CN115248197A - Three-dimensional imaging device and imaging method - Google Patents
Three-dimensional imaging device and imaging method Download PDFInfo
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- CN115248197A CN115248197A CN202111506363.9A CN202111506363A CN115248197A CN 115248197 A CN115248197 A CN 115248197A CN 202111506363 A CN202111506363 A CN 202111506363A CN 115248197 A CN115248197 A CN 115248197A
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- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
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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
Technical Field
The embodiment of the invention relates to the technical field of imaging, in particular to a three-dimensional imaging device and an imaging method.
Background
The acquisition of three-dimensional structural information of a sample by imaging is one of the important techniques for biological and medical research and testing. Obtaining more image pixels through imaging in a short time to extract more information of the sample is an important direction for the development of three-dimensional imaging technology.
In order to specifically recognize specific signals at different positions in biological and medical samples, fluorescent molecular markers and the like are usually adopted so that different molecules in the samples can generate specific fluorescent signals under the illumination of specific lasers. Collecting these signals in a full coverage manner for all regions within the sample allows the reconstruction of the three-dimensional structure that generated the sample. The three-dimensional sample imaging throughput depends on which full coverage approach is used to cover all sample areas.
The existing imaging technology usually obtains a planar image of a sample detection area, then images parallel sample planes, and superimposes the parallel planes to reconstruct a three-dimensional image of the sample. Taking scanning confocal imaging or photo-section imaging as an example, the imaging scheme generally adopted by the existing imaging technology is as follows: firstly, point-by-point imaging (confocal imaging) is carried out on a plane vertical to the imaging direction (namely the main axis z direction of the detection light path), or part of the whole visual field is imaged simultaneously (optical section imaging); then, translating in the z direction according to the resolution requirement to reach a new plane, and imaging the new plane; the above motion and imaging are repeated until the sample is covered in the z-direction, completing three-dimensional imaging in one field of view. Since imaging while translating motion in the z-direction tends to produce motion blur, exposure imaging is typically performed only when the sample is stationary at the target location. These ineffective imaging times reduce the imaging throughput since the time it takes for the sample to move between adjacent z-positions is typically much greater than the exposure time.
In the multi-angle light section imaging technology, a sample is rotated by a certain angle, then the scanning is started to complete the imaging of one layer, then the scanning is performed after the scanning is stopped by a certain angle, and then the imaging is performed, and the three-dimensional reconstruction can be performed on the sample through the multi-angle imaging of a plurality of angles (usually not more than 8 different angles). But also involves a number of processes that interrupt the imaging during the rotation between different angles. After the vibration caused by each rotation and waiting starting and stopping is dissipated, the imaging can be continued, so that long-time imaging interruption is generated, the imaging time and the imaging workload are increased, and the imaging flux is reduced.
Disclosure of Invention
The invention provides a three-dimensional imaging device and an imaging method, which are used for realizing multi-view continuous imaging in the three-dimensional imaging process, reducing unnecessary imaging interruption and improving the imaging speed and the imaging efficiency.
In a first aspect, the present invention provides a three-dimensional imaging apparatus comprising:
the lighting device comprises a lighting source and a lighting light path and is used for lighting or exciting a sample to be tested;
at least one detection device comprising a detector and a detection light path and used for detecting a signal generated by the excitation of the sample to be detected by the illumination device in the region to be detected;
at least one rotating mechanism, which is used for enabling the sample to be detected to rotate relative to the illuminating device and the detecting device in the imaging process;
the illumination light path excites the sample to be detected by an illumination light source in a scanning beam mode; the main shaft of the detection light path is intersected with the main shaft of the illumination light path, and the included angle is not zero; the plane of the main shaft of the illumination light path and the plane of the main shaft of the detection light path are coincident with, parallel to or perpendicular to the rotating shaft of the rotating mechanism.
Optionally, at least one scanning mechanism is included in the illumination device for continuously moving the illumination source to form the scanning beam.
