CN217639723U - Multi-focus structured light illumination microscopic imaging system - Google Patents
Multi-focus structured light illumination microscopic imaging system Download PDFInfo
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Abstract
The utility model provides a little imaging system of multifocal structural light illumination, include: the system comprises an illumination module, a first super lens array, a carrying platform and a receiving module; the lighting module is configured to emit light; the first super lens array is positioned on the light-emitting side of the lighting module, is configured to modulate incident light into a light spot array and project the light spot array to the carrying platform; the carrying platform can rotate along the circumferential direction of the light spot array; the receiving module is configured to receive the fluorescent signal emitted by the object platform. Through the multi-focus structured light illumination microscopic imaging system provided by the embodiment of the utility model, the surface of the sample can be penetrated while the imaging precision is higher, the interior of the sample is detected, the detection depth is deeper, and the super-resolution imaging of the thick sample can be realized; the first super lens array is utilized to generate the multi-focus light spot array, the structure is simple, the size is small, the overall structure of the system is simplified, the movement is convenient, and the real-time detection can be realized.
Description
Technical Field
The utility model relates to a super-resolution microscopic imaging technical field particularly, relates to a multifocal structure light illumination microscopic imaging system.
Background
An optical microscope is an important research tool in the fields of biomedicine and the like at present, and structured light illumination microscopy (SIM) is a common super-resolution microscopic imaging technology based on the optical microscope, which can break through the limit of the diffraction limit of the traditional optical microscope and has higher imaging resolution.
Although wide field structured light illumination microscopy does not have any limitations on fluorescent dyes, almost all commonly used dyes can be used for imaging, and wide field imaging techniques simultaneously meet the requirements for large-scale, high-speed imaging. However, the application of the wide-field imaging to thick tissue samples is limited, and the power density of the wide-field excitation light is weak, so that the wide-field excitation light cannot penetrate through the surface of the tissue to perform three-dimensional imaging, and therefore, the thick tissue cannot be subjected to super-resolution imaging.
In recent years, a technique of multifocal structured light illumination microscopy (MSIM) and instantaneous structured light illumination microscopy (iSIM) have been proposed. However, the existing SIM system adopts a DMD (Digital Micromirror Device) or a microlens array to generate a multi-focus array, which results in a large overall size of the system and the SIM system cannot be popularized and applied well.
SUMMERY OF THE UTILITY MODEL
To solve the above problem, an object of the embodiments of the present invention is to provide a multifocal structured light illumination microscopic imaging system.
The embodiment of the utility model provides a multifocal structure light illumination microscopic imaging system, include: the system comprises an illumination module, a first super lens array, a carrying platform and a receiving module;
the lighting module is configured to emit light;
the first super lens array is positioned on the light-emitting side of the illumination module, is configured to modulate incident light into a light spot array, and projects the light spot array to the carrying platform;
the object carrying platform is positioned on the light emergent side of the first superlens array, is configured for placing a sample and can rotate along the circumferential direction of the light spot array;
the receiving module is located on the light-emitting side of the fluorescence signal emitted by the object carrying platform and is configured to be capable of receiving the fluorescence signal, and the fluorescence signal is an optical signal generated by projecting the light spot array onto the sample.
In one possible implementation, the stage is located at a focal plane of the first superlens array.
In one possible implementation, the system further comprises: a spectroscope and a detection hole array;
the beam splitter is positioned between the illumination module and the first super lens array and is configured to adjust at least part of light rays emitted by the illumination module to be emitted to the first super lens array;
the spectroscope is also positioned between the receiving module and the first super lens array and is configured to adjust at least part of the fluorescent signal transmitted through the first super lens array to be emitted to the receiving module;
the detection hole array is positioned on one side of the receiving module close to the spectroscope and is configured to allow the fluorescent signals focused on the focal plane of the first super lens array to pass through.
In one possible implementation, the beam splitter is a dichroic mirror.
In a possible implementation manner, the first superlens array has the same modulation effect on the light emitted by the illumination module and the fluorescent signal.
In one possible implementation, the system further includes: an image integration device;
the image integration device is positioned on the light incident side of the receiving module and is configured to emit the fluorescent signals corresponding to the object platform when the object platform rotates to different angles to different positions of the receiving module, so that the fluorescent signals at multiple angles can be integrated into one image.
