution of optical microscopes is limited by the diffraction limit and is difficult to exceed 200nm. Existing super-resolution microscopic imaging technologies such as stimulated emission depletion microscopy (STED), single molecule localization super-resolution microscopy (SMLM) and structured illumination microscopy (SIM) continue to label samples and only provide morphological information of samples in the spatial dimension, which cannot meet the research needs of complex life science issues such as the physical and chemical characteristics of samples.

[0003] Although spectral imaging can simultaneously obtain spatial and spectral information of the target and has the characteristic of "image and spectrum in one", traditional spectral imaging cannot break through the diffraction limit and obtain spectral information on the spatial scale of a single molecule or even an atomic scale. Therefore, there is an urgent need to combine super-resolution imaging technology with spectral detection technology to achieve super-resolution analysis of deep information such as the physical and chemical properties of samples at the molecular scale and even the atomic scale. Summary of the invention

[0004] The technical problem to be solved by the present invention is how to overcome the difficulty that the existing super-resolution microscopic imaging system cannot obtain the spectral information of the target physical and chemical properties. In order to overcome the defects of the above-mentioned prior art, the present invention provides a label-free far-field super-resolution microscopic spectral imaging system and an implementation method, including a label-free far-field super-resolution microscopic spectral imaging system and a label-free far-field super-resolution microscopic spectral imaging implementation method.

[0005] The present invention provides a label-free far-field super-resolution micro-spectral imaging system, which comprises:

[0006] Planar achromatic super-resolution lens chip, used to provide far-field super-resolution point light source;

[0007] A stage, provided with a sample displacement stage for placing the sample to be imaged;

[0008] A beam splitter, used for splitting the light path to obtain a first light beam and a second light beam;

[0009] A dynamically adjustable spectral filtering module, used for dynamically splitting the first light beam;

[0010] A spectral signal acquisition module, used for acquiring spectral information output by the dynamically adjustable spectral filtering module;

[0011] An imaging objective lens, used for amplifying the second light beam in real time to form an amplified super-resolution image;

[0012] An image signal acquisition module, used for acquiring the super-resolution image in real time;

[0013] in,

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

[0015] The non-labeled far-field super-resolution micro-spectral imaging system disclosed in the present invention adopts a planar achromatic super-resolution lens chip to provide a far-field super-resolution point light source. The planar achromatic super-resolution lens chip can achieve focusing that breaks the diffraction limit in the far field, and has shown significant technical advantages in the field of super-resolution imaging. The planar achromatic super-resolution lens chip uses micro-nano structures to finely control the light field behind the lens, achieve specific interference at a specific position, and then produce a super-oscillation phenomenon. Therefore, the planar achromatic super-resolution lens chip can achieve focusing that breaks the diffraction limit in the far field. Furthermore, the use of a planar achromatic super-resolution lens chip as a lighting source for imaging has the advantages of being able to break the diffraction limit, being customizable, and being able to be mass-produced by silicon microprocessing.

[0016] At the same time, the present invention uses the far-field super-resolution focus generated by the planar super-oscillating lens chip as the incident light source to build a non-labeled super-resolution micro-spectral imaging system. Considering the problem of very small spatial resolution, the spectral splitting method selects a dynamically adjustable spectral filter module as a spectral splitting method, which can realize the online spectral information acquisition of molecular and even atomic scale samples. Combined with the image signal acquisition module for real-time acquisition of super-resolution images, it can achieve the physical and chemical characteristics of micro-scale samples while breaking through the spatial diffraction limit. In addition, the dynamically adjustable spectral filter device is selected for the high spatial spectral resolution of the system without sacrificing the spatial resolution of the system.

[0017] In a possible implementation, the planar achromatic super-resolution lens chip is a planar super-oscillation lens chip; the planar super-resolution lens chip has far-field super-resolution characteristics, thereby breaking through the working distance limitation of far-field non-label super-resolution micro-spectroscopy imaging.

[0018] In a 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 including one of the above devices can ensure smooth spectral information acquisition without sacrificing the spatial resolution of the system.

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

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

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

[0022] S2: Select dynamically adjustable spectral filter modules with different channel numbers and working bands as system spectroscopic elements;

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

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

[0025] S5: Using an image stitching algorithm and a spectral image processing method to process the spectral image obtained in step S4 to obtain a final sample super-resolution microscopic spectral image.

