CN113176662A - Microscopic imaging device - Google Patents
Microscopic imaging device Download PDFInfo
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- CN113176662A CN113176662A CN202110606043.4A CN202110606043A CN113176662A CN 113176662 A CN113176662 A CN 113176662A CN 202110606043 A CN202110606043 A CN 202110606043A CN 113176662 A CN113176662 A CN 113176662A
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- imaging
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
- G02B21/365—Control or image processing arrangements for digital or video microscopes
- G02B21/367—Control or image processing arrangements for digital or video microscopes providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/02—Objectives
- G02B21/025—Objectives with variable magnification
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/02—Objectives
- G02B21/04—Objectives involving mirrors
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides
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- Microscoopes, Condenser (AREA)
Abstract
The present invention provides a microscopic imaging apparatus comprising: the device comprises an object stage, a magnifying assembly, an imaging assembly, a plurality of miniature cameras and an image processing module. Under the condition of fixed visual field, the invention obtains the images of a plurality of imaging surfaces by one-time imaging, ensures that the image content, the position and the image size equivalent of the image fusion are consistent, greatly shortens the acquisition time of the image to be fused, and is matched with an effective depth of field fusion algorithm to improve the accuracy of depth of field fusion.
Description
Technical Field
The invention relates to the field of microscopic imaging, in particular to a microscopic imaging device.
Background
Due to the limitation of an optical principle, the microscopic imaging system has the condition that the resolution and the depth of field range are limited, and the high resolution means that the depth of field is shallow. When a sample with depth feeling is detected, and the depth span of the sample exceeds the depth of field of a microscope, objects at different distances in the same scene cannot be imaged clearly at the same time, namely the multi-focal-plane problem. In biological and medical research and application, the sample is usually required to be subjected to thin section treatment, so that observation and clear imaging can be carried out under a microscope, and when the sample structure cannot be modified for thick sections or body fluid visible component detection, living cell culture and the like, the traditional microscope cannot meet the requirement of single clear imaging.
The traditional depth-of-field fusion methods are generally a zoom method, a variable aperture method, a defocusing method, a depth-of-field superposition method and the like, and although images with ultra depth of field can be obtained, the focal length needs to be changed for many times and the images need to be collected. At present, the modes for obtaining multi-focal plane images are all manual focusing or rely on a motor to drive an objective table to move up and down or a piezoelectric objective actuator to drive an objective lens to move so as to adjust the distance between the objective lens and a sample, namely, the focal length is adjusted, but the modes are limited by a mechanical structure, the modes spend different degrees of time in the focusing process, cannot meet the requirement of rapid focusing, and are difficult to realize real-time depth-of-field fusion in the true sense. At present, there is also a method for realizing real-time fast focusing, in which a liquid lens is added to the rear end surface of an objective lens, and the diopter of the liquid lens is continuously changed within an exposure time of a camera, thereby realizing fast focal length adjustment and possibly realizing real-time on-line depth-of-field fusion. However, this method may cause a change in magnification, which is linear and repeatable in the z-axis direction, and this may increase difficulty in image registration in subsequent image processing, and increase difficulty and calculation time for performing size registration and then fusion on the acquired images of multiple focal planes. ) In addition, for the digital pathological microscopic imaging system, the objective table needs to continuously move in two dimensions, namely scanning, and finally each visual field is spliced into a digital pathological picture, so that a group of pictures shot in a period of changing the diopter of the liquid lens are not the same visual field but deviate along with the movement of the platform, the difficulty, the calculation time and the accuracy of a subsequent depth-of-field fusion algorithm are increased, and the real-time depth-of-field fusion is difficult to realize.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to overcome the problem that the time for collecting the image is too long when depth of field fusion is carried out.
In a first aspect, the present invention provides a microscopic imaging apparatus comprising: the objective table is used for bearing a sample; the amplifying assembly is arranged above the objective table; the imaging assembly is arranged above the amplifying assembly and used for receiving the parallel light emitted by the amplifying assembly and converging the parallel light into a point, and the imaging assembly comprises at least two focal planes; the miniature cameras are respectively arranged at the imaging surfaces corresponding to the focal surfaces and shoot images of the imaging surfaces; and the image processing module is used for acquiring the images of the plurality of imaging surfaces and fusing the images into one image.