In one possible implementation, the image integration apparatus includes: a scanning galvanometer and a second superlens array; the scanning galvanometer is configured to adjust the incident fluorescence signal to be emitted to the second super lens array and change the emitting direction; the second superlens array is configured to focus the incident fluorescence signal to the receiving module;
or,
the image integration device comprises an adjustable superlens array; the adjustable super lens array comprises a plurality of adjustable super lens units arranged in an array, and the adjustable super lens units are configured to focus the incident fluorescence signals to the receiving module and focus the fluorescence signals to different positions under the action of different excitations.
In a possible implementation, in a case where the image integration apparatus includes an adjustable superlens array, the adjustable superlens unit includes: the phase change material comprises a first electrode layer, a second electrode layer, a substrate, a nano structure and a phase change material layer;
the nano structures and the first electrode layer are arranged on the same side of the substrate, the nano structures are arranged in a periodic array mode, and the first electrode layer is filled among the nano structures; the height of the first electrode layer is less than the height of the nanostructures;
the phase change material layer is positioned on one side of the first electrode layer, which is far away from the substrate, and is filled between the nano structures; the sum of the heights of the first electrode layer and the phase change material layer is greater than the height of the nano structure;
the second electrode layer is positioned on one side, far away from the first electrode layer, of the phase change material layer; the first electrode layer and the second electrode layer are configured to be capable of applying voltages of different magnitudes.
In one possible implementation, the carrier platform is configured to be movable along a plane.
In one possible implementation, the lighting module includes: a light source and a collimating metalens;
the light source is configured to emit light;
the collimating metalens is positioned on the light-emitting side of the light source and is configured to collimate light emitted by the light source.
In one possible implementation, the lighting module further includes: a beam expanding lens;
the beam expanding lens is positioned on the light outlet side of the collimating metalens and is configured to expand the collimated light.
In one possible implementation, the light source includes a plurality of light emitting units arranged in an array, and the collimating metalens includes a plurality of collimating metalens units arranged in an array; the light-emitting units correspond to the collimating metalens units in position one to one.
In one possible implementation, the system further includes: a processing device;
the processing device is connected with the receiving module and is configured to reconstruct a super-resolution image of the sample based on the fluorescence signals received by the receiving module.
In the scheme provided by the embodiment of the utility model, the imaging characteristic of the multi-focus SIM is utilized, the surface of the sample can be penetrated, the interior of the sample is detected, the deep detection depth is realized, and the super-resolution imaging of the thick sample can be realized; and moreover, the first super lens array is utilized to generate the multi-focus light spot array, the structure is simple, the size is small, the overall structure of the multi-focus structured light illumination microscopic imaging system is simplified, the movement is convenient, and the instant detection can be realized.
In order to make the aforementioned and other objects, features and advantages of the present invention comprehensible, preferred embodiments accompanied with figures are described in detail below.
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, it is obvious that the drawings in the description below are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
Fig. 1 is a schematic diagram illustrating a first structure of a multi-focus structured light illumination microscopic imaging system according to an embodiment of the present invention;
fig. 2 is a schematic diagram illustrating a second structure of a multi-focus structured light illumination microscopic imaging system according to an embodiment of the present invention;
fig. 3 is a schematic diagram illustrating a third structure of a multi-focus structured light illumination micro-imaging system provided by an embodiment of the present invention;
fig. 4 shows a fourth structural diagram of the multifocal structured light illuminated microscopic imaging system provided by the embodiment of the present invention;
fig. 5 is a schematic diagram illustrating a fifth structure of a multi-focus structured light illumination microscopic imaging system according to an embodiment of the present invention;
fig. 6 is a schematic structural diagram illustrating an image integration apparatus according to an embodiment of the present invention;
FIG. 7 is a schematic diagram illustrating a tunable superlens unit provided by an embodiment of the present invention;
fig. 8 shows a sixth structural diagram of a multifocal structured light illuminated microscopy imaging system provided by an embodiment of the present invention;
fig. 9 shows a seventh structural diagram of the multifocal structured light illuminated microscopy imaging system according to the embodiment of the present invention.