Optionally, the scanning mechanism comprises one or more of a scanning galvanometer, a resonant scanning mirror, a rotating polygon mirror, or an acousto-optic modulator.
Optionally, the illumination device comprises one or more lasers, light emitting diodes or X-ray generators.
Optionally, the signal generated by the sample to be detected excited by the illumination device is one or more of fluorescence, elastic scattering light, raman scattering, second harmonic signal, third harmonic signal or laser raman scattering signal.
Optionally, the detection device and the illumination device work synchronously.
Optionally, the detector uses a charge coupled device or a complementary metal oxide semiconductor array as an imaging device.
Optionally, the rotating mechanism comprises at least one rotating device, or a combination of a rotating device and a linear motion device.
Optionally, the rotating mechanism further comprises at least one control device for controlling the movement of the rotating device and the linear motion device.
In a second aspect, an embodiment of the present invention further provides a three-dimensional imaging method, where the method includes:
controlling at least one lighting device to excite a sample to be detected in a sample detection area along the main shaft direction of a lighting light path to generate a detected signal;
controlling at least one detection device to detect a signal generated in the sample detection region along a main axis direction of a detection optical path;
controlling at least one rotating mechanism to enable the sample to be detected to rotate relative to the illuminating device and the detecting device in the imaging process, and acquiring signals generated by the sample to be detected in different areas to be detected through the at least one detecting device;
converting the collected signals into digital signals;
carrying out image reconstruction based on the digital signal to obtain a three-dimensional image of the sample to be detected;
the illumination light path excites the sample to be detected by an illumination light source in a scanning beam mode; the main shaft of the detection light path is intersected with the main shaft of the illumination light path; the rotating mechanism rotates along a rotating shaft which is parallel to or vertical to the plane where the main shaft of the illumination light path and the main shaft of the detection light path are located; the scanning direction of the scanning light beam is parallel to or vertical to the direction of the rotating shaft.
Optionally, the controlling at least one rotating mechanism to make the sample to be detected rotate relative to the illumination device and the detection device in the imaging process includes:
and controlling at least one rotating mechanism to enable the sample to be detected to continuously rotate in the imaging process, or to continuously move in a mode of combining linear motion and rotary motion, so that the sample to be detected and the lighting device and the detection device generate relative rotation.
Optionally, the sample to be detected is excited by the lighting device within a preset time; and in the preset time, the displacement of the sample to be detected in the relative rotation process is respectively less than or equal to the longitudinal resolution and the transverse resolution of the three-dimensional imaging device.
According to the technical scheme of the embodiment of the invention, the sample, the illuminating device and the detecting device perform uniform relative motion in the process of detecting the sample signal, namely, the sample rotates and simultaneously performs continuous signal acquisition, so that the problems of long imaging interruption time and low imaging efficiency in the three-dimensional imaging process are solved, and the imaging flux is improved; the imaging interruption in the three-dimensional imaging process can be reduced, the blurring caused by sample rotation in the imaging in the existing three-dimensional imaging technology is eliminated, and the aberration is improved.
Drawings
Fig. 1 is a schematic structural diagram of a three-dimensional imaging apparatus according to an embodiment of the present invention;
FIG. 2 is a schematic three-dimensional structure diagram of a sample rotation mechanism according to an embodiment of the present invention;
FIG. 3 is a schematic diagram of a three-dimensional imaging scanning process according to an embodiment of the present invention;
FIG. 4 is a schematic view of an imaging scan provided by an embodiment of the present invention;
FIG. 5 is a schematic diagram of a principle of motion blur elimination in continuous scanning according to an embodiment of the present invention;
FIG. 6 is a schematic diagram of an example of fluorescence imaging of a mouse brain according to an embodiment of the present invention;
fig. 7 is a schematic diagram of a moving track of an imaging sample during an imaging signal acquisition process according to an embodiment of the present invention;
FIG. 8 is a schematic diagram of a movement track of an imaging sample during an imaging signal acquisition process according to an embodiment of the present invention;
FIG. 9 is a schematic diagram of a movement track of an imaging sample during an imaging signal acquisition process according to an embodiment of the present invention;
fig. 10 is a schematic diagram of a three-dimensional imaging effect of an imaging sample according to an embodiment of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings and examples. It is to be understood that the specific embodiments described herein are merely illustrative of the invention and are not limiting of the invention. It should be further noted that, for the convenience of description, only some of the structures related to the present invention are shown in the drawings, not all of the structures.
