CN105301794B - Super-resolution imaging device for fast moving objects - Google Patents

Abstract

The invention relates to a super high resolution imaging device for fast moving objects. The device includes: a conventional resolution microscopic imaging module, which is used to display moving object samples and super-high resolution imaging targets to provide both position and motion information; a fast image acquisition and processing module, which is used to obtain ultra-high resolution imaging targets The motion velocity V _i , the motion velocity V _r and the position of the region of interest ROI and the size of the imaging shooting area Sp are provided to the position feedback control module and the super-high resolution imaging module; the position feedback control module is used to adjust the imaging shooting The position of the area Sp or the proposed imaging area Si, so that the intended shooting area Si and the imaging shooting area Sp keep coincident; the super-resolution imaging module is used to adjust the size of the imaging shooting area Sp according to the requirements of the super-resolution imaging rate, for moving object samples Ultra-high-resolution imaging on the super-high-resolution imaging target. The invention can eliminate the influence of object movement on super-high resolution imaging.

Description

Ultra-high-resolution imaging device for fast-moving objects

technical field

The invention relates to the field of image acquisition technology, in particular to a super-high-resolution imaging device for fast-moving objects.

Background technique

In general, the size of the smallest object that the human eye can distinguish is about 0.1 mm. If you want to see smaller objects, you need to use microscopy. In 1873, Ernst Abbe, a German microtechnical expert, revealed the principle that optical microscopes have limitations due to light diffraction effects and limited aperture resolution. It is this principle that "constrains" the application of traditional optical microscopes in the nanometer world.

When there is only a single fluorescent molecule in the field of view of the objective lens of the microscope, it is easy to exceed the optical resolution limit through specific algorithm fitting. In order to explore the microscopic world, ultra-high-resolution microscopy technology, which breaks through the optical limit of optical microscopes, came into being. In 1981, Barak and Webb first introduced single-molecule tracking technology into life science. Although single-molecule localization can be achieved at the nanometer level, it does not improve the resolution of light microscopy when resolving two or more point sources of light.

In 2002, Patterson and Lippincott-Schwartz first used a variant of green fluorescent protein (GFP) (PA-GFP) to observe the trajectory of specific proteins in cells. Germany's Eric Bezig keenly realized that the application of single-molecule fluorescence imaging technology, combined with the luminescent properties of this fluorescent protein, can break through the limit of optical resolution—photoactivated localization microscopy (PALM) was born. The imaging method of PALM can only be used to observe exogenously expressed proteins, but it is powerless for endogenous proteins in cells. In 2006, the experimental group of Xiaowei Zhuang, a Chinese scientist at the Howard Hughes Institute in the United States, discovered that different wavelengths can control the switching of the chemical fluorescent molecule Cy5 between the fluorescent excited state and the dark state. In view of this, stochastic optical reconstruction microscopy (STORM) was developed. Regardless of the PALM or STORM super-resolution microscopy method, its point spread function imaging is still consistent with traditional microscopy imaging, requiring repeated activation-quenching of fluorescent molecules, so most experiments are done on fixed cells.

In 2000, German scientist Stefan Hell proposed to reduce the spot size of the excitation light through physical processes, directly reduce the FWHM of the point spread function to improve the resolution, and successfully developed the stimulated emission depletion microscopy (STED). Another way to change the point spread function to break through the optical diffraction limit is Saturated Structured Illumination Microscopy (SSIM). In 2005, Gustafsson first introduced nonlinear structural optical illumination components to traditional microscopes, and obtained images with a resolution of 50nm. However, the existing ultra-high-resolution technology has a slow imaging speed, and it is difficult to photograph moving (especially fast-moving) samples.

Contents of the invention

One of the objectives of the present invention is to provide a super-high-resolution imaging device for fast-moving objects, so as to solve the technical problem in the prior art that the imaging speed is slow and it is difficult to photograph moving objects, especially living organisms.

In order to achieve the purpose of the above invention, an embodiment of the present invention provides a super-high-resolution imaging device for fast-moving objects, including:

The conventional resolution microscopic imaging module is used to display the moving object sample and the super high resolution imaging target to provide the position and motion information of the moving object sample and the super high resolution imaging target;

The fast image acquisition and processing module is used to obtain the moving speed V _i of the super-high resolution imaging target, the moving speed V _r and the position of the region of interest ROI, and the size of the imaging shooting area Sp, and provide them to the position feedback control module and the super high-resolution resolution imaging module;

A position feedback control module, configured to adjust the position of the imaging shooting area Sp or the proposed imaging area Si, so that the intended shooting area Si and the imaging shooting area Sp keep overlapping;

The ultra-high-resolution imaging module is used to adjust the size of the imaging shooting area Sp according to the super-resolution imaging rate requirement, and perform ultra-high-resolution imaging on the ultra-high-resolution imaging target on the moving object sample.

