CN115248197A - A three-dimensional imaging device and imaging method - Google Patents

Abstract

The invention discloses a high-throughput three-dimensional imaging device and an imaging method. Wherein, three-dimensional image device includes: at least one lighting device for exciting the sample to be tested in the form of light beam scanning; at least one detection device, wherein the main shaft of the detection light path and the main shaft of the illumination light path intersect at an included angle which is not zero; and the rotating shaft of the at least one rotating mechanism is not parallel to the main shafts of the detection light path and the illumination light path. The three-dimensional imaging method comprises the steps that different angle sections of a sample to be detected are continuously placed in a field of view to be detected through a rotating mechanism, different section signals of the sample are obtained through a detection device, and the signals are stacked to form a three-dimensional image; the time for switching the visual field in imaging is reduced through continuous rotation, so that high-flux imaging is achieved; through the synchronous cooperation of the lighting device, the detection device and the rotating mechanism, the motion blur can be ignored, and clear imaging is realized. The embodiment of the invention solves the problems of long imaging interruption time and low efficiency in the three-dimensional imaging process, and can realize multi-view high-flux three-dimensional imaging.

Description

A three-dimensional imaging device and imaging method

technical field

Embodiments of the present invention relate to the field of imaging technologies, and in particular, to a three-dimensional imaging device and an imaging method.

Background technique

Obtaining the three-dimensional structure information of samples by imaging is one of the important technologies for biological and medical research and detection. Obtaining more image pixels through imaging in a short time to extract more information about samples is an important direction for the development of 3D imaging technology.

In order to specifically identify specific signals from different parts of biological and medical samples, fluorescent molecular markers are usually used to enable different molecules in the sample to generate specific fluorescent signals under the illumination of a specific laser. Collecting these signals from all regions within the sample in full coverage allows the reconstruction of the resulting three-dimensional structure of the sample. 3D sample imaging throughput depends on which full coverage method is used to cover all sample areas.

Existing imaging techniques usually obtain the plane imaging in the sample detection area first, then image the parallel sample planes, and then superimpose these mutually parallel planes to reconstruct the three-dimensional image of the sample. Taking scanning confocal imaging or optical slice imaging as an example, the imaging scheme usually adopted by the existing imaging technology is as follows: first, point-by-point imaging (confocal imaging) of a plane perpendicular to the imaging direction (ie, the z direction of the main axis of the detection optical path), Or image part of the entire field of view at the same time (optical slice imaging); then translate in the z direction according to the resolution requirements, reach a new plane, and image the new plane; repeat the above movement and imaging until the sample is covered in the z direction, Complete three-dimensional imaging within a field of view. Because imaging while translational in the z-direction is prone to motion blur, exposure imaging is typically performed only when the sample is stationary at the target position. Since the time it takes for the sample to move between adjacent z-positions is typically much greater than the exposure time, these ineffective imaging times reduce imaging throughput.

In the multi-angle optical slice imaging technology, the sample is rotated at a certain angle, and then stops to start scanning to complete the imaging of a layer, and then rotates at a certain angle to stop before scanning and imaging through multiple angles (usually no more than 8 different angles) ) multi-view imaging, which can perform three-dimensional reconstruction of the sample. However, the process of rotating between different angles also includes a large number of interrupted imaging processes. Imaging can only be continued after each rotation and waiting for the vibration caused by starting and stopping to dissipate, resulting in long-term imaging interruptions, increasing imaging time and imaging workload, and reducing imaging throughput.

SUMMARY OF THE INVENTION

The invention provides a three-dimensional imaging device and an imaging method to realize multi-view continuous imaging during the three-dimensional imaging process, reduce unnecessary imaging interruptions, and improve imaging speed and imaging efficiency.

In a first aspect, the present invention provides a three-dimensional imaging device, the device comprising:

At least one illuminating device, including an illuminating light source and an illuminating light path, for illuminating or exciting the sample to be tested;

At least one detection device, including a detector and a detection optical path, used to detect the signal generated by the sample to be tested excited by the illumination device in the area to be tested;

at least one rotating mechanism, used to make the sample under test rotate relative to the illuminating device and the detecting device during the imaging process;

Wherein, the illumination light path excites the sample to be measured by the illumination light source in the form of a scanning beam; the main axis of the detection optical path intersects with the main axis of the illumination optical path and the included angle is not zero; the main axis of the illumination optical path and the The plane where the main axis of the detection optical path is located coincides with, is parallel to or perpendicular to the rotation axis of the rotation mechanism.

