CN113552710B - Multi-plane microscopic imaging system based on gradient variable refractive index lens - Google Patents
Multi-plane microscopic imaging system based on gradient variable refractive index lens Download PDFInfo
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Abstract
The invention discloses an arbitrary depth multi-plane microscopic imaging system and method based on a gradient variable refractive index lens, wherein the system comprises: the gradient variable refractive index lens is used for generating a virtual image of the sample to be imaged in an observation area of the gradient variable refractive index lens on a microscope focal plane; the image plane correction module is used for generating a virtual image of the sample to be imaged outside an observation region of the gradient variable refractive index lens and adjusting the position of an image plane outside the observation region of the gradient variable refractive index lens; the microscopic optical amplification module is used for collecting optical signals emitted from a focal plane of the microscope and carrying out optical amplification; and the information acquisition module is used for receiving the optical signal output by the microscopic optical amplification module and imaging. The system enables the traditional optical microscope to realize simultaneous observation of surface information, any deep layer or section information of an observation sample, improves the imaging speed and expands the depth area capable of imaging.
Description
Technical Field
The invention relates to the field of optical microscopic imaging, in particular to an arbitrary-depth multi-plane microscopic imaging system and method based on a gradient variable refractive index lens.
Background
The optical microscopic imaging technology is a technology for acquiring a high-resolution image of a surrounding object by an optical method, is widely applied to structural imaging and functional signal detection of microorganisms such as cells, bacteria, viruses and the like at present, and has become a common method for biological research at present. In the research of brain science, in combination with a chemical indicator, an optical microscope can detect changes in neuronal activity by using changes in fluorescence intensity caused by changes in calcium concentration, chemical transmitter concentration, voltage changes, and the like. It will be appreciated that the brain is a three-dimensional structure with layers, different layers having different structures and functions, and different layers having functional connections to each other. For example, the hippocampus of the brain of a mouse is usually located below about 1mm of the brain surface, and is directly related to information exchange, memory formation and disease onset of the cerebral cortex, so in order to study the above problems, we need to observe the hippocampus and the cortex of the brain simultaneously. However, the traditional optical microscopy technology can only clearly image the tissue structure at the focal plane by single shooting, and cannot image the target beyond the focal plane. In addition, the traditional optical microscopy is influenced by brain tissue scattering, and the tissues below 300 mu m cannot be imaged. The above two disadvantages greatly limit the application of optical microscopy in brain imaging.
In the related art, in order to realize the observation of the multi-plane sample, the following three schemes are generally adopted: the first uses mechanical means to move the objective lens to achieve different depth positions, but mechanical inertia limits the speed of imaging; the second method adopts the electric-adjusting zoom lens to change the focal length of the objective lens, and compared with the first method, the method improves the imaging speed, but is limited by the principle of the electric-adjusting lens, and the field range is limited; the third is a light field microscopic imaging method, which realizes the acquisition and reconstruction of a three-dimensional light field by sacrificing the transverse resolution of an imaging system, cannot obtain information of brain tissue fine structures, can realize three-dimensional reconstruction only by needing a large amount of data algorithms and time, and cannot observe the brain tissue conditions in real time. In addition, in order to realize deep imaging, a confocal microscope and a multiphoton scanning microscope are the most commonly used methods at present, but the time resolution of the system is greatly reduced by adopting a scanning mode to acquire images, and the problem is more serious in large-field imaging.
Disclosure of Invention
The present invention is directed to solving, at least to some extent, the technical problems in the related art.
Therefore, the first objective of the present invention is to provide an arbitrary depth multi-plane micro-imaging system based on a gradient variable refractive index lens, which adds the gradient variable refractive index lens between an objective lens and a sample surface of a conventional optical microscope to realize simultaneous imaging of different depth regions in a field of view, and also adds a high refractive index substance as an image plane correction module in a region other than the gradient variable refractive index lens to adjust the position of an image plane, so that the image planes inside and outside an observation region of the gradient variable refractive index lens are at the same height, thereby realizing imaging of arbitrary depth, improving imaging speed, and expanding the field of view.
The second purpose of the invention is to provide an arbitrary depth multi-plane microscopy imaging method based on the gradient variable refractive index lens.
A third object of the invention is to propose a non-transitory computer-readable storage medium.