Optionally, the position feedback control module includes an electronically controlled sample stage and a first lens, a second lens, and an electronically controlled rotating mirror sequentially arranged on the first imaging optical path, wherein:

One side of the first lens is provided with an electronically controlled sample stage, and the other side is provided with a second lens;

The electric control rotating mirror is arranged on the side of the second lens away from the electric control sample stage.

Optionally, by setting the positions of the first lens and the second lens, the electronically controlled rotating mirror is in a conjugate relationship with the entrance pupil of the imaging objective lens in the conventional resolution microscopic imaging module.

Optionally, the fast image acquisition and processing module includes a fast image acquisition device and a computing device, wherein:

The fast image acquisition device is used to collect moving object samples and super-high-resolution imaging targets to provide position and motion information of the moving object samples and super-high-resolution imaging targets;

The computing device is used to obtain the moving velocity V _i of the super-high-resolution imaging target, the moving velocity V _r and the position of the region of interest ROI and the imaging shooting area according to the position and motion information of the moving object sample and the super-high-resolution imaging target The size of Sp.

Optionally, the fast image acquisition device uses an image sensor or a position sensitive detector.

Optionally, the computing device obtains the imaging frame rate f _s according to the following formula:

|V _r −V _i |/PPI _s ≤ f _s .

Optionally, the conventional resolution microscopic imaging module includes: an illumination unit and an imaging unit:

The illumination unit includes an illumination device, a third lens, and a condenser lens arranged sequentially on the second imaging optical path; one side of the third lens is provided with the illumination device, and the other side is provided with the condenser lens; The side of the condenser far away from the lighting equipment is provided with an electronically controlled sample stage;

The imaging unit includes an imaging objective lens, a beam splitter, and an imaging lens and a filter mirror arranged on the second imaging optical path in sequence; one side of the imaging objective lens is provided with an electronically controlled sample stage, and the other side The optical splitter is provided; the optical splitter changes the light on the second imaging optical path to form a third imaging optical path; the imaging lens is arranged on one side of the optical splitter, and the optical filter is arranged on the other side.

In the embodiment of the present invention, the super-resolution imaging rate is determined by acquiring the region of interest of the moving object sample and the position and moving speed of the super-resolution imaging target, and combining the resolution of the super-resolution imaging module, and adjusting the area of the imaging shooting area based on this , to ensure the ultra-high resolution imaging rate; by adjusting the position of the imaging shooting area or the position of the intended shooting area on the sample, so that the imaging shooting area and the position of the intended shooting area on the sample keep coincident, thereby eliminating the impact of the movement of the moving object sample on the super high Resolve the effects of imaging. The invention can quickly and automatically perform ultra-high-resolution imaging on fast-moving objects, and can also automatically analyze the morphological structure of the moving objects, and is especially suitable for rapid ultra-high-resolution imaging of various moving objects such as sperm cells and living tissues.

Description of drawings

The features and advantages of the present invention will be more clearly understood by referring to the accompanying drawings, which are schematic and should not be construed as limiting the invention in any way. In the accompanying drawings:

Fig. 1 is a schematic flow chart of a super-high-resolution imaging method for fast-moving objects provided by an embodiment of the present invention;

Fig. 2 is a schematic structural diagram of a super-high-resolution imaging device for fast-moving objects provided by an embodiment of the present invention;

FIG. 3 is a schematic diagram of the process of adjusting the overlapping of the imaging shooting area and the intended shooting area in an embodiment of the present invention.

detailed description

The specific implementation manners of the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

In the first aspect, an embodiment of the present invention provides a super-resolution imaging method for fast-moving objects, as shown in FIG. 1 , including:

Setting a region of interest ROI containing the super-resolution imaging target according to the position of the super-resolution imaging target on the moving object sample;

Acquiring the motion velocity V _i of the super-high resolution imaging target and the motion velocity V _r and position of the region of interest ROI;

