CN114236801B - Light sheet generating device and microscope system with same - Google Patents
Light sheet generating device and microscope system with same Download PDFInfo
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- CN114236801B CN114236801B CN202111243238.3A CN202111243238A CN114236801B CN 114236801 B CN114236801 B CN 114236801B CN 202111243238 A CN202111243238 A CN 202111243238A CN 114236801 B CN114236801 B CN 114236801B
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- 239000013307 optical fiber Substances 0.000 claims abstract description 128
- 238000001514 detection method Methods 0.000 claims abstract description 84
- 239000000835 fiber Substances 0.000 claims abstract description 78
- 238000005286 illumination Methods 0.000 claims abstract description 75
- 239000000758 substrate Substances 0.000 claims abstract description 37
- 238000006243 chemical reaction Methods 0.000 claims description 30
- 230000003287 optical effect Effects 0.000 claims description 13
- 239000012780 transparent material Substances 0.000 claims description 3
- 238000000034 method Methods 0.000 description 8
- 238000010586 diagram Methods 0.000 description 7
- 230000010354 integration Effects 0.000 description 7
- 230000005540 biological transmission Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 238000003384 imaging method Methods 0.000 description 4
- 230000007547 defect Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000004806 packaging method and process Methods 0.000 description 3
- 206010034972 Photosensitivity reaction Diseases 0.000 description 2
- 239000000428 dust Substances 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 208000007578 phototoxic dermatitis Diseases 0.000 description 2
- 231100000018 phototoxicity Toxicity 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/06—Means for illuminating specimens
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
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Abstract
The embodiment of the application provides a light sheet generating device and have its microscope system, this light sheet generating device includes: the substrate, the top surface of the substrate has recesses as the detection area of the sample to be measured, the bottom surface of the detection area is parallel to top surface of the substrate; the multi-core optical fiber is used for transmitting illumination light, a plurality of fiber cores of the multi-core optical fiber are arranged on the top surface of the substrate in a parallel and closely arranged mode, and the illumination light emitted by the emitting end of the multi-core optical fiber passes through the position right above the detection area. According to the light sheet generating device provided by the embodiment of the application, the illumination light beams emitted by the multiple fiber cores of the multi-core optical fiber are arranged in parallel right above the bottom surface of the detection area and are approximately a sheet-shaped light sheet, and the light sheet is parallel to the bottom surface of the detection area and can be used for illuminating a sample to be detected in the detection area, namely, the light sheet generating device converts light rays emitted by the light source into the light sheet by utilizing the multi-core optical fiber, so that the volume and the structural complexity of the whole device are reduced, and the light sheet generating device is easier to integrate.
Description
Technical Field
The present disclosure relates to the field of light sheet microscopy, and in particular, to a light sheet generating device and a microscope system having the same.
Background
This section provides only background information related to the present application and is not necessarily prior art.
A light sheet microscope is a microscope that utilizes light sheet illumination. The light sheet is approximately lamellar illumination light formed by a light beam parallel to the imaging plane of the microscope, which illuminates only the sample located at the focal plane. Compared with the traditional wide-field illumination microscope, the light sheet illumination ensures that the samples above and below the focal plane are not illuminated, has optical slicing capability similar to that of a confocal microscope, improves the contrast and axial resolution of images and backgrounds, reduces photobleaching and phototoxicity, and can detect living organisms for a long time.
Light sheet microscopes have received a great deal of attention for their excellent performance, with the formation of light sheets being critical to the microstructure of the light sheet. In some related art, various ways of generating an optical sheet are proposed, for example: firstly, a cylindrical lens is introduced into a light path, a Gaussian beam is compressed into a plane in one dimension to form a light sheet, but the light sheet is as bright in the middle and dark at two sides as the Gaussian beam, so that a relatively complex light path compensation structure is generally added in the light sheet generating device; secondly, the light sheet is formed by utilizing a light beam scanning mode, such as a Gaussian beam scanning light sheet, and the Gaussian beam is moved at a high speed in one direction, so that the Gaussian beam is equivalent to one light sheet in a short time, and the light sheet is more uniform than the first light sheet formed by a lens, but the light power requirement is higher, and a moving device capable of meeting the precision high speed is required to be arranged; third, the lattice light sheet is to use the spatial light modulator to generate parallel light beams, so that the light beam is similar to a light sheet, but the light sheet is limited by the resolution and modulation capability of the current spatial light modulator, and the quality and intensity of the generated light beams are poor. Besides, the method has the defects of complex structure, large volume and difficult integration, and under the trend that the biological detection equipment is automatically, integrally and miniaturized nowadays, the light sheet generating device in the related technology is difficult to meet the development requirement of the future biological detection equipment. Therefore, how to reduce the volume and the structural complexity of the light sheet generating device, so that the light sheet generating device is easy to integrate is a technical problem to be solved in the field.