Example one
Fig. 1 is a schematic structural diagram of a three-dimensional imaging apparatus according to an embodiment of the present invention, which is applicable to a case of performing three-dimensional imaging using the apparatus, in particular, performing three-dimensional imaging on biological and medical samples.
As shown in fig. 1, the three-dimensional imaging apparatus specifically includes: at least one lighting device 1, at least one detection device 2; at least one turning mechanism 3. The illumination device 1 consists of an illumination light source and an illumination light path, the illumination light source is exemplified by a laser 4 with a wavelength of 488 nanometers, the illumination light path comprises a scanning mechanism exemplified by a scanning galvanometer 5 and a relay lens 9, and the scanning galvanometer 5 excites a sample to be detected in a scanning light beam mode; the detection device 2 is composed of a detector and a detection optical path, wherein the detector is a Complementary Metal Oxide Semiconductor (CMOS) array camera 6 as an example, and the detection optical path comprises an objective lens 7 and a tube lens 8; the main axis of the illumination light path intersects with the main axis of the detection light path at an included angle that is not zero, and here, the perpendicular intersection is taken as an example.
Wherein the sample detection area is located in the rotating mechanism 3, reference can be made to the detection area shown in the three-dimensional schematic view of the rotating mechanism shown in fig. 2. The plane where the main axis of the illumination light path and the main axis of the detection light path are located is parallel to or perpendicular to the rotating shaft of the rotating mechanism, and the vertical arrangement is taken as an example here; the rotating mechanism is internally provided with a rotating motor and a linear motion motor for example, and is used for enabling the sample to rotate relative to the lighting device and the detection device in the imaging process so as to obtain different regions to be detected of the sample; wherein the rotating motor and the linear motion motor move at a constant speed.
It should be noted here that fig. 1 shows only one lighting device 1, one detection device 2, and one rotation mechanism 3. Taking the illumination device 1 as an example, when the three-dimensional imaging device includes a plurality of illumination devices 1, the excitation light sources of the illumination devices 1 are different, and the excitation light sources with different wavelengths can illuminate and excite different samples to be detected. In the course of performing imaging, the respective illumination device 1 may be selected according to the desired wavelength range of the excitation light.
The three-dimensional imaging process can be seen with reference to fig. 3. In fig. 3, a top view, a front view and a left side view of the sample detection area during imaging are shown, respectively. The arrows in the top view indicate the direction of movement during rotation of the sample, and the dots or lines of different gray values indicate the sample cells sampled by illumination. The sample and the imaging device keep continuous clockwise and uniform spiral upward rotation, and the axial direction of the relative rotation is respectively vertical to the main shaft direction of the illumination light path and the main shaft direction of the detection light path. The arrows in the image plane of the front view indicate the laser beam scanning direction modulated by the galvanometer scan. The sample areas corresponding to two images continuously acquired 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 the sample moves a distance of one field of view in the direction of the rotating shaft of the relative rotation when the imaging plane is modulated for one period. The imaging plane is modulated for one period, namely the imaging plane rotates for one circle along the direction of the rotating shaft rotating relatively, and the imaging plane at the initial position are on the same plane.
Because the imaging sample rotates and simultaneously acquires signals for imaging basically and continuously, and the relative rotation has the characteristics of uniform speed and basically no pause, the unnecessary imaging interruption is reduced. When the imaged sample unit is excited by the illumination device within a specific time (limited excitation time) in the effective detection area of the detection device, the motion-induced blur is negligible, i.e. the irregular jitter caused by speed variation or start-up and pause can be eliminated. Wherein, in the specific 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 the motion still exists, the motion blur can be eliminated by calculation through a deconvolution method and the like.