Obtain the imaging frame rate f _s of the ultra-high resolution imaging module according to the resolution PPI _s of the ultra-high resolution imaging module, the motion velocity V _r of the region of interest ROI, and the motion speed V _i of the ultra-high resolution imaging target ;

Calculate the size of the imaging shooting area Sp according to the relationship between the imaging frame rate f _s and the imaging shooting area Sp; define a proposed shooting area Si that includes a super-resolution imaging target, and the size of the intended shooting area Si is related to the imaging The shooting areas Sp have the same size and have the same moving speed V _r ;

Adjust the position of the intended shooting area Si or the imaging shooting area Sp so that the intended shooting area Si and the imaging shooting area Sp keep overlapping; Resolve imaging targets for super-resolution imaging.

In the embodiment of the present invention, the area of the imaging shooting area is obtained by obtaining the position and moving speed of the moving object sample and the ultra-high-resolution imaging target in combination with the resolution of the ultra-high-resolution imaging module, so as to ensure the ultra-high-resolution imaging rate; by adjusting the imaging shooting area Or the position of the sample is kept coincident with the position of the intended shooting area of the imaging shooting area, so as to eliminate the influence of the movement of the moving object sample on the super-resolution imaging. The invention can quickly and automatically perform ultra-high-resolution imaging on moving objects, and can also automatically analyze the morphological structure of the moving objects, and is especially suitable for rapid ultra-high-resolution imaging of various moving objects such as sperm cells and living tissues.

Optionally, the imaging of the super-high-resolution imaging module is acquired according to the resolution PPI _s of the super-high-resolution imaging module, the moving velocity V _r of the region of interest ROI, and the moving velocity V _i of the super-high-resolution imaging target In the step of frame frequency f _s , the following formula is used to obtain the imaging frame frequency f _s :

|V _r −V _i |/PPI _s ≤ f _s .

Optionally, acquiring the motion velocity V _i of the super-high resolution imaging target and the motion velocity V _r and the position of the region of interest ROI are acquired in the following manner:

Obtain the positions of moving object samples and super-resolution imaging targets in at least two images;

Acquiring at least two images of the ROI of the region of interest within a preset time;

Comparing the positions of the moving object sample and the super-high-resolution imaging target in at least two images to obtain the displacement of the moving object sample and the super-high-resolution imaging target;

The motion velocity V _r of the region of interest ROI and the motion velocity V _i of the super-resolution imaging target are acquired according to the displacement of the moving object sample and the super-resolution imaging target and the imaging frame rate.

Optionally, the motion velocity V _r and position of the region of interest ROI are acquired in the following manner:

Select a first representative point that can represent its motion characteristics in the region of interest ROI, and select a second representative point that can represent its motion characteristics on the super-high resolution imaging target;

collecting the light intensities of the first representative point and the second representative point respectively at different times;

Comparatively analyze the relationship between the light intensity of the first representative point and the second representative point over time, so as to obtain the ROI of the region of interest of the moving object sample and the displacement and velocity of the super-resolution imaging target.

In practical application, the present invention can also obtain the motion speed and position of the point through the position detector.

Optionally, keeping the intended shooting area Si and the imaging shooting area Sp coincident can be achieved by the following methods:

The position of the imaging shooting region Sp is adjusted according to the moving speed V _r of the region of interest ROI, so that the movement direction of the imaging shooting region Sp is the same as that of the region of interest ROI, and the speed is equal.

or,

The imaging shooting area Sp remains unchanged, and the sample cell is adjusted according to the moving speed V _r of the region of interest ROI, so that the sample produces a moving speed -V _r that is opposite to and equal to the moving direction of the ROI of the region of interest.

In order to reflect the superiority of a super-high-resolution imaging method for fast-moving objects provided by the embodiment of the present invention, the embodiment of the present invention also provides a super-high-resolution imaging device for fast-moving objects, as shown in Figure 2 shown, including:

The conventional resolution microscopic imaging module 10 is used to display the moving object sample and the super high resolution imaging target to provide the position and motion information of the moving object sample and the super high resolution imaging target;

The fast image acquisition and processing module 20 is used to obtain the moving velocity V of the moving object sample, the moving velocity V _i of the ultra-high-resolution imaging target, the moving velocity V _r and the position of the intended shooting area Si, and the size of the imaging shooting area Sp, and Provided to the position feedback control module 40 and the super-resolution imaging module 30;

A position feedback control module 40, configured to adjust the position of the imaging shooting area Sp or the proposed imaging area Si, so that the intended shooting area Si and the imaging shooting area Sp keep overlapping;

The ultra-high-resolution imaging module 30 is configured to adjust the size of the imaging shooting area Sp according to the super-resolution imaging rate requirement, and perform ultra-high-resolution imaging on the ultra-high-resolution imaging target on the moving object sample.

