CN111624756A - Depth-of-field extended microscopic imaging three-dimensional reconstruction device and method - Google Patents
Depth-of-field extended microscopic imaging three-dimensional reconstruction device and method Download PDFInfo
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- CN111624756A CN111624756A CN202010594981.2A CN202010594981A CN111624756A CN 111624756 A CN111624756 A CN 111624756A CN 202010594981 A CN202010594981 A CN 202010594981A CN 111624756 A CN111624756 A CN 111624756A
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Abstract
The invention provides a depth of field extended microscopic imaging three-dimensional reconstruction device, which is characterized by comprising the following components: line structured light unit: the system is used for generating linear structured light and three-dimensionally reconstructing the surface appearance of a target object based on the linear structured light; a microscopic imaging unit: for microscopic imaging; a light splitting unit: the light source is used for introducing the line structure light generated by the line structure light unit into the microscopic imaging unit and dividing the line structure light into transmitted light and reflected light; electric objective table: the device is used for placing and moving a target object; mechanical fixing: used for fixing and mounting; the invention also provides a microscopic imaging three-dimensional reconstruction method, which specifically comprises the following steps: s1: camera calibration, namely directly calibrating coordinates of a camera through a calibration plate and establishing a mapping relation between world coordinates and pixel coordinates; s2: collecting three-dimensional information, and collecting the three-dimensional information of a target object; s3: and (3) three-dimensional reconstruction, namely performing three-dimensional reconstruction on the acquired three-dimensional information to obtain the three-dimensional appearance of the target.
Description
Technical Field
The invention relates to the field of microscopic imaging, in particular to a depth of field extended microscopic imaging three-dimensional reconstruction device and method.
Background
In the field of microscopic research, such as material surface reconstruction, biomedicine, etc., a microscope is an indispensable powerful tool, which can help researchers to study the characteristics of objects from microscopic forms by observing and displaying the microstructure through physical magnification of an optical system, and particularly in the centuries of birth of the microscope, every major breakthrough of biomedicine is closely related to the microscope.
With the development of computer technology, microelectronic technology and imaging technology, the development of optical microscopy technology enters into the acquisition and reconstruction of three-dimensional information from two-dimensional images, and the development of microscopic three-dimensional reconstruction technology as an important research branch of image analysis is rapid and the application value is continuously increased.
However, microscopic three-dimensional reconstruction still has certain problems, as shown below:
1. the existing microscopic three-dimensional reconstruction technology can not realize the real-time, high-efficiency and rapid measurement of a large number of samples under the high three-dimensional reconstruction precision;
2. in practical application, the existing microscopic three-dimensional reconstruction technology has the limitation of depth of field, and cannot realize high magnification and high resolution under large depth of field, so that the range of microscopic detection is influenced;
3. the existing microscopic three-dimensional reconstruction technology has high requirements on a detection instrument and high cost.
Disclosure of Invention
Aiming at the defects of the prior art, the invention introduces a three-dimensional reconstruction technology based on line structured light into microscopic imaging, realizes the three-dimensional reconstruction and imaging of the surface appearance of a tiny object in the field of a microscope, adds digital zoom to realize scanning type depth of field expansion, and realizes high-precision, real-time and high-efficiency three-dimensional reconstruction by the method of the recomposition of a three-dimensional reconstruction algorithm and the calibration of a camera.
The invention provides a depth of field extended microscopic imaging three-dimensional reconstruction device on one hand, which specifically comprises
Line structured light unit: the system is used for generating linear structured light and three-dimensionally reconstructing the surface appearance of a target object based on the linear structured light;
a microscopic imaging unit: for microscopic imaging;
a light splitting unit: the light source is used for introducing the line structure light generated by the line structure light unit into the microscopic imaging unit and dividing the line structure light into transmitted light and reflected light;
electric objective table: the device is used for placing and moving a target object;
mechanical fixing: used for fixing and mounting.
Furthermore, the line structured light unit comprises a laser light source, a light beam expanding and collimating assembly, an adjustable line structured light generator and a reflector assembly which are coaxially connected in sequence.
Further, the microscopic imaging unit comprises a microscopic objective group, a digital zoom lens and an image acquisition device.
Further, the wavelength output by the laser light source is one of 380-780 nm.
Further, the beam expanding and collimating assembly comprises a laser beam expander and a collimator.