Icon:
10-an illumination module, 20-a first super lens array, 30-an object carrying platform, 40-a receiving module, 50-a processing device, 60-a spectroscope, 70-a detection hole array, 80-an image integration device, 101-a light source, 102-a collimation super lens, 103-a beam expanding lens, 81-a scanning galvanometer, 82-a second super lens array, 831-a first electrode layer, 832-a second electrode layer, 833-a substrate, 834-a nanostructure and 835-a phase change material layer.
Detailed Description
In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, and are only for convenience of description and simplification of the description, but do not indicate or imply that the device or element referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore, should not be construed as limiting the present 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 one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically limited otherwise.
In the present invention, unless otherwise expressly specified or limited, the terms "mounted," "connected," and "fixed" are to be construed broadly and may, for example, be fixedly connected, detachably connected, or integrally connected; 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 meaning of the above terms in the present invention can be understood according to specific situations by those skilled in the art.
The embodiment of the utility model provides a multifocal structure light illumination microscopic imaging system, it is shown with reference to fig. 1 that this system includes: an illumination module 10, a first superlens array 20, a stage 30 and a receiving module 40.
The lighting module 10 is configured to emit light; the first superlens array 20 is located on the light exit side of the illumination module 10, and configured to modulate the incident light into a light spot array and project the light spot array to the object stage 30; the object stage 30 is positioned on the light-emitting side of the first superlens array 20, is configured to place a sample, and can rotate along the circumferential direction of the light spot array; the receiving module 40 is located at the light emitting side of the stage 30, and is configured to receive the fluorescent signal, which is the light signal generated by projecting the spot array onto the sample.
In the embodiment of the present invention, the lighting module 10 can emit light to provide light to the first superlens array 20 located at the light emitting side thereof; the light emitted from the illumination module 10 is an excitation light for exciting a fluorescence signal of the sample. As shown in fig. 1, the lighting module 10 can emit light from top to bottom, that is, the light emitting side of the lighting module 10 is the lower side of fig. 1, and accordingly, the first superlens array 20 is located at the lower side of the lighting module 10.
The first superlens array 20 includes a plurality of superlens units arranged in an array, each superlens unit is a superlens manufactured based on a super-surface technology, and has a small thickness and a light weight, and can modulate light emitted from the illumination module 10, so as to focus the light emitted from the illumination module 10, and form a light focus (e.g., airy spot). Wherein each superlens unit can form an optical focus, and a light spot array is formed by using the first superlens array 20 in an array form; alternatively, each superlens unit may form an array comprising a plurality of optical foci, such that the first superlens array 20 is capable of forming a larger array of spots.
The object stage 30 is located at the light-emitting side of the first superlens array 20, and the light spot array can be projected onto the object stage 30. As shown in fig. 1, the first superlens array 20 is a transmissive super-surface capable of emitting an array of spots to an underlying stage 30. The stage 30 is used to position a sample (e.g. animal tissue, plant cells, etc.) so that an array of spots can be projected onto the sample. Furthermore, the stage 30 can rotate along the circumferential direction of the spot array, so that the relative angle between the spot array and the stage 30 can be changed, that is, the relative angle between the spot array and the sample can be changed (the sample is generally fixedly placed on the stage 30). The circumferential direction of the light spot array refers to the direction corresponding to the periphery of the light spot array; in particular, the stage 30 may be rotatable about the major axis of the array of spots, which is the axis perpendicular to the plane of the array of spots (e.g. the plane of the stage 30) and passing through a middle position of the array of spots (e.g. the centre of the array of spots). For example, the stage 30 can rotate around the main optical axis of the first superlens array 20, so that the rotation along the circumferential direction of the spot array can be realized, and fluorescence signals at different angles can be formed.
Optionally, the stage 30 is located at the focal plane of the first superlens array 20 so that the spot array can better excite the fluorescence signal of the sample. Compared with wide-field excitation light, the light spot array has higher power density, can penetrate through the surface of a sample (such as a tissue surface) to carry out three-dimensional imaging, has deeper detection depth, can detect the interior of the sample, and thus can realize super-resolution imaging of a thick sample. Further optionally, the carrier platform 30 is configured to be movable along a plane. For example, the stage 30 can move in a direction perpendicular to the main optical axis of the first superlens array 20, so that the spot array can irradiate different positions of the stage 30, and scanning of samples at different positions on the stage 30 is realized.