Further, limiting the excitation time may be achieved by a pulsed light source of suitable pulse width, or a continuous light source may be used for illumination by suitable switching modulation. But more efficiently, the continuous light source can also realize the short-time excitation of the point by a scanning mode, one specific scanning mode is shown in fig. 4, namely, the continuous light beam rapidly scans on the photosensitive element of the camera, the scanned photosensitive element starts to expose and record at the moment, and the scanning and the exposure are synchronized until one frame of picture is completed. One implementation is shown in fig. 5, where the microscope field of view, referenced to the imaging camera, has a width d of 2000 pixels. Accordingly, the excitation light width of the line shape is about 10 pixels. When the excitation light is swept across the entire field of view in 0.01 seconds, the time each pixel is actually excited is only about 0.01 seconds divided by 2000 times 10, i.e., 50 microseconds. In order to realize the synchronization of the rotation scanning and the imaging, the motion blurring caused by the rotation in the imaging process is eliminated. The camera pixel count is 2000 × 2000, the pixel size is 6.5 × 6.5 microns, and the full frame rate is 100Hz, i.e., 10 milliseconds per frame. The galvanometer 6 is synchronously scanned with a unidirectional 10 millisecond sawtooth wave. The maximum linear velocity of the sample rotation is less than 100 microns/second, i.e. 1 micron/frame. The motion-induced blur is less than 1/200, and can be ignored. Meanwhile, in order to eliminate motion blur caused by uniform motion during rotation, the time of one circle of rotation is equal to the time of one imaging width which is moved along the rotating shaft at a uniform speed, namely pi d/omega = w/upsilon (imaging width d, scanning height w, rotating speed omega, and vertical motion speed upsilon).
In a specific example, according to the above scanning imaging method, the contrast depended on by imaging is fluorescence, the rotation speed of the rotating mechanism is 3.6 seconds/rotation, and the fluorescence of the mouse brain is imaged by moving at a constant speed of 0.2 μm/ms along the rotation axis, and the imaging result can be referred to the content shown in fig. 6. In which figure 6 shows an imaging view taken at every 200ms rotation of 20, but with continuous imaging and motion.
In one embodiment, the environment in which the sample is imaged is filled with a transparent substance, and the difference between the refractive index of the transparent substance and the refractive index of the sample is smaller than a preset value. The refractive index of the transparent substance is similar to that of the imaging sample, so that refracted light can be reduced, more light rays enter the lens, and the resolution is improved.
In one embodiment, the excitation light source of the illumination device 1 is at least one of one or more light emitting diodes, a continuous laser, a pulsed laser, and an X-ray generator.
In one embodiment, the contrast to be detected may be one or more of fluorescence, elastically scattered light, raman scattering, second harmonic signal, third harmonic signal, or stimulated raman scattering signal.
In one embodiment, the illumination means uses pulses having a pulse time less than or equal to the specific time, and accordingly, the detection means uses a matrix photosensitive device as the imaging device. The displacement generated by the relative rotation is not greater than the resolution of the three-dimensional imaging device within the specific time. 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 pulsed light source may be operated in synchronization with the imaging device.
In one embodiment, the scanning mechanism comprises a combination of one or more scanning galvanometer (galvometer scanner), resonant scanning mirror, rotating polygon mirror, and acousto-optic modulator.
In one embodiment, the detection device is a matrix photosensitive device such as a CCD or CMOS camera as the imaging device, which is synchronized with the scanning mechanism. The detection means may comprise an array or matrix light sensitive device such as an LED array, CCD, CMOS camera as the imaging device.
