Imaging system, imaging method, and gene sequencer

Abstract

The present application discloses an imaging system, an imaging method, and a gene sequencer. The imaging system comprises a light source assembly, a sample stage, a first light beam rotating assembly, an imaging assembly, and a second light beam rotating assembly; the sample stage bears a sample under test; the first light beam rotating assembly is configured to have a first state and a second state; in the first state, excitation light emitted by the light source assembly irradiates said sample to excite first line-region fluorescence extending in a first direction; in the second state, the excitation light emitted by the light source assembly passes through the first light beam rotating assembly and then irradiates said sample to excite second line-region fluorescence extending in a second direction; and the imaging assembly is used for receiving the first line-region fluorescence at multiple positions to generate a first area array fluorescence image set of a surface region, receiving the second line-region fluorescence at multiple positions to generate a second area array fluorescence image set of a surface region, and synthesizing the first area array fluorescence image set and the second area array fluorescence image set into a super-resolution image.

Description

Imaging system, imaging method and gene sequencer

[0001] The present invention claims priority to Chinese patent application number 202311669425.7, filed with the Patent Office of China on December 7, 2023, entitled “IMAGING SYSTEM AND IMAGING METHOD,” the entire contents of which are incorporated herein by reference. Technical Field

[0002] The present invention relates to the field of gene sequencing and biological sample technology, and more specifically, to an imaging system, an imaging method and a gene sequencer. Background Art

[0003] During the gene sequencing process, a gene sequencer detects the four bases (ATCG) on the chip. Generally, higher data density on the chip leads to lower detection costs. However, detection on high-density chips requires a higher-resolution gene sequencer. If the density of DNA clusters on the chip exceeds the resolution of the microscope objective, the gene sequencer must use super-resolution optical methods to achieve this.

[0004] In related technologies, the imaging system of a gene sequencer can use structured light illumination super-resolution microscopy to achieve higher-resolution imaging. However, traditional structured light illumination solutions require a diffraction device to diffract the laser, and a large amount of energy is wasted in the diffraction process. At the same time, the stripes generated on the focal plane of the microscope objective (sample surface) are all distributed in two dimensions. When the structured light illumination super-resolution microscope performs line scanning imaging, the ratio of the laser energy actually used for imaging (single line) to the total laser energy passing through the objective (two-dimensional plane) is low, and it is difficult to ensure sufficient laser energy. If sufficient laser energy cannot be guaranteed, the signal-to-noise ratio of the imaging cannot be guaranteed, resulting in poor image quality.

[0005] Summary of the Invention

[0006] Embodiments of the present application provide an imaging system, an imaging method using the imaging system, and a gene sequencer.

[0007] An imaging system according to an embodiment of the present application is used in a gene sequencer, comprising:

[0008] A light source assembly, configured to emit excitation light;

[0009] A stage, disposed on the optical path of the light source assembly, and carrying a sample to be tested;

[0010] The first beam rotation assembly is configured to have a first state and a second state, wherein the second state is located in an optical path between the light source assembly and the stage; in the first state, the excitation light emitted by the light source assembly is irradiated onto the sample to be tested to excite fluorescence in a first linear region extending along a first direction; in the second state, the excitation light emitted by the light source assembly is irradiated onto the sample to be tested after passing through the first beam rotation assembly to excite fluorescence in a second linear region extending along a second direction;

[0011] an imaging component, the imaging component being configured to receive a first area array fluorescence image group of a surface area generated by the first line area fluorescence at a plurality of positions, and a second area array fluorescence image group of a surface area generated by the second line area fluorescence at a plurality of positions, wherein the first area array fluorescence image group and the second area array fluorescence image group are used to reconstruct a two-dimensional super-resolution image;

[0012] The second beam rotation assembly is configured to have a first imaging state and a second imaging state, wherein the second imaging state is located on the optical path between the imaging assembly and the stage; in the first imaging state, the imaging assembly receives the first line area fluorescence from multiple positions to generate a first area array fluorescence image group of the surface area; in the second imaging state, the second beam rotation assembly rotates the second line area fluorescence by 90 degrees and enables the imaging assembly to receive the second line area fluorescence from multiple positions to generate a second area array fluorescence image group of the surface area.

[0013] In some embodiments, in the first state, the first beam rotation assembly is in a first position, and the beam rotation assembly is outside the optical path between the light source assembly and the object stage in the first position. In the second state, the first beam rotation assembly is in a second position, and the beam rotation assembly is in the optical path between the light source assembly and the object stage in the second position.

[0014] In some embodiments, the first light beam rotation assembly includes a plurality of reflective mirrors therein, and the plurality of reflective mirrors form a preset angle with the optical axis of the excitation light.

[0015] In some embodiments, the first beam rotation assembly includes a beam entrance and a beam exit; when the beam rotation assembly is in the first position, the reflectors near the beam entrance and the beam exit are moved outside the optical path between the light source assembly and the object stage; when the beam rotation assembly is in the second position, the reflectors near the beam entrance and the beam exit are moved into the optical path between the light source assembly and the object stage.

[0016] In some embodiments, the reflective mirrors near the light beam inlet and the light beam outlet move by rotation, flipping or translation.

[0017] In some embodiments, the reflector includes a first reflector, a second reflector, a third reflector, a fourth reflector, a fifth reflector and a sixth reflector. In the second state, the excitation light passes through the first reflector, the second reflector, the third reflector, the fourth reflector, the fifth reflector and the sixth reflector in sequence. The first reflector is perpendicular to the first plane and forms an angle of 45 degrees with the second plane. The second reflector is perpendicular to the third plane and forms an angle of 22.5 degrees with the first plane. The angles formed by the third reflector and the first plane, the second plane and the third plane are all 45 degrees. The fourth reflector is perpendicular to the first plane and forms an angle of 45 degrees with the second plane. The fifth reflector is perpendicular to the third plane and forms an angle of 22.5 degrees with the second plane. The angles formed by the first plane, the second plane and the third plane of the sixth reflector are all 45 degrees. The first plane, the second plane and the third plane are perpendicular to each other.

[0018] In some embodiments, in the first state, the first reflector and the sixth reflector are out of the optical path of the excitation light.

[0019] In some embodiments, the second beam rotation assembly has the same structure as the first beam rotation assembly.

[0020] In some embodiments, when the first beam rotation assembly is in a first state, the second beam rotation assembly is in a first imaging state; when the first beam rotation assembly is in a second state, the second beam rotation assembly is in a second imaging state.

