CN108227233B - Microscopic tomography super-resolution imaging method and system based on light sheet structured light - Google Patents
Microscopic tomography super-resolution imaging method and system based on light sheet structured light Download PDFInfo
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- CN108227233B CN108227233B CN201711447792.7A CN201711447792A CN108227233B CN 108227233 B CN108227233 B CN 108227233B CN 201711447792 A CN201711447792 A CN 201711447792A CN 108227233 B CN108227233 B CN 108227233B
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Abstract
The invention discloses a microscopic tomography super-resolution imaging method and a system based on light sheet structured light, wherein the method comprises the following steps: spatially modulating incident light at the illumination end to generate a light sheet to illuminate the sample from the side; modulating the optical sheet at the illumination end into a plurality of pattern patterns with different frequencies and different directions, and correspondingly shooting a plurality of images at the detection end; and improving the image resolution by a structured light super-resolution method according to the plurality of images to obtain a super-resolution microscopic tomography image. The method can effectively improve the imaging resolution and eliminate the background noise.
Description
Technical Field
The invention relates to the technical fields of physical optics, computational photography and life science, in particular to a microscopic tomography super-resolution imaging method and system based on light sheet structured light.
Background
Optical microscopes are optical instruments that are used primarily to magnify tiny objects (on the nano-micron scale) so that one can observe microscopic structures (e.g., cells, viruses, etc.) that are not visible to the naked eye. At present, the optical microscope has become an indispensable instrument in the field of life sciences. Optical microscopes include many types for various research purposes, and common optical microscopes and fluorescence microscopes are common. Wherein the fluorescence microscope utilizes immunofluorescence techniques and uses fluorescein labeling techniques to calibrate targets. When incident light of a certain wavelength impinges on the sample, these fluoresceins fluoresce due to the energy level transitions of the atoms, making the sample visible, which is not visible to the naked eye. With the aid of the rapid development in the field of chemical materials, almost all visible spectral regions are now available for selection of corresponding fluorescent markers. Therefore, fluorescence microscopy has become an important imaging tool for biologists, enabling widespread use in the field of life sciences.
The light sheet microscopy is a three-dimensional chromatography fluorescence microscopic imaging method. Technically, light sheet microscopy achieves a thin layer of light sheet illuminating the sample from the side by spatially modulating the incident light at the illumination end, thus exciting only a thin layer of sample depth. The fluorescence emitted by the thin layer after excitation is collected above or below the sample along an optical axis perpendicular to the illumination plane. By such an illumination method, fluorescent molecules in both the upper and lower parts of the illumination plane are not excited to generate fluorescence. And finally, scanning images at different depths to realize microscopic three-dimensional depth tomography. The technology adopts wide-field illumination, so that the imaging time resolution is higher; the light sheet illumination is adopted, so that the depth resolution is high, and meanwhile, the biological sample is subjected to extremely small light damage and light bleaching. Therefore, the light sheet microscopy is widely applied to observing three-dimensional samples in the field of life science, and the microscopic tomography is realized.
Disclosure of Invention
The present invention is directed to solving, at least to some extent, one of the technical problems in the related art.
Therefore, one objective of the present invention is to provide a microscopic tomographic super-resolution imaging method based on light sheet structured light, which can effectively improve tomographic resolution and eliminate background noise.
The invention also aims to provide a micro-tomography super-resolution imaging system based on light sheet structure light.
In order to achieve the above object, an embodiment of the invention provides a micro-tomography super-resolution imaging method based on light sheet structure light, which includes the following steps: spatially modulating incident light at the illumination end to generate a light sheet to illuminate the sample from the side; modulating the optical sheets at the illumination end into a plurality of pattern patterns with different frequencies and different directions, and correspondingly shooting a plurality of images at the detection end; and improving the image resolution by a structured light super-resolution method according to the plurality of images to obtain a super-resolution microscopic tomography image.
According to the microscopic tomography super-resolution imaging method based on the light sheet structured light, disclosed by the embodiment of the invention, the light sheet microscopic tomography technology and the structured light super-resolution technology are combined, the robustness of medium scattering at an illumination end by using the light sheet microscopic technology is utilized, and the structured light super-resolution technology is combined, so that the resolution of the traditional microscopic three-dimensional tomography is improved, the image resolution is effectively improved, and the super-resolution microscopic tomography is realized.