The receiving module 40 is located at the light-emitting side of the fluorescence signal emitted from the object platform 30, so as to receive the fluorescence signal generated by projecting the light spot array onto the sample. As shown in fig. 1, the carrier 30 is transparent to fluorescent signals, and the receiving module 40 may be located on a side of the carrier 30 away from the lighting module 10. Moreover, the receiving module 40 can receive the fluorescence signals at different angles, and reconstruct an image of the sample based on the fluorescence signals. The receiving module 40 may include a single photon avalanche diode array (SPAD).
Optionally, referring to fig. 2, the microscopic imaging system further comprises a processing device 50; the processing device 50 is connected to the receiving module 40 and is configured to reconstruct a super-resolved image of the sample based on the fluorescence signals received by the receiving module 40. In this embodiment, the processing device 50 may reconstruct the fluorescence signals at a plurality of different angles, so as to reconstruct a super-resolution image of the sample. For example, the processing device 50 may reconstruct a super-resolution wide-field image of the sample via pixel repositioning and deconvolution algorithms. The process of reconstructing an image of a sample based on multiple fluorescence signals is a mature technology in the existing multi-focus SIM system, and is not described in detail here.
The embodiment of the utility model provides a multifocal structured light illumination microscopic imaging system utilizes the formation of image characteristics of multifocal SIM, can pierce through the sample surface, detects the sample inside, has darker detection depth, can realize the super-resolution formation of image to thick sample; moreover, the first super lens array 20 is used for generating the multi-focus light spot array, the structure is simple, the size is small, the overall structure of the multi-focus structured light illumination microscopic imaging system is simplified, the movement is convenient, and the instant detection can be realized.
Optionally, the multifocal structured light illuminated microscopy imaging system is a confocal structure. Referring to fig. 3 and 4, the multifocal structured light illuminated microscopy imaging system further comprises: a beamsplitter 60 and a detection aperture array 70. The beam splitter 60 is located between the illumination module 10 and the first superlens array 20, and is configured to adjust at least a portion of the light emitted by the illumination module 10 to be emitted toward the first superlens array 20; the beam splitter 60 is also located between the receiving module 40 and the first superlens array 20, and is configured to adjust at least a portion of the fluorescence signal transmitted through the first superlens array 20 to be directed to the receiving module 40. The detection aperture array 70 is located on a side of the receiving module 40 close to the beam splitter 60, and is configured to allow the fluorescence signal focused on the focal plane of the first superlens array 20 to pass through.
In the embodiment of the present invention, the spectroscope 60 is an optical element with a transflective function, which can transmit and reflect light. For example, referring to fig. 3, the beam splitter 60 is configured to transmit at least a portion of the light emitted from the illumination module 10 to the first superlens array 20 and reflect at least a portion of the fluorescence signal transmitted through the first superlens array 20 to the receiving module 40; at this time, the illumination module 10 and the first superlens array 20 may be coaxial. Alternatively, referring to fig. 4, the beam splitter 60 is configured to reflect at least a portion of the light emitted by the illumination module 10 to the first superlens array 20 and transmit at least a portion of the fluorescence signal transmitted by the first superlens array 20 to the receiving module 40; at this time, the receiving module 40 and the first superlens array 20 may be coaxial.
And, there is a detection hole array 70 on the light incident side of the receiving module 40, and the detection hole array 70 is a small hole array, which can filter the optical signal out of the focal plane to allow the fluorescent signal focused on the focal plane of the first superlens array 20 to pass through. Utilize the detection hole battle array 70 for the point beyond the focal plane is kept off and can not be imaged outside detection hole battle array 70, and the fluorescence signal that receiving module 40 obtained is the optical tangent plane of sample, can realize the effect of optical section, has avoided the interference of stray light on the non-focal plane, can overcome the blurred shortcoming of ordinary microscope image, can obtain clear confocal image on the whole focal plane.
Taking the microscopic imaging system shown in fig. 3 as an example, the working process of the microscopic imaging system is as follows: the illumination module 10 emits an excitation light, and the excitation light is emitted to the beam splitter 60, passes through the beam splitter 60, and then is incident to the first super-lens array 20; the first super lens array 20 converges the excitation light into a spot array and emits the spot array to a sample on a stage 30; the light spot array can excite the fluorescent molecules of the sample labeling part to generate a fluorescent signal. The fluorescence signal passes through the first superlens array 20 and is emitted to the beam splitter 60, the beam splitter 60 reflects the fluorescence signal again, so that the portion of the fluorescence signal focused on the focal plane of the first superlens array 20 can pass through the detection hole array 70, and the receiving module 40 can receive the filtered fluorescence signal.