Further, the adjustable line structured light generator is an adjustable slit of 0-6mm for generating line structured light of different widths.
Further, the reflector assembly comprises a reflector and a fixing frame, and the light propagation direction of the line structure is changed to enter the microscopic imaging unit.
Furthermore, the microscopic imaging unit comprises a microscopic objective group, a digital zoom lens, a connecting pipe and an image acquisition device which are coaxially connected in sequence.
Further, the microscope objective group is a long-working-distance objective lens with the magnification of 1-20 times.
Furthermore, the digital zoom lens can continuously change the focal length under the driving of current, change the equivalent focal length of the microscopic imaging unit and realize real-time in-focus tomography.
Further, the light splitting unit comprises a light splitting prism and is arranged in the light path of the microscopic imaging unit.
The invention also provides a depth of field extended microscopic imaging three-dimensional reconstruction method, which is based on the depth of field extended microscopic imaging three-dimensional reconstruction device and specifically comprises the following steps:
s1: camera calibration, namely directly calibrating coordinates of a camera through a calibration plate and establishing a mapping relation between world coordinates and pixel coordinates;
s2: collecting three-dimensional information, and collecting the three-dimensional information of a target object;
s3: and (3) three-dimensional reconstruction, namely performing three-dimensional reconstruction on the acquired three-dimensional information to obtain the three-dimensional appearance of the target.
Further, in the step S1, the world coordinates are coordinates in a real three-dimensional space, and the pixel coordinates are coordinates in a two-dimensional image acquired by the image acquisition device.
Further, the step S2 specifically includes:
s21: the longitudinal tomography of the target object is realized by continuously changing the focal length of the digital zoom lens, and a tomography image sequence is collected;
s22: and (4) transversely moving the electric object stage, wherein the moving distance is 1um-10um, different moving distances are selected according to the target object for the specific moving distance, and the step S21 is repeated until the whole target object is scanned.
Further, the step S3 specifically includes:
s31, center extraction, namely extracting the center point information of the tomographic image sequence to obtain the pixel coordinates of each position of the target object;
s32: coordinate conversion, namely converting the two-dimensional pixel coordinate of the target object into a three-dimensional world coordinate according to the mapping relation between the world coordinate and the pixel coordinate;
s33: and carrying out three-dimensional shape reconstruction on the target object according to the three-dimensional world coordinates.
The invention has the beneficial effects that:
1. the invention combines the microscopic technology with the line structured light scanning three-dimensional reconstruction technology to realize the three-dimensional reconstruction of microscopic objects in the microscopic field;
2. by introducing the digital zoom lens, the in-focus scanning of the longitudinal fault is realized, the limitation of the depth of a microscope scene is solved, and the three-dimensional reconstruction of the surface appearance of a large-size target object with high magnification and high resolution is realized;
3. compared with the traditional microscopic three-dimensional reconstruction, such as a confocal microscope, the system provided by the invention has a simple structure and high reconstruction speed, so that the cost of the instrument and the three-dimensional reconstruction period are reduced.
Drawings
FIG. 1 is a schematic diagram of the system architecture.
Figure 2 a digital zoom lens tomography schematic.
Reference numerals:
1. the device comprises a laser light source 2, a laser beam expander 3, a collimator 4, an adjustable line structured light generator 5, a reflector assembly 6, a light splitting unit 7, a digital zoom lens 8, a microscope objective group 9, a connecting pipe 10, an image acquisition device 11 and an electric objective table.
Detailed Description
For the purpose of facilitating an understanding of the embodiments of the present invention, the following description will be further explained in terms of several specific embodiments with reference to the accompanying drawings, each of which is not intended to limit the embodiments of the present invention, and the drawings are schematic drawings, so that the apparatus and equipment of the present invention are not limited by the size or scale of the schematic drawings.
It is to be noted that in the claims and the description of the present patent, relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, the use of the verb "comprise a" to define an element does not exclude the presence of another, same element in a process, method, article, or apparatus that comprises the element.
Example 1
In this embodiment, a foramen is used as a detection target, and as shown in fig. 1, the depth-of-field extended microscopic imaging three-dimensional reconstruction apparatus of the present invention includes:
line structured light unit, microscopic imaging unit, light splitting unit 6, electric objective table 11 and mechanical firmware.