In order to achieve the above object, a first aspect of the present invention provides an arbitrary depth multi-plane micro-imaging system based on a gradient variable refractive index lens, comprising:
the gradient variable refractive index lens is used for generating a virtual image of a sample to be imaged in an observation area of the gradient variable refractive index lens on a microscope focal plane;
the objective lens window is used for fixing the gradient variable refractive index lens and maintaining the smoothness of an observation visual field of the imaging system;
the image plane correction module is used for generating a virtual image of a sample to be imaged outside an observation area of the gradient variable refractive index lens and adjusting the position of an image plane outside the observation area of the gradient variable refractive index lens so as to enable the image planes inside and outside the observation area of the gradient variable refractive index lens to be at the same height;
the microscopic optical amplification module is used for collecting optical signals emitted from the focal plane of the microscope and carrying out optical amplification;
and the information acquisition module is used for receiving the optical signal output by the microscopic optical amplification module and imaging.
According to the multi-plane micro-imaging system with any depth based on the gradient variable refractive index lens, the gradient variable refractive index lens is added between the objective lens and the sample, and meanwhile, the height of the image surface inside and outside the observation area of the gradient variable refractive index lens is adjusted by using the high-refractive-index medium, so that the image surfaces inside and outside the gradient variable refractive index lens are at the same height, images formed by different depth areas at the same time can be obtained in the microscope visual field, effective and rapid multi-plane micro-imaging with any depth can be realized, and the capacity of simultaneously imaging different depths in a large range in the sample can be realized.
In addition, the multi-plane micro imaging system with any depth based on the gradient variable refractive index lens according to the above embodiment of the invention can also have the following additional technical features:
optionally, in an embodiment of the present invention, when the length of the gradient variable refractive index lens is greater than or equal to the tissue depth of the sample to be imaged, the gradient variable refractive index lens and the image plane correction module map the sample to be imaged inside and outside the observation area of the gradient variable refractive index lens to the same virtual image plane; when the length of the gradient variable refractive index lens is smaller than the tissue depth of the sample to be imaged, the gradient variable refractive index lens maps the sample to be imaged in the observation area to a first virtual image surface, and the image surface correction module maps a virtual image on the first virtual image surface and the sample to be imaged outside the observation area to a second virtual image surface.
Optionally, in an embodiment of the present invention, the image plane correction module is made of a high refractive index material, and the image plane correction module includes: BK7 glass.
Optionally, in an embodiment of the present invention, the micro-optical amplification module is specifically configured to implement size amplification of focal plane information through refraction of an optical signal in a medium, and couple the amplified optical signal to the information acquisition module.
Optionally, in an embodiment of the present invention, the information acquisition module is an area array light intensity detector, and the information acquisition module is further configured to convert the two-dimensional optical signal into an electrical signal.
In order to achieve the above object, a second aspect of the present invention provides a method for arbitrary depth multi-plane micro-imaging based on a gradient variable refractive index lens, comprising the following steps:
installing a large view field window on the surface of a sample to be imaged;
implanting a gradient variable refractive index lens module at a preset depth in the sample to be imaged;
placing an image plane correction module on the large view field window;
adjusting the axial position of the gradient variable refractive index lens and performing image surface correction to image and observe the sample to be imaged, comprising: generating a virtual image of a sample to be imaged in an observation area of the gradient variable refractive index lens on a microscope focal plane through the gradient variable refractive index lens; generating a virtual image of a sample to be imaged outside an observation region of the gradient variable refractive index lens through an image plane correction module, and adjusting the position of an image plane outside the observation region of the gradient variable refractive index lens; and coupling the optical signal emitted from the focal plane of the microscope to an information acquisition module of the microscope for imaging.
Optionally, in an embodiment of the present invention, the method further includes: when the length of the gradient variable refractive index lens is larger than or equal to the preset depth, controlling the gradient variable refractive index lens and the image plane correction module to map the samples to be imaged inside and outside the observation area of the gradient variable refractive index lens to the same virtual image plane; when the length of the gradient variable refractive index lens is smaller than the tissue depth of the sample to be imaged, the gradient variable refractive index lens is controlled to map the sample to be imaged in the observation area to a first virtual image surface, and the image surface correction module is controlled to map a virtual image on the first virtual image surface and the sample to be imaged outside the observation area to a second virtual image surface.
Optionally, in an embodiment of the present invention, the image plane correction module is made of a high refractive index material, and the image plane correction module includes: BK7 glass.