Further, the magnification assembly includes an objective lens.
Further, the light splitting module comprises a barrel mirror, and a central axis of the barrel mirror is collinear with a central axis of the amplifying assembly.
Further, the microscopic imaging device further comprises: a beam splitting module for separating the imaging plane from the central axis; the light splitting module includes: and the light splitting element is arranged above the imaging assembly and corresponds to the converged light of the imaging surface.
Further, the light splitting element comprises a semi-reflecting and semi-transparent mirror or a light splitting prism.
Further, the microscopic imaging device further comprises: the light splitting module is used for separating the imaging surface at the central axis; the light splitting module comprises a light splitting element, parallel rays are arranged in the light splitting module and correspond to at least one focus, and the light splitting element is used for refracting the parallel rays to the side part of the light splitting module; and the light condensing assembly is arranged on the side part of the light splitting module and is used for condensing the refracted parallel light into the imaging surface.
Further, the light splitting element comprises a semi-reflecting and semi-transparent mirror or a light splitting prism; the light condensing assembly includes a barrel mirror.
Further, the miniature camera comprises a CCD camera.
Furthermore, the miniature cameras are respectively arranged at the imaging surfaces corresponding to the upper part and the lower part of each focal plane.
Further, the microscopic imaging device further comprises: the condenser lens is arranged below the objective table; the reflecting mirror is arranged below the collecting mirror; and the motion control system is used for controlling the object stage to move.
The technical scheme of the invention has the following advantages:
the invention provides a microscopic imaging device, which obtains images of a plurality of imaging surfaces through one-time imaging under the condition of fixed visual field, ensures that the image content, the position and the image size equivalent of image fusion are consistent, greatly shortens the acquisition time of the image to be fused, is matched with an effective depth-of-field fusion algorithm, is used for improving the accuracy of depth-of-field fusion, shortens the time of depth-of-field fusion and can realize real-time depth-of-field fusion.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a schematic structural view of a microscopic imaging apparatus provided in example 1 of the present invention;
fig. 2 is a schematic structural diagram of a spectroscopy module provided in embodiment 1 of the present invention;
fig. 3 is a schematic structural diagram of a spectroscopic element and an imaging assembly provided in embodiment 1 of the present invention;
fig. 4 is a schematic structural diagram of a spectroscopic element and an imaging assembly provided in embodiment 2 of the present invention;
an object stage 101; an amplifying assembly 102; a light splitting module 103;
a miniature camera 104; an image processing module 200; a condenser lens 106; a mirror 107;
a motion control system 108; a slide 105; a light splitting element 110.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the accompanying drawings, and it should be understood that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the description of the present invention, it should be noted that the terms "upper", "lower", "inside", "outside", and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, and are only for convenience in describing the present invention and simplifying the description, but do not indicate or imply that the referred device or element must have a specific orientation, be constructed in a specific orientation, and be operated, and thus, should not be construed as limiting the present invention. Furthermore, the terms "first," "second," and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
In addition, the technical features involved in the different embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
Example 1
As shown in fig. 1, embodiment 1 of the present invention provides a microscopic imaging apparatus including: a stage 101, a magnification assembly 102, an imaging assembly 111, a plurality of miniature cameras 104, and an image processing module 200.
The stage 101 is used for carrying a sample.
The magnifying assembly 102 is disposed above the stage 101.
The imaging assembly 111 is disposed above the amplifying assembly 102, and is configured to receive the parallel light emitted from the amplifying assembly 102 and converge into a point, where the imaging assembly 102 includes at least two focal planes.
The plurality of miniature cameras 104 are respectively arranged at the imaging surfaces corresponding to the focal plane and shoot images of the imaging surfaces.
The image processing module 200 is used to acquire images of a plurality of imaging planes and fuse the images into one image.