The line-structured light unit comprises a laser light source 1, a laser beam expander 2, a collimator 3, an adjustable line-structured light generator 4 and a reflector component 5 which are coaxially connected in sequence; the laser beam emitted by the laser light source 1 sequentially passes through the laser beam expander 2 and the collimator 3 to obtain parallel light, the parallel light irradiates the adjustable line structure light generator 4 to generate line structure light, the line structure light with different widths is generated by adjusting the size of a slit of the adjustable line structure light generator 4, the size range of the slit is 0-6mm, the line structure light reaches the reflector group, and the reflector reflects the line structure light to enter the light splitting unit 6.
The light splitting unit 6 is coaxially and fixedly connected with the microscopic imaging unit and comprises a light splitting prism, the light splitting prism divides the entering line structure light into transmission light and reflection light, and the reflection light enters the microscopic imaging unit.
The microscopic imaging unit comprises a microscopic objective group 8, a digital zoom lens 7, a connecting pipe 9 and an image acquisition device 10 which are coaxially connected in sequence; the reflected light split by the light splitting unit 6 sequentially passes through the digital zoom lens 7 and the microscope objective group 8 to irradiate on the holed worms, the holed worms reflect the light back to the original light path, the reflected light reaches the light splitting unit 6 to be split into transmitted light and reflected light, and the transmitted light enters the image acquisition device 10 to obtain image information.
The digital zoom lens 7 can change the focal length so as to change the integral focal length of the microscopic imaging unit, as shown in fig. 2, the surface of the holed insect sample has fluctuant height, the depth of field of the microscopic objective lens group 8 is shallow, the holed insect sample is not focused at every position, the longitudinal focusing position of the microscopic objective lens group 8 is changed by changing the focal length of the digital zoom lens 7, and the focusing point of the microscopic objective lens group 8 shown in the figure can be O1-O-O2 so as to realize longitudinal tomographic scanning of the holed insect sample, thereby avoiding that the light beam can be focused on the holed insect even if the sample has fluctuant height.
The electric objective table 11 is provided with a hole worm sample, and the hole worm sample moves along the transverse position to realize transverse scanning of the hole worm.
Example 2
In this embodiment, the method for three-dimensional reconstruction of depth-of-field extended microscopic imaging still uses a foramen as a detection target object, and specifically includes the following steps:
(1) a system is built according to the depth-of-field extended microscopic imaging three-dimensional reconstruction device, and the specific system structure can refer to the depth-of-field extended microscopic imaging three-dimensional reconstruction device provided in embodiment 1, which is not described herein again;
(2) calibrating a camera, namely directly calibrating coordinates of the camera by using a calibration plate, and establishing a mapping relation between world coordinates and pixel coordinates, wherein the world coordinates are coordinates in a real three-dimensional space, and the pixel coordinates are coordinates in a two-dimensional image acquired by the image acquisition device 10;
(3) placing a holed insect sample on an electric objective table 11, moving the sample to an initial position, and collecting three-dimensional information of the holed insect;
the step (3) specifically comprises:
(31): changing the focal length of the digital zoom lens 7 to realize longitudinal tomography of the holed worms and collecting a tomography image sequence;
(32): moving the electric object stage 11, setting the moving distance to be 1um-10um, selecting different moving distances according to the target object, and repeating the step (31) until a complete sample of the porous insects is scanned;
(4) three-dimensional reconstruction, namely performing three-dimensional reconstruction on the acquired three-dimensional information to obtain the three-dimensional appearance of the target;
the step (4) specifically comprises:
(41): center extraction, namely extracting center point information of a tomography image sequence to obtain pixel coordinates of each position of a target object, wherein the center point information is in-focus tomography image information of a polyporous;
(42): coordinate conversion, namely converting the two-dimensional pixel coordinate of the central point information of the holed worms into a three-dimensional world coordinate according to the mapping relation between the world coordinate and the pixel coordinate;
(43): and carrying out three-dimensional shape reconstruction on the holed worms according to the three-dimensional world coordinates.
Claims (11)
1. A depth of field extended microscopy imaging three-dimensional reconstruction device is characterized by comprising:
line structured light unit: the system is used for generating linear structured light and three-dimensionally reconstructing the surface appearance of a target object based on the linear structured light;
a microscopic imaging unit: for microscopic imaging;
spectroscopic unit (6): the light source is used for introducing the line structure light generated by the line structure light unit into the microscopic imaging unit and dividing the line structure light into transmitted light and reflected light;
electric stage (11): the device is used for placing and moving a target object;
mechanical fixing: used for fixing and mounting;
the line-structured light unit comprises a laser light source (1), a laser beam expander (2), a collimator (3), an adjustable line-structured light generator (4) and a reflector component (5) which are coaxially connected in sequence; the microscopic imaging unit comprises a microscopic objective group (8), a digital zoom lens (7) and an image acquisition device (10).