[0026] The method for realizing label-free far-field super-resolution microscopic spectral imaging provided by the present invention utilizes the characteristic of a planar achromatic super-resolution lens chip to produce a far-field super-resolution focus, combined with the advantage of a dynamically adjustable spectral filter device without sacrificing the spatial resolution of spectral imaging, to build a label-free far-field super-resolution microscopic spectral imaging system, and then obtains a scanning image sequence and spectral images under different spectral channels, which can realize the online spectral information acquisition of molecular and even atomic scale samples, and overcomes the problem that existing super-resolution microscopic imaging systems cannot obtain the spectral information of the target physical and chemical properties, and provides a reliable tool for the fields of biological monitoring, new drug development, life medicine, etc.

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

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

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

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

[0031] S34: splitting the light beam passing through the stage by a beam splitter to obtain a first light beam and a second light beam;

[0032] S35: arrange the beam splitter, the dynamically adjustable spectral filtering module, and the spectral signal acquisition module in sequence along the propagation direction of the first light beam, and arrange the beam splitter, the imaging objective lens, and the image signal acquisition module in sequence along the propagation direction of the second light beam.

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

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

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

[0036] S43: determining whether the dynamically adjustable spectral filter module has switched all channels; if not, returning to step S42; if yes, triggering the sample displacement stage to move a first unit length along the axis X direction, and executing the next step;

[0037] S44: determining whether the displacement value of the sample displacement stage along the axis X direction reaches the preset range limit; if not, returning to the step S42; if yes, triggering the sample displacement stage to return to the initial position and move a second unit length along the axis Y direction, and executing the next step;

[0038] S45: determining whether the displacement value of the sample displacement stage along the axis Y direction reaches the preset range limit, if not, executing the next step; if yes, stopping image acquisition;

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

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

[0041] This solution can control the dynamically adjustable spectral filtering module to traverse the spectral images under all spectral channels until the scanning of all target points within the entire imaging range is completed and the acquisition can be stopped. Finally, the spectral image is obtained to ensure that the image is accurate and clear.

[0042] In a possible implementation, the spectral signal acquisition module and the image signal acquisition module are both sCMOS sensors; this helps to acquire the spectrum and image information of the target to be measured in real time, and obtain the scanning image sequence and the spectral images under different spectral channels. BRIEF DESCRIPTION OF THE DRAWINGS

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

[0044] Figure 2 A flow chart of the method disclosed in the embodiment of the present invention;

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

[0046] Description of reference numerals:

[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 DESCRIPTION

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

[0049] In the embodiments of the present application, unless otherwise clearly specified and limited, a first feature being "above" or "below" a second feature may mean that the first and second features are in direct contact, or the first and second features are in indirect contact through an intermediate medium. Moreover, a first feature being "above", "above" or "above" a second feature may mean that the first feature is directly above or obliquely above the second feature, or simply means that the first feature is higher in level than the second feature. A first feature being "below", "below" or "below" a second feature may mean that the first feature is directly below or obliquely below the second feature, or simply means that the first feature is lower in level than the second feature.

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

[0051] The present application discloses a non-labeling far-field super-resolution micro-spectral imaging system, see 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 a sample 2 to be imaged 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 to see Figure 1 As shown, in this system, the plane 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 light beam and a second light beam; the dynamically adjustable spectral filter module 4 is used to dynamically split the first light beam; the spectral signal acquisition module 5 is used to collect the spectral information output by the dynamically adjustable spectral filter module 4; the imaging objective lens 7 is used to amplify the second light beam in real time to form an amplified super-resolution image; the image signal acquisition module 8 is used to collect the super-resolution image in real time. Among them, the plane achromatic super-resolution lens chip 1, the stage 3, and the beam splitter 6 are arranged in sequence along the propagation direction of the light signal emitted by the point light source, the beam splitter 6, the dynamically adjustable spectral filter module 4, and the spectral signal acquisition module 5 are arranged in sequence along the propagation direction of the first light beam, and the beam splitter 6, the imaging objective lens 7, and the image signal acquisition module 8 are arranged in sequence along the propagation direction of the second light beam.

[0053] Please continue to see 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 filter module 4 includes a liquid crystal tunable filter, an acousto-optic tunable filter or a dynamically adjustable spectral filter chip. In the imaging objective lens 7: the magnification of the objective lens is set to 100 times, and the numerical aperture of the air lens is set to 0.9.

[0054] Please continue to see Figure 1As shown, after the incident plane wave is vertically incident on the upper surface of the plane achromatic super-resolution lens chip 1, it is focused by the far-field super-resolution of the lens, and the generated point light source hits the sample 2 to be imaged. The sample 2 to be imaged is placed on the hollow stage 3. After 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, and the spectral information after the splitting is collected and photographed by the spectral signal acquisition module 5; after the splitting, the other beam of light is amplified by the imaging objective lens 7, and the obtained image is collected and photographed by the image signal acquisition module 8. Finally, the two modules synchronously obtain the image and spectral signal at the same super-resolution point focusing incident sample.