In addition, the micro-tomography super-resolution imaging method based on the light sheet structure light according to the above embodiment of the invention may further have the following additional technical features:
further, in an embodiment of the present invention, the spatially modulating incident light of the illumination end further includes: modulating the incident light into the light sheet using a lenticular, Bessel (Bessel) light modulation, or grid (lattice) light modulation method to laterally illuminate a depth layer of the sample.
Further, in one embodiment of the invention, the light sheet is modulated into different patterns by changing the scan function, DMD modulation, acousto-optic tunable filter modulation, or rotating the sample.
Further, in one embodiment of the present invention, the sample image is reconstructed by a frequency domain stitching algorithm, an alternating projection algorithm, or the like.
In order to achieve the above object, another embodiment of the present invention provides a micro-tomography super-resolution imaging system based on light sheet structure light, including: the first modulation module is used for carrying out spatial modulation on incident light of the illumination end so as to generate a light sheet to illuminate the sample from the side; the second modulation module is used for modulating the optical sheet at the illumination end into a plurality of pattern patterns with different frequencies and different directions and correspondingly shooting a plurality of images at the detection end; and the acquisition module is used for improving the image resolution by a structured light super-resolution method according to the plurality of images so as to obtain a super-resolution microscopic tomography image.
According to the microscopic tomography super-resolution imaging system based on the light sheet structured light, disclosed by the embodiment of the invention, the light sheet microscopic tomography technology is combined with the structured light super-resolution technology, the robustness of medium scattering at an illumination end is improved by using the light sheet microscopic technology, and the structured light super-resolution technology is combined, so that the resolution of the traditional microscopic three-dimensional tomography is improved, the image resolution is effectively improved, and the super-resolution microscopic tomography is realized.
In addition, the micro-tomography super-resolution imaging system based on light sheet structure light according to the above embodiment of the invention may also have the following additional technical features:
further, in an embodiment of the present invention, the first modulation module further includes:
a modulation unit for modulating the incident light into the optical sheet using a lenticular, Bessel (Bessel) light modulation, or lattice (lattice) light modulation method to laterally illuminate a certain depth layer of the sample.
Further, in one embodiment of the invention, the light sheet is modulated into different patterns by changing the scan function, DMD modulation, acousto-optic tunable filter modulation, or rotating the sample.
Further, in one embodiment of the present invention, the sample image is reconstructed by a frequency domain stitching algorithm, an alternating projection algorithm, or the like.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
The foregoing and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:
FIG. 1 is a flow chart of a micro-tomography super-resolution imaging method based on light sheet structured light according to an embodiment of the invention;
FIG. 2 is a schematic structural diagram of a micro-tomography super-resolution imaging system based on light sheet structured light according to an embodiment of the present invention;
fig. 3 is a schematic structural diagram of a micro-tomography super-resolution imaging system based on light sheet structure light according to an embodiment of the invention.
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the drawings are illustrative and intended to be illustrative of the invention and are not to be construed as limiting the invention.
The method and system for microscopic tomographic super-resolution imaging based on optical sheet structured light according to the embodiments of the present invention will be described below with reference to the accompanying drawings.
FIG. 1 is a flow chart of a microscopic tomography super-resolution imaging method based on light sheet structured light according to an embodiment of the present invention.
As shown in FIG. 1, the microscopic tomography super-resolution imaging method based on light sheet structured light comprises the following steps:
in step S101, incident light of the illumination end is spatially modulated to generate a light sheet to illuminate the sample from the side.
Further, in an embodiment of the present invention, the spatially modulating incident light of the illumination end further includes: modulating the incident light into the light sheet using a lenticular, Bessel (Bessel) light modulation, or grid (lattice) light modulation method to laterally illuminate a depth layer of the sample.
It is understood that, in conjunction with fig. 1 and fig. 2, the embodiment of the present invention may illuminate a certain depth layer of the sample from the side by using a method such as a lenticular lens, Bessel (Bessel) light modulation, or grid (lattice) light modulation, wherein modulating the incident light into a light sheet is not limited to the above method, and includes a method such as lenticular direct modulation, Bessel (Bessel) light modulation, or grid (lattice) light modulation, which is specifically modulated according to actual situations by those skilled in the art, and is not limited thereto.