Optionally, in an embodiment of the present invention, the information acquisition module is an area array light intensity detector, and the coupling of the optical signal emitted from the focal plane of the microscope to the information acquisition module of the microscope for imaging includes: and after the two-dimensional optical signal is converted into an electric signal through the information acquisition module, imaging is carried out according to the converted electric signal.
According to the multi-plane microimaging method based on the gradient variable refractive index lens, the gradient variable refractive index lens is added between the objective lens and the sample, the axial position of the gradient variable refractive index lens is adjusted, and meanwhile, the height of the image surface inside and outside the observation area of the gradient variable refractive index lens is adjusted by using a high refractive index medium, so that the image surfaces of the inside and outside areas of the gradient variable refractive index lens are at the same height, images formed by different depth areas at the same time can be obtained in the microscope visual field, effective and rapid multi-plane and multi-plane microimaging at any depth is realized, and the capability of imaging different depths in a large range in the sample at the same time is realized.
To achieve the above object, a non-transitory computer-readable storage medium is provided in an embodiment of a third aspect of the present invention, and a computer program is stored thereon, and when being executed by a processor, the computer program implements an arbitrary depth multi-plane micro-imaging method based on a gradient variable refractive index lens according to an embodiment of the second aspect of the present application.
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 schematic structural diagram of an arbitrary depth multi-plane micro-imaging system based on gradient variable index lenses according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of light rays for arbitrary depth multi-plane wide-field microscopy imaging based on a gradient variable refractive index lens according to an embodiment of the present invention;
FIG. 3 is a schematic diagram of arbitrary depth multi-plane imaging based on a gradient index lens according to an embodiment of the present invention;
FIG. 4 is a schematic diagram of light rays for arbitrary depth multi-plane wide-field microscopy imaging based on gradient variable index lens according to an embodiment of the present invention;
FIG. 5 is a schematic diagram of an arbitrary depth multi-plane imaging based on a gradient index lens according to an embodiment of the present invention;
FIG. 6 is a flow chart of a method for providing arbitrary depth multi-plane micro-imaging based on gradient index lens according to one embodiment of the present 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 monolithic photonic integrated chip for the externally tuned laser array proposed according to the embodiments of the present invention is described below with reference to the accompanying drawings.
FIG. 1 is a schematic structural diagram of an arbitrary depth multi-plane micro-imaging system based on a gradient index lens according to an embodiment of the invention.
As shown in FIG. 1, the wide-field arbitrary depth multi-plane micro-imaging system based on the gradient variable refractive index lens comprises:
the device comprises a gradient variable refractive index lens 10, an objective lens window 20, an image plane correction module 30, a micro-optical amplification module 40 and an information acquisition module 50. The connection relationship of the modules is shown in fig. 1.
The gradient variable refractive index lens 10 is used for generating a virtual image of a sample to be imaged in an observation area of the gradient variable refractive index lens 10 on a microscope focal plane.
The observation region of the gradient refractive index lens 10 is a region of a sample to be imaged, which can be observed through an observation window below the gradient refractive index lens 10, and the sample to be imaged can be any sample with a multilayer structure, which needs to be imaged and observed through a microscope in practical application, for example, in the research of brain science, the sample to be imaged can be brain tissue including a cerebral cortex and a hippocampus located under the cortex.
Specifically, in an embodiment of the present invention, the gradient-index lens 10 may be implanted inside a sample to be imaged, and an optical signal of a deep tissue of the sample to be imaged below the deep tissue is input into a microscope and moved to a focal plane of the microscope, for example, a deep brain tissue structure (e.g., a hippocampus) first forms a virtual image of the hippocampus on the focal plane of the microscope through the gradient-index lens 10, so that the virtual image of the hippocampus is subsequently imaged on the information acquisition module through an optical amplification module of the microscope.
And an objective lens window 20 for fixing the gradient index lens 10 and maintaining the flatness of the observation field of the imaging system of the present invention.
Specifically, the objective lens window 20 keeps the observation field of the microscopic imaging system flat, so that the fluorescence of the sample to be imaged, such as cerebral cortex, is transmitted to the microscopic imaging light path, and at the same time, the objective lens window plays a role of fixing the gradient variable refractive index lens 10
And the image plane correction module 30 is configured to generate a virtual image of the sample to be imaged outside the observation region of the gradient variable refractive index lens 10, and adjust the image plane position outside the observation region of the gradient variable refractive index lens so that the image planes inside and outside the observation region of the gradient variable refractive index lens 10 are at the same height.