Disclosure of Invention
An objective of the embodiments of the present application is to provide a light sheet generating device and a microscope system with the same, so as to reduce the volume and the structural complexity of the light sheet generating device, and make it easy to integrate. The specific technical scheme is as follows:
an embodiment of a first aspect of the present application provides an optical sheet generating device, including:
the device comprises a substrate, wherein the top surface of the substrate is provided with a groove as a detection area of a sample to be detected, and the bottom surface of the detection area is parallel to the top surface of the substrate;
the multi-core optical fiber is used for transmitting illumination light, a plurality of fiber cores of the multi-core optical fiber are arranged on the top surface of the substrate in a parallel and closely arranged mode, and the illumination light emitted by the emergent end of the multi-core optical fiber passes through the position right above the detection area.
According to the light sheet generating device provided by the embodiment of the first aspect of the application, the multi-core optical fiber is utilized to transmit illumination light, the multiple fiber cores of the multi-core optical fiber are arranged on the top surface of the substrate in a parallel and closely arranged mode, the illumination light emitted by the emitting end of the multi-core optical fiber passes through the position right above the detection area, the top surface of the substrate is parallel to the bottom surface of the detection area, it can be understood that the illumination light beams emitted by the multiple fiber cores of the multi-core optical fiber are arranged in parallel right above the bottom surface of the detection area, the light sheet is approximately a sheet-shaped light sheet, and the light sheet is parallel to the bottom surface of the detection area and can be used for illuminating a sample to be detected in the detection area. Therefore, the light sheet generating device provided by the embodiment of the application can convert the light rays emitted by the light source into the light sheets by utilizing the multi-core optical fibers, a complex light path formed by the multi-lens combination in the related technology is not needed, the size and the structural complexity of the whole device are reduced, and the integration is easier.
In some embodiments of the present application, the multicore fibers include a first multicore fiber and a second multicore fiber; the multiple fiber cores of the first multi-core optical fiber are located on a first side of the detection area, the multiple fiber cores of the second multi-core optical fiber are located on a second side of the detection area, the first side and the second side are oppositely arranged, and the first multi-core optical fiber and the second multi-core optical fiber are located on the same plane.
In some embodiments of the present application, the exit end face of each core of the multicore fiber has a light converging portion.
In some embodiments of the present application, the light sheet generating device further comprises a cover plate, wherein the cover plate is matched with the top surface of the substrate and is used for packaging the multi-core optical fiber.
Embodiments of the second aspect of the present application provide a microscope system comprising a light sheet generating device provided according to embodiments of the first aspect of the present application.
According to the microscope system provided by the embodiment of the second aspect of the application, the light sheet generating device transmits illumination light by using the multi-core optical fiber, the multiple fiber cores of the multi-core optical fiber are arranged on the top surface of the substrate in a parallel and closely arranged mode, the illumination light emitted by the emitting end of the multi-core optical fiber passes through the position right above the detection area, the top surface of the substrate is parallel to the bottom surface of the detection area, it can be understood that the illumination light beams emitted by the multiple fiber cores of the multi-core optical fiber are arranged in parallel and are approximately a sheet-shaped light sheet, and the light sheet is parallel to the bottom surface of the detection area and can be used for illuminating the sample to be detected positioned on the bottom surface of the detection area. Therefore, compared with the light sheet microscope in the related art, in the microscope system provided by the embodiment of the application, the light sheet generating device can convert the light rays emitted by the light source into the light sheets by using the multi-core optical fiber, a complex light path formed by the combination of multiple lenses in the related art is not needed, the volume and the structural complexity of the whole device are reduced, and further the volume and the structural complexity of the microscope system provided by the embodiment of the application are also reduced, so that the microscope system is easier to integrate.
In some embodiments of the present application, the microscope system further comprises:
a light source device for emitting uniform light;
the optical fiber conversion head is connected with the multi-core optical fiber and is used for receiving the uniform light as the illumination light and uniformly dividing the illumination light to each fiber core of the multi-core optical fiber.
In some embodiments of the present application, the light source apparatus includes a light emitter and a light homogenizing sheet, wherein the light homogenizing sheet is disposed between an exit end of the light emitter and a receiving end of the fiber optic adapter.
In some embodiments of the present application, the multicore fibers include a first multicore fiber and a second multicore fiber; the multiple fiber cores of the first multi-core optical fiber are positioned on a first side of the detection area, the multiple fiber cores of the second multi-core optical fiber are positioned on a second side of the detection area, the first side and the second side are arranged opposite to each other, and the first multi-core optical fiber and the second multi-core optical fiber are positioned on the same plane;
the microscope system further comprises a half-mirror, wherein the half-mirror is arranged between the optical fiber conversion head and the light homogenizing sheet and is used for equally dividing the uniform light into a first beam and a second beam;
the optical fiber conversion head comprises a first optical fiber conversion head and a second optical fiber conversion head, wherein the first optical fiber conversion head is connected with the first multi-core optical fiber, and the first optical fiber conversion head is used for receiving the first beam as part of the illumination light and equally dividing the part of the illumination light to each fiber core of the first multi-core optical fiber; the second optical fiber conversion head is connected with the second multi-core optical fiber and is used for receiving the second beam as another part of the illumination light and equally dividing the other part of the illumination light to each fiber core of the second multi-core optical fiber.
