Myocardial cell function detection light path structure for multi-mode synchronous imaging

Abstract

The invention relates to the technical field of multi-modal synchronous imaging, in particular to a myocardial cell function detection light path structure for multi-modal synchronous imaging, which is characterized in that emitted light excited by a shot sample is filtered by a first fluorescent filter block, and the emitted light is respectively filtered and reflected by a second fluorescent filter block; the first camera shooting port and the second camera shooting port are arranged, so that two paths of emitted light are formed at the first camera shooting port and the second camera shooting port, incident light of the first camera shooting port and incident light of the second camera shooting port are collected through the first camera and the second camera respectively, mechanical data and calcium imaging of a shot sample are collected at the same time, and therefore the problems existing in the prior art are solved.

Description

Technical Field

[0001] The present invention relates to the technical field of multimodal synchronous imaging, and in particular to a myocardial cell function detection optical path structure of multimodal synchronous imaging. Background Art

[0002] The combination of calcium concentration imaging technology and nano-hydrogel mechanical analysis is a cutting-edge research direction in the fields of biomedical engineering and materials science. Calcium imaging technology uses fluorescent probes or genetically encoded calcium indicators (such as GCaMP) to monitor changes in intracellular calcium ion concentration in real time, and is widely used in neuroscience, cardiovascular disease research and other fields. Its core equipment relies on high-speed, high-sensitivity imaging systems, such as high-frame rate cameras and large-field optical modules, to capture dynamic cell activities. Nano-hydrogels are used in drug delivery, tissue engineering and biosensing due to their high water content, biocompatibility and controllable mechanical properties. In recent years, researchers have tried to use hydrogels as mechanical response carriers to analyze the mechanical properties of the microenvironment (such as hardness and viscoelasticity) through their deformation or conductivity changes.

[0003] However, in the prior art, calcium imaging equipment and mechanical analysis systems usually operate independently, leading to the following problems: Data asynchrony: There is a time difference between the acquisition of calcium signals and mechanical signals, making it difficult to correlate dynamic biological processes with changes in the mechanical microenvironment; Equipment redundancy: Multiple systems take up a lot of space, are complex to operate, and increase experimental costs; Single function: Calcium imaging equipment (such as) focuses on optical signal capture, lacks mechanical sensing modules, and cannot synchronously obtain material deformation or stress data. Mechanical analysis equipment (such as atomic force microscopes) requires contact measurement, which may interfere with cell activity or destroy hydrogel structures. Summary of the invention

[0004] In view of this, an object of the present invention is to provide a multi-modal synchronous imaging myocardial cell function detection optical path structure to overcome the problems existing in the current prior art.

[0005] To achieve the above objectives, the present invention adopts the following technical solutions: The present application provides a multi-modal synchronous imaging myocardial cell function detection optical path structure, including: a fluorescent light source, a microscope objective lens, a first fluorescent filter block, a reflector, a second fluorescent filter block, a first camera, a second camera and a supporting structure; The microscope objective lens is arranged at the sample focusing port of the support structure, and is used to focus and photograph the sample; wherein the photographed sample is a cell sample spread on a hydrogel containing nano fluorescent microbeads, and the cell sample is treated with fluorescent dyeing; The fluorescent light source is aligned with the light source entrance of the supporting structure to provide excitation light; The first fluorescent filter block is arranged in the supporting structure and at the entrance of the light source. The excitation light is filtered and reflected by the first fluorescent filter block and passes through the objective lens of the microscope to be emitted toward the photographed sample. The reflector is arranged at a corner of the support structure, and is used to reflect the emission light excited by the photographed sample after being irradiated by the excitation light; wherein the emission light is filtered by the first fluorescence filter block; The support structure has a first camera shooting port and a second camera shooting port; The second fluorescent filter block is arranged at the intersection of the first camera shooting port and the second camera shooting port, and is used to receive the emission light reflected by the reflector, and filter and reflect the emission light so that the emission light is respectively emitted into the first camera shooting port and the second camera shooting port; The first camera is aligned with the first camera shooting port to receive the emission light filtered by the second fluorescent filter block to complete the collection of mechanical data; The second camera is aligned with the second camera shooting port to receive the emission light reflected and filtered by the second fluorescent filter block to complete calcium imaging.

[0006] Furthermore, in the above-mentioned optical path structure, the fluorescent light source is a mercury lamp or a multi-color LED light source.

[0007] Furthermore, in the above-mentioned optical path structure, the first fluorescent filter block includes: a first filter block holder, a DM505 filter, a BA510-550 filter and a BP460-495 filter; The DM505 filter, the BA510-550 filter and the BP460-495 filter are arranged in the first filter block holder, and the DM505 filter is arranged at a preset angle with the BA510-550 filter and the BP460-495 filter respectively; The excitation light is filtered by the BP460-495 filter, and then reflected by the DM505 filter, passes through the microscope objective lens and is emitted to the photographed sample; The emitted light is filtered by the DM505 filter, and then filtered by the BA510-550 filter to be emitted to the reflector.