In step S102, the optical sheet at the illumination end is modulated into a plurality of pattern patterns with different frequencies and different directions, and a plurality of images are correspondingly captured at the detection end.
Further, in one embodiment of the invention, the light sheet is modulated into different patterns by changing the scan function, DMD modulation, acousto-optic tunable filter modulation, or rotating the sample.
It is to be understood that the embodiment of the present invention may adjust the optical sheet into different patterns by changing the scanning function, DMD modulation, acousto-optic tunable filter modulation, or rotating the sample, which is not limited to the above methods, including changing the scanning function, using DMD modulation, using acousto-optic tunable filter modulation, rotating the sample, and the like, and the method is specifically performed by those skilled in the art according to the actual situation, and is not limited to this.
In step S103, the image resolution is improved by a structured light super-resolution method according to the plurality of images to obtain a super-resolution microscopic tomographic image.
Further, in one embodiment of the present invention, the sample image is reconstructed by a frequency domain stitching algorithm, an alternating projection algorithm, or the like.
According to the microscopic tomography super-resolution imaging method based on the light sheet structure light, which is provided by the embodiment of the invention, the light sheet microscopic tomography technology is combined with the structure light super-resolution technology, the robustness of medium scattering at an illumination end by the light sheet micro-technology is utilized, and the structure light super-resolution technology is combined, so that the resolution of the traditional microscopic three-dimensional tomography is improved, the image resolution is effectively improved, and the super-resolution microscopic tomography is realized.
Next, a micro-tomography super-resolution imaging system based on light sheet structured light according to an embodiment of the present invention will be described with reference to the drawings.
FIG. 3 is a schematic structural diagram of a micro-tomography super-resolution imaging system based on light sheet structured light according to an embodiment of the present invention
As shown in fig. 3, the micro-tomography super-resolution imaging system 10 based on light sheet structured light includes:
the first modulation module 100 is configured to spatially modulate incident light at the illumination end to generate a light sheet to illuminate the sample from the side. The second modulation module 200 is configured to modulate the optical sheet at the illumination end into a plurality of pattern patterns with different frequencies and different directions, and correspondingly shoot a plurality of images at the detection end. The obtaining module 300 is configured to improve the image resolution according to the multiple images by using a structured light super-resolution method to obtain a super-resolution microscopic tomography image. The system 10 of the embodiment of the invention can effectively improve the resolution of tomography, eliminate background noise and have stronger robustness.
Further, in an embodiment of the present invention, the first modulation module further includes: a modulation unit for modulating the incident light into the light sheet using a cylindrical lens, Bessel (Bessel) light modulation, or lattice (lattice) light modulation to laterally illuminate a certain depth layer of the sample.
Further, in one embodiment of the present invention, the light sheet is modulated into different patterns by changing scanning function, DMD modulation, acousto-optic tunable filter modulation, rotating sample, etc.
Further, in one embodiment of the present invention, the sample image is reconstructed by a frequency domain stitching algorithm, an alternating projection algorithm.
It should be noted that the foregoing explanation of the embodiment of the optical sheet structured light-based micro-tomography super-resolution imaging method is also applicable to the optical sheet structured light-based micro-tomography super-resolution imaging system of the embodiment, and is not repeated here.
According to the microscopic tomography super-resolution imaging system based on the light sheet structure light, which is provided by the embodiment of the invention, the light sheet microscopic tomography technology is combined with the structure light super-resolution technology, the robustness of medium scattering at an illumination end by using the light sheet micro-technology is combined with the structure light super-resolution technology, the resolution of the traditional microscopic three-dimensional tomography is improved, the image resolution is effectively improved, and the super-resolution microscopic tomography is realized.
In the description of the present invention, it is to be understood that the terms "central," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," "circumferential," and the like are used in the orientations and positional relationships indicated in the drawings for convenience in describing the invention and to simplify the description, and are not intended to indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and are therefore not to be considered limiting of the invention.
Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In the description of the present invention, "a plurality" means at least two, e.g., two, three, etc., unless specifically limited otherwise.
In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," "secured," and the like are to be construed broadly and can, for example, be fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; they may be directly connected or indirectly connected through intervening media, or they may be connected internally or in any other suitable relationship, unless expressly stated otherwise. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.