In an embodiment of the present invention, the image plane correction module 30 is a high refractive index material, for example, the image plane correction module 30 may be BK7 glass, and the image plane correction module 30 may form a virtual image from a surface tissue of a plane different from the observation area of the gradient refractive index lens 10, and adjust the position of the image plane to be the same as the image plane in the observation area of the gradient refractive index lens 10, that is, make the image planes inside and outside the observation area of the gradient refractive index lens 10 at the same height, which may be a microscope focal plane.
And the microscopic optical amplification module 40 is used for collecting optical signals emitted on the focal plane of the microscope and carrying out optical amplification. In an embodiment of the present invention, the micro-optical amplifying module 40 is specifically configured to implement size amplification of focal plane information through refraction of an optical signal in a medium, and couple the amplified optical signal to the information collecting module 50
And the information acquisition module 50 is used for receiving the optical signal output by the micro optical amplification module and imaging.
Specifically, in an embodiment of the present invention, the information acquisition module is an area array light intensity detector, for example, it may be a CMOS or CCD detector, and the information acquisition module is specifically configured to convert the amplified two-dimensional optical signal into an electrical signal and perform imaging according to the converted electrical signal.
The system aims to improve the performance of the traditional microscopic imaging light path, so that the simultaneous observation of different depth signals at different view field positions is realized on the premise of keeping the traditional microscopic imaging light path. Wherein, the observation surface of gradient variable refractive index lens 10, the image plane of image plane correction module 30 and the microscope focal plane constitute the imaging light path of microscope to guarantee that the observation surface of waiting to image the sample deep layer and shallow tissue can both image on the focal plane of microscope, thereby the sample of different planes can both clearly image.
It should be noted that, in an embodiment of the present invention, the image plane correction module 30 may not be limited to a solid substance, and may also be used to place a liquid and a gas substance. Also, the shape of the image plane correction module 20 may not be limited to a cube, but may be any shape, for example, it may be a convex lens and a concave lens to realize simultaneous observation of multiple planes.
In an embodiment of the present invention, in practical implementation, an optical signal of an observation window of the gradient refractive index lens 10 forms a virtual image on a focal plane of a microscope objective through the gradient refractive index lens 10 and the objective window 20, the image plane correction module 30 maps a signal of a shallow brain region outside the observation window of the gradient refractive index lens 10 onto the focal plane of the microscope objective to form a virtual image, and in addition, an optical signal of a wide field observation region of the microscope at the focal plane of the microscope passes through the wide field observation window and passes through the microscopic optical amplification module 40 together with the virtual image of the gradient refractive index lens observation window to form an image on the information acquisition module 50.
It can be understood that, in practical application, the length of the gradient variable refractive index lens may be greater than or less than the tissue depth of a sample to be imaged, in an embodiment of the present invention, when the length of the gradient variable refractive index lens is greater than or equal to the tissue depth of the sample to be imaged, the gradient variable refractive index lens and the image plane correction module map the sample to be imaged inside and outside an observation region of the gradient variable refractive index lens to the same virtual image plane, when the length of the gradient variable refractive index lens is less than the tissue depth of the sample to be imaged, the gradient variable refractive index lens maps the sample to be imaged in the observation region to a first virtual image plane, and the image plane correction module maps a virtual image on the first virtual image plane and the sample to be imaged outside the observation region to a second virtual image plane.
To sum up, the many planes of arbitrary degree of depth microscopic imaging system based on gradient variable refractive index lens of this application, through add gradient variable refractive index lens between traditional optical microscope's objective and sample face, form images when realizing the different depth zone in the field of vision, and, still, high refractive index material is added as image plane correction module in the region beyond gradient variable refractive index lens, with the position of adjustment image plane, make the image plane of gradient variable refractive index lens's observation region inside and outside at same height, realized the formation of image to arbitrary degree of depth, imaging speed has been improved, the field of view scope has been enlarged.
In order to more clearly illustrate the imaging principle and the connection relationship of the components of the multi-plane imaging system with arbitrary depth based on the gradient variable refractive index lens, two specific embodiments are described below.
Specific example 1:
fig. 2 is a light ray diagram of a multi-planar wide-field microscopic imaging at any depth based on a gradient variable refractive index lens according to an embodiment of the present invention, which is suitable for multi-planar imaging for realizing simultaneous observation of the surface of a hippocampus and a cortex, and the length of the gradient variable refractive index lens is longer than the depth of tissue in an observation area.