Alternatively, the light (excitation light) emitted by the illumination module 10 and the fluorescence signal excited by the sample are signals with different wavelengths, and the dichroic mirror 60 may be specifically a dichroic mirror, which has different processing effects on the light of the excitation light wave band and the light of the fluorescence signal wave band; for example, as shown in fig. 3, the dichroic mirror may transmit light in the excitation light band and reflect light in the fluorescence signal band; alternatively, as shown in fig. 4, the dichroic mirror may reflect light of the excitation light band and transmit light of the fluorescence signal band.
Alternatively, the multifocal structured light illumination microscopic imaging system may adopt nonlinear imaging, such as two-photon fluorescence microscopic imaging, where the first superlens array 20 has the same modulation effect on the light emitted by the illumination module 10 and the fluorescence signal, that is, the phase modulation effect of the first superlens array 20 on the light in the excitation light band and the light in the fluorescence signal band is the same.
Optionally, when the object platform 30 rotates along the circumferential direction of the light spot array, the object platform 30 rotates to different angles, and corresponding fluorescent signals are generated; in order to reconstruct an image of a sample, fluorescence signals at different angles need to be processed respectively, and the imaging speed is low and the real-time observation capability is not provided. In the embodiment of the utility model provides an in, utilize instantaneous structured light illumination micro-imaging (iSIM) technique to improve imaging speed. Specifically, referring to fig. 5, the multi-focal structured light illumination microscopy imaging system further comprises: the image integration device 80. The image integration device 80 is located at the light incident side of the receiving module 40 and configured to emit the fluorescence signals corresponding to the different angles of the stage 30 to different positions of the receiving module 40, so that the fluorescence signals at multiple angles can be integrated into one image.
In the embodiment of the present invention, the image integration device 80 is disposed on the light incident side of the receiving module 40, and the image integration device 80 can adjust the position of the fluorescent signal to the receiving module 40. Specifically, for the fluorescence signals of different angles, the image integration device 80 adjusts them to be directed to different positions of the receiving module 40, so that the receiving module 40 can distinguish the fluorescence signals of different angles, and the fluorescence signals of multiple angles can be naturally integrated into one image. After the integrated image is determined, the image of the sample can be generated by reconstructing the image, and the process does not need to process and carry out super-resolution reconstruction on a plurality of images, so that the calculation power can be saved, and the processing speed can be improved.
Alternatively, referring to fig. 6, the image integration apparatus 80 includes: a scanning galvanometer 81 and a second superlens array 82; the scanning galvanometer 81 is configured to adjust the incident fluorescence signal to be directed to the second superlens array 82, and to be able to change the exit direction; the second superlens array 82 is configured to focus the incident fluorescence signal to the receiving module 40.
In the embodiment of the present invention, the second superlens array 82 can focus the fluorescence signal to form a fluorescence array (the fluorescence array is different from the above-mentioned light spot array); and, the fluorescence array is focused to the receiving module 40, so that the receiving module 40 can receive the fluorescence signal in an array form, i.e. the fluorescence array. The focal length of the second superlens array 82 may be smaller than that of the first superlens array 20, for example, the former is one-half of the latter. The scanning galvanometer 81 can coordinate with the rotation of the object carrying platform 30 to adjust the fluorescent signals of different angles to have different emitting directions, so that the fluorescent signals of different angles can form fluorescent arrays at different positions of the receiving module 40 after penetrating through the second superlens array 82, that is, an integrated image with a plurality of points represented by lines can be formed; one form of the integrated image is shown on the right side of fig. 6. The scanning galvanometer 81 may be a MEMS galvanometer.
Alternatively, the image integration device 80 may not change the direction of the fluorescence signal by using the mechanical scanning galvanometer 81. In particular, the image integration device 80 includes a tunable superlens array; the tunable superlens array includes a plurality of tunable superlens units arranged in an array, and the tunable superlens units are configured to focus the incident fluorescence signal to the receiving module 40 and focus the fluorescence signal to different positions under different excitations.