[0021] In some embodiments, the stage is connected to a drive assembly, and when the first beam rotation assembly is in the first state, the drive assembly drives the stage to move along the second direction, and when the first beam rotation assembly is in the second state, the drive assembly drives the stage to move along the first direction.

[0022] In some embodiments, the imaging system further includes a control component, which, when the first beam rotation component is in the first state, controls the drive component to drive the stage to move along the second direction and controls the light source component to emit the excitation light with a preset switching sequence, so that the excitation light irradiates the sample to be tested along the second direction to excite multiple first line area fluorescence, and there is at least one unirradiated line area between the multiple irradiated line areas; when the first beam rotation component is in the second state, the control component controls the drive component to drive the stage to move along the first direction, and controls the light source component to emit the excitation light with a preset switching sequence, so that the excitation light irradiates the sample to be tested along the second direction to excite multiple second line area fluorescence, and there is at least one unirradiated line area between the multiple irradiated line areas.

[0023] In some embodiments, the imaging system further includes a first dichroic mirror and an objective lens, wherein the objective lens is used to collect the fluorescence and transmit the fluorescence to the first dichroic mirror, and the first dichroic mirror is used to reflect the excitation light to the sample to be measured and to transmit the fluorescence to the imaging component.

[0024] In some embodiments, the imaging assembly includes a plurality of second dichroic mirrors, which distribute the fluorescence to a plurality of imaging channels. The second dichroic mirrors are located between the imaging channels and the first dichroic mirrors. Each of the imaging channels includes a converging lens and a line array camera. The converging lens is used to converge the fluorescence to the line array camera, and the line array camera is used to receive the fluorescence.

[0025] In some embodiments, the number of the second dichroic mirrors is three, and each second dichroic mirror has a different coating, so as to separate four types of fluorescence corresponding to the four base types of ATCG.

[0026] The present application provides an imaging method for an imaging system, wherein the imaging system includes a light source assembly, a stage, a first beam rotation assembly, and an imaging assembly, wherein a sample to be measured is carried on the stage, and the imaging method includes:

[0027] Adjusting the first beam rotation assembly to a first state, and emitting excitation light through the light source assembly to illuminate the sample to be tested and excite fluorescence in a first linear region extending along a first direction;

[0028] Receiving the first line-area fluorescence through the imaging component to generate a first area array fluorescence image group of an area;

[0029] Adjusting the first light beam rotating assembly to a second state, and irradiating the sample to be tested with excitation light to excite fluorescence in a second linear region extending along a second direction, wherein the second direction is perpendicular to the first direction;

[0030] receiving the second linear region fluorescence again through the imaging component to generate a second area array fluorescence image group of an area;

[0031] The imaging component combines the first area array fluorescence image group and the second area array fluorescence image group into a super-resolution image.

[0032] In some embodiments, the method for generating the first area array fluorescence image includes:

[0033] The stage is driven by a driving component to move the sample to be tested along the second direction, and the light source component is controlled to make the excitation light irradiate the sample to be tested along the second direction and excite multiple first line area fluorescences. There is at least one unirradiated first line area among the multiple first line area fluorescences. Each time a first line area fluorescence is excited, the imaging component can generate the first area array fluorescence image of a surface area.

[0034] In some embodiments, the method for generating the second area array fluorescence image includes:

[0035] The stage is driven by a driving component to move the sample to be tested along the first direction, and the light source component is controlled to make the excitation light irradiate the sample to be tested along the first direction and excite multiple second-line area fluorescences. There is at least one unirradiated second line area among the multiple second-line area fluorescences. Each time a second-line area fluorescence is excited, the imaging component can generate a second area array fluorescence image of a surface area.

[0036] A gene sequencer according to an embodiment of the present application includes the imaging system described in any embodiment of the present application.

[0037] In the imaging system of the embodiment of the present application, excitation light is emitted by the light source component, and the excitation light is rotated by the beam rotation unit to form orthogonal excitation light, and then images are collected respectively by the imaging components to form super-resolution imaging. Compared with traditional super-resolution imaging, this scheme uses line scanning to achieve super-resolution imaging with high energy utilization, no energy waste, and can ensure sufficient laser energy, which is beneficial to improving image quality. At the same time, this scheme uses excitation light in two orthogonal directions to form super-resolution imaging, which can greatly improve the resolution in the orthogonal directions.

[0038] Additional aspects and advantages of the embodiments of the present application will be given in part in the description below, and in part will become obvious from the description below, or will be learned through practice of the embodiments of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] The above and / or additional aspects and advantages of the present application will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

[0040] FIG1 is a schematic diagram of the main structure of an imaging system according to an embodiment of the present application;

[0041] FIG2 is a schematic diagram of a module of an imaging system according to an embodiment of the present application;

[0042] FIG3 is a schematic structural diagram of an imaging system according to an embodiment of the present application;

[0043] 4 to 6 are schematic diagrams of the excitation light rotation process according to an embodiment of the present application;

[0044] 7 and 8 are schematic diagrams of the internal structure of the first light beam rotation assembly at different angles according to an embodiment of the present application;

[0045] 9 to 11 are schematic diagrams of two-dimensional imaging of structured light illumination stripes in the X direction according to an embodiment of the present application;

[0046] 12 to 17 are schematic diagrams of the first light beam rotation assembly in the first state irradiating the excitation light according to an embodiment of the present application;

[0047] 18 to 20 are schematic diagrams of two-dimensional imaging of structured light illumination stripes in the Y direction according to an embodiment of the present application;

[0048] 21 to 26 are schematic diagrams of the irradiation of the excitation light by the first light beam rotation assembly in the second state according to an embodiment of the present application;

[0049] FIG27 is a flow chart of an imaging method according to an embodiment of the present application.

[0050] Component symbol explanation: imaging system 100, light source assembly 10, excitation light 11, laser emitter 12, beam shaper 13, multimode optical fiber 14, stage 20, first beam rotation assembly 30, first reflector 31, second reflector 32, third reflector 33, fourth reflector 34, fifth reflector 35, sixth reflector 36, beam entrance 37, beam exit 38, imaging assembly 40, second dichroic mirror 41, imaging channel 42, converging lens 421, line array camera 422, filter 423, sample to be measured 50, first line area fluorescence 51, second line area fluorescence 52, imaging area 53, first line area 54, second line area 55, first dichroic mirror 61, objective lens 62, drive assembly 70, control assembly 80, second beam rotation assembly 90. DETAILED DESCRIPTION

[0051] The following further describes the embodiments of the present application in conjunction with the accompanying drawings. The same or similar reference numerals in the accompanying drawings represent the same or similar elements or elements with the same or similar functions.