[0055] The following discloses a spectral imaging implementation method corresponding to the system, see Figure 2 and Figure 3 As shown, the method comprises the following steps:

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

[0057] S2: Select a MEMS-FP dynamically adjustable spectrum filter chip as the dynamically adjustable spectrum filter module 4. This dynamically adjustable spectrum filter chip is a spectroscopic element, and its operating band is 400nm-650nm, and its minimum spectrum bandwidth is 20nm.

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

[0059] S31: A plane wave is vertically incident on 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 in sequence along the propagation direction of the light signal emitted by the point light source.

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

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

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

[0064] S4: Use the spectral signal acquisition module 5 and the image signal acquisition module 8 to collect the spectrum and image information of the imaged sample 2 in real time, and obtain the scanning image sequence and the spectral images under different spectral channels. The spectral signal acquisition module 5 and the image signal acquisition module 8 of this embodiment are both high quantum efficiency sCMOS sensors, which are conducive to the real-time acquisition of the spectrum and image information of the target to be measured, and the acquisition of the scanning image sequence and the spectral images under different spectral channels.

[0065] See also Figure 3 As shown, in step S4, the process of collecting the spectral image by the spectral signal collection module 5 is as follows:

[0066] S41: placing the sample 2 to be imaged on the sample displacement stage of the stage 3, and setting 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 is controlled to change a wavelength channel to trigger the spectral signal acquisition module 5 to acquire a spectral image under the channel.

[0068] S43: Determine whether the MEMS-FP dynamically adjustable spectral filter module 4 has switched all channels. If not, return to step S42; if yes, trigger the sample translation stage to move a first unit length along the axis X direction, and execute the next step.

[0069] S44: Determine whether the displacement value of the sample displacement stage along the axis X direction reaches the preset range limit. If not, return to step S42; if yes, trigger the sample displacement stage to return to the initial position and move a second unit length along the axis Y direction, and execute the next step.

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

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

[0072] S47: Determine whether the MEMS-FP dynamically adjustable spectral filter module 4 has switched all channels. If not, return to step S46; if yes, trigger the sample translation 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, first, 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 dynamic adjustable spectral filter module is controlled by a high-precision timing control algorithm to change a wavelength channel, that is, switch from 400nm to 420nm; the spectral signal acquisition module 5 is triggered to acquire a spectral image under the channel; it is determined whether all channels have been switched to perform spectral image acquisition. If all spectral channels have not been acquired, the timing control of the dynamic adjustable spectral filter module 4 is continued to change a wavelength channel, and the channel switching interval is 20nm; if all spectral channels have been acquired, the sample displacement stage is triggered to move 10nm along the X-axis direction, and then the image information at the corresponding position is acquired. After the acquisition, it is determined whether the displacement value in the X direction reaches the preset value. Set a range limit, the preset range is 0-20μm; if it is not reached, continue to control the MEMS-FP dynamically adjustable spectral filter module 4 to change one wavelength channel, traverse all the filter channels at this position and collect the corresponding spectral images, and collect a spectral image every 20nm in the wavelength range of 400nm-650nm; until the X direction moves to the preset range limit, the translation stage returns to the initial position in the X direction and moves 1 unit length (20nm) in the Y direction, and sequentially, the translation stage collects the image information at the corresponding position every step it moves and adjusts the MEMS-FP dynamically adjustable spectral filter module 4 to traverse the spectral images under all spectral channels until the scanning of all target points in the entire imaging range is completed and the collection can be stopped.

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

[0075] The system disclosed in this embodiment can obtain the physical and chemical characteristics of micro-scale samples while breaking through the spatial diffraction limit. The planar super-resolution lens chip has far-field super-resolution characteristics, breaking through the working distance limit of far-field non-label super-resolution micro-spectral imaging. At the same time, the spectroscopic element involved in this embodiment is a MEMS-FP dynamically adjustable spectral filter device, which is selected based on the high spatial spectral resolution of the system without sacrificing the spatial resolution of the system.

[0076] In the description of the embodiments of the present application, it should be noted that in the description of the present application, terms such as "inside" and "outside" indicating directions or positional relationships are based on the directions or positional relationships 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, be constructed and operated in a specific orientation. Therefore, it cannot be understood as a limitation on the present application.