Optionally, at least one scanning mechanism is included in the illuminating device, which is used to continuously move the illuminating light source to form a scanning light beam.

Optionally, the scanning mechanism includes one or more scanning galvanometers, resonant scanning mirrors, rotating polygonal mirrors or acousto-optic modulators.

Optionally, the illuminating device includes one or more lasers, light emitting diodes or X-ray generators.

Optionally, the signal generated by the sample to be tested excited by the illumination device is one of fluorescence, elastic scattered light, Raman scattering, second harmonic signal, third harmonic signal or Exciting Raman scattering signal or Various.

Optionally, the detection device works synchronously with the lighting device.

Optionally, the detector uses a charge-coupled element or a complementary metal-oxide-semiconductor array as an imaging device.

Optionally, the rotating mechanism includes at least one rotating device, or a combined device of a rotating device and a linear motion device.

Optionally, the rotation mechanism further includes at least one control device for controlling the movement of the rotation device and the linear motion device.

In a second aspect, the embodiment of the present invention also provides a three-dimensional imaging method, the method comprising:

Controlling at least one lighting device to excite the sample to be tested in the sample detection area along the main axis direction of the illumination light path to generate a detected signal;

Controlling at least one detection device to detect signals generated in the sample detection area along the main axis of the detection optical path;

Controlling at least one rotating mechanism, so that the sample to be tested rotates relative to the illumination device and the detection device during the imaging process, and the sample to be tested is collected in different regions to be tested by the at least one detection device the signal generated;

Convert the collected signal into a digital signal;

performing image reconstruction based on the digital signal to obtain a three-dimensional image of the sample to be tested;

Wherein, the illumination light path excites the sample to be measured by the illumination light source in the form of a scanning beam; the main axis of the detection optical path intersects with the main axis of the illumination light path; The plane where the main axis of the detection optical path is located rotates parallel to or perpendicular to the rotation axis; the scanning direction of the scanning light beam is parallel to or perpendicular to the direction of the rotation axis.

Optionally, the controlling at least one rotating mechanism to cause the sample to be tested to rotate relative to the illuminating device and the detecting device during the imaging process includes:

At least one rotating mechanism is controlled so that the sample to be tested rotates continuously during the imaging process, or continuously moves in a combination of linear motion and rotary motion, and generates relative rotation with the illumination device and the detection device.

Optionally, the sample to be tested is excited by the lighting device within a preset time; wherein, within the preset time, the displacements of the sample to be tested during the relative rotation are less than or equal to Longitudinal resolution and lateral resolution of 3D imaging device.

The technical solution of the embodiment of the present invention solves the problem of imaging interruption time in the three-dimensional imaging process by making the sample, the illumination device and the detection device move relative to each other at a uniform speed during the process of detecting the sample signal, that is, performing continuous signal acquisition while the sample is rotating. Long, low imaging efficiency, improve imaging throughput; can reduce imaging interruption in the process of three-dimensional imaging, eliminate blur caused by sample rotation during imaging in existing three-dimensional imaging technology, and improve aberrations.

Description of drawings

FIG. 1 is a schematic structural diagram of a three-dimensional imaging device provided by Embodiment 1 of the present invention;

Fig. 2 is a three-dimensional structural schematic diagram of a sample rotating mechanism provided by Embodiment 1 of the present invention;

Fig. 3 is a schematic diagram of a three-dimensional imaging scanning process provided by Embodiment 1 of the present invention;

FIG. 4 is a schematic diagram of an imaging scan provided by Embodiment 1 of the present invention;

Fig. 5 is a schematic diagram of the principle of continuous scanning to eliminate motion blur in imaging provided by Embodiment 1 of the present invention;

6 is a schematic diagram of a mouse brain fluorescence imaging example provided by Embodiment 1 of the present invention;

Fig. 7 is a schematic diagram of the moving track of an imaging sample in the process of acquiring imaging signals provided by Embodiment 1 of the present invention;

Fig. 8 is a schematic diagram of the moving track of an imaging sample in the process of acquiring imaging signals provided by Embodiment 1 of the present invention;

FIG. 9 is a schematic diagram of the moving track of an imaging sample in the process of acquiring imaging signals provided by Embodiment 1 of the present invention;

FIG. 10 is a schematic diagram of a three-dimensional imaging effect of an imaging sample provided by Embodiment 1 of the present invention.