The invention provides a microscopic imaging device, which obtains images of a plurality of imaging surfaces through one-time imaging under the condition of fixed visual field, ensures that the image content, the position and the image size equivalent of image fusion are consistent, greatly shortens the acquisition time of the image to be fused, is matched with an effective depth-of-field fusion algorithm, is used for improving the accuracy of depth-of-field fusion, shortens the time of depth-of-field fusion and can realize real-time depth-of-field fusion.
In this embodiment, the stage 101 is provided with a slide 105, and the sample is disposed on the slide 105.
In this embodiment, the magnifying assembly 102 comprises an objective lens. The objective lens is a lens group formed by combining a plurality of lenses. The combined use aims to overcome the imaging defects of a single lens and improve the optical quality of the objective lens. The magnification component 102 is configured to magnify the sample image.
In this embodiment, the light splitting module 103 includes a cylindrical mirror, and a central axis of the cylindrical mirror is collinear with a central axis of the amplifying assembly 102.
In this embodiment, the microscopic imaging apparatus further includes: the light splitting module 103 is used for separating the imaging plane from the central axis, so that a plurality of miniature cameras 104 can be arranged, and the miniature cameras 104 can be better arranged.
The light splitting module 103 includes: and a light splitting element 110 disposed above the imaging assembly and corresponding to the converged light of the imaging surface. The light splitting element 110 includes a half-reflecting half-mirror or a light splitting prism. The number of the light splitting elements 110 is equal to the number of the imaging planes minus 1.
As shown in fig. 2 and 3, the focal planes 1,2,3, the number and positions of which are customized, correspond to the imaging planes 1 ', 2 ', 3 '. The light splitting element 110 in fig. 3 is folded for the optical path of the converging light of the imaging planes 2 ', 3' to be used for separating the imaging planes from the central axis, and the camera can be conveniently arranged. And then the miniature cameras can be arranged on the imaging surfaces corresponding to the upper part and the lower part of each focal plane respectively. I.e., one miniature camera 104 for each imaging plane. And one micro camera 104 is arranged on the imaging plane of the current focusing plane, so that 2n +1 micro cameras 104 are included in fig. 1.
The barrel mirror has an imaging surface of a current focusing surface, for example, the focal surface 1 may be the current focusing surface, and 1' is the imaging surface of the current focusing surface.
The miniature camera 104 comprises a CCD camera. The CCD is a charge coupled device (charge coupled device) for short, which can convert light into electric charge and store and transfer the electric charge, and can also take out the stored electric charge to change the voltage, so it is an ideal CCD camera element.
In this embodiment, the microscopic imaging apparatus further includes: a collection mirror 106, a mirror 107, and a motion control system 108.
The condenser 106 is arranged below the stage 101, and the condenser 106 is used for condensing light; the reflector 107 is arranged below the condenser 106, and the reflector 107 is used for reflecting light; and the motion control system 108 is configured to control the movement of the stage 101.
In the present embodiment, the image processing module 200 includes: an acquisition unit and a fusion unit.
The acquisition unit is used for acquiring images of a plurality of imaging surfaces; the fusion unit is used for fusing the images of the plurality of imaging surfaces into one image. The image processing module 200 and the motion control system 108 are controlled by a computer.
According to the microscopic imaging device provided by the invention, when the objective table 101 is fixed, light is collected by the condenser 106 and emitted to the surface of the reflector 107, the light is reflected to a sample by the reflector 107, an image of the sample is amplified by the objective lens, a plurality of focal planes and corresponding imaging planes are formed at the light splitting module 103, and the images of the imaging planes are collected by the miniature camera 104 and then fused to form an image.
The method can eliminate time consumption of focusing movement caused by the traditional manual focusing mode, the focusing mode of driving the objective table 101 by a motor and the focusing mode of a piezoelectric objective lens, is particularly suitable for application scenes such as digital pathological scanning and splicing imaging, and can avoid the problem that the difficulty and the calculation time are increased for subsequent image processing due to the simultaneous change of the field of view and the focal length in the scanning process. The method can provide original picture materials for subsequent depth-of-field fusion, and provides powerful assistance for realizing real-time depth-of-field fusion.