In the embodiment of the present invention, the adjustable super lens array is similar to the above-mentioned second super lens array 82, and both of them comprise a plurality of super lens units arranged in an array, and the difference lies in that the super lens unit in the adjustable super lens array is phase adjustable, i.e. the super lens unit is an adjustable super lens unit. Each adjustable superlens unit can converge the fluorescence signal, so that the adjustable superlens array can converge the fluorescence signal into a fluorescence array and focus the fluorescence array on the receiving module 40; in addition, the adjustable superlens unit can focus the fluorescence signals at different angles to different positions under different phase modulations, so that the receiving module 40 can integrate the fluorescence signals at multiple angles into one image, thereby realizing image integration.
Optionally, referring to fig. 7, the tunable superlens unit includes: a first electrode layer 831, a second electrode layer 832, a substrate 833, a nanostructure 834, and a phase change material layer 835.
The nanostructures 834 and the first electrode layer 831 are both arranged on the same side of the substrate 833, the nanostructures 834 are arranged in a periodic array, and the first electrode layer 831 is filled between the nanostructures 834; the height of the first electrode layer 831 is less than the height of the nanostructures 834; the phase change material layer 835 is located on one side of the first electrode layer 831 far away from the substrate 833 and is filled between the nanostructures 834; the sum of the heights of the first electrode layer 831 and the phase change material layer 835 is greater than the height of the nanostructure 834; the second electrode layer 832 is located on a side of the phase change material layer 835 away from the first electrode layer 831; the first electrode layer 831 and the second electrode layer 832 are configured to be able to apply voltages of different magnitudes.
In the embodiment of the present invention, the substrate 833 and the plurality of nanostructures 834 arranged periodically on one side thereof constitute a basic super surface, and the first electrode layer 831 and the second electrode layer 832 are disposed on both sides of the phase change material layer 835, and different voltages are applied to the first electrode layer 831 and the second electrode layer 832 to form a voltage difference, so that the phase change material layer 835 made of the phase change material can be electrically excited, and then the phase change state of the phase change material layer 835 is changed. Optionally, the phase change material is a material capable of realizing crystalline state and amorphous state conversion; for example, the phase change material may be germanium antimony telluride (Ge) X SB Y TE Z ) Germanium telluride (Ge) X TE Y ) Antimony telluride (Sb) X TE Y ) Silver antimony telluride (Ag) X SB Y TE Z ) And the like. For example, the phase change material is GST (Ge) 2 SB 2 TE 5 ) By applying voltage and the like, the crystalline state of the phase-change material can be realized
Fast conversion of the amorphous state; also, partial crystallization may be achieved so that the phase change material can be in one state between the crystalline and amorphous states.
The first electrode layer 831 and the phase change material layer 835 are filled around the nanostructure 834, and the equivalent refractive index of the position of the nanostructure 834 can be changed by changing the phase change state of the phase change material layer 835, so that the modulation effect of the adjustable superlens unit is changed. The sum of the heights of the first electrode layer 831 and the phase-change material layer 835 is greater than the height of the nanostructure 834, so that the second electrode layer 832 is spaced from the nanostructure 834 by a certain distance, the nanostructure 834 can be prevented from contacting the second electrode layer 832, and electric leakage can be prevented.
The embodiment of the present invention provides different voltages to the phase change material layer 835, which can change the phase modulated by the first superlens array 20 to realize different modulation effects. As shown in FIG. 7, the fluorescence signal is at an incident angle θ i The fluorescence signal is emitted into the adjustable super lens unit, and the emergent angle of the fluorescence signal after the phase modulation is carried out on the emitted fluorescence signal is theta o . When different voltages are applied to the phase change material layer 835 by the two electrode layers, the emergent angles of the emergent rays of the adjustable superlens unit are different; as shown in FIG. 7, a voltage V is applied to the phase change material layer 835 2 The emergent angle is larger than the voltage V applied to the phase-change material layer 835 1 The time-lapse emergent angle enables the adjustable superlens unit to adjust the fluorescent signal to irradiate to different positions, and different phase modulation is realized.