Specifically, as shown in fig. 2, deep hippocampal tissue is mapped to an imaginary image plane through a gradient variable index lens, and meanwhile, shallow tissue is also mapped to the same imaginary image plane through a BK7 glass column. As shown in FIG. 3, the depth of observation from the surface layer of the brain tissue to the deep hippocampus was 1.5 mm. The gradient index lens has a diameter of 0.5mm and a length of 2 mm. The diameter of the cover glass is 8mm, the thickness of the cover glass is 0.17mm, and a hole with the diameter of 0.5mm is arranged at the position 1mm away from the center of the circle. The image plane correcting device adopts a BK7 glass column, the thickness is 1.5mm, the diameter is 6mm, the recording center of the punching position is 1mm, and the diameter is 0.8 mm. The upper surface of the cover glass coincides with the lower surface of the BK glass column. The spatial position of the gradient index lens can be precisely adjusted by a mechanical structure. The microscopic magnification imaging adopts a wide-view-field high-resolution objective lens imaging system, the magnification is 10X, the numerical aperture is 0.3, the view field size is 1 cm, and the resolution is 0.8 mu m. The detector adopts an sCMOS chip of Fairychild company in the United states, and the pixel size is 6.5 mu m.
Specific example 2:
fig. 4 is another light ray diagram for multi-plane wide-field microscopy imaging at any depth based on a gradient variable refractive index lens according to an embodiment of the present invention, which is suitable for multi-plane imaging for simultaneous observation of the surface of the hippocampus and the cortex, and the length of the gradient variable refractive index lens is smaller than the depth of the tissue in the observation region.
Specifically, as shown in fig. 4, deep hippocampal tissue is mapped to an virtual image plane 1 through a gradient variable index lens and is further mapped to an virtual image plane 2 through a BK7 glass column. The position of the virtual image plane 2 at this time coincides with the cortical position. As shown in FIG. 5, the depth of observation from the surface layer of the brain tissue to the deep hippocampus was 1.5 mm. The gradient index lens has a diameter of 0.5mm and a length of 1 mm. The inner diameter of the sleeve is 0.5mm, and the wall thickness is 0.1 mm. The outer wall of the gradient refractive index-changing lens is bonded with the overlapped part of the inner wall of the sleeve to ensure that the total length is 1.5mm, and then the gradient refractive index-changing lens is bonded with the lower surface of the cover glass. The diameter of the cover glass is 8mm, and the thickness of the cover glass is 0.17 mm. The image plane correcting device adopts a BK7 glass column, the thickness is 1.5mm, the diameter is 0.8 mm, and the upper surface of the cover glass column is superposed with the lower surface of the BK glass column. The spatial position of the gradient index lens can be precisely adjusted by a mechanical structure. The microscopic magnification imaging adopts a wide-view-field high-resolution objective lens imaging system, the magnification is 10X, the numerical aperture is 0.3, the view field size is 1 cm, and the resolution is 0.8 mu m. The detector adopts an sCMOS chip of Fairychild company in the United states, and the pixel size is 6.5 mu m.
Based on the description of the gradient variable index lens-based arbitrary depth multi-plane micro-imaging system of the embodiment of the above aspect, the following describes an arbitrary depth multi-plane micro-imaging method based on a gradient variable index lens according to another aspect of the embodiment of the invention with reference to the accompanying drawings.
FIG. 6 is a flow chart of a method for any depth multi-plane micro-imaging based on gradient index lens according to an embodiment of the present invention, as shown in FIG. 6, the method includes the following steps:
in step S1, a large field of view window is installed on the surface of the sample to be imaged.
Specifically, taking a sample to be imaged as brain tissue of a mouse, performing imaging observation as an example, a craniotomy is performed before a window is installed, a skull which cannot be penetrated by the optics is carefully removed by using a cranial drill or other tools, and a punched cover glass is installed on the upper surface of the brain of the mouse.
Step S2, implanting the gradient index lens module into the sample to be imaged at a predetermined depth.