In one embodiment, the illumination device may comprise a beam splitting mechanism for splitting an illumination beam into a plurality of illumination beams to illuminate or excite a sub-region of the sample to be measured, thereby reducing the overall time for imaging the sample. The beam splitting mechanism may comprise a combination of one or more lens arrays, mirror arrays, half-mirror arrays, or comprise fixed or adjustable phase filters or digital micromirror arrays (DMDs).
In one embodiment, the illumination device and the detection device may comprise a pair of conjugated apertures, which constitute a scanning confocal imaging structure.
In one embodiment, the illumination device may include at least one wavefront sensor and a wavefront corrector for correcting wavefront distortion of illumination light of the region to be measured, thereby improving imaging quality.
According to the technical scheme of the embodiment, the sample, the illuminating device and the detecting device perform uniform spiral motion in the process of detecting the sample signal, namely, continuous signal acquisition is performed while the sample rotates, so that the problems of long imaging interruption time and low imaging efficiency in the three-dimensional imaging process are solved; the imaging interruption in the three-dimensional imaging process can be reduced, the blurring caused by sample rotation in the imaging in the existing three-dimensional imaging technology is eliminated, and the aberration is improved.
In a specific embodiment, fig. 7 shows a process of performing imaging scanning on an imaging sample, in which different cross sections of the full height of the sample are continuously placed in an excitation region to be measured through uniform rotation of a rotating mechanism, uniform linear movement in a vertical direction, and movement in a horizontal direction, an innermost side of the sample is a 2 d-cylinder, and an outer side of the sample is a hollow circular tube with a thickness of 2d, sequentially from inside to outside, or from outside to inside, until the whole sample is covered. After the sample is excited, the detection device can be used for detecting the signal generated by the illumination or excitation of the illumination device, and finally, a three-dimensional image of the large sample is 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 period, the sample moves in the direction of the rotating shaft of the relative rotation by a distance of scanning height w, wherein d and w are the same as the imaging width d and the scanning height w in fig. 5.
For the same imaging sample, during the imaging process, the movement track shown in fig. 8 can also be adopted. In the imaging process of fig. 8, a cylinder with an imaging height w is obtained in a spiral manner by the uniform rotation of the rotating mechanism and the uniform linear motion in the horizontal direction; and then sequentially increasing the height by moving in the vertical direction until a three-dimensional image of the whole sample is obtained. In a preferred embodiment, the horizontal direction 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 by the imaging field width d.
Further, during the imaging process, the movement track shown in fig. 9 can be adopted. In the imaging process of fig. 9, the rotating mechanism rotates at a constant speed, and moves in the horizontal direction and the vertical direction at the same time, a cylinder with the scanning height w is imaged, and then an annular region with the view field width d is sequentially imaged by moving in the horizontal direction; and then moving in the vertical direction, sequentially increasing the height, and repeating the scanning and imaging processes until a three-dimensional image of the whole sample is obtained. In a preferred embodiment, when the imaging plane is modulated for one period, then the sample is moved in the horizontal and vertical directions by the imaging field width d.
In this embodiment, the illumination device, the rotation mechanism and the detection device operate synchronously, the rotation mechanism is used to make the imaging sample rotate relative to the illumination device and the detection device, and the relative rotation is a uniform motion, and the axial direction of the relative rotation is perpendicular to the main axis direction of the illumination light path and the main axis direction of the detection light path respectively. The imaging sample rotates at a constant speed and simultaneously detects signals, namely, imaging is carried out while rotating, so that imaging interruption can be reduced.
In another embodiment, motion blur can be suppressed by limiting the excitation time, which is to ensure that the relative resolution of motion at each point in the sample over the excitation time is negligible, i.e., the excitation time multiplied by the motion velocity is significantly less than the minimum resolution. If the resolution requirement is 1 micron and the motion speed is 1 mm per second, then the excitation time is required to be less than 1 millisecond. A pulsed light source with a suitable pulse width can directly meet this requirement. The continuous light source can also be used for illumination by means of suitable switching modulation, but more efficiently, the continuous light source can also achieve a short excitation of the dots by means of a scanning mode. When the motion blur is negligible, the contrast signal can be converted into a digital signal, and then the digital signal is directly reconstructed to obtain a three-dimensional image. All the images obtained by imaging are arranged according to the corresponding sample space positions, and a three-dimensional image of the sample can be reconstructed, as shown in fig. 10.