In an embodiment of the present invention, the first electrode layer 831 and the second electrode layer 832 of the tunable superlens unit can be independently controlled, i.e., different tunable superlens units independently apply excitation to the phase change material layer 835. Alternatively, a plurality of adjustable superlens units in the adjustable superlens array share the same first electrode layer 831 and the same second electrode layer 832, that is, the plurality of adjustable superlens units can be synchronously controlled, so that the change degrees of the phase modulations of the plurality of adjustable superlens units are the same, and thus the position of the fluorescence array at the receiving module 40 can be integrally changed.
Alternatively, if the multi-focus structured light illumination microscopic imaging system is a confocal structure, it can also be provided with an image integration device 80, and the specific structure of the system can be seen in fig. 8; for details of the working principle of the image integration apparatus 80, reference is made to the above contents, which are not described herein again. In this embodiment, the image integration device 80 may be specifically disposed between the detection hole array 70 and the receiving module 40 (fig. 8 does not show the detection hole array 70), and the detection hole array 70 is firstly used to block the fluorescence signals from the focal plane, so as to improve the contrast of the fluorescence, and then the fluorescence signals at different angles are converged to different positions of the receiving module 40.
On the basis of any of the above embodiments, referring to fig. 9, the lighting module 10 includes: a light source 101 and a collimating metalens 102. The light source 101 is configured to emit light; the collimating metalens 102 is located on the light exit side of the light source 101 and is configured to collimate the light emitted by the light source 101.
The embodiment of the utility model provides an in, the super lens 102 of collimation can carry out the collimation to the light that light source 101 sent for the first super lens array 20's of directive light is collimated, cooperates the first phase modulation effect of super lens array 20, conveniently generates clear facula array. The light source 101 may be a Light Emitting Diode (LED) or a laser, which is not limited in this embodiment.
Further optionally, as shown in fig. 9, the lighting module 10 further includes: a beam expanding lens 103; the beam expanding lens 103 is located on the light-emitting side of the collimating metalens 102 and is configured to expand the collimated light. The embodiment of the utility model provides an in, through the beam expanding effect of beam expanding lens 103, can expand light to wider, do benefit to and realize facula array on a wider scale. The beam expanding lens 103 may be a superlens.
Optionally, the light source 101 includes a plurality of light emitting units arranged in an array, and correspondingly, the collimating metalens 102 includes a plurality of collimating metalens units arranged in an array; the light-emitting units correspond to the collimating metalens units one by one. The light source 101 and the collimating metalens 102 in an array form are utilized to facilitate the subsequent formation of a light spot array. When the illumination module 10 further includes the expander lens 103, the expander lens 103 may be a super lens array including a plurality of super lenses arranged in an array.
The above description is only for the specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto, and any person skilled in the art can easily think of the technical solutions of the changes or replacements within the technical scope of the present invention, and all should be covered within the protection scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (13)
1. A multi-focal structured light illuminated microscopy imaging system, comprising: the system comprises an illumination module (10), a first super lens array (20), an object carrying platform (30) and a receiving module (40);
the lighting module (10) is configured to emit light;
the first super lens array (20) is positioned on the light-emitting side of the illumination module (10), and is configured to modulate incident light rays into a light spot array and project the light spot array to the object stage (30);
the object carrying platform (30) is positioned on the light outlet side of the first super lens array (20), is configured for placing a sample and can rotate along the circumferential direction of the light spot array;
the receiving module (40) is located at the light-emitting side of the fluorescence signal emitted from the object stage (30) and configured to receive the fluorescence signal, wherein the fluorescence signal is a light signal generated by projecting the light spot array onto the sample.
2. The system of claim 1, wherein the stage platform (30) is located at a focal plane of the first superlens array (20).
3. The system of claim 1, further comprising: a spectroscope (60) and a detection hole array (70);
the beam splitter (60) is positioned between the illumination module (10) and the first superlens array (20) and is configured to adjust at least part of the light emitted by the illumination module (10) to be directed to the first superlens array (20);
the beam splitter (60) is also positioned between the receiving module (40) and the first superlens array (20) and is configured to adjust at least part of the fluorescence signal transmitted through the first superlens array (20) to be directed to the receiving module (40);
the detection hole array (70) is positioned on one side of the receiving module (40) close to the spectroscope (60) and is configured to allow the fluorescent signals focused on the focal plane of the first super lens array (20) to pass through.