In some embodiments of the invention, a gradient index lens module is mounted into brain tissue. In the specific implementation, the cortical tissue covering the surface layer of the deep brain tissue is sucked out by using the holes on the cover glass of the vacuum liquid sucker, and the brain tissue is stopped being sucked out after the depth reaches the designated depth. And then inserting the gradient variable refractive index lens into the brain tissue of the mouse, and accurately adjusting the insertion depth of the gradient variable refractive index lens through a stereotaxic instrument.
Step S3, the image plane correction module is placed on the large field of view window.
Specifically, a perforated glass column of appropriate thickness is placed on the cover glass to ensure that the hole is directly opposite the gradient variable index lens.
Step S4, adjusting the axial position of the gradient variable refractive index lens and performing image plane correction to image and observe a sample to be imaged, including: generating a virtual image of a sample to be imaged in an observation area of the gradient variable refractive index lens on a microscope focal plane through the gradient variable refractive index lens; generating a virtual image of a sample to be imaged outside an observation area of the gradient variable refractive index lens through an image plane correction module, and adjusting the position of an image plane outside the observation area of the gradient variable refractive index lens; and coupling the optical signal emitted from the focal plane of the microscope to an information acquisition module of the microscope for imaging.
In specific implementation, focusing of different planes can be realized by adjusting the axial position of the gradient variable-refractive-index lens, and the brain region is observed by using a multi-plane micro-imaging system based on the gradient variable-refractive-index lens.
In one embodiment of the present invention, the image plane correction module is made of a high refractive index material, and includes: BK7 glass.
In one embodiment of the present invention, the information acquisition module is an area array light intensity detector, and coupling the optical signal emitted from the focal plane of the microscope to the information acquisition module of the microscope for imaging includes: after the two-dimensional optical signal is converted into an electric signal through the information acquisition module, imaging is carried out according to the converted electric signal.
In one embodiment of the invention, step S4 further includes performing size amplification of the focal plane information by refraction of the optical signal in the medium by the micro-optical amplification module, and coupling the amplified optical signal to the information acquisition module.
It should be noted that the above description of the embodiment of the multi-plane micro-imaging system with any depth based on the gradient variable refractive index lens is also applicable to the multi-plane micro-imaging method with any depth based on the gradient variable refractive index lens, and the implementation principle is similar, and is not described herein again.
According to the multi-plane microimaging method based on the gradient variable refractive index lens, an effective and rapid multi-plane microimaging mode is created by adding the gradient variable refractive index lens and the image surface correction module between the objective lens and the sample, the functions of the existing microscope can be expanded economically and efficiently, and the capability of realizing simultaneous imaging of different depths in a large range in the sample is achieved.
In order to achieve the above embodiments, the present invention further proposes a non-transitory computer-readable storage medium, on which a computer program is stored, which, when executed by a processor, implements an arbitrary depth multi-plane micro-imaging method based on a gradient variable index lens according to the embodiment of the second aspect of the present invention.
Although the present application has been disclosed in detail with reference to the accompanying drawings, it is to be understood that such description is merely illustrative and not restrictive of the application of the present application. The scope of the present application is defined by the appended claims and may include various modifications, adaptations, and equivalents of the invention without departing from the scope and spirit of the application.
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 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.
The logic and/or steps represented in the flowcharts or otherwise described herein, e.g., an ordered listing of executable instructions that can be considered to implement logical functions, can be embodied in any computer-readable medium for use by or in connection with an instruction execution system, apparatus, or device, such as a computer-based system, processor-containing system, or other system that can fetch the instructions from the instruction execution system, apparatus, or device and execute the instructions. For the purposes of this description, a "computer-readable medium" can be any means that can contain, store, communicate, propagate, or transport the program for use by or in connection with the instruction execution system, apparatus, or device. More specific examples (a non-exhaustive list) of the computer-readable medium would include the following: an electrical connection (electronic device) having one or more wires, a portable computer diskette (magnetic device), a Random Access Memory (RAM), a read-only memory (ROM), an erasable programmable read-only memory (EPROM or flash memory), an optical fiber device, and a portable compact disc read-only memory (CDROM). Additionally, the computer-readable medium could even be paper or another suitable medium upon which the program is printed, as the program can be electronically captured, via for instance optical scanning of the paper or other medium, then compiled, interpreted or otherwise processed in a suitable manner if necessary, and then stored in a computer memory.