In some embodiments of the present application, the microscope system further comprises an observation device comprising an objective lens and a camera, wherein the objective lens is used for collecting optical signals generated by illumination of the sample to be tested in the detection area by illumination light of the light sheet generating device; the camera is used for receiving the optical signals and recording sample images of the sample to be detected.
In some embodiments of the present application, the observation device further comprises a filter disposed between the objective lens and the camera.
In some embodiments of the present application, the substrate on the bottom surface of the detection area is made of transparent material, and the number of the observation devices is two, where a first one of the observation devices is disposed directly above the detection area, and a second one of the observation devices is disposed directly below the detection area.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the following description will briefly introduce the drawings that are required to be used in the embodiments or the description of the prior art, and it is obvious that the drawings in the following description are only some embodiments of the present application, and other embodiments may also be obtained according to these drawings to those skilled in the art.
Fig. 1 is an orthogonal view of a light sheet generating device according to an embodiment of the present disclosure;
FIG. 2 is a front view of a light sheet generating device according to an embodiment of the present disclosure;
FIG. 3 is a top view of FIG. 2;
fig. 4 is a schematic structural diagram of a core of a multi-core optical fiber according to an embodiment of the present application;
FIG. 5 is a schematic diagram of a microscope system according to an embodiment of the present application;
FIG. 6 is a schematic diagram of another embodiment of a microscope system according to the present disclosure;
FIG. 7 is a schematic view of a structure of an observation device of a microscope system according to an embodiment of the present disclosure;
FIG. 8 is a schematic diagram of a microscope system integrated into a cell sorting apparatus according to an embodiment of the present disclosure;
fig. 9 is a schematic structural diagram of an application device of a microscope system integrated in a microfluidic chip according to an embodiment of the present application.
Detailed Description
The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, in which like parts are denoted by like reference numerals. It will be apparent that the described embodiments are only some, but not all, of the embodiments of the present application. Based on the embodiments herein, a person of ordinary skill in the art would be able to obtain all other embodiments based on the disclosure herein, which are within the scope of the disclosure herein.
It will be appreciated that in the description of the present application, terms such as "center," "length," "width," "height," "upper," "lower," "front," "rear," "top," "bottom," "inner," "outer," and the like are used for convenience in describing the present application and simplifying the description based on the orientation or positional relationship shown in the drawings, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present application.
A light sheet microscope is a microscope that utilizes light sheet illumination. The light sheet is approximately lamellar illumination light formed by a light beam parallel to the imaging plane of the microscope, which illuminates only the sample located at the focal plane. Compared with the traditional wide-field illumination microscope, the light sheet illumination can ensure that the samples above and below the focal plane are not illuminated, has optical slicing capability similar to that of a confocal microscope, can improve the contrast and axial resolution of images and backgrounds, reduces photobleaching and phototoxicity, can detect living organisms for a long time, and has excellent performance, so that the light sheet microscope is attracting more attention.
Among them, the generation of the light sheet is the key of the microstructure of the light sheet, and in some related technologies, various ways of generating the light sheet are proposed, for example: firstly, a cylindrical lens is introduced into a light path, a Gaussian beam is compressed into a plane in one dimension to form a light sheet, but the light sheet is as bright in the middle and dark at two sides as the Gaussian beam, so that a relatively complex light path compensation structure is generally added in the light sheet generating device; secondly, the light sheet is formed by utilizing a light beam scanning mode, such as a Gaussian beam scanning light sheet, and the Gaussian beam is moved at a high speed in one direction, so that the Gaussian beam is equivalent to one light sheet in a short time, and the light sheet is more uniform than the first light sheet formed by a lens, but the light power requirement is higher, and a moving device capable of meeting the precision high speed is required to be arranged; third, the lattice light sheet is to use the spatial light modulator to generate parallel light beams, so that the light beam is similar to a light sheet, but the light sheet is limited by the resolution and modulation capability of the current spatial light modulator, and the quality and intensity of the generated light beams are poor. Besides, the method for generating the light sheet has certain defects, and also has the defects of complex structure, large volume and difficult integration. With the current trend of the development of the biological detection device towards automation, integration and miniaturization, the light sheet generating device in the related technology is difficult to meet the development requirement of the future biological detection device.
In view of this, as shown in fig. 1 to 3, an embodiment of a first aspect of the present application provides a light sheet generating device 10. The light sheet generating device 10 comprises a substrate 11 and a multi-core optical fiber 12, wherein the top surface of the substrate 11 is provided with a groove as a detection area 101 of a sample to be detected, and the bottom surface of the detection area 101 is parallel to the top surface of the substrate 11; the multi-core fiber 12 is used for transmitting illumination light, the multiple cores 200 of the multi-core fiber 12 are arranged on the top surface of the substrate 11 in parallel and closely arranged, and the illumination light emitted from the emitting end of the multi-core fiber 12 passes through the position right above the detection area 101.