4. A system according to claim 3, characterized in that the beam splitter (60) is a dichroic mirror.
5. The system according to claim 3, characterized in that said first superlens array (20) has the same modulating effect on the light emitted by said lighting module (10) and on said fluorescent signal.
6. The system of claim 1, further comprising: an image integration device (80);
the image integration device (80) is located at the light incident side of the receiving module (40) and configured to emit the fluorescence signals corresponding to different angles of the object stage (30) to different positions of the receiving module (40), so that the fluorescence signals at multiple angles can be integrated into one image.
7. The system according to claim 6, characterized in that said image integration means (80) comprise: a scanning galvanometer (81) and a second superlens array (82); the scanning galvanometer (81) is configured to adjust the incident fluorescence signal to be directed to the second superlens array (82) and can change the emergent direction; the second superlens array (82) is configured to focus the incident fluorescence signal to the receiving module (40);
or,
the image integration device (80) comprises an adjustable superlens array; the adjustable superlens array comprises a plurality of adjustable superlens units arranged in an array, and the adjustable superlens units are configured to focus the incident fluorescence signals to the receiving module (40) and focus the fluorescence signals to different positions under the action of different excitations.
8. The system according to claim 7, wherein in case the image integration means (80) comprises an array of adjustable superlenses, the adjustable superlens unit comprises: a first electrode layer (831), a second electrode layer (832), a substrate (833), a nanostructure (834), and a phase change material layer (835);
the nano structures (834) and the first electrode layer (831) are arranged on the same side of the substrate (833), the nano structures (834) are arranged in a periodic array mode, and the first electrode layer (831) is filled between the nano structures (834); the height of the first electrode layer (831) is less than the height of the nanostructures (834);
the phase change material layer (835) is positioned on one side, far away from the substrate (833), of the first electrode layer (831) and is filled between the nanostructures (834); the sum of the heights of the first electrode layer (831) and the phase change material layer (835) is greater than the height of the nanostructure (834);
the second electrode layer (832) is positioned on one side of the phase change material layer (835) far away from the first electrode layer (831); the first electrode layer (831) and the second electrode layer (832) are configured to be capable of applying voltages of different magnitudes.
9. The system of claim 1, wherein the carrier platform (30) is configured to be movable along a plane.
10. The system according to claim 1, characterized in that said lighting module (10) comprises: a light source (101) and a collimating metalens (102);
the light source (101) is configured to emit light;
the collimating metalens (102) is located at a light exit side of the light source (101) and is configured to collimate light emitted by the light source (101).
11. The system of claim 10, wherein the lighting module (10) further comprises: a beam expanding lens (103);
the beam expanding lens (103) is positioned on the light outlet side of the collimating metalens (102) and is configured to expand collimated light.
12. The system according to claim 10, wherein the light source (101) comprises a plurality of arrayed light emitting units, and the collimating metalens (102) comprises a plurality of arrayed collimating metalens units; the light-emitting units correspond to the collimating metalens units in position one to one.
13. The system of claim 1, further comprising: a processing device (50);
the processing device (50) is connected to the receiving module (40) and is configured to reconstruct a super-resolved image of the sample based on the fluorescence signals received by the receiving module (40).
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11927769B2 (en) | 2022-03-31 | 2024-03-12 | Metalenz, Inc. | Polarization sorting metasurface microlens array device |
| US11978752B2 (en) | 2019-07-26 | 2024-05-07 | Metalenz, Inc. | Aperture-metasurface and hybrid refractive-metasurface imaging systems |
| US11988844B2 (en) | 2017-08-31 | 2024-05-21 | Metalenz, Inc. | Transmissive metasurface lens integration |
| US12140778B2 (en) | 2019-07-02 | 2024-11-12 | Metalenz, Inc. | Metasurfaces for laser speckle reduction |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11988844B2 (en) | 2017-08-31 | 2024-05-21 | Metalenz, Inc. | Transmissive metasurface lens integration |
| US12140778B2 (en) | 2019-07-02 | 2024-11-12 | Metalenz, Inc. | Metasurfaces for laser speckle reduction |
| US11978752B2 (en) | 2019-07-26 | 2024-05-07 | Metalenz, Inc. | Aperture-metasurface and hybrid refractive-metasurface imaging systems |
| US11927769B2 (en) | 2022-03-31 | 2024-03-12 | Metalenz, Inc. | Polarization sorting metasurface microlens array device |
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