It should be understood that portions of the present invention may be implemented in hardware, software, firmware, or a combination thereof. In the above embodiments, the various steps or methods may be implemented in software or firmware stored in memory and executed by a suitable instruction execution system. If implemented in hardware, as in another embodiment, any one or combination of the following techniques, which are known in the art, may be used: a discrete logic circuit having a logic gate circuit for implementing a logic function on a data signal, an application specific integrated circuit having an appropriate combinational logic gate circuit, a Programmable Gate Array (PGA), a Field Programmable Gate Array (FPGA), or the like.
It will be understood by those skilled in the art that all or part of the steps carried by the method for implementing the above embodiments may be implemented by hardware related to instructions of a program, which may be stored in a computer readable storage medium, and when the program is executed, the program includes one or a combination of the steps of the method embodiments.
In addition, functional units in the embodiments of the present invention may be integrated into one processing module, or each unit may exist alone physically, or two or more units are integrated into one module. The integrated module can be realized in a hardware mode, and can also be realized in a software functional module mode. The integrated module, if implemented in the form of a software functional module and sold or used as a stand-alone product, may also be stored in a computer readable storage medium.
The storage medium mentioned above may be a read-only memory, a magnetic or optical disk, etc. 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 (4)
1. An arbitrary depth multi-plane microscopic imaging system based on a gradient variable refractive index lens, comprising:
the gradient variable refractive index lens is used for generating a virtual image of a sample to be imaged in an observation area of the gradient variable refractive index lens on a microscope focal plane;
the objective lens window is used for fixing the gradient variable refractive index lens and maintaining the smoothness of an observation visual field of the imaging system;
the image plane correction module is used for generating a virtual image of a sample to be imaged outside an observation region of the gradient variable refractive index lens and adjusting the position of the image plane outside the observation region of the gradient variable refractive index lens so that the image planes inside and outside the observation region of the gradient variable refractive index lens are at the same height, wherein the image plane correction module is made of a high-refractive-index substance, and when the length of the gradient variable refractive index lens is greater than or equal to the tissue depth of the sample to be imaged, the gradient variable refractive index lens and the image plane correction module map the sample to be imaged inside and outside the observation region of the gradient variable refractive index lens to the same virtual image plane;
the microscopic optical amplification module is used for collecting optical signals emitted from the focal plane of the microscope and carrying out optical amplification;
and the information acquisition module is used for receiving the optical signal output by the microscopic optical amplification module and imaging.
2. The microscopic imaging system according to claim 1, wherein the image plane correction module is a high refractive index substance, the image plane correction module comprising: BK7 glass.
3. The microscopic imaging system according to claim 1, wherein the microscopic optical amplification module is specifically configured to achieve size amplification of focal plane information through refraction of an optical signal in a medium, and couple the amplified optical signal to the information acquisition module.
4. The microscopic imaging system according to claim 1, wherein the information acquisition module is an area array light intensity detector, and the information acquisition module is further configured to convert a two-dimensional optical signal into an electrical signal.
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| CN109061860A (en) * | 2018-08-16 | 2018-12-21 | 上海理工大学 | A kind of portable high-resolution micro imaging system |
| CN111722391A (en) * | 2019-03-18 | 2020-09-29 | 苏州溢博伦光电仪器有限公司 | Three-dimensional head-mounted microscope |
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| CN113341551A (en) * | 2021-05-19 | 2021-09-03 | 哈尔滨工业大学 | Multi-eye miniature three-dimensional microscopic imaging device and method based on gradient refractive index lens |
| CN113341554A (en) * | 2021-05-19 | 2021-09-03 | 哈尔滨工业大学 | Endoscopic three-dimensional microscopic imaging device and method based on gradient refractive index lens |
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| EP3004847B1 (en) * | 2013-06-03 | 2019-04-10 | Ventana Medical Systems, Inc. | Fluorescence imaging system for tissue detection |
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| CN109061860A (en) * | 2018-08-16 | 2018-12-21 | 上海理工大学 | A kind of portable high-resolution micro imaging system |
| CN112930492A (en) * | 2018-09-28 | 2021-06-08 | 香港大学 | Apparatus and method for fast volumetric fluorescence microscopy using time multiplexed light sheets |
| CN111722391A (en) * | 2019-03-18 | 2020-09-29 | 苏州溢博伦光电仪器有限公司 | Three-dimensional head-mounted microscope |
| CN113341551A (en) * | 2021-05-19 | 2021-09-03 | 哈尔滨工业大学 | Multi-eye miniature three-dimensional microscopic imaging device and method based on gradient refractive index lens |
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