2. The depth-of-field extended microscopy imaging three-dimensional reconstruction apparatus according to claim 1, characterized in that: the wavelength output by the laser light source (1) is one of 380-780 nm.
3. The depth-of-field extended microscopy imaging three-dimensional reconstruction apparatus according to claim 1, characterized in that: the adjustable line structured light generator (4) is a slit with adjustable width and is used for generating line structured light with different widths.
4. The depth-of-field extended microscopy imaging three-dimensional reconstruction apparatus according to claim 3, characterized in that: the width of the slit ranges from 0 mm to 6 mm.
5. The depth-of-field extended microscopy imaging three-dimensional reconstruction apparatus according to claim 1, characterized in that: the microscope objective group (8) is a long-working-distance objective lens, and the magnification is any magnification of 1-20 times.
6. The depth-of-field extended microscopy imaging three-dimensional reconstruction apparatus according to claim 1, characterized in that: the digital zoom lens (7) can continuously change the focal length under the driving of current, change the equivalent focal length of the microscopic imaging unit and realize real-time in-focus tomography.
7. The depth-of-field extended microscopy imaging three-dimensional reconstruction apparatus according to claim 1, characterized in that: the light splitting unit (6) comprises a light splitting prism and is arranged in the light path of the microscopic imaging unit.
8. A depth of field extended microscopic imaging three-dimensional reconstruction method is realized on the basis of a depth of field extended microscopic imaging three-dimensional reconstruction device, and is characterized by comprising the following steps:
s1: camera calibration, namely directly calibrating coordinates of a camera through a calibration plate and establishing a mapping relation between world coordinates and pixel coordinates;
s2: collecting three-dimensional information, and collecting the three-dimensional information of a target object;
s3: and (3) three-dimensional reconstruction, namely performing three-dimensional reconstruction on the acquired three-dimensional information to obtain the three-dimensional appearance of the target.
9. The depth-of-field extended microscopy imaging three-dimensional reconstruction method of claim 8, characterized in that: in step S1, the world coordinates are coordinates in a real three-dimensional space, and the pixel coordinates are coordinates in a two-dimensional image acquired by the image acquisition device (10).
10. The depth-of-field extended microscopy imaging three-dimensional reconstruction method of claim 8, characterized in that: the step S2 specifically includes:
s21: the longitudinal tomography of the target object is realized by continuously changing the focal length of the digital zoom lens (7), and a tomography image sequence is acquired;
s22: and (3) transversely moving the electric object stage (11) for a moving distance of 1um-10um, selecting different moving distances according to the target object for the specific moving distance, and repeating the step S21 until the whole target object is scanned.
11. The depth-of-field extended microscopy imaging three-dimensional reconstruction method of claim 8, characterized in that: the step S3 specifically includes the steps of,
s31, center extraction, namely extracting the center point information of the tomographic image sequence to obtain the pixel coordinates of each position of the target object;
s32: converting coordinates, namely converting the obtained two-dimensional pixel coordinates of the target object into three-dimensional world coordinates according to the mapping relation between the world coordinates and the pixel coordinates;
s33: and carrying out three-dimensional shape reconstruction on the target object according to the three-dimensional world coordinates.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112903236A (en) * | 2021-01-29 | 2021-06-04 | 上海交通大学 | Focal plane scanning-based aerodynamic thermal parameter optical test device and method |
| CN114562958A (en) * | 2022-04-27 | 2022-05-31 | 板石智能科技(深圳)有限公司 | Self-built microscopic imaging system based on optical energy transmission equation |
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2020
- 2020-06-28 CN CN202010594981.2A patent/CN111624756A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112903236A (en) * | 2021-01-29 | 2021-06-04 | 上海交通大学 | Focal plane scanning-based aerodynamic thermal parameter optical test device and method |
| CN114562958A (en) * | 2022-04-27 | 2022-05-31 | 板石智能科技(深圳)有限公司 | Self-built microscopic imaging system based on optical energy transmission equation |
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