According to the light sheet generating device 10 provided in the embodiment of the first aspect of the present application, the multi-core optical fiber 12 is used to transmit illumination light, the multiple cores 200 of the multi-core optical fiber 12 are arranged on the top surface of the substrate 11 in a parallel and closely arranged manner, the illumination light emitted from the emitting end of the multi-core optical fiber 12 passes through the top surface of the detection area 101 and is parallel to the bottom surface of the detection area 101, it can be understood that the illumination light beams emitted from the multiple cores 200 of the multi-core optical fiber 12 are arranged in parallel and are approximately a sheet-shaped light sheet, and the light sheet is parallel to the bottom surface of the detection area 101 and can be used to illuminate the sample to be measured located on the bottom surface of the detection area 101. Therefore, the light sheet generating device 10 provided in the embodiment of the present application can convert the light emitted by the light source into the light sheet by using the multi-core optical fiber 12, and does not need to use the complex light path formed by the multi-lens combination in the related art, so that the volume and the structural complexity of the whole device are reduced, and the integration is easier.
In some embodiments of the present application, the multi-core optical fiber 12 includes a first multi-core optical fiber and a second multi-core optical fiber; the multiple cores 200 of the first multi-core optical fiber are located on a first side of the detection area 101, the multiple cores 200 of the second multi-core optical fiber are located on a second side of the detection area 101, the first side is opposite to the second side, and the first multi-core optical fiber and the second multi-core optical fiber are located on the same plane. That is, two opposite sides of the detection area 101 are respectively provided with a row of fiber cores 200, and the two rows of fiber cores 200 are located on the same plane. It can be understood that the direction of the illumination light emitted from the emitting end of the fiber core 200 located at the first side of the detection area 101 is opposite to the direction of the illumination light emitted from the emitting end of the fiber core 200 located at the second side of the detection area 101, so that two-sided illumination is implemented directly above the detection area 101, and the two-sided illumination light compensates for uneven light intensity and reduced brightness caused by shielding of the sample to be detected, so that the uniformity of illumination can be improved, and the quality of the formed light sheet is higher.
In some embodiments of the present application, as shown in fig. 4, the exit end face of each core 200 of the multi-core optical fiber 12 has a light converging portion 201. For example, the exit end surface of each fiber core 200 may be processed into a conical surface, a circular table surface, a spherical surface, a sphere-like surface, a wedge surface, or the like, as the light converging portion 201, after two rows of fiber cores 200 that are closely arranged in parallel and are placed opposite to each other are connected to each other, the light of the illumination light is converged at a middle position between each fiber core 200 and the fiber core 200 opposite to the fiber core, and a light spot effect formed near the position is as shown in a dashed line frame of fig. 4, and may be approximately a sheet-like strip-like light sheet, in which a crossing line at the front end of the light converging portion 201 of each fiber core 200 in fig. 4 represents a light transmission path of the illumination light.
Further, by adjusting the light converging capability of the light converging portion 201 and adjusting the distance between every two oppositely disposed fiber cores 200, the width and thickness of the light sheet formed by the light sheet generating device 10 can be controlled, so that the suitability of the light sheet formed by the light sheet generating device 10 to practical application needs is stronger.
In some embodiments of the present application, the light sheet generating device 10 further includes a cover plate 13, where the cover plate 13 is adapted to the top surface of the substrate 11 and is used to encapsulate the multicore optical fiber 12. As shown in fig. 1 and 2, by using the cooperation between the cover 13 and the substrate 11, an optical fiber accommodating layer may be formed between the cover 13 and the substrate 11, so as to encapsulate the parallel and closely arranged portions of the multiple cores 200 of the multi-core optical fiber 12 in the optical fiber accommodating layer, thereby preventing the influence of impurities such as external dust on the light transmission of the illumination light, and improving the quality of the light sheet formed by the light sheet generating device 10.
In some embodiments of the present application, as shown in fig. 1, the cover 13 includes a first block and a second block, where the first block is adapted to a top surface of the substrate 11 located on a first side of the detection area 101, for packaging a first multi-core optical fiber; the second block is adapted to the top surface of the substrate 11 on the second side of the detection area 101 for encapsulating a second multicore fiber. In this case, two optical fiber accommodating layers are formed on both sides of the detection region 101, and a sample cell for inflow of a sample to be measured can be formed by being surrounded by the middle region between the two optical fiber accommodating layers, the middle region between the first block and the second block of the cover plate 13, and the detection region 101.
Embodiments of the second aspect of the present application provide a microscope system 1. As shown in fig. 1 to 7, the microscope system 1 includes a light sheet generating device 10 provided according to an embodiment of the first aspect of the present application.