In the present invention, unless otherwise expressly stated or limited, the first feature "on" or "under" the second feature may be directly contacting the first and second features or indirectly contacting the first and second features through an intermediate. Also, a first feature "on," "over," and "above" a second feature may be directly or diagonally above the second feature, or may simply indicate that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature may be directly under or obliquely under the first feature, or may simply mean that the first feature is at a lesser elevation than the second feature.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.
Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
Claims (6)
1. A microscopic tomography super-resolution imaging method based on light sheet structured light is characterized by comprising the following steps:
spatially modulating incident light at the illumination end to generate a light sheet to illuminate the sample from the side;
modulating the optical sheets at the illumination end into a plurality of pattern patterns with different frequencies and different directions, and correspondingly shooting a plurality of images at the detection end; and
improving the image resolution by a structured light super-resolution method according to the multiple images to obtain a super-resolution microscopic tomography image;
and the optical sheets are modulated into different patterns by changing scanning functions, DMD modulation, acousto-optic tunable filter modulation, sample rotation and the like.
2. The light sheet structured light-based micro-tomography super-resolution imaging method according to claim 1, wherein the spatially modulating incident light at the illumination end further comprises:
modulating the incident light into the light sheet using a lenticular, Bessel (Bessel) light modulation, or grid (lattice) light modulation method to laterally illuminate a depth layer of the sample.
3. The light-sheet structured light-based microscopy tomography super-resolution imaging method according to claim 1, wherein the sample image is reconstructed by a frequency domain stitching algorithm, an alternative projection algorithm, or the like.
4. A microscopic tomography super-resolution imaging system based on light sheet structured light is characterized by comprising:
the first modulation module is used for carrying out spatial modulation on incident light of the illumination end so as to generate a light sheet to illuminate the sample from the side;
the second modulation module is used for modulating the optical sheet at the illumination end into a plurality of pattern patterns with different frequencies and different directions and correspondingly shooting a plurality of images at the detection end; and
the acquisition module is used for improving the image resolution by a structured light super-resolution method according to the plurality of images so as to obtain a super-resolution microscopic tomography image;
the sheets are modulated into different patterns by changing the scanning function, DMD modulation, acousto-optic tunable filter modulation, or rotating the sample.
5. The light sheet structured light-based micro-tomography super-resolution imaging system according to claim 4, wherein the first modulation module further comprises:
a modulation unit for modulating the incident light into the optical sheet using a lenticular, Bessel (Bessel) light modulation, or lattice (lattice) light modulation method to laterally illuminate a certain depth layer of the sample.
6. The light-sheet structured light-based micro-tomography super-resolution imaging system according to claim 4, wherein the sample image is reconstructed by a frequency domain stitching algorithm, an alternative projection algorithm, or the like.
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| CN110109239B (en) * | 2019-04-30 | 2020-08-14 | 华中科技大学 | Light sheet illumination microscopic imaging method and system for simultaneous multilayer imaging |
| CN112485230B (en) * | 2019-09-11 | 2022-03-18 | 复旦大学 | Super-resolution microscopic imaging method and device based on active time modulation frequency mixing excitation irradiation |
| CN111781733A (en) * | 2020-06-09 | 2020-10-16 | 北京理工大学 | Multilayer complex field imaging method and device based on light wave modulation and phase recovery |
| CN112651884B (en) * | 2021-01-19 | 2023-01-24 | 佛山职业技术学院 | Method and device for acquiring chromatography super-resolution image and electronic equipment |
| CN113917677B (en) * | 2021-09-09 | 2023-05-05 | 北京纳析光电科技有限公司 | Three-dimensional super-resolution light sheet microscopic imaging method and microscope |
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| CN106290284B (en) * | 2016-09-19 | 2023-03-10 | 浙江大学 | Two-photon fluorescence microscope system and method with structured light illumination |
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| CN106885796B (en) * | 2017-04-01 | 2023-09-01 | 北京工业大学 | Super-resolution fluorescence digital holographic tomography microscopic imaging system and method |
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| WO2017064629A1 (en) * | 2015-10-12 | 2017-04-20 | Crestoptics S.R.L. | Confocal microscopy apparatus and related process for acquiring and processing images |
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