[0008] Furthermore, in the above-mentioned optical path structure, the reflector is a total reflection reflector.

[0009] Furthermore, in the above-mentioned optical path structure, the second fluorescence filter block includes: a second filter block holder, a BA510IF filter, a BA578-633 filter and a BA510-550 filter; The BA510IF filter, the BA578-633 filter and the BA510-550 filter are arranged in the second filter block holder, and the BA510IF filter is arranged at a preset angle with the BA578-633 filter and the BA510-550 filter respectively; The emitted light reflected by the reflector is filtered by the BA510IF filter and then filtered by the BA578-633 filter to be emitted to the shooting port of the first camera; The emitted light reflected by the reflector is also reflected by the BA510IF filter and filtered by the BA510-550 filter before being emitted to the shooting port of the second camera.

[0010] Furthermore, in the above-mentioned optical path structure, the first camera is: a high quantum efficiency CMOS or SCMOS camera, which is used to photograph fluorescent microbeads.

[0011] Furthermore, in the above optical path structure, the second camera is a high quantum efficiency CMOS or SCMOS camera used for calcium imaging.

[0012] The beneficial effects of the present invention are: The present application comprises a fluorescent light source, a microscope objective lens, a first fluorescent filter block, a reflector, a second fluorescent filter block, a first camera, a second camera and a supporting structure; the microscope objective lens is arranged at a sample focusing port of the supporting structure, and is used to focus and photograph the sample; wherein the photographed sample is a cell sample spread on a hydrogel containing nano fluorescent microbeads, and the cell sample is treated with fluorescent dyeing; the fluorescent light source is aligned with a light source entrance of the supporting structure, and is used to provide excitation light; the first fluorescent filter block is arranged in the supporting structure, and is arranged at the light source entrance; the excitation light is filtered and reflected by the first fluorescent filter block, and passes through the microscope objective lens to shoot toward the photographed sample; the reflector is arranged at a corner of the supporting structure, and is used to reflect the excited light. After luminous irradiation, the emission light excited by the sample is photographed; wherein the emission light is filtered by the first fluorescent filter block; the supporting structure has a first camera shooting port and a second camera shooting port; the second fluorescent filter block is arranged at the intersection of the first camera shooting port and the second camera shooting port, for receiving the emission light reflected by the reflector, and filtering and reflecting the emission light, so that the emission light is respectively shot into the first camera shooting port and the second camera shooting port; the first camera is aimed at the first camera shooting port, for receiving the emission light filtered by the second fluorescent filter block, and completing the collection of mechanical data; the second camera is aimed at the second camera shooting port, for receiving the emission light reflected and filtered by the second fluorescent filter block, and completing calcium imaging. In the present application, the emission light excited by the sample is filtered by the first fluorescent filter block, and the emission light is filtered and reflected by the second fluorescent filter block, respectively, so that two emission lights are formed at the first camera shooting port and the second camera shooting port, and the incident light of the first camera shooting port and the second camera shooting port is respectively collected by the first camera and the second camera, so as to simultaneously complete the collection of mechanical data and calcium imaging of the sample, thereby solving the problems existing in the prior art. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0014] Figure 1 It is a structural schematic diagram provided by an embodiment of a multi-modal synchronous imaging myocardial cell function detection optical path structure of the present invention; Figure 2 It is a schematic diagram of the structure of a first fluorescent filter block provided in an embodiment of a multi-modal synchronous imaging myocardial cell function detection optical path structure of the present invention; Figure 3It is a schematic diagram of the second fluorescent filter block structure provided in an embodiment of a multi-modal synchronous imaging myocardial cell function detection optical path structure of the present invention. DETAILED DESCRIPTION

[0015] To make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described in detail below. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other implementation methods obtained by ordinary technicians in this field without creative work belong to the scope of protection of the present invention.

[0016] like Figure 1 As shown, Figure 1 This is a structural schematic diagram of an embodiment of a multi-modal synchronous imaging myocardial cell function detection optical path structure provided by the present invention. This embodiment may include: Fluorescent light source 1, microscope objective lens 2, first fluorescent filter block 4, reflector 9, second fluorescent filter block 6, first camera 7, second camera 8 and supporting structure 5; The microscope objective lens 2 is arranged at the sample focusing port of the support structure 5, and is used to focus and photograph the sample 3; wherein the photographed sample 3 is a cell sample spread on a hydrogel containing nano fluorescent microbeads, and the cell sample is treated with fluorescent dyeing; The fluorescent light source 1 is aligned with the light source entrance of the support structure 5 to provide excitation light 10; The first fluorescent filter block 4 is arranged in the support structure 5 and at the entrance of the light source. The excitation light 10 is filtered and reflected by the first fluorescent filter block 4 and passes through the microscope objective lens 2 to be emitted to the photographed sample 3. The reflector 9 is disposed at the corner of the support structure 5, and is used to reflect the emission light 11 excited by the sample 3 after being irradiated by the excitation light 10; wherein the emission light 11 is filtered by the first fluorescence filter block 4; The support structure 5 has a first camera 7 shooting port and a second camera 8 shooting port; The second fluorescent filter block 6 is disposed at the intersection of the first camera 7 shooting port and the second camera 8 shooting port, and is used to receive the emission light 11 reflected by the reflector 9, and filter and reflect the emission light 11, so that the emission light 11 is respectively emitted into the first camera 7 shooting port and the second camera 8 shooting port; The first camera 7 is aligned with the shooting port of the first camera 7 to receive the emission light 11 filtered by the second fluorescent filter block 6 to complete the collection of mechanical data; The second camera 8 is aligned with the shooting port of the second camera 8 to receive the emission light 11 reflected and filtered by the second fluorescent filter block 6 to complete calcium imaging.