According to the microscope system 1 provided in the embodiment of the second aspect of the present application, the light sheet generating device 10 uses the multi-core optical fiber 12 to transmit illumination light, the multiple cores 200 of the multi-core optical fiber 12 are arranged on the top surface of the substrate 11 in parallel and closely arranged, and the illumination light emitted from the emitting end of the multi-core optical fiber 12 passes through the top surface of the detection area 101 and is parallel to the bottom surface of the detection area 101, it can be understood that the illumination light beams emitted from the multiple cores 200 of the multi-core optical fiber 12 are arranged in parallel and are approximately a sheet-shaped light sheet, and the light sheet is parallel to the bottom surface of the detection area 101 and can be used to illuminate the sample to be measured located in the detection area 101. Therefore, compared with the light sheet microscope in the related art, in the microscope system 1 provided in the embodiment of the present application, the light sheet generating device 10 can convert the light emitted by the light source into the light sheet by using the multi-core optical fiber, and the complex light path formed by the multi-lens combination in the related art is not required, so that the volume and the structural complexity of the whole device are reduced, and further, the volume and the structural complexity of the microscope system provided in the embodiment of the present application are also reduced, and the integration is easier.
In some embodiments of the present application, as shown in fig. 5 and 6, the microscope system 1 further comprises a light source device 20 and a fiber optic adapter 30. Wherein the light source device 20 is for emitting uniform light; the optical fiber conversion head 30 is connected to the multi-core optical fiber 12, and the optical fiber conversion head 30 is configured to receive the uniform light as illumination light and to equally divide the illumination light into each of the cores 200 of the multi-core optical fiber 12. The uniform light is provided by the light source device 20, and then the uniform light is uniformly distributed as illumination light to each core 200 of the multi-core optical fiber 12 by the optical fiber conversion head 30, so that the illumination light beams transmitted by the multiple cores 200 closely arranged in parallel by the multi-core optical fiber 12 are more uniform, thereby improving the quality of the light sheet formed by the light sheet generating device 10 of the microscope system 1.
In some embodiments of the present application, as shown in fig. 5 and 6, the light source apparatus 20 includes a light emitter 21 and a light homogenizing sheet 22, wherein the light homogenizing sheet 22 is disposed between an exit end of the light emitter 21 and a receiving end of the light conversion head 30. The light source light can be provided for the microscope system 1 through the light emitter 21, and then the light source light emitted by the light emitter 21 is converted into uniform light by utilizing the light homogenizing sheet arranged at the emitting end of the light emitter 21, so that the light received by the optical fiber conversion head 30 is uniform light to serve as illumination light transmitted by the multi-core optical fiber 12, and the uniformity of the illumination light forming the light homogenizing sheet is ensured.
In some embodiments of the present application, as shown in fig. 5 and 6, the multicore fiber 12 includes a first multicore fiber 121 and a second multicore fiber 122; the multiple cores 200 of the first multi-core optical fiber 121 are located on a first side of the detection area 101, the multiple cores 200 of the second multi-core optical fiber 122 are located on a second side of the detection area 101, the first side is opposite to the second side, and the first multi-core optical fiber 121 and the second multi-core optical fiber 122 are located on the same plane. That is, two opposite sides of the detection area 101 are respectively provided with a row of fiber cores 200, and the two rows of fiber cores 200 are located on the same plane. It can be understood that the direction of the illumination light emitted from the emitting end of the fiber core 200 located at the first side of the detection area 101 is opposite to the direction of the illumination light emitted from the emitting end of the fiber core 200 located at the second side of the detection area 101, so that two-sided illumination is implemented directly above the detection area 101, and the illumination light on two sides compensates for uneven light intensity and reduced brightness caused by shielding of the sample to be detected, so as to improve the uniformity of illumination, and make the quality of the formed light sheet higher.
In one case, as shown in fig. 5, the microscope system 1 may further include a half mirror 40, where the half mirror 40 is disposed between the optical fiber conversion head 30 and the light homogenizing sheet 22, and the half mirror 30 is used to divide the uniform light into a first beam and a second beam; in this case, the optical fiber conversion head 30 includes a first optical fiber conversion head 31 and a second optical fiber conversion head 32, wherein the first optical fiber conversion head 31 is connected to the first multi-core optical fiber 121, and the first optical fiber conversion head 31 is configured to receive the first beam as a part of illumination light and equally divide the part of illumination light to each core 200 of the first multi-core optical fiber 121; second fiber optic conversion head 32 is coupled to second multicore fiber 122, and second fiber optic conversion head 32 is configured to receive the second beam as another portion of illumination light and to divide the other portion of illumination light equally into each of cores 200 of second multicore fiber 122. Thus, uniform light can be equally divided to two opposite sides of the detection area 101, so as to improve the uniformity of illumination of the light sheet right above the detection area 101.
In another case, as shown in fig. 6, a thicker (i.e., a larger number of cores 200 are included) multi-core fiber 12 may be used, and after the uniform light generated by the light source device 20 is equally divided into two parts as illumination light by one fiber conversion head 30 to each core 200 in the thicker multi-core fiber 12, the multiple cores 200 in the thicker multi-core fiber 12 are equally divided into two parts, wherein one part is disposed as a first multi-core fiber 121 on the first side of the detection area 101 and the other part is disposed as a second multi-core fiber 122 on the second side of the detection area 101. In this case, the overall structure of the light sheet generating device 10 is simpler than in the case of the above-described one, but the requirements for the light source device 20 and the multi-core optical fiber 12 are increased, the light source device 20 is required to generate uniform light of a larger diameter, and the manufacturing process of the multi-core optical fiber 12 is also more complicated.