As a specific example of a conventional resolution microscopic imaging module 10 , as shown in FIG. 2 , the conventional resolution microscopic imaging module 10 provided by the embodiment of the present invention includes an illumination unit and an imaging unit. The illuminating unit comprises an illuminating device 101, a third lens 102 and a condenser lens 103 which are sequentially arranged on the second imaging optical path, wherein the illuminating device 101 is arranged on one side of the third lens 102, and the condenser lens 103 is arranged on the other side; An electrically controlled sample stage 104 is provided on the side of 103 away from the lighting device 101 . The imaging unit includes an imaging objective lens 105, a beam splitter 106, and an imaging lens 107 and a filter lens 108 arranged on the second imaging optical path in turn; one side of the imaging objective lens 105 is provided with an electronically controlled sample stage 104, and the other A beam splitter 106 is set on one side; the beam splitter 106 changes the light on the second imaging optical path to form a third imaging optical path; the imaging lens 107 is set on one side of the beam splitter 106, and a filter 108 is set on the other side.

In practical applications, those skilled in the art can select a conventional resolution microscopic imaging module with appropriate parameters according to specific usage scenarios to realize the display of fast-moving object samples and super-high-resolution imaging targets, which is not limited by the present invention.

As a specific example of a fast image acquisition and processing module 20, as shown in FIG. 2, the fast image acquisition and processing module 20 provided by the embodiment of the present invention includes a fast image acquisition device 201 and a computing device 202, wherein:

The fast image acquisition device 201 is used to acquire the moving object sample and the super high resolution imaging target to provide the position and motion information of the moving object sample and the super high resolution imaging target;

The computing device 202 is used to obtain the moving velocity V of the moving object sample, the moving velocity V _i of the super-high resolution imaging target, and the moving velocity V of the area Si to be photographed according to the position and movement information of the moving object sample and the super-high resolution imaging target _r is related to the position and the size of the imaging shooting area Sp.

In practical applications, the fast image acquisition device 201 may use an image sensor such as a charge-coupled device (Charge-coupled Device, CCD) or a complementary metal oxide semiconductor (Complementary Metal Oxide Semiconductor, CMOS), or a position-sensitive detector. Those skilled in the art can choose according to the specific usage scenario, which is not limited in the present invention.

In practical applications, the computing device 202 adopts different background modeling, target recognition, trajectory generation and other technologies to calculate For the speed of motion, those skilled in the art can select an appropriate processing method according to a specific usage scenario.

As a specific example of a position feedback control module 40, as shown in FIG. 2, the position feedback control module 40 provided by the embodiment of the present invention includes a first lens 403, a second lens 402 and an electric Control rotating mirror 401, wherein:

One side of the first lens 403 is provided with an electronically controlled sample stage 104, and the other side is provided with a second lens 402;

The side of the second lens 402 away from the electronically controlled sample stage 104 is provided with an electronically controlled rotating mirror 401 .

In the embodiment of the present invention, by adjusting the positions of the first lens 403 and the second lens 402 , the electrically controlled rotating mirror 401 is in a conjugate relationship with the entrance pupil of the imaging objective lens 105 . Adjust the electronically controlled rotating mirror 401 in time to make the imaging shooting area Sp move rapidly on the moving object sample, so that the imaging shooting area Sp and the intended shooting area Si always keep coincident, thereby eliminating the influence of the movement of the moving object sample on super-resolution imaging. In the embodiment of the present invention, it is also possible to quickly change the position of the intended imaging area Si by controlling the electronically controlled sample stage, so as to achieve the purpose of overlapping the positions of the imaging imaging area Sp and the intended imaging area Si. Those skilled in the art can make selections according to specific occasions, which are not limited in the present invention.

As a specific example of a super-resolution imaging module 30, as shown in FIG. Adjust the first imaging optical path to change the imaging shooting area Sp of the super-high-resolution imaging module 30; meanwhile, it is also connected to the computing device 202 to receive the imaging frame rate f s transmitted from the computing device 202, and simulate the imaging frame rate f _s according to the imaging frame rate f _s The imaging shooting area Si performs ultra-high resolution imaging, so that clear images of moving objects and samples can be taken during the movement process.