Detailed ways

The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, but not to limit the present invention. In addition, it should be noted that, for the convenience of description, only some structures related to the present invention are shown in the drawings but not all structures.

Example 1

FIG. 1 is a schematic structural diagram of a three-dimensional imaging device provided by Embodiment 1 of the present invention. This embodiment is applicable to the situation of using the device for three-dimensional imaging, especially for three-dimensional imaging of biological and medical samples.

As shown in FIG. 1 , the three-dimensional imaging device specifically includes: at least one illuminating device 1 , at least one detecting device 2 ; and at least one rotating mechanism 3 . Wherein, the illuminating device 1 is composed of an illuminating light source and an illuminating light path, the illuminating light source is a laser 4 with a wavelength of 488 nanometers as an example, and the illuminating light path includes a scanning mechanism, a relay lens 9, a scanning vibrating mirror 5, and a scanning vibrating mirror 5 for example. The sample to be tested is excited in the form of a scanning beam; wherein, the detection device 2 is composed of a detector and a detection optical path, and the detector is a complementary metal oxide semiconductor (CMOS) array camera 6 as an example, and the detection optical path includes an objective lens 7 and a barrel lens 8 ; Wherein, the main axis of the illumination optical path and the main axis of the detection optical path intersect at a non-zero angle, and the vertical intersection is taken as an example here.

Wherein, the sample detection area is located in the rotating mechanism 3 , and reference may be made to the detection area shown in the three-dimensional schematic diagram of the rotating mechanism shown in FIG. 2 . Wherein, the plane where the main shaft of the illumination optical path and the main shaft of the detection optical path are located is parallel or perpendicular to the rotating shaft of the rotating mechanism, and the vertical arrangement is taken as an example here; wherein, a rotating motor and a linear motion motor are arranged in the rotating mechanism as For example, it is used to cause the sample to rotate relative to the illumination device and the detection device during the imaging process to obtain different regions of the sample to be measured; wherein, the rotating motor and the linear motion motor move at a uniform speed.

It should be noted here that only one lighting device 1 , one detection device 2 and one rotating mechanism 3 are shown in FIG. 1 . Taking the illumination device 1 as an example, when the three-dimensional imaging device includes multiple illumination devices 1 , the excitation light sources of each illumination device 1 are different, and excitation light sources with different wavelengths can illuminate and excite different samples to be tested. During the imaging process, the corresponding illuminating device 1 can be selected according to the required wavelength range of the excitation light.

The three-dimensional imaging process can be referred to as shown in Figure 3. In FIG. 3 , the top view, front view and left view of the sample detection area during the imaging process are respectively shown. Arrows in the top view indicate the direction of motion during sample rotation, and dots or lines of different gray values indicate sample units that are sampled by illumination. The sample and the imaging device keep a continuous clockwise and spiral upward rotation at a constant speed, and the directions of the relative rotation axes are respectively perpendicular to the main axis direction of the illumination optical path and the main axis direction of the detection optical path. The arrows in the imaging plane of the front view indicate the scanning direction of the laser beam modulated by the galvanometer scan. The sample areas corresponding to the two images continuously collected by the three-dimensional imaging device are not coplanar. In a preferred embodiment, the moving speed of the sample can be controlled by the rotating mechanism, so that when the imaging surface is modulated for one period, the sample moves a distance of one field of view in the direction of the relative rotation axis. Wherein, one period of modulation of the imaging surface means that the imaging surface rotates one cycle along the direction of the relative rotation axis, and is on the same plane as the imaging surface at the initial position.

Since the imaging sample is rotated while collecting signals for imaging, and the relative rotation has a uniform speed and basically no pause, unnecessary interruption of imaging is reduced. When in the effective detection area of the detection device, the imaged sample unit is excited by the illumination device for a certain time (limited excitation time), the blurring caused by motion is negligible, that is, speed changes or start-up, Irregular jitters caused by pauses. Wherein, within the specified time, the displacement generated by the relative rotation is not greater than the resolution of the three-dimensional imaging device. If the blur caused by motion still exists, it can be calculated and eliminated by methods such as deconvolution.