In some embodiments of the present application, as shown in fig. 5 to 7, the microscope system 1 further comprises an observation device 50, the observation device 50 comprising an objective lens 51 and a camera 52, wherein the objective lens 51 is used for collecting an optical signal generated by illumination of the sample to be detected in the detection area 101 by illumination light of the light sheet generating device 10; the camera 52 is used to receive the light signal and record a sample image of the sample to be measured. For example, the objective lens 51 may be disposed directly above the detection area 101 of the light sheet generating device 10, collect the light signal generated by the light sheet formed by the light sheet generating device 10 irradiating the sample to be tested, and further dispose the camera 52 on the side of the objective lens 51 far away from the light sheet generating device 10, receive the light signal collected by the objective lens 51 through the objective lens 51 and record the sample image of the sample to be tested irradiated by the light sheet, so that the staff can analyze the sample to be tested by observing the sample image recorded by the camera 52.
Further, since the light intensity of the light sheet formed by the light sheet generating device 10 is periodic, that is, has structured light characteristics, in some embodiments of the present application, a structured light microscopic imaging algorithm may be incorporated to enhance the lateral resolution of the image of the sample captured by the camera 52.
In some embodiments of the present application, as shown in fig. 5 to 7, the observation device 50 may further include a filter 53, the filter 53 being disposed between the objective lens 51 and the camera 52. The optical filter 53 may be set according to practical application requirements, for example, if the microscope system 1 is applied to fluorescence measurement, the optical filter 53 needs to be set between the objective lens 51 and the camera 52, and the optical filter 53 should be a bandpass filter with fluorescence wavelength, so as to screen a wavelength band of an optical signal, so that the camera 52 can better capture a sample image of a sample to be measured.
In some embodiments of the present application, as shown in fig. 7, the substrate of the bottom surface of the detection area 101 is made of transparent material, and the number of observation devices 50 is two, where a first one of the observation devices 50 is disposed directly above the detection area 101, and a second one of the observation devices 50 is disposed directly below the detection area 101. Thus, the condition of the bottom of the sample to be detected can be observed through the observation device 50 under the detection area 101, the operation that the sample to be detected needs to be rotated 180 degrees in practical application to observe the bottom of the sample to be detected is reduced, the practicability and convenience are improved, and the sample images of the sample to be detected are recorded simultaneously through the cameras 52 in the upper observation device 50 and the lower observation device 50, so that a worker can clearly observe the complete imaging condition of the light sheet of the sample to be detected in situ, and the detection effect is improved.
In some embodiments of the present application, as shown in fig. 4, the exit end face of each core 200 of the multi-core optical fiber 12 has a light converging portion 201. For example, the exit end surface of each fiber core 200 may be processed into a conical surface, a circular table surface, a spherical surface, a sphere-like surface, a wedge surface, or the like, as the light converging portion 201, after two rows of fiber cores 200 that are closely arranged in parallel and are placed opposite to each other are connected to each other, the light of the illumination light is converged at a middle position between each fiber core 200 and the fiber core 200 opposite to the fiber core, and a light spot effect formed near the position is as shown in a dashed line frame of fig. 4, and may be approximately a sheet-like strip-like light sheet, in which a crossing line at the front end of the light converging portion 201 of each fiber core 200 in fig. 4 represents a light transmission path of the illumination light.
Further, by adjusting the light converging capability of the light converging portion 201 and adjusting the distance between every two oppositely disposed fiber cores 200, the width and thickness of the light sheet formed by the light sheet generating device 10 can be controlled, so that the suitability of the light sheet formed by the light sheet generating device 10 to practical application needs is stronger.
In some embodiments of the present application, the light sheet generating device 10 further includes a cover plate 13, where the cover plate 13 is adapted to the top surface of the substrate 11 and is used to encapsulate the multicore optical fiber 12. As shown in fig. 1 and 2, by using the cooperation between the cover 13 and the substrate 11, an optical fiber accommodating layer may be formed between the cover 13 and the substrate 11, so as to encapsulate the parallel and closely arranged portions of the multiple cores 200 of the multi-core optical fiber 12 in the optical fiber accommodating layer, thereby preventing the influence of impurities such as external dust on the light transmission of the illumination light, and improving the quality of the light sheet formed by the light sheet generating device 10.
In some embodiments of the present application, as shown in fig. 1, the cover 13 includes a first block and a second block, where the first block is adapted to a top surface of the substrate 11 located on a first side of the detection area 101, for packaging a first multi-core optical fiber; the second block is adapted to the top surface of the substrate 11 on the second side of the detection area 101 for encapsulating a second multicore fiber. In this case, two optical fiber accommodating layers are formed on both sides of the detection region 101, and a sample cell for inflow of a sample to be measured can be formed by being surrounded by the middle region between the two optical fiber accommodating layers, the middle region between the first block and the second block of the cover plate 13, and the detection region 101.