Example two
The second embodiment of the invention provides a three-dimensional imaging method, which is applicable to the three-dimensional imaging condition, in particular to the three-dimensional imaging of biological and medical samples.
The three-dimensional imaging method comprises the following steps:
and S110, controlling at least one lighting device, and exciting the sample to be detected in the sample detection area along the main shaft direction of the lighting light path to generate a detected signal.
After a light source of the lighting device emits irradiation light, the sample to be tested is excited by the lighting device within a preset time; and in a preset time, the displacement of the sample to be detected in the relative rotation process is respectively less than or equal to the longitudinal resolution and the transverse resolution of the three-dimensional imaging device.
And S120, controlling at least one detection device to detect signals generated in the sample detection area along the main axis direction of the detection light path.
S130, controlling at least one rotating mechanism to enable the sample to be detected to rotate relative to the lighting device and the detection device in the imaging process, and collecting signals generated by the sample to be detected in different areas to be detected through the at least one detection device.
Specifically, the above steps are applicable to the three-dimensional imaging apparatus described in any of the above embodiments. The illumination light path excites the sample to be detected (namely an imaging sample) by the illumination light source in a scanning beam mode; the main shaft of the detection light path is intersected with the main shaft of the illumination light path; the rotating mechanism rotates along a rotating shaft which is parallel to or vertical to the plane where the main shaft of the illumination light path and the main shaft of the detection light path are positioned; the scanning direction of the scanning beam is parallel or vertical to the direction of the rotating shaft.
In the process of acquiring imaging signals, at least one rotating mechanism is controlled, so that a sample to be detected rotates relative to the illuminating device and the detecting device in the imaging process, namely, at least one rotating mechanism is controlled, so that the sample to be detected rotates continuously in the imaging process, or the sample to be detected moves continuously in a combined mode of linear motion and rotary motion and rotates relative to the illuminating device and the detecting device.
The illuminating device, the rotating mechanism and the detecting device of the three-dimensional imaging device work synchronously, the rotating mechanism is utilized to enable the imaging sample, the illuminating device and the detecting device to rotate relatively, the relative rotation is uniform motion, and the axial direction of the relative rotation is respectively vertical to the main shaft direction of the illuminating light path and the main shaft direction of the detecting light path. The imaging sample rotates at a constant speed and simultaneously detects signals, namely, imaging is carried out while rotating, so that imaging interruption can be reduced. In the process of imaging sample rotation, signals are collected, and then the collected signals are used as the basis of three-dimensional image reconstruction.
Further, by setting different motion parameters for the rotating mechanism, different motion trajectories (refer to fig. 7, 8 and 9) of the imaging sample can be generated, that is, different imaging data can be acquired, so as to obtain a final imaging result.
And S140, converting the acquired signals into digital signals.
The acquired signals are electrical signals and need to be further converted into digital signals for image reconstruction.
And S150, carrying out image reconstruction based on the digital signal to obtain a three-dimensional image of the sample to be detected.
In particular, the process of image reconstruction according to the data signal may employ any preset algorithm that can achieve three-dimensional reconstruction. In a preferred embodiment, the digital signal is also subjected to denoising processing to eliminate motion artifacts generated in the imaging process, so as to obtain a clearer imaging result.
According to the technical scheme of the embodiment, by using the three-dimensional imaging device in the embodiment, the sample, the illuminating device and the detecting device perform uniform spiral motion in the process of detecting the sample signal, namely, continuous signal acquisition is performed while the sample rotates, so that the problems of long imaging interruption time and low imaging efficiency in the three-dimensional imaging process are solved; the imaging interruption in the three-dimensional imaging process can be reduced, the blurring caused by the rotation of the sample in the existing three-dimensional imaging technology can be eliminated, and the aberration can be improved.