Example 2
Specifically, the light splitting module includes a light splitting element, which is disposed in the light splitting module and corresponds to the parallel light of the at least one focus, and is configured to refract the parallel light to a side of the light splitting module. The light condensing assembly is arranged on the side part of the light splitting module and used for condensing the refracted parallel light into the imaging surface. The light splitting element comprises a semi-reflecting and semi-transparent mirror or a light splitting prism; the light condensing assembly includes a barrel mirror.
Referring to fig. 4, focal planes 1,2 and 3, in this embodiment, optical paths of focal planes 1 and 3 are modified, a light splitting element is disposed inside the optical path to lead out parallel optical paths to a side portion of the imaging component, and then parallel light is converged into imaging planes 1 'and 3'. This has the advantages that: the distance between the objective lens and the cylindrical lens is long, so that a plurality of light splitting elements can be added, and the light splitting elements are matched with the light condensing assembly, so that a plurality of imaging surfaces can be obtained, and the number of images shot by the miniature camera can be increased; besides, the influence of the light splitting element additionally arranged in the parallel light path on the imaging quality is small, so that the image quality of the obtained imaging surface is high.
The improved place of the embodiment 1 is above the imaging assembly, while the embodiment 2 is improved in the imaging assembly, so that more imaging positions and better imaging quality can be obtained; and further the subsequent depth-of-field fusion effect is better.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (10)
1. A microscopic imaging apparatus, comprising:
the objective table is used for bearing a sample;
the amplifying assembly is arranged above the objective table;
the imaging assembly is arranged above the amplifying assembly and used for receiving the parallel light emitted by the amplifying assembly and converging the parallel light into a point, and the imaging assembly comprises at least two focal planes;
the miniature cameras are respectively arranged at the imaging surfaces corresponding to the focal surfaces and shoot images of the imaging surfaces; and
and the image processing module is used for acquiring the images of the plurality of imaging surfaces and fusing the images into one image.
2. A microscopic imaging apparatus according to claim 1,
the magnification assembly includes an objective lens.
3. A microscopic imaging apparatus according to claim 1,
the light splitting module comprises a cylindrical mirror, and the central axis of the cylindrical mirror is collinear with the central axis of the amplifying assembly.
4. A microscopic imaging apparatus according to claim 1 or 3, characterized by further comprising:
a beam splitting module for separating the imaging plane from the central axis;
the light splitting module includes: and the light splitting element is arranged above the imaging assembly and corresponds to the converged light of the imaging surface.
5. A microscopic imaging apparatus according to claim 4,
the light splitting element comprises a semi-reflecting and semi-transparent mirror or a light splitting prism.
6. A microscopic imaging apparatus according to claim 1 or 3, characterized by further comprising: the light splitting module is used for separating the imaging surface at the central axis;
the light splitting module comprises a light splitting element, parallel rays are arranged in the light splitting module and correspond to at least one focus, and the light splitting element is used for refracting the parallel rays to the side part of the light splitting module;
and the light condensing assembly is arranged on the side part of the light splitting module and is used for condensing the refracted parallel light into the imaging surface.
7. A microscopic imaging apparatus according to claim 6,
the light splitting element comprises a semi-reflecting and semi-transparent mirror or a light splitting prism; the light condensing assembly includes a barrel mirror.
8. A microscopic imaging apparatus according to claim 1,
the miniature camera includes a CCD camera.
9. A microscopic imaging apparatus according to claim 1,
the miniature cameras are respectively arranged on the imaging surfaces corresponding to the upper part and the lower part of each focal surface.
10. The microscopic imaging apparatus according to claim 1, further comprising:
the condenser lens is arranged below the objective table; and
and the reflecting mirror is arranged below the collecting mirror.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110606043.4A CN113176662A (en) | 2021-05-31 | 2021-05-31 | Microscopic imaging device |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202110606043.4A CN113176662A (en) | 2021-05-31 | 2021-05-31 | Microscopic imaging device |
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| CN113176662A true CN113176662A (en) | 2021-07-27 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117572625A (en) * | 2024-01-15 | 2024-02-20 | 深圳安侣医学科技有限公司 | Multipath microscopic imaging device and method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117572625A (en) * | 2024-01-15 | 2024-02-20 | 深圳安侣医学科技有限公司 | Multipath microscopic imaging device and method |
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