In practical applications, the ultra-high-resolution imaging module 30 can be obtained from devices such as photoactivated localization microscopy (PALM), stochastic optical reconstruction microscopy (STORM) or structured illumination microscopy (SIM). High-resolution imaging equipment such as laser scanning light focusing, two (multi) photon fluorescence microscopy imaging, etc., can be selected by those skilled in the art according to specific usage scenarios, which are not limited by the present invention.

The working process of a super-high-resolution imaging device for a fast-moving object provided by an embodiment of the present invention is described below.

As shown in FIG. 3 , the moving object sample A is placed in the sample cell in the sample stage 104 .

The light emitted by the illuminating device 101 passes through the third lens 102 and the condenser lens 103 and then projects on the bottom of the sample pool. The images formed by the moving object sample A and the ultra-high resolution imaging targets B1, B2 and B3 pass through the imaging objective lens 105 and the beam splitter 106, then pass through the imaging lens 107 and the filter 108, and then are transmitted to the fast image acquisition device 201.

The fast image acquisition device 201 quickly collects at least two images and transmits them to the computing device 202, and the computing device 202 includes super-resolution imaging targets B1, B2 and B3 according to the positions of the super-resolution imaging targets B1, B2 and B3 on the moving object sample. ROI of B2 and B3, and perform calibration.

The computing device 202 acquires the positions of the moving object sample and the super-high resolution imaging target in at least two images; respectively compares the positions of the moving object sample and the super high resolution imaging target in the at least two images to obtain the moving object sample and the super high resolution imaging target. The displacement of the high-resolution imaging target; according to the displacement of the moving object sample and the ultra-high resolution imaging target and the imaging frame rate f _s when at least two images are acquired, the moving speed V of the moving object sample A and the moving speed V of the ROI of the region of interest are obtained _r and the moving velocity V _i of the ultra-high resolution imaging target. Among them, the imaging frame frequency f _s and the moving speed V of the moving object sample A, the moving speed V _r of the area Si to be photographed, the moving speed V _i of the ultra-high resolution imaging target, and the resolution PPI _s of the ultra-high resolution imaging module need to meet The following conditions:

|V _r -V _i |/f _s ≤ PPI _s

The computing device 202 obtains the imaging rate according to the above formula and transmits it to the super-high resolution imaging module 30, and obtains the moving speed V of the moving object sample A, the moving speed V _r of the area Si to be photographed, and the moving speed V of the super-high resolution imaging target _i Obtain the angle of the electronically controlled rotating mirror 401 and transmit it to the position feedback control module 40 .

The position feedback control module 40 adjusts the angle of the electronically controlled rotating mirror 401 according to the received angle information, and adjusts the electrically controlled rotating mirror 401 in time to make the imaging shooting area Sp move rapidly on the moving object sample, so that the imaging shooting area Sp is in line with the virtual object. The shooting areas Si always keep coincident.

In the embodiment of the present invention, the position of the sample can also be adjusted by transmitting the position information to the electronically controlled sample stage 104, so that the imaging shooting area Sp and the intended shooting area Si keep overlapping, thereby eliminating the influence of the moving speed V _r of the intended shooting area Si.

The imaging rate of the high-resolution imaging module 30 is usually related to the size of its imaging area Sp; when the resolution is constant, the larger the imaging area Sp is, the smaller the imaging frame rate f _s is. The ultra-high-resolution imaging module 30 acquires the imaging frame rate f _s of the ultra-high-resolution imaging module according to the resolution PPI _s of the ultra-high-resolution imaging module, the motion velocity V _r of the area Si to be photographed, and the motion velocity V _i of the ultra-high-resolution imaging target And imaging the shooting area Sp, thereby reducing or eliminating the influence of (V _i -V _r ). Then the high-resolution imaging module 30 performs super-resolution imaging on the super-resolution imaging target on the moving object sample A according to the size of the imaging shooting area Sp.