It is to be noted that the foregoing is only illustrative of the preferred embodiments of the present invention and the technical principles employed. It will be understood by those skilled in the art that the present invention is not limited to the particular embodiments described herein, but is capable of various obvious changes, rearrangements and substitutions as will now become apparent to those skilled in the art without departing from the scope of the invention. Therefore, although the present invention has been described in greater detail by the above embodiments, the present invention is not limited to the above embodiments, and may include other equivalent embodiments without departing from the spirit of the present invention, and the scope of the present invention is determined by the scope of the appended claims.
It will be understood by those skilled in the art that the modules or steps of the invention described above may be implemented by a general purpose computing device, they may be centralized on a single computing device or distributed across a network of computing devices, and optionally they may be implemented by program code executable by a computing device, such that it may be stored in a memory device and executed by a computing device, or it may be separately fabricated into various integrated circuit modules, or it may be fabricated by fabricating a plurality of modules or steps thereof into a single integrated circuit module. Thus, the present invention is not limited to any specific combination of hardware and software.
It is to be noted that the foregoing is only illustrative of the preferred embodiments of the present invention and the technical principles employed. It will be understood by those skilled in the art that the present invention is not limited to the particular embodiments described herein, but is capable of various obvious changes, rearrangements and substitutions as will now become apparent to those skilled in the art without departing from the scope of the invention. Therefore, although the present invention has been described in greater detail by the above embodiments, the present invention is not limited to the above embodiments, and may include other equivalent embodiments without departing from the spirit of the present invention, and the scope of the present invention is determined by the scope of the appended claims.
Claims (12)
1. A three-dimensional imaging apparatus, comprising:
the lighting device comprises a lighting source and a lighting light path and is used for lighting or exciting a sample to be tested;
at least one detection device comprising a detector and a detection light path and used for detecting a signal generated by the excitation of the sample to be detected by the illumination device in the region to be detected;
at least one rotating mechanism, which is used for enabling the sample to be detected to rotate relative to the illuminating device and the detecting device in the imaging process;
the illumination light path excites the sample to be detected by an illumination light source in a scanning beam mode; the main shaft of the detection light path is intersected with the main shaft of the illumination light path, and the included angle is not zero; the plane of the main shaft of the illumination light path and the plane of the main shaft of the detection light path are coincident with, parallel to or perpendicular to the rotating shaft of the rotating mechanism.
2. The three-dimensional imaging apparatus according to claim 1, wherein at least one scanning mechanism is included in the illumination apparatus for continuously moving the illumination light source to form the scanning light beam.
3. The three-dimensional imaging device according to claim 2, wherein the scanning mechanism comprises one or more of a scanning galvanometer, a resonant scanning mirror, a rotating polygon mirror, or an acousto-optic modulator.
4. The three-dimensional imaging apparatus according to claim 1, wherein the illumination apparatus 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 tested excited by the illumination device is one or more of fluorescence, elastic scattered light, raman scattering, second harmonic signal, third harmonic signal or laser raman scattering signal.
6. The three-dimensional imaging apparatus according to claim 1, wherein the detecting means operates in synchronization with the illuminating means.
7. The three-dimensional imaging apparatus according to claim 1, wherein the detector has a charge coupled device or a complementary metal oxide semiconductor array as an imaging device.
8. The three-dimensional imaging apparatus according to claim 1, wherein the rotating mechanism comprises at least one rotating device, or a combination of a rotating device and a linear motion device.
9. The three-dimensional imaging apparatus according to claim 8, wherein said rotating mechanism further comprises at least one control device for controlling the movement of said rotating device and said linear motion device.