The microscope system 1 according to the embodiment of the second aspect of the present application has a smaller overall structure volume and a smaller structural complexity than those of the light sheet microscope in the related art, and is easier to integrate, so that various integrated devices applied to biological detection can be realized based on the microscope system 1. For example, as shown in fig. 8 and 9, two integrated device application embodiments are implemented for the microscope system 1 provided according to the embodiment of the second aspect of the present application. Wherein fig. 8 is a schematic structural diagram of a microscope system 1 integrated in a cell sorting apparatus according to an embodiment of the second aspect of the present application, where the cell sorting apparatus includes the microscope system 1, a sorting channel 2, and a sorting control unit (not shown in the drawing) provided according to an embodiment of the second aspect of the present application, and the sorting channel 2 includes one total channel and a plurality of sub-channels (such as A, B, C sub-channels shown in fig. 8); in practical application, the discrete sample cell flows through the sample cell of the microscope system 1 via the total channel, the staff can analyze and judge the attribute classification of each sample cell according to the sample image shot by the camera 52, and then sort the sample cells obtained with the judgment result to the corresponding channels in the multiple sorting channels via the sorting control unit, in this case, the structure of the total channel of the sorting channel 2 is made to be matched with the sample cell, so that the microscope system 1 provided by the embodiment of the second aspect of the present application can be integrated in the cell sorting device. Fig. 9 is a schematic structural diagram of an application device of integrating a microscope system 1 into a microfluidic chip according to an embodiment of a second aspect of the present application, where the application device includes the microscope system 1 and a sample enrichment structure 3 to be tested provided according to an embodiment of the second aspect of the present application, and the sample enrichment structure 3 to be tested includes a channel 31 through which a sample to be tested flows and a photographing region 32; in practical application, the sample to be measured flows into and is enriched in the photographing region 32 through the channel 31, and assuming that the photographing region 32 has a hexagonal structure as shown in fig. 9, in this case, the sample to be measured flows into and is enriched in the sample cell through the channel 31, that is, the photographing region 32, by modifying the structure of the sample cell to a structure matching the size and shape of the photographing region 32 and arranging the flow valves on both sides of the sample cell. In summary, the microscope system 1 according to the second aspect of the present application is conveniently applied to integration.
It is noted that relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising one … …" does not exclude the presence of other like elements in a process, method, article, or apparatus that comprises the element.
The various embodiments of the present application are described in a related manner, and identical and similar parts of the various embodiments are all mutually referred to, and each embodiment is mainly described in the differences from the other embodiments.
The foregoing description is only of the preferred embodiments of the present application and is not intended to limit the scope of the present application. Any modifications, equivalent substitutions, improvements, etc. that fall within the spirit and principles of the present application are intended to be included within the scope of the present application.
Claims (10)
1. A light sheet generating apparatus, comprising:
the device comprises a substrate, wherein the top surface of the substrate is provided with a groove as a detection area of a sample to be detected, and the bottom surface of the detection area is parallel to the top surface of the substrate;
the multi-core optical fiber is used for transmitting illumination light emitted by the light source equipment, a plurality of fiber cores of the multi-core optical fiber are arranged on the top surface of the substrate in a parallel and closely arranged mode, and the illumination light emitted by the emergent end of the multi-core optical fiber passes through the position right above the detection area;
the end face of the emergent end of each fiber core of the multi-core optical fiber is provided with a light converging part;
the light converging part is formed by processing the end face of the emergent end of the fiber core into a conical surface, a round table surface, a spherical surface, a sphere-like surface or a wedge surface.
2. The light sheet generating device of claim 1, wherein the multicore fibers comprise a first multicore fiber and a second multicore fiber; the multiple fiber cores of the first multi-core optical fiber are located on a first side of the detection area, the multiple fiber cores of the second multi-core optical fiber are located on a second side of the detection area, the first side and the second side are oppositely arranged, and the first multi-core optical fiber and the second multi-core optical fiber are located on the same plane.
3. The light sheet generating device of any one of claims 1-2, further comprising a cover plate adapted to the top surface of the substrate and configured to encapsulate the multicore fiber.
4. A microscope system comprising a light sheet generating device according to any one of claims 1 to 3.
5. The microscope system according to claim 4, further comprising:
a light source device for emitting uniform light;
the optical fiber conversion head is connected with the multi-core optical fiber and is used for receiving the uniform light as the illumination light and uniformly dividing the illumination light to each fiber core of the multi-core optical fiber.
6. The microscope system according to claim 5, wherein the light source device comprises a light emitter and a light homogenizing sheet, wherein the light homogenizing sheet is disposed between an exit end of the light emitter and a receiving end of the fiber optic adapter.
7. The microscope system according to claim 6, wherein the multicore optical fibers comprise a first multicore optical fiber and a second multicore optical fiber; the multiple fiber cores of the first multi-core optical fiber are positioned on a first side of the detection area, the multiple fiber cores of the second multi-core optical fiber are positioned on a second side of the detection area, the first side and the second side are arranged opposite to each other, and the first multi-core optical fiber and the second multi-core optical fiber are positioned on the same plane;
the microscope system further comprises a half-mirror, wherein the half-mirror is arranged between the optical fiber conversion head and the light homogenizing sheet and is used for equally dividing the uniform light into a first beam and a second beam;
the optical fiber conversion head comprises a first optical fiber conversion head and a second optical fiber conversion head, wherein the first optical fiber conversion head is connected with the first multi-core optical fiber, and the first optical fiber conversion head is used for receiving the first beam as part of the illumination light and equally dividing the part of the illumination light to each fiber core of the first multi-core optical fiber; the second optical fiber conversion head is connected with the second multi-core optical fiber and is used for receiving the second beam as another part of the illumination light and equally dividing the other part of the illumination light to each fiber core of the second multi-core optical fiber.