Further, limiting the excitation time can be achieved by a pulsed light source with an appropriate pulse width, or a continuous light source can also be used for illumination by proper on-off modulation. But more efficiently, the continuous light source can also achieve short-term excitation of the point by scanning. A specific scanning method is shown in Figure 4, that is, the continuous light beam quickly scans on the photosensitive element of the camera, and the scanned photosensitive element starts at this time. Expose and record, scanning and exposure are synchronized until a frame is completed. An implementation manner is shown in FIG. 5 as an example, the field of view of the microscope, taking the imaging camera as a reference, and the width d is 2000 pixels. Correspondingly, the width of the line-shaped excitation light is about 10 pixels. When the excitation light sweeps across the entire field of view within 0.01 second, the actual time for each pixel to be excited is only about 0.01 second divided by 2000 times 10 or 50 microseconds. In order to realize the synchronization of rotation scanning and imaging, the motion blur caused by rotation during imaging can be eliminated. The number of pixels of the camera is 2000×2000, the pixel size is 6.5×6.5 microns, and the full-frame frame rate is 100Hz, that is, each frame is 10 milliseconds. The vibrating mirror 6 scans synchronously with a one-way 10 millisecond sawtooth wave. The maximum linear speed at which the sample rotates is less than 100 microns/second or 1 micron/frame. Motion-induced blur is less than 1/200 and can be ignored. At the same time, in order to eliminate the motion blur caused by moving at a constant speed during rotation, the time for one revolution is equal to the time for moving an imaging width at a constant speed along the rotation axis, that is, πd/ω=w/υ (imaging width d, scanning height w, rotational speed is ω, Vertical motion speed υ).

In a specific example, according to the above scanning imaging method, the contrast on which the imaging depends is fluorescence, the rotation speed of the rotating mechanism is 3.6 seconds/rev, and the mouse brain fluorescence imaging is performed by moving at a constant speed of 0.2 μm/ms along the rotation axis, The imaging results can refer to the content shown in Figure 6. Among them, Fig. 6 shows the imaging images rotated by 20° every 200ms, but the imaging and motion are continuous.

In one embodiment, the environment where the imaging sample is located is filled with a transparent substance, and the difference in refractive index between the transparent substance and the sample is smaller than a preset value. That is, choosing a transparent material with a refractive index similar to that of the imaging sample can reduce refracted light, allow more light to enter the lens, and improve resolution.

In one embodiment, the excitation light source of the illuminating device 1 is at least one of one or more light emitting diodes, continuous lasers, pulsed lasers, and X-ray generators.

In one embodiment, the detected contrast may be one or more of fluorescence, elastic scattered light, Raman scattering, second harmonic signal, third harmonic signal or stimulated Raman scattering signal.

In one embodiment, the illumination device uses pulses with a pulse time less than or equal to the specified time, and correspondingly, the detection device uses a matrix photosensitive device as an imaging device. During the specified time, the displacement caused by the relative rotation is not greater than the resolution of the three-dimensional imaging device. The pulsed light source may comprise one or more pulsed lasers, pulsed light-emitting diodes or a combination of continuous light sources modulated in different ways. And the pulse light source can work synchronously with the imaging device.

In one embodiment, the scanning mechanism includes a combination of one or more galvanometer scanners, resonant scanning mirrors, rotating polygonal mirrors, and acousto-optic modulators.

In one embodiment, the detection device uses a matrix photosensitive device such as a CCD or a CMOS camera as an imaging device, and the imaging device works synchronously with the scanning mechanism. The detection device may include array or matrix photosensitive devices such as LED arrays, CCD, CMOS cameras as imaging devices.

In one embodiment, the illuminating device may include a beam splitting mechanism for splitting one illuminating beam into multiple illuminating beams to illuminate or excite sub-regions of the sample to be tested, so as to shorten the total imaging time of the entire sample. The beam splitting mechanism may comprise a combination of one or more lens arrays, mirror arrays, half-mirror arrays, or a fixed or adjustable phase filter or a digital micromirror array (DMD).

In one embodiment, the illuminating device and the detecting device may include a pair of conjugated small holes, forming a scanning confocal imaging structure.