[0052] In addition, the embodiments of the present application described below in conjunction with the accompanying drawings are exemplary and are only used to explain the embodiments of the present application and should not be understood as limiting the present application.

[0053] In this application, unless otherwise expressly specified or limited, when a first feature is "above" or "below" a second feature, it 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. Furthermore, when a first feature is "above," "above," or "above" a second feature, it may mean that the first feature is directly above or diagonally above the second feature, or simply means that the first feature is at a higher level than the second feature. When a first feature is "below," "below," or "below" a second feature, it may mean that the first feature is directly below or diagonally below the second feature, or simply means that the first feature is at a lower level than the second feature.

[0054] During the sequencing process, gene sequencers require fluorescence imaging of each of the four bases (ATCG). Because each target point is located at the submicron scale, gene sequencers inevitably employ microscopic imaging to achieve detection. A key metric for gene sequencers is the density of the dots on the chip, which determines the amount of data on a chip and, consequently, the cost per unit of data. Because the amount of reagents used per unit area is minimally affected by dot density, higher dot density results in lower detection costs.

[0055] High-density chips require higher-resolution microscopes. However, traditional wide-field microscopes are diffraction-limited, with resolutions typically capped at around 200nm. To achieve higher resolution and thus higher chip point density, "super-resolution" optical techniques are required.

[0056] In the field of gene sequencing, imaging speed is an important indicator that determines the time it takes to output a sequencing report. In order to further improve the speed of imaging detection, the use of line scanning imaging solutions is the future trend. Traditional SIM solutions require diffraction and spectroscopy through devices such as gratings, spatial light modulators, and digital micromirrors (DMDs). The diffracted ±1-order light beams interfere on the focal plane of the objective lens (sample surface) to produce sinusoidal fringes to form structured light illumination. Alternatively, non-diffraction spectroscopy solutions implemented by later-appearing optical devices such as beam splitters and prisms can also form interference fringes on the focal plane of the objective lens (sample surface).

[0057] However, the biggest problem with traditional solutions is their extremely low energy utilization. First, the energy utilization of the diffraction spectrometry solution is limited by the diffraction efficiency of the diffraction device, and it cannot be fully transmitted to the back focal plane of the objective lens, resulting in a huge waste of energy. Secondly, whether it is a diffraction spectrometry or a non-diffraction spectrometry solution, the stripes generated on the focal plane of the objective lens (sample surface) are distributed in two dimensions. For line scanning imaging systems, the ratio of the laser energy actually used for imaging (single line) to the total laser energy passing through the objective lens (two-dimensional plane) is also extremely low. The imaging speed of the line scanning imaging system is much faster than that of the traditional surface scanning imaging system. As a result, the exposure time of a single pixel of the line array camera is much shorter than that of a single pixel of the area array camera. If sufficient laser energy cannot be guaranteed, the signal-to-noise ratio of the imaging cannot be guaranteed, resulting in poor image quality.

[0058] In view of this, please refer to Figures 1 to 27. An embodiment of the present application provides an imaging system 100, which is used for a gene sequencer. The imaging system 100 includes a light source assembly 10, a stage 20, a first beam rotation assembly 30, an imaging assembly 40 and a second beam rotation assembly 90.

[0059] The light source assembly 10 is used to emit excitation light 11, the stage 20 is arranged on the optical path of the light source assembly 10, and the stage 20 carries a sample to be tested 50. The first light beam rotation assembly 30 is configured to have a first state and a second state. The second state is located on the optical path between the light source assembly 10 and the stage 20. In the first state, the excitation light 11 emitted by the light source assembly 10 is irradiated to the sample to be tested 50 to excite a first line area fluorescence 51 extending along a first direction. In the second state, the excitation light 11 emitted by the light source assembly 10 is irradiated to the sample to be tested 50 after passing through the first light beam rotation assembly 30 to excite a second line area fluorescence 52 extending along a second direction. The imaging assembly 40 is used to receive the first line area fluorescence 51 at multiple positions to generate a first area array fluorescence image of the surface area. The imaging assembly 40 is configured to receive the second line-area fluorescence 52 at multiple positions to generate a second area array fluorescence image group of the surface area, and reconstruct a two-dimensional super-resolution image based on the first area array fluorescence image group and the second area array fluorescence image group. The second beam rotation assembly 90 is configured to have a first imaging state and a second imaging state, and the second imaging state is located on the optical path between the imaging assembly 40 and the stage 20. In the first imaging state, the imaging assembly 40 receives the first line-area fluorescence 51 at multiple positions to generate a first area array fluorescence image of the surface area. In the second imaging state, the second beam rotation assembly 90 rotates the second line-area fluorescence 52 by 90 degrees and enables the imaging assembly 40 to receive the second line-area fluorescence 52 at multiple positions to generate a second area array fluorescence image of the surface area.

[0060] In the imaging system 100 of the embodiment of the present application, the excitation light 11 is emitted by the light source component 10, and the excitation light 11 is rotated by the beam rotation unit to form orthogonal excitation light 11, and then images are collected respectively by the imaging component 40 to form super-resolution imaging. Compared with traditional super-resolution imaging, the present solution uses line scanning to achieve high energy utilization rate of super-resolution imaging, without energy waste, and can ensure sufficient laser energy, which is beneficial to improving the image signal-to-noise ratio. At the same time, the present solution uses excitation light 11 in two orthogonal directions and forms super-resolution imaging, which can greatly improve the resolution in the orthogonal directions.

[0061] Specifically, the sample to be tested 50 is a biological sample with a fluorescent dye, which is set on a gene sequencing chip, and the gene sequencing chip is placed on the stage 20. When the first light beam rotation component 30 is in the first state, the excitation light 11 emitted by the light source component 10 is irradiated on the sample to be tested 50 to form a first light spot. The first light spot excites the fluorescent dye on the biological sample to form a first line area fluorescence 51. The light spot and the first line area fluorescence 51 both extend in the first direction X. When the first light beam rotation component 30 is in the second state, the excitation light 11 emitted by the light source component 10 is irradiated on the sample to be tested 50 to form a second light spot. The second light spot excites the fluorescent dye on the biological sample to form a second line area fluorescence 52. The second light spot and the second area fluorescence both extend in the second direction Y.