To sum up, the ultra-high-resolution imaging device for fast-moving objects provided by the embodiment of the present invention obtains the position and moving speed of the moving object sample and the ultra-high-resolution imaging target, and combines the resolution of the ultra-high-resolution imaging module Obtain the area of the imaging shooting area to ensure the super-high resolution imaging rate; adjust the position of the imaging shooting area to keep coincident with the position of the intended shooting area on the sample, thereby eliminating the influence of the movement of the moving object sample on the super-high resolution imaging. The invention can quickly and automatically perform ultra-high-resolution imaging on moving objects, and can also automatically analyze the morphological structure of the moving objects, and is especially suitable for rapid ultra-high-resolution imaging of various moving objects such as sperm cells and living tissues.

It should also be noted that in this article, relational terms such as first and second etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply that these entities or operations Any such actual relationship or order exists between. Furthermore, the term "comprises", "comprises" or any other variation thereof is intended to cover a non-exclusive inclusion such that a process, method, article, or apparatus comprising a set of elements includes not only those elements, but also includes elements not expressly listed. other elements of or also include elements inherent in such a process, method, article, or device.

Although the embodiments of the present invention have been described in conjunction with the accompanying drawings, those skilled in the art can make various modifications and variations without departing from the spirit and scope of the present invention. within the bounds of the requirements.

Claims (7)

1. A super-high-resolution imaging device for fast-moving objects, characterized in that, comprising: The conventional resolution microscopic imaging module is used to display the moving object sample and the super high resolution imaging target to provide the position and motion information of the moving object sample and the super high resolution imaging target; The fast image acquisition and processing module is used to obtain the moving speed V _i of the super-high resolution imaging target, the moving speed V _r and the position of the region of interest ROI, and the size of the imaging shooting area Sp, and provide them to the position feedback control module and the super high-resolution resolution imaging module; A position feedback control module, configured to adjust the position of the imaging shooting area Sp or the proposed imaging area Si, so that the intended shooting area Si and the imaging shooting area Sp keep overlapping; The ultra-high-resolution imaging module is used to adjust the size of the imaging shooting area Sp according to the super-resolution imaging rate requirement, and perform ultra-high-resolution imaging on the ultra-high-resolution imaging target on the moving object sample. 2. The ultra-high resolution imaging device according to claim 1, wherein the position feedback control module includes an electronically controlled sample stage and a first lens, a second lens, and an electronically controlled sample stage that are sequentially arranged on the first imaging optical path. mirror, where: One side of the first lens is provided with an electronically controlled sample stage, and the other side is provided with a second lens; The electric control rotating mirror is arranged on the side of the second lens away from the electric control sample stage. 3. The super-high resolution imaging device according to claim 2, characterized in that, by setting the positions of the first lens and the second lens so that the electronically controlled rotating mirror and the conventional resolution microscopic imaging module The entrance pupil of the imaging objective is in a conjugate relationship. 4. The ultra-high resolution imaging device according to claim 1, wherein the fast image acquisition and processing module includes a fast image acquisition device and a computing device, wherein: The fast image acquisition device is used to collect moving object samples and super-high-resolution imaging targets to provide position and motion information of the moving object samples and super-high-resolution imaging targets; The computing device is used to obtain the moving velocity V _i of the super-high-resolution imaging target, the moving velocity V _r and the position of the region of interest ROI and the imaging shooting area according to the position and motion information of the moving object sample and the super-high-resolution imaging target The size of Sp. 5. The ultra-high resolution imaging device according to claim 4, wherein the fast image acquisition device adopts an image sensor or a position sensitive detector. 6. The ultra-high resolution imaging device according to claim 4, wherein the computing device obtains the imaging frame rate f _s according to the following formula: |V _r -V _i |/PPI _s ≤ f _s ; Wherein, PPI _s is the resolution of the super-high resolution imaging module. 7. The ultra-high resolution imaging device according to claim 1, wherein the conventional resolution microscopic imaging module comprises: an illumination unit and an imaging unit: The illumination unit includes an illumination device, a third lens, and a condenser lens arranged sequentially on the second imaging optical path; one side of the third lens is provided with the illumination device, and the other side is provided with the condenser lens; The side of the condenser far away from the lighting equipment is provided with an electronically controlled sample stage; The imaging unit includes an imaging objective lens, a beam splitter, and an imaging lens and a filter mirror arranged on the second imaging optical path in sequence; one side of the imaging objective lens is provided with an electronically controlled sample stage, and the other side The optical splitter is provided; the optical splitter changes the light on the second imaging optical path to form a third imaging optical path; the imaging lens is arranged on one side of the optical splitter, and the optical filter is arranged on the other side.