10. A method of high throughput three-dimensional imaging, comprising:
controlling at least one lighting device to excite a sample to be detected in a sample detection area along the main shaft direction of a lighting light path to generate a detected signal;
controlling at least one detection device to detect a signal generated in the sample detection region along a main axis direction of a detection optical path;
controlling at least one rotating mechanism to enable the sample to be detected to rotate relative to the illuminating device and the detecting device in the imaging process, and acquiring signals generated by the sample to be detected in different areas to be detected through the at least one detecting device;
converting the collected signals into digital signals;
carrying out image reconstruction based on the digital signal to obtain a three-dimensional image of the sample to be detected;
the illumination light path excites the sample to be detected by an illumination light source in a scanning beam mode; the main shaft of the detection light path is intersected with the main shaft of the illumination light path; the rotating mechanism rotates along a rotating shaft which is parallel to or vertical to the plane where the main shaft of the illumination light path and the main shaft of the detection light path are located; the scanning direction of the scanning light beam is parallel to or vertical to the direction of the rotating shaft.
11. The method of claim 10, wherein controlling the at least one rotation mechanism to rotate the sample to be tested relative to the illumination device and the detection device during the imaging process comprises:
and controlling at least one rotating mechanism to enable the sample to be detected to continuously rotate in the imaging process, or to continuously move in a mode of combining linear motion and rotary motion, so that the sample to be detected and the lighting device and the detection device generate relative rotation.
12. The method of claim 11, wherein the sample to be tested is excited by the illumination device for a preset time during the imaging process; and in the preset time, the displacement of the sample to be detected in the relative rotation process is respectively less than or equal to the longitudinal resolution and the transverse resolution of the three-dimensional imaging device.
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| CN202111506363.9A CN115248197A (en) | 2021-12-10 | 2021-12-10 | Three-dimensional imaging device and imaging method |
| PCT/CN2022/136589 WO2023103960A1 (en) | 2021-12-10 | 2022-12-05 | Three-dimensional imaging apparatus and imaging method |
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| WO2023103960A1 (en) * | 2021-12-10 | 2023-06-15 | 中国科学院深圳先进技术研究院 | Three-dimensional imaging apparatus and imaging method |
| CN117434084A (en) * | 2023-12-06 | 2024-01-23 | 四川万圣通实业有限公司 | Digital detection device and detection method for steel pipe |
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| CN117893695B (en) * | 2024-03-15 | 2024-06-04 | 浙江托普云农科技股份有限公司 | Three-dimensional reconstruction method, system and device for arbitrary plant height |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| DE102010013223B4 (en) * | 2010-03-29 | 2016-05-12 | Lavision Biotec Gmbh | Method and arrangement for microscopy |
| US8575570B2 (en) * | 2010-08-25 | 2013-11-05 | California Institute Of Technology | Simultaneous orthogonal light sheet microscopy and computed optical tomography |
| CN107991766A (en) * | 2016-10-26 | 2018-05-04 | 中国科学技术大学 | A kind of microscope and imaging method with three-dimensional imaging ability |
| CN108061964B (en) * | 2017-10-25 | 2020-03-27 | 中国科学技术大学 | High-speed three-dimensional microscopic imaging device and method for large sample |
| EP3748414A1 (en) * | 2019-06-07 | 2020-12-09 | Koninklijke Philips N.V. | A simple and efficient biopsy scanner with improved z-axis resolution |
| CN110638424B (en) * | 2019-09-19 | 2022-05-13 | 哈尔滨工业大学 | Scanning light sheet harmonic wave microscopic imaging method and device |
| CN113624731B (en) * | 2021-09-09 | 2024-09-27 | 深圳技术大学 | Super-resolution fluorescence hyperspectral microscopic imaging system |
| CN115248197A (en) * | 2021-12-10 | 2022-10-28 | 中国科学院深圳先进技术研究院 | Three-dimensional imaging device and imaging method |
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| WO2023103960A1 (en) * | 2021-12-10 | 2023-06-15 | 中国科学院深圳先进技术研究院 | Three-dimensional imaging apparatus and imaging method |
| CN117434084A (en) * | 2023-12-06 | 2024-01-23 | 四川万圣通实业有限公司 | Digital detection device and detection method for steel pipe |
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