[0062] Please refer to Figure 3. In some embodiments, the imaging system 100 also includes a first dichroic mirror 61 and an objective lens 62. The objective lens 62 is used to collect fluorescence and transmit the fluorescence to the first dichroic mirror 61. The first dichroic mirror 61 is used to reflect the excitation light 11 to the sample to be measured 50 and to transmit the fluorescence to the imaging component 40.

[0063] In some embodiments, the light source assembly 10 includes a laser emitter 12 and a beam shaper 13. The laser emitter 12 is used to emit laser light, and the beam shaper 13 is used to shape the laser light into excitation light 11. Specifically, the laser emitter 12 can emit laser light of a specific wavelength to excite the sample 50 to produce fluorescence. The light source assembly 10 can also include a multimode optical fiber 14. The multimode optical fiber 14 is located between the laser emitter 12 and the beam shaper 13. The laser emitter 12 is connected to the beam shaper 13 via the multimode optical fiber 14.

[0064] In some embodiments, the imaging assembly 40 includes multiple second dichroic mirrors 41, which distribute fluorescence to multiple imaging channels 42. The second dichroic mirrors 41 are located between the imaging channels 42 and the first dichroic mirror 61. Each imaging channel 42 includes a converging lens 421 and a line array camera 422. The converging lens 421 is used to converge fluorescence to the line array camera 422, and the line array camera 422 is used to receive fluorescence.

[0065] Specifically, since the photosensitive element of the line array camera 422 is one-dimensional, it can only image one dimension of the sample when not scanning. When the first beam rotation component 30 rotates the excitation light 11 90°, the second line area fluorescence 52 formed extends along the second direction Y. At this time, the line array camera 422 needs to be rotated. Otherwise, the second line area fluorescence 52 at this time cannot be identified and imaged. Therefore, a second beam rotation component 90 is set in the optical path between the imaging component 40 and the stage 20. The second beam rotation component 90 rotates the second line area fluorescence 52 and transmits it to the line array camera 422. The extension direction of the second line area fluorescence 52 after rotation is the first direction X. At this time, the line array camera 422 can perform detection and identification without rotating the line array camera 422, thereby simplifying the operation difficulty.

[0066] In some embodiments, the imaging assembly 40 includes a filter 423 . The filter 423 is located between the converging lens 421 and the linear array camera 422 . The filter 423 is used to filter fluorescence.

[0067] Please refer to Figure 3. In some embodiments, in the first state, the first beam rotation assembly 30 is in a first position, and the first position is outside the optical path between the light source assembly 10 and the object stage 20. In the second state, the first beam rotation assembly 30 is in a second position, and the second position is located on the optical path between the light source assembly 10 and the object stage 20. Specifically, the first beam rotation assembly 30 can be movably set between the first position and the second position. When the first beam rotation assembly 30 is in the first position, the excitation light 11 does not enter the first beam rotation assembly 30 and directly shines on the object stage 20. When the first beam rotation assembly 30 is in the second position, the excitation light 11 enters the first beam rotation assembly 30, and the first beam rotation assembly 30 rotates the excitation light 11 90° before shining it on the object stage 20.

[0068] In some embodiments, multiple reflectors are disposed inside the first beam rotation assembly 30, and the multiple reflectors and the optical axis of the excitation light 11 form a preset angle. Specifically, the excitation light 11 enters from the beam entrance 37 of the first beam rotation assembly 30, is reflected and rotated by the multiple reflectors, and then is emitted from the beam exit 38.

[0069] Please refer to Figures 7 and 8. In some embodiments, the reflector includes a first reflector 31, a second reflector 32, a third reflector 33, a fourth reflector 34, a fifth reflector 35 and a sixth reflector 36. In the second state, the excitation light 11 passes through the first reflector 31, the second reflector 32, the third reflector 33, the fourth reflector 34, the fifth reflector 35 and the sixth reflector 36 in sequence. The first reflector 31 is perpendicular to the first plane and forms an angle of 45 degrees with the second plane. The second reflector 32 is perpendicular to the third plane and forms an angle of 22.5 degrees with the first plane. The angles formed by the third reflector 33 with the first plane, the second plane and the third plane are all 45 degrees. The fourth reflector 34 is perpendicular to the first plane and forms an angle of 45 degrees with the second plane. The fifth reflector 35 is perpendicular to the third plane and forms an angle of 22.5 degrees with the second plane. The angles formed by the first plane, the second plane and the third plane of the sixth reflector 36 are all 45 degrees. Among them, the first plane, the second plane and the third plane are perpendicular to each other.

[0070] Please refer to Figures 4 to 6 as well as Figures 7 and 8. Specifically, the direction of the optical axis of the excitation light 11 is the third direction Z, the first plane is denoted as XOY, the second plane is denoted as ZOY, and the third plane is denoted as XOZ. As shown in Figure 4, when the light shaped by the beam shaper 13 enters the beam rotation optical path system through the beam entrance 37, its light intensity distribution is an ellipse with the major axis along the X-axis direction. The excitation light 11 enters the first beam rotation component 30 from the beam entrance 37 and is first reflected by the first reflector 31. The angle between the normal of the plane where the first reflector 31 is located and the optical axis of the incident excitation light 11 on the XOZ plane is 45°. After reflection, the excitation light 11 reaches the second reflector 32. The angle between the normal of the plane where the second reflector 32 is located and the optical axis of the incident excitation light 11 on the XOY plane is 22.5°. Then, the excitation light 11 is reflected by the second reflector 32 to the third reflector 33, wherein the normal of the plane where the third reflector 33 is located is 22.5°. The angle between the excitation light 11 and the optical axis of the incident excitation light 11 on the XOZ plane is 45°; as shown in Figure 5, the shape of the light spot intensity distribution after the excitation light 11 is reflected is still an ellipse, but the direction of the major axis of the ellipse has changed, forming an angle of 45° with the X-axis; the excitation light 11 is then reflected by the fourth reflector 34, wherein the angle between the normal of the plane where the fourth reflector 34 is located and the optical axis of the incident excitation light 11 on the XOZ plane is 45°, and then the excitation light 11 reaches the fifth reflector 35, and the angle between the normal of the plane where the fifth reflector 35 is located and the optical axis of the incident excitation light 11 on the XOY plane is 22.5°; then the fifth reflector 35 reflects the excitation light 11 to the sixth reflector 36, and the angle between the normal of the plane where the sixth reflector 36 is located and the optical axis of the incident excitation light 11 on the XOZ plane is 45°, and finally the sixth reflector 36 reflects the excitation light 11 along the beam outlet 38 out of the first beam rotation component 30. As shown in Figure 6 , the intensity of the light spot after reflection by the sixth reflector 36 remains elliptical, with the major axis of the ellipse parallel to the Y-axis. This means that the intensity distribution of the excitation light 11 at the beam exit 38 is rotated 90° about the Z-axis compared to the intensity distribution of the excitation light 11 at the beam entrance 37. Furthermore, by rationally designing the spatial positions of the individual reflectors, it is ensured that the excitation light 11 at the beam exit 38 is emitted directly in front of the beam entrance 37 in the Z-axis direction. This ensures that the rotation of the excitation light 11 in and out of the optical path system will not affect the original directionality of the excitation light 11.