In one embodiment, the illuminating device may include at least one wavefront sensor and a wavefront corrector, which are used to correct wavefront distortion of illumination light in the area to be measured and improve imaging quality.

The technical solution of this embodiment solves the problem of long imaging interruption time in the process of three-dimensional imaging by making the sample, the illumination device, and the detection device perform uniform spiral motion during the process of detecting the sample signal, that is, performing continuous signal acquisition while the sample is rotating. , the problem of low imaging efficiency; it can reduce the imaging interruption during the three-dimensional imaging process, eliminate the blur caused by the sample rotation during imaging in the existing three-dimensional imaging technology, and improve the aberration.

In a specific embodiment, FIG. 7 shows the process of imaging and scanning the imaging sample. In this process, through the uniform rotation of the rotating mechanism, the uniform linear movement in the vertical direction and the horizontal movement, the full height of the sample is Different sections are continuously placed in the area to be tested, the innermost imaging diameter is a 2d cylinder, and the outermost imaging is a hollow tube with a thickness of 2d, from inside to outside, or from outside to inside, until the entire sample is covered. After the sample is excited, the detection device can be used to detect the signal generated by its illumination or excitation by the illumination device, and finally a three-dimensional image of the large sample can be obtained. In a preferred embodiment, the vertical speed of the sample can be controlled by the rotating mechanism, so that when the imaging surface is modulated for one cycle, the sample moves a distance of the scanning height w in the direction of the relative rotation axis, where d and w is the same as the imaging width d and scanning height w in Fig. 5.

For the same imaging sample, during the imaging process, the moving trajectory as shown in FIG. 8 can also be adopted. In the imaging process in Figure 8, through the uniform rotation of the rotating mechanism and the uniform linear motion in the horizontal direction, the cylinder with the imaging height w is obtained in a spiral manner; then the height is increased sequentially by moving in the vertical direction until the three-dimensional image of the entire sample is obtained. In a preferred embodiment, the horizontal speed of the sample can be controlled by the rotating mechanism, so that when the imaging surface is modulated for one cycle, the sample moves in the horizontal direction at a distance d of the imaging field of view.

Further, during the imaging process, a moving trajectory as shown in FIG. 9 may also be adopted. In the imaging process in Figure 9, the rotation mechanism rotates at a constant speed while moving in the horizontal direction and the vertical direction. First, the cylinder whose height is the scanning height w is imaged, and then the circular area with the field of view width d is sequentially imaged by moving in the horizontal direction; and then By moving in the vertical direction, the height is increased sequentially, and then the aforementioned scanning and imaging process is repeated until a three-dimensional image of the entire sample is obtained. In a preferred embodiment, when the imaging surface is modulated for one period, then the sample is moved in the horizontal direction and the vertical direction by a distance of imaging field width d.

In this embodiment, the illuminating device, the rotating mechanism and the detecting device work synchronously, and the rotating mechanism is used to make the imaging sample rotate relative to the illuminating device and the detecting device, and the relative rotation is a uniform motion. The main axis direction and the main axis direction of the detection optical path are vertical. Signal detection is performed while the imaging sample is rotating at a constant speed, that is, imaging is performed while rotating, thereby reducing imaging interruption.

In another embodiment, the method of limiting the excitation time can also be used to suppress motion blur, the main point of which is to ensure that the relative resolution of the motion of each point in the sample within the excitation time can be ignored, that is, the excitation time multiplied by the motion speed Significantly smaller than the minimum resolution scale. If the resolution requirement is 1 micron and the motion speed is 1 mm/s, the excitation time is required to be less than 1 millisecond. A pulsed light source with an appropriate pulse width can directly meet this requirement. CW light sources can also be modulated by appropriate switches for lighting, but more efficiently, CW light sources can also be scanned to achieve short-term excitation of points. When the motion blur is negligible, after the contrast signal is converted into a digital signal, the 3D image can be obtained by directly reconstructing the digital signal. All images obtained by imaging are arranged according to their corresponding sample spatial positions, and a three-dimensional image of the sample can be reconstructed, as shown in Figure 10.

Embodiment 2

Embodiment 2 of the present invention provides a three-dimensional imaging method, and this embodiment is applicable to three-dimensional imaging, especially for three-dimensional imaging of biological and medical samples.