[0077] In the description of the present application, the description with reference to the terms "one embodiment", "some embodiments", "in the present embodiment", "specific example", or "some examples" etc. means that the specific features, mechanisms, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present application. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, mechanisms, materials or characteristics described may be combined in a suitable manner in any one or more embodiments or examples. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0078] The above is only a specific implementation of the present application, but the protection scope of the present application is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed in the present application should be included in the protection scope of the present application. Therefore, the protection scope of the present application shall be based on the protection scope of the claims.

Claims

1. A label-free far-field super-resolution micro-spectral imaging system, characterized in that: include: A planar achromatic super-resolution lens chip (1), used for providing a far-field super-resolution point light source; The object stage (3) is provided with a sample displacement stage for placing the sample (2) to be imaged; A beam splitter (6) is used for splitting the light path to obtain a first light beam and a second light beam; A dynamically adjustable spectral filtering module (4), used for dynamically splitting the first light beam; A spectral signal acquisition module (5), used for acquiring spectral information output by the dynamically adjustable spectral filtering module (4); An imaging objective lens (7) is used to amplify the second light beam in real time to form an amplified super-resolution image; An image signal acquisition module (8), used for acquiring 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 in sequence along the propagation direction of the signal 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 in sequence along the propagation direction of the first light beam; and the beam splitter (6), the imaging objective lens (7), and the image signal acquisition module (8) are arranged in sequence along the propagation direction of the second light beam.

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

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

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

5. A method for realizing label-free far-field super-resolution micro-spectral imaging, characterized in that: The label-free far-field super-resolution micro-spectral imaging system according to any one of claims 1 to 4 comprises the following steps: S1: Design and select planar achromatic super-resolution lens chips for different working distances, focal lengths and working bandwidths (1); S2: Selecting dynamically adjustable spectral filter modules (4) with different numbers of channels and working bands as system spectroscopic elements; S3: constructing the super-resolution microscopic spectral imaging system based on the planar achromatic super-resolution lens chip (3) and the dynamically adjustable spectral filtering module (4); S4: using the spectral signal acquisition module (5) and the image signal acquisition module (8) to collect the spectrum and image information of the sample to be imaged (2) in real time, and obtaining a scanning image sequence and spectral images under different spectral channels; S5: Using an image stitching algorithm and a spectral image processing method to process the spectral image obtained in step S4 to obtain a final sample super-resolution microscopic spectral image.

6. The method for realizing label-free far-field super-resolution micro-spectral imaging according to claim 5, characterized in that: The step S3 comprises the following steps: S31: vertically incident a plane wave onto the upper surface of the planar achromatic super-resolution lens chip (1) to provide a far-field super-resolution point light source; S32: arranging the planar achromatic super-resolution lens chip (1), the stage (3), and the beam splitter (6) in sequence along the propagation direction of the light signal emitted by the point light source; S33: placing the sample to be imaged (2) on the sample displacement stage of the stage (3) so that the point light source is projected onto the sample to be imaged (2); S34: splitting the light beam passing through the stage (3) by a beam splitter (6) to obtain a first light beam and a second light beam; S35: arrange the beam splitter (6), the dynamically adjustable spectral filtering module (4), and the spectral signal acquisition module (5) in sequence along the propagation direction of the first light beam, and arrange the beam splitter (6), the imaging objective lens (7), and the image signal acquisition module (8) in sequence along the propagation direction of the second light beam.

7. The method for realizing label-free far-field super-resolution micro-spectral imaging according to claim 6, characterized in that: In step S4, the process of collecting the spectral image by the spectral signal collection module (5) comprises the following steps: S41: placing the sample to be imaged (2) on the sample displacement stage of the stage (3), and setting the initial position of the sample displacement stage; S42: using a high-precision timing control algorithm to control the 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 the channel; S43: judging whether the dynamically adjustable spectral filtering module (4) has switched all channels; if not, returning to the step S42; if yes, triggering the sample displacement stage to move a first unit length along the axis X direction, and executing the next step; S44: determining whether the displacement value of the sample displacement stage along the axis X direction reaches the preset range limit; if not, returning to the step S42; if yes, triggering the sample displacement stage to return to the initial position and move a second unit length along the axis Y direction, and executing the next step; S45: determining whether the displacement value of the sample displacement stage along the axis Y direction reaches the preset range limit, if not, executing the next step; if yes, stopping image acquisition; S46: using a high-precision timing control algorithm to control the 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 the channel; S47: Determine whether the dynamically adjustable spectral filtering module (4) has switched all channels. If not, return to execute the step S46; if so, trigger the sample displacement stage to move along the Y axis by the second unit length, and then return to execute the step S45.

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

Images