[0071] Referring to Figures 7 and 8, in some embodiments, in a first state, the first reflector 31 and the sixth reflector 36 are out of the optical path of the excitation light 11. Specifically, the first beam rotation assembly 30 is fixedly disposed in the optical path between the light source assembly 10 and the stage 20. When the first beam rotation assembly 30 is in the first state, the first reflector 31 and the sixth reflector 36 move out of the optical path, and the excitation light 11 enters the first beam rotation assembly 30 from the beam inlet 37 and is directly emitted from the beam outlet 38 without being reflected. When the first beam rotation assembly 30 is in the second state, the first reflector 31 and the sixth reflector 36 move into the optical path, and the excitation light 11 enters the first beam rotation assembly 30 from the beam inlet 37 and is emitted from the beam outlet 38 after being reflected. Optionally, the first reflector 31 and the sixth reflector 36 can move by rotation, flipping, or translation. This configuration eliminates the need to set two positions for the first beam rotation assembly 30, which helps reduce the size of the imaging system 100.

[0072] In certain embodiments, the length of the images of the first and second line-area fluorescence 51, 52 on the photosensitive element of the imaging assembly 40 is less than or equal to the length of the photosensitive element, and the width of the images of the first and second line-area fluorescence 51, 52 on the photosensitive element of the imaging assembly 40 is less than or equal to the width of a pixel of the photosensitive element of the imaging assembly 40. Specifically, taking the first beam rotation assembly 30 in the first state as an example, if the width of the first line-area fluorescence 51 is greater than the width of a pixel of the photosensitive element, the first line-area fluorescence 51 will interfere with the imaging effect of adjacent areas, thereby reducing the frequency of fringes in the first area array fluorescence image and the resolution of the synthesized super-resolution image. Therefore, when the width of the images of the first and second line-area fluorescence 51, 52 on the photosensitive element of the imaging assembly 40 is less than or equal to the width of a pixel of the photosensitive element of the imaging assembly 40, the imaging effect is optimal.

[0073] Please refer to Figures 3 and 7 to 8. In some embodiments, the internal structure of the second beam rotation assembly 90 is the same as that of the first beam rotation assembly 30. Specifically, six reflectors are also provided inside the second beam rotation assembly 90, and the installation positions of the six reflectors are the same as those of the first beam rotation assembly 30. In other embodiments, the second beam rotation assembly 90 can also be other structures that can achieve the same effect.

[0074] In some embodiments, when the first beam rotation assembly 30 is in the first state, the second beam rotation assembly 90 is in the first imaging state; when the first beam rotation assembly 30 is in the second state, the second beam rotation assembly 90 is in the second imaging state. Specifically, the first beam rotation assembly 30 and the second beam rotation assembly 90 are linked, and the first beam rotation assembly 30 and the second beam rotation assembly 90 can be driven by the same driving method, which helps to reduce the difficulty of operation.

[0075] Please refer to Figure 2. In some embodiments, the worktable 20 is connected to the driving component 70. When the first beam rotation component 30 is in the first state, the driving component 70 drives the worktable 20 to move along the second direction. When the first beam rotation component 30 is in the second state, the driving component 70 drives the worktable 20 to move along the first direction.

[0076] Please refer to Figures 2 and 3. In some embodiments, the imaging system 100 further includes a control component 80. When the first beam rotation component 30 is in the first state, the control component 80 controls the drive component 70 to drive the stage 20 to move along the second direction and controls the light source component 10 to emit the excitation light 11 with a preset switching sequence, so that the excitation light 11 irradiates the sample to be tested 50 along the second direction to excite multiple first line area fluorescence 51, and there is at least one unirradiated line area between the multiple irradiated line areas; when the first beam rotation component 30 is in the second state, the control component 80 controls the drive component 70 to drive the stage 20 to move along the first direction, and controls the light source component 10 to emit the excitation light 11 with a preset switching sequence, so that the excitation light 11 irradiates the sample to be tested 50 along the second direction to excite multiple second line area fluorescence 52, and there is at least one unirradiated line area between the multiple irradiated line areas.

[0077] Please refer to Figures 2 and 3. Specifically, the control component 80 can control the movement of the stage 20 through the drive component 70, thereby controlling the movement of the sample to be tested 50. While keeping the light source component 10 stationary, the control component 80 controls the movement of the stage 20 along the second direction Y, that is, the sample to be tested 50 moves relative to the excitation light 11 along the second direction Y, so that the multiple line areas sweep across the irradiation area along the second direction Y. It will be understood that the excitation light 11 can irradiate the sample to be tested 50 in a direction perpendicular to the surface of the sample to be tested 50, or can irradiate the sample to be tested 50 in other directions, while keeping the light spot of the excitation light 11 on the sample to be tested 50 extending along the first direction X.

[0078] When the preset switch timing is in the on period, the light source assembly 10 emits excitation light 11 to excite the sample to be tested 50 to emit first line area fluorescence 51 or second line area fluorescence 52; when the preset switch timing is in the off period, the light source assembly 10 turns off the excitation light 11 to form an un-irradiated first line area 54 or second line area 55.

[0079] Please refer to FIG3 . In some embodiments, the number of imaging channels 42 is four. Specifically, the gene sequencer needs to perform fluorescence imaging of the four ATCG bases during the sequencing process, and the four imaging channels 42 correspond to the four ATCG bases respectively.

[0080] In some embodiments, there are three second dichroic mirrors 41 . Specifically, the three second dichroic mirrors 41 are of different types. Furthermore, the three second dichroic mirrors 41 have different coatings. The four ATCG bases form four types of fluorescence. At least three second dichroic mirrors 41 are required to separate the four fluorescences one by one.