8. The microscope system according to any one of claims 4 to 7, further comprising an observation device comprising an objective lens and a camera, wherein the objective lens is used to collect optical signals generated by illumination of the sample to be examined in the detection area by illumination of the light sheet generating device; the camera is used for receiving the optical signals and recording sample images of the sample to be detected.
9. The microscope system according to claim 8, wherein the observation device further comprises a filter disposed between the objective lens and the camera.
10. The microscope system according to claim 8, wherein the substrate of the bottom surface of the detection region is made of transparent material, and the number of the observation devices is two, wherein a first one of the observation devices is disposed directly above the detection region, and a second one of the observation devices is disposed directly below the detection region.
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE102012211780A1 (en) * | 2012-07-05 | 2014-01-09 | Carl Zeiss Microscopy Gmbh | Device for supporting and lighting i.e. biological specimens on slide of microscope, has movement generation unit generating relative movement between sample and light sheet such that relative movement is generated between belt and sample |
| CN107942530A (en) * | 2017-11-24 | 2018-04-20 | 浙江大学 | Integrated light guide super-resolution micro imaging system |
| CN110023812A (en) * | 2016-10-28 | 2019-07-16 | 卡尔蔡司显微镜有限责任公司 | Monoplane illuminates microscope |
| CN110799878A (en) * | 2017-06-23 | 2020-02-14 | 莱卡微系统Cms有限责任公司 | Microscope system with functional units for light sheet microscopy |
| CN210140580U (en) * | 2019-06-27 | 2020-03-13 | 杭州安塔生物科技有限公司 | Imaging optical system for PCR reaction device |
| CN111307075A (en) * | 2019-10-30 | 2020-06-19 | 桂林电子科技大学 | Roughness measuring device capable of identifying texture direction |
| DE102019214929A1 (en) * | 2019-09-27 | 2021-04-01 | Leica Microsystems Cms Gmbh | Compact light sheet microscope |
| WO2021121911A1 (en) * | 2019-12-18 | 2021-06-24 | Universität Bielefeld | Light sheet microscopy assembly and method |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10539772B2 (en) * | 2013-10-09 | 2020-01-21 | Howard Hughes Medical Institute | Multiview light-sheet microscopy |
| ITUB20153920A1 (en) * | 2015-09-28 | 2017-03-28 | Milano Politecnico | Optofluidic device. |
| GB201617089D0 (en) * | 2016-10-07 | 2016-11-23 | University Court Of The University Of St Andrews The | Light sheet imaging with acoustic trapping |
| JP2019032437A (en) * | 2017-08-08 | 2019-02-28 | オリンパス株式会社 | Light sheet microscope |
-
2021
- 2021-10-25 CN CN202111243238.3A patent/CN114236801B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE102012211780A1 (en) * | 2012-07-05 | 2014-01-09 | Carl Zeiss Microscopy Gmbh | Device for supporting and lighting i.e. biological specimens on slide of microscope, has movement generation unit generating relative movement between sample and light sheet such that relative movement is generated between belt and sample |
| CN110023812A (en) * | 2016-10-28 | 2019-07-16 | 卡尔蔡司显微镜有限责任公司 | Monoplane illuminates microscope |
| CN110799878A (en) * | 2017-06-23 | 2020-02-14 | 莱卡微系统Cms有限责任公司 | Microscope system with functional units for light sheet microscopy |
| CN107942530A (en) * | 2017-11-24 | 2018-04-20 | 浙江大学 | Integrated light guide super-resolution micro imaging system |
| CN210140580U (en) * | 2019-06-27 | 2020-03-13 | 杭州安塔生物科技有限公司 | Imaging optical system for PCR reaction device |
| DE102019214929A1 (en) * | 2019-09-27 | 2021-04-01 | Leica Microsystems Cms Gmbh | Compact light sheet microscope |
| CN111307075A (en) * | 2019-10-30 | 2020-06-19 | 桂林电子科技大学 | Roughness measuring device capable of identifying texture direction |
| WO2021121911A1 (en) * | 2019-12-18 | 2021-06-24 | Universität Bielefeld | Light sheet microscopy assembly and method |
Non-Patent Citations (2)
| Title |
|---|
| Multimode fibre: Light-sheet microscopy at the tip of a needle;Zbynek Dostal 等;Scientific Reports;第1-7页 * |
| 长线列密排光纤阵列高精度设计制作;蔡易平;胡孟春;彭星宇;;光纤与电缆及其应用技术(第06期);第4-13页 * |
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|---|---|
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