[0017] It can be understood that the present embodiment comprises a fluorescent light source 1, a microscope objective lens 2, a first fluorescent filter block 4, a reflector 9, a second fluorescent filter block 6, a first camera 7, a second camera 8 and a support structure 5; the microscope objective lens 2 is arranged at the sample focusing port of the support structure 5, and is used to focus and photograph the sample 3; wherein the photographed sample 3 is a cell sample spread on a hydrogel containing nano fluorescent microbeads, and the cell sample has been subjected to fluorescent dyeing; the fluorescent light source 1 is aligned with the light source entrance of the support structure 5, and is used to provide excitation light 10; the first fluorescent filter block 4 is arranged in the support structure 5, and is arranged at the light source entrance, and the excitation light 10 is filtered and reflected by the first fluorescent filter block 4, passes through the microscope objective lens 2 and is emitted to the photographed sample 3; the reflector 9 is arranged at the corner of the support structure 5, and is used to reflect the excited light 10. After the irradiation of the light 10, the emission light 11 excited by the sample 3 is photographed; wherein, the emission light 11 is filtered by the first fluorescent filter block 4; the support structure 5 has a first camera 7 shooting port and a second camera 8 shooting port; the second fluorescent filter block 6 is arranged at the intersection of the first camera 7 shooting port and the second camera 8 shooting port, and is used to receive the emission light 11 reflected by the reflector 9, and filter and reflect the emission light 11, so that the emission light 11 is respectively incident on the first camera 7 shooting port and the second camera 8 shooting port; the first camera 7 is aligned with the first camera 7 shooting port, and is used to receive the emission light 11 filtered by the second fluorescent filter block 6, and completes the collection of mechanical data; the second camera 8 is aligned with the second camera 8 shooting port, and is used to receive the emission light 11 reflected and filtered by the second fluorescent filter block 6, and completes calcium imaging. In this embodiment, the emission light 11 excited by the photographed sample 3 is filtered by the first fluorescent filter block 4, and the emission light 11 is filtered and reflected respectively by the second fluorescent filter block 6, so that two emission lights 11 are formed at the shooting port of the first camera 7 and the shooting port of the second camera 8, and the incident light at the shooting port of the first camera 7 and the shooting port of the second camera 8 are collected by the first camera 7 and the second camera 8 respectively, so as to complete the collection of mechanical data and calcium imaging of the photographed sample 3 at the same time, thereby solving the problems existing in the prior art.

[0018] Preferably, the fluorescent light source 1 is a mercury lamp or a multi-color LED light source.

[0019] Preferably, the first fluorescent filter block 4 comprises: a first filter block support, a DM505 filter, a BA510-550 filter and a BP460-495 filter; The DM505 filter, the BA510-550 filter and the BP460-495 filter are arranged in the first filter block holder, and the DM505 filter is arranged at a preset angle with the BA510-550 filter and the BP460-495 filter respectively; The excitation light 10 is filtered by the BP460-495 filter, then reflected by the DM505 filter and passes through the microscope objective lens 2 to the photographed sample 3; The emitted light 11 is filtered by the DM505 filter, and then filtered by the BA510-550 filter before being directed to the reflector 9.

[0020] Understandably, Figure 2 FIG. 4 is a schematic diagram of the structure of a first fluorescent filter block 4 provided in an embodiment of a multi-modal synchronous imaging myocardial cell function detection optical path structure of the present invention, as shown in FIG. Figure 2 As shown, 401 is the first filter block holder, 402 is the DM505 filter, 403 is the BA510-550 filter, and 404 is the BP460-495 filter.

[0021] DM505 filter 402, used to reflect light below 505nm and transmit light above 505. But it is not limited to this model to meet the needs; BA510-550 filter 403, used to transmit light in the range of 510-550nm and cut off light of other wavelengths. However, it is not limited to this model to meet the needs; BP460-495 filter 404 is used to transmit light in the range of 460-495nm and cut off light of other wavelengths. However, it is not limited to this model to meet the needs.

[0022] The excitation light 10 emitted by the fluorescent light source 1 can be a broad spectrum light or a narrow band light, which is filtered by the BP460-495 filter 404 and