The three-dimensional imaging method includes the following steps:

S110. Control at least one illuminating device to excite the sample to be tested in the sample detection area along the main axis of the illumination light path to generate a detected signal.

After the light source of the lighting device emits light, the sample to be tested is excited by the lighting device within a preset time; within the preset time, the displacement of the sample to be tested during the relative rotation process is less than or equal to the longitudinal resolution of the three-dimensional imaging device rate and horizontal resolution.

S120. Control at least one detection device to detect signals generated in the sample detection area along the main axis direction of the detection optical path.

S130. Control at least one rotating mechanism so that the sample to be tested rotates relative to the illuminating device and the detection device during the imaging process, and collect the sample to be tested at different positions through the at least one detection device. Signals generated in the measurement area.

Specifically, the above steps are applicable to the three-dimensional imaging device described in any of the above embodiments. The illuminating light path excites the illuminating light source in the form of scanning light beams to excite the sample to be measured (i.e. the imaging sample); the main axis of the detection optical path intersects with the main axis of the illuminating optical path; The vertical shaft rotates; the scanning direction of the scanning beam is parallel to or perpendicular to the direction of the shaft.

In the process of collecting imaging signals, at least one rotating mechanism will be controlled so that the sample to be tested will rotate relative to the illumination device and the detection device during the imaging process, that is, at least one rotating mechanism will be controlled to make the sample to be tested rotate in the imaging process. The continuous rotation during the imaging process, or the continuous movement in the form of a combination of linear motion and rotary motion, produces relative rotation with the illumination device and the detection device.

The illuminating device, the rotating mechanism and the detecting device of the three-dimensional imaging device work synchronously, and the imaging sample is relatively rotated with the illuminating device and the detecting device by using the rotating mechanism, and the relative rotation is a uniform motion. , The direction of the main axis of the detection optical path is vertical. Signal detection is performed while the imaging sample is rotating at a constant speed, that is, imaging is performed while rotating, thereby reducing imaging interruption. That is, during the rotation of the imaging sample, the signal is collected, and then the collected signal is used as the basis for three-dimensional image reconstruction.

Furthermore, by setting different motion parameters for the rotating mechanism, different motion trajectories of the imaging sample (refer to FIG. 7, FIG. 8 and FIG. 9) will be generated, that is, different imaging data will be obtained to obtain the final imaging result.

S140. Convert the collected signal into a digital signal.

The collected signal is an electrical signal, which needs to be further converted into a digital signal for image reconstruction.

S150. Perform image reconstruction based on the digital signal to obtain a three-dimensional image of the sample to be tested.

In the specific process of image reconstruction based on the data signal, any preset algorithm capable of realizing three-dimensional reconstruction may be used. In a preferred implementation manner, denoising processing is also performed on the digital signal to eliminate motion artifacts generated during the imaging process and obtain clearer imaging results.

In the technical solution of this embodiment, by using the three-dimensional imaging device in the above embodiment, the sample, the lighting device, and the detection device perform a uniform spiral motion during the process of detecting the sample signal, that is, continuous signal acquisition is performed while the sample is rotating, The problem of long imaging interruption time and low imaging efficiency in the three-dimensional imaging process is solved; the imaging interruption in the three-dimensional imaging process can be reduced, the blur caused by the sample rotation during imaging in the existing three-dimensional imaging technology can be eliminated, and the aberration can be improved.

Note that the above are only preferred embodiments of the present invention and applied technical principles. Those skilled in the art will understand that the present invention is not limited to the specific embodiments described herein, and that various obvious changes, readjustments and substitutions can be made by those skilled in the art without departing from the protection scope of the present invention. Therefore, although the present invention has been described in detail through the above embodiments, the present invention is not limited to the above embodiments, and can also include more other equivalent embodiments without departing from the concept of the present invention, and the present invention The scope is determined by the scope of the appended claims.

Those of ordinary skill in the art should understand that each module or each step of the present invention described above can be realized by a general-purpose computing device, and they can be concentrated on a single computing device, or distributed on a network formed by multiple computing devices. Optionally, they can be implemented with executable program codes of computer devices, so that they can be stored in storage devices and executed by computing devices, or they can be made into individual integrated circuit modules, or a plurality of modules in them Or the steps are fabricated into a single integrated circuit module to realize. As such, the present invention is not limited to any specific combination of hardware and software.