[0081] Referring to Figure 3 , specifically, the optical path of the excitation light 11 is as follows: laser light emitted by the laser emitter 12 is transmitted through the multimode optical fiber 14 into the beam shaper 13 to form the excitation light 11. The excitation light 11 is reflected by the first dichroic mirror 61 and illuminates the sample 50 through the objective lens 62. The fluorescence stimulated by the excitation light 11 is collected by the objective lens 62, passes through the first dichroic mirror 61, and is then split into four paths by three second dichroic mirrors 41 and transmitted to the four line scan cameras 422, where images of the four ATCG bases are respectively generated. The first beam rotation assembly 30 is then adjusted to rotate the excitation light 11. The excitation light 11 is reflected by the first dichroic mirror 61 and illuminates the sample 50 through the objective lens 62. The fluorescence stimulated by the excitation light 11 is collected by the objective lens 62, passes through the first dichroic mirror 61, and is then split into four paths by three second dichroic mirrors 41 and transmitted to the four line scan cameras 422, where images of the four ATCG bases are again generated.

[0082] Referring to FIG. 2 and FIG. 27 , an embodiment of the present application provides an imaging method for an imaging system 100 . The imaging system 100 includes a light source assembly 10 , a stage 20 , a first beam rotation assembly 30 , and an imaging assembly 40 . A sample 50 to be measured is carried on the stage 20 . The imaging method includes:

[0083] Step 01: Adjust the first beam rotating assembly to a first state, and emit excitation light through the light source assembly to illuminate the sample to be tested to excite fluorescence in a first linear region extending along a first direction;

[0084] Step 02: Receive a first line area fluorescence through an imaging component to generate a plurality of first area array fluorescence images of a surface area;

[0085] Step 03: Adjust the first beam rotating assembly to a second state, and emit excitation light to illuminate the sample to be tested to excite fluorescence in a second linear region extending along a second direction; the second direction is perpendicular to the first direction;

[0086] Step 04: receiving the second line area fluorescence again through the imaging component to generate a plurality of second area array fluorescence images of a surface area;

[0087] Step 05: Synthesize a super-resolution image based on the multiple first array fluorescence images and the multiple second array fluorescence images through an imaging component.

[0088] Specifically, the imaging system 100 also includes a control component 80, which adjusts the first beam rotation component 30 to a first state, and emits excitation light 11 through the light source component 10 to irradiate the sample to be tested 50 to excite a first line area fluorescence 51 extending along a first direction, and then the control component 80 controls the imaging component 40 to receive the first line area fluorescence 51 to generate multiple first area array fluorescence images of a surface area; then the control component 80 adjusts the first beam rotation component 30 to a second state, and irradiates the sample to be tested 50 by emitting excitation light 11 to excite a second line area fluorescence 52 extending along a second direction; the second direction is perpendicular to the first direction; the control component 80 again receives the second line area fluorescence 52 through the imaging component 40 to generate multiple second area array fluorescence images of a surface area; finally, the imaging component 40 synthesizes a super-resolution image based on the multiple first area array fluorescence images and the multiple second area array fluorescence images.

[0089] Furthermore, the method for generating the first area array fluorescence image includes:

[0090] The driving component 70 drives the stage 20 to move the sample to be tested 50 along the second direction, and at the same time controls the light source component 10 so that the excitation light 11 irradiates the sample to be tested 50 along the second direction and excites multiple first line area fluorescences 51. There is at least one unirradiated first line area 54 between the multiple first line area fluorescences 51. Each time a first line area fluorescence 51 is excited, a first area array fluorescence image of a surface area can be generated by the imaging component 40.

[0091] The method for generating the second array fluorescence image includes:

[0092] The driving component 70 drives the stage 20 to move the sample to be tested 50 along the first direction, and at the same time controls the light source component 10 so that the excitation light 11 irradiates the sample to be tested 50 along the first direction and excites multiple second line area fluorescences 52. There is at least one unirradiated second line area 55 between the multiple second line area fluorescences 52. Every time a second line area fluorescence 52 is excited, a second area array fluorescence image of a surface area can be generated by the imaging component 40.

[0093] Referring to Figures 9 through 11 , in certain embodiments, imaging requires illuminating the sample with periodic stripes that alternate between light and dark. To achieve this imaging effect, the stripes are phase-shifted three times in the dimension that improves imaging resolution. Images captured at these three different illumination stripe phases are then reconstructed to produce a super-resolution image.

[0094] Specifically, as shown in Figures 9 to 11, they are schematic diagrams of two-dimensional imaging of structured light illumination stripes in the X direction, including the bright area of ​​the structured light stripes, that is, the illuminated part, and the dark area of ​​the structured light stripes, that is, the unilluminated part. First, the structured light illumination stripes are first irradiated on columns A, B, C and G, H, and I to complete the first image. After that, the structured light illumination stripes change their phase and illuminate columns C, D, E and I, J, and K to complete the second image. Finally, the structured light illumination stripes change their phase again and illuminate columns E, F, G and K, L, and A to complete the third image. After reconstructing the above three images, the resolution improvement in the dimension perpendicular to the image in the stripe direction can be obtained.

[0095] Referring to Figures 9 to 11 , to produce the fringe imaging effect shown, we need to control the light source assembly 10 with a preset on / off sequence. Specifically, the laser emitter 12 is controlled by the control assembly 80 with a preset on / off sequence. Figures 12 to 17 illustrate how fringe formation occurs when the first beam rotation assembly 30 is in the first state and the excitation light 11 is irradiated onto the sample 50 to be tested. First line region 51 is the illuminated area, while first line region 54 is the unilluminated area.

[0096] Because the detector of line scan camera 422 is one-dimensional, it can only image one dimension of the sample when not scanning. As shown in Figures 12 to 17, line scan camera 422 can only image in the X direction, and then scans and completes two-dimensional imaging in conjunction with the movement of motorized stage 20 in the Y direction.

[0097] Please refer to FIG. 9 to FIG. 11 . In order to achieve the same illumination effect as the illumination stripe period under the illumination mode of the excitation light 11 , it is necessary to preset the on-off timing of the laser.

[0098] Please refer to Figures 12 to 17. Specifically, the control component 80 controls the laser emitter 12 to emit excitation light 11 to illuminate column A so that column A excites the first line area fluorescence 51. At this time, the imaging area 53 of the linear array camera 422 is also located in column A, and a first planar array fluorescence image is generated. After imaging, the control component 80 controls the drive component 70 to drive the stage 20 to move along the Y direction, and repeats the above operation in columns B and C. When the stage 20 moves to column D, the control component 80 controls the laser emitter 12 to turn off the emission and continues to move the stage 20. At the same time, the linear array camera 422 images in columns D, E, and F. At this point, the imaging of the first planar array fluorescence image within a complete fringe cycle is completed. The above steps are repeated until the imaging of the first planar array fluorescence image of the entire sample to be tested is completed. In addition, the three-step phase shift of the fringes can also be achieved by changing the laser preset switch timing through the above method.

[0099] Please refer to Figures 18 to 20. After completing the above imaging content, it is necessary to rotate the structured light illumination stripes 90° to form structured light illumination stripes in the Y direction and perform imaging again to ultimately achieve orthogonal line scanning super-resolution imaging. The structured light illumination stripes in the Y direction also need to undergo three-step phase shifting.

[0100] Specifically, when the first beam rotation component 30 completes imaging of all first array fluorescence images in the first state, the first beam rotation component 30 is adjusted to the second state. At this time, the stripe generation method when the excitation light 11 is irradiated on the sample to be tested 50 is shown in Figures 18 to 20, wherein the gray area is the illumination area, that is, the second line area fluorescence 52, and the white area is the unilluminated area, that is, the second line area 55.

[0101] Since the second beam rotation assembly 90 is also in the second state when the first beam rotation assembly 30 is in the second state, the second beam rotation assembly 90 will rotate the second line area fluorescence 52 and then image it by the linear array camera 422. Therefore, there is no need to rotate the linear array camera 422. At this time, the linear array camera 422 performs imaging in the Y direction, and cooperates with the movement of the electric stage 20 in the X direction to complete the two-dimensional imaging by scanning.

[0102] Please refer to FIG. 18 to FIG. 20 . In order to achieve the same lighting effect as one lighting stripe cycle in the excitation light 11 illumination mode, it is necessary to achieve this through a laser preset switching timing.

[0103] Please refer to Figures 21 to 26. Specifically, the control component 80 controls the laser emitter 12 to emit excitation light 11 to illuminate column A, so that column A excites the first line area fluorescence 51. At this time, the imaging area 53 of the line array camera 422 is also located in row A, and a second area array fluorescence image is generated. After imaging, the control component 80 controls the drive component 70 to drive the stage 20 to move along the X direction, and repeats the above operation for rows B and C. When the stage 20 moves to row D, the control component 80 controls the laser emitter 12 to turn off emission and continues to move the stage 20. At the same time, the line array camera 422 images in rows D, E, and F, thereby completing the imaging of the second area array fluorescence image within a complete fringe cycle. The above steps are repeated until the imaging of the second area array fluorescence image of the entire sample to be tested is completed. In addition, the three-step phase shift of the fringe can also be achieved by changing the preset laser on and off timing using the above method.

[0104] At this point, the first area array fluorescence image and the second area array fluorescence image of the entire sample to be tested are obtained, and all the first area array fluorescence images and the second area array fluorescence images are synthesized to obtain the final super-resolution imaging.

[0105] Specifically, referring to Figures 9 to 11 and 12 to 17, or Figures 18 to 20 and 21 to 26, in the preset switching sequence, one on period and one off period constitute a minimum cycle of the preset switching sequence. The duration T of the on period and the off period is the same and can correspond to the time required for the illumination area to scan across three line areas.

[0106] For example, the duration T of an on period or an off period may satisfy:

[0107] T=3*S / M / V,

[0108] Wherein, S is the pixel size of the imaging component 40 , M is the magnification of the imaging system 100 , and V is the moving speed of the sample 50 to be measured.

[0109] In certain embodiments of the present invention, the module involved may be a single-chip microcomputer chip that integrates a processor, memory, communication module, etc. The processor may be a central processing unit (CPU), or other general-purpose processors, digital signal processors (DSP), application-specific integrated circuits (ASIC), field programmable gate arrays (FPGA), or other programmable logic devices, discrete gate or transistor logic devices, discrete hardware components, etc.

[0110] Any process or method description in a flowchart or otherwise described herein may be understood to represent a module, segment or portion of code comprising one or more executable instructions for implementing the steps of a specific logical function or process, and the scope of the preferred embodiments of the present invention includes alternative implementations in which functions may be performed out of the order shown or discussed, including performing functions in a substantially simultaneous manner or in the reverse order depending on the functions involved, which should be understood by those skilled in the art to which the embodiments of the present invention pertain.

[0111] The logic and / or steps represented in the flowchart or otherwise described herein, for example, can be considered as an ordered list of executable instructions for implementing logical functions, and can be embodied in any computer-readable medium for use by an instruction execution system, apparatus or device (such as a computer-based system, a system including a processing module, or other system that can fetch instructions from an instruction execution system, apparatus or device and execute instructions), or used in conjunction with such instruction execution systems, apparatuses or devices.

[0112] Throughout this specification, reference to the terms "certain embodiments," "one embodiment," "some embodiments," "illustrative embodiments," "examples," "specific examples," or "some examples" means that a specific feature, structure, material, or characteristic described in conjunction with the embodiment or example is 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. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

[0113] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be understood to indicate or imply relative importance or implicitly specify the number of technical features indicated. Thus, a feature specified as "first" or "second" may explicitly or implicitly include at least one of the features. In the description of this application, "plurality" means at least two, for example, two or three, unless otherwise specifically defined.

[0114] Although the embodiments of the present application have been shown and described above, it can be understood that the above embodiments are illustrative and cannot be understood as limitations on the present application. Ordinary technicians in this field can change, modify, replace and modify the above embodiments within the scope of the present application. The scope of the present application is defined by the claims and their equivalents.

Claims

1. An imaging system for a gene sequencer, characterized in that: The imaging system comprises: A light source assembly, used for emitting excitation light; A stage, arranged on the optical path of the light source assembly, the stage carrying a sample to be tested; The first light beam rotating assembly is configured to have a first state and a second state, wherein the second state is located on the optical path between the light source assembly and the stage; in the first state, the excitation light emitted by the light source assembly is irradiated to the sample to be tested to excite a first line area fluorescence extending along a first direction, and in the second state, the excitation light emitted by the light source assembly is irradiated to the sample to be tested after passing through the first light beam rotating assembly to excite a second line area fluorescence extending along a second direction; An imaging component, the imaging component is used to receive a first area array fluorescence image group of a surface area generated by the first line area fluorescence at multiple positions, and receive a second area array fluorescence image group of a surface area generated by the second line area fluorescence at multiple positions, the first area array fluorescence image group and the second area array fluorescence image group are used to reconstruct a two-dimensional super-resolution image; The second beam rotation component is configured to have a first imaging state and a second imaging state, wherein the second imaging state is located on the optical path between the imaging component and the stage; in the first imaging state, the imaging component receives the first line area fluorescence at multiple positions to generate a first area array fluorescence image group of the surface area, and in the second imaging state, the second beam rotation component rotates the second line area fluorescence by 90 degrees and enables the imaging component to receive the second line area fluorescence at multiple positions to generate a second area array fluorescence image group of the surface area.

2. The imaging system according to claim 1, characterized in that In the first state, the first beam rotation assembly is in a first position, and the beam rotation assembly is outside the optical path between the light source assembly and the stage in the first position. In the second state, the first beam rotation assembly is in a second position, and the beam rotation assembly is located in the optical path between the light source assembly and the stage in the second position.

3. The imaging system according to claim 2, characterized in that The first light beam rotation assembly includes a plurality of reflectors, and the plurality of reflectors and the optical axis of the excitation light form a preset angle.

4. The imaging system according to claim 3, characterized in that The first beam rotation assembly includes a beam inlet and a beam outlet; When the light beam rotation assembly is in the first position, the reflectors close to the light beam entrance and the light beam exit are both moved outside the light path between the light source assembly and the stage; when the light beam rotation assembly is in the second position, the reflectors close to the light beam entrance and the light beam exit are both moved into the light path between the light source assembly and the stage.

5. The imaging system according to claim 3, characterized in that: The reflectors close to the light beam entrance and the light beam exit move in the form of rotation, flipping or translation.

6. The imaging system according to claim 3, characterized in that: The reflectors include a first reflector, a second reflector, a third reflector, a fourth reflector, a fifth reflector and a sixth reflector. In the second state, the excitation light passes through the first reflector, the second reflector, the third reflector, the fourth reflector, the fifth reflector and the sixth reflector in sequence. The first reflector is perpendicular to the first plane and forms an angle of 45 degrees with the second plane. The second reflector is perpendicular to the third plane and forms an angle of 22.5 degrees with the first plane. The angles formed by the third reflector and the first plane, the second plane and the third plane are all 45 degrees. The fourth reflector is perpendicular to the first plane and forms an angle of 45 degrees with the second plane. The fifth reflector is perpendicular to the third plane and forms an angle of 22.5 degrees with the second plane. The angles formed by the sixth reflector, the first plane, the second plane and the third plane are all 45 degrees. The first plane, the second plane and the third plane are perpendicular to each other.

7. The imaging system according to claim 6, characterized in that: In the first state, the first reflector and the sixth reflector are separated from the optical path of the excitation light.

8. The imaging system according to claim 4, characterized in that: The second light beam rotation assembly has the same structure as the first light beam rotation assembly.

9. The imaging system according to claim 1, characterized in that: When the first beam rotation component is in the first state, the second beam rotation component is in the first imaging state, and when the first beam rotation component is in the second state, the second beam rotation component is in the second imaging state.

10. The imaging system according to claim 1, characterized in that The stage is connected to a driving component, and when the first light beam rotation component is in the first state, the driving component drives the stage to move along the second direction, and when the first light beam rotation component is in the second state, the driving component drives the stage to move along the first direction.

11. The imaging system according to claim 10, characterized in that: The imaging system also includes a control component, which controls the drive component to drive the stage to move along the second direction and controls the light source component to emit the excitation light with a preset switching sequence when the first light beam rotation component is in the first state, so that the excitation light irradiates the sample to be tested along the second direction to excite multiple first line area fluorescence, and there is at least one un-irradiated line area between the multiple irradiated line areas; when the first light beam rotation component is in the second state, the control component controls the drive component to drive the stage to move along the first direction, and controls the light source component to emit the excitation light with a preset switching sequence, so that the excitation light irradiates the sample to be tested along the second direction to excite multiple second line area fluorescence, and there is at least one un-irradiated line area between the multiple irradiated line areas.

12. The imaging system according to claim 1, characterized in that The imaging system also includes a first dichroic mirror and an objective lens, wherein the objective lens is used to collect the fluorescence and transmit the fluorescence to the first dichroic mirror, and the first dichroic mirror is used to reflect the excitation light to the sample to be tested and to transmit the fluorescence to the imaging component.

13. The imaging system according to claim 12, characterized in that: The imaging component includes a plurality of second dichroic mirrors, which distribute the fluorescence to a plurality of imaging channels. The second dichroic mirrors are located between the imaging channels and the first dichroic mirrors. Each of the imaging channels includes a converging lens and a line array camera. The converging lens is used to converge the fluorescence to the line array camera, and the line array camera is used to receive the fluorescence.

14. The imaging system according to claim 13, characterized in that There are three second dichroic mirrors, each of which has a different coating, and is used to separate four types of fluorescence corresponding to the four base types of ATCG.

15. An imaging method of an imaging system, characterized in that: The imaging system comprises a light source assembly, a stage, a first light beam rotation assembly and an imaging assembly, the stage carries a sample to be measured, and the imaging method comprises: The first light beam rotating assembly is adjusted to a first state, and the excitation light is emitted by the light source assembly to irradiate the sample to be tested and excite the fluorescence of a first line region extending along a first direction; Receiving the first line area fluorescence through the imaging component to generate a first area array fluorescence image group of an area; The first light beam rotating assembly is adjusted to a second state, and the sample to be tested is illuminated by emitting excitation light to excite fluorescence in a second linear region extending along a second direction, wherein the second direction is perpendicular to the first direction; Receiving the second line area fluorescence again through the imaging component to generate a second area array fluorescence image group of an area; The imaging component combines the first area array fluorescence image group and the second area array fluorescence image group into a super-resolution image.

16. The imaging method according to claim 15, characterized in that: The method for generating the first array fluorescence image comprises: The stage is driven by a driving component to move the sample to be tested along the second direction, and the light source component is controlled to make the excitation light illuminate the sample to be tested along the second direction and excite multiple first line area fluorescences, there is at least one un-irradiated first line area among the multiple first line area fluorescences, and each time a first line area fluorescence is excited, the imaging component can generate the first surface array fluorescence image of a surface area.

17. The imaging method according to claim 16, characterized in that: The method for generating the second array fluorescence image includes: The stage is driven by a driving component to move the sample to be tested along the first direction, and the light source component is controlled to make the excitation light illuminate the sample to be tested along the first direction and excite a plurality of second line area fluorescences, wherein there is at least one un-irradiated second line area among the plurality of second line area fluorescences, and each time a second line area fluorescence is excited, the imaging component can generate a second surface array fluorescence image of a surface area.

18. A gene sequencer, characterized in that: Comprising the imaging system of any one of claims 1 to 14.

Images