Note that the above are only preferred embodiments of the present invention and applied technical principles. Those skilled in the art will understand that the present invention is not limited to the specific embodiments described herein, and that various obvious changes, readjustments and substitutions can be made by those skilled in the art without departing from the protection scope of the present invention. Therefore, although the present invention has been described in detail through the above embodiments, the present invention is not limited to the above embodiments, and can also include more other equivalent embodiments without departing from the concept of the present invention, and the present invention The scope is determined by the scope of the appended claims.

Claims (12)

1. A three-dimensional imaging device, characterized in that, comprising: At least one illuminating device, including an illuminating light source and an illuminating light path, for illuminating or exciting the sample to be tested; At least one detection device, including a detector and a detection optical path, used to detect the signal generated by the sample to be tested excited by the illumination device in the area to be tested; at least one rotating mechanism, used to make the sample under test rotate relative to the illuminating device and the detecting device during the imaging process; Wherein, the illumination light path excites the sample to be measured by the illumination light source in the form of a scanning beam; the main axis of the detection optical path intersects with the main axis of the illumination optical path and the included angle is not zero; the main axis of the illumination optical path and the The plane where the main axis of the detection optical path is located coincides with, is parallel to or perpendicular to the rotation axis of the rotation mechanism. 2 . The three-dimensional imaging device according to claim 1 , wherein at least one scanning mechanism is included in the illuminating device for continuously moving the illuminating light source to form a scanning light beam. 3 . 3. The three-dimensional imaging device according to claim 2, wherein the scanning mechanism comprises one or more scanning galvanometers, resonant scanning mirrors, rotating polygonal mirrors or acousto-optic modulators. 4. The three-dimensional imaging device according to claim 1, wherein the illumination device comprises one or more lasers, light emitting diodes or X-ray generators. 5. The three-dimensional imaging device according to claim 1, wherein the signal generated by the sample to be measured excited by the illumination device is fluorescence, elastic scattered light, Raman scattering, second harmonic signal, third harmonic signal One or more of wave signals or Stimulated Raman scattering signals. 6. The three-dimensional imaging device according to claim 1, wherein the detection device works synchronously with the illumination device. 7 . The three-dimensional imaging device according to claim 1 , wherein the detector uses a charge-coupled device or a complementary metal-oxide-semiconductor array as an imaging device. 8. The three-dimensional imaging device according to claim 1, wherein the rotating mechanism comprises at least one rotating device, or a combined device of a rotating device and a linear motion device. 9. The three-dimensional imaging device according to claim 8, wherein the rotating mechanism further comprises at least one control device for controlling the movement of the rotating device and the linear moving device. 10. A high-throughput three-dimensional imaging method, comprising: Controlling at least one lighting device to excite the sample to be tested in the sample detection area along the main axis direction of the illumination light path to generate a detected signal; Controlling at least one detection device to detect signals generated in the sample detection area along the main axis of the detection optical path; Controlling at least one rotating mechanism, so that the sample to be tested rotates relative to the illumination device and the detection device during the imaging process, and the sample to be tested is collected in different regions to be tested by the at least one detection device the signal generated; Convert the collected signal into a digital signal; performing image reconstruction based on the digital signal to obtain a three-dimensional image of the sample to be tested; Wherein, the illumination light path excites the sample to be measured by the illumination light source in the form of a scanning beam; the main axis of the detection optical path intersects the main axis of the illumination light path; The plane where the main axis of the detection optical path is located rotates parallel to or perpendicular to the rotation axis; the scanning direction of the scanning light beam is parallel to or perpendicular to the direction of the rotation axis. 11. The method according to claim 10, wherein the controlling at least one rotating mechanism to make the sample under test rotate relative to the illuminating device and the detecting device during the imaging process comprises: At least one rotating mechanism is controlled so that the sample to be tested rotates continuously during the imaging process, or continuously moves in a combination of linear motion and rotary motion, and generates relative rotation with the illuminating device and the detecting device. 12. The method according to claim 11, characterized in that, during the imaging process, the sample to be tested is excited by the illumination device within a preset time; wherein, within the preset time, the sample to be tested The displacement of the test sample during the relative rotation process is less than or equal to the longitudinal resolution and lateral resolution of the three-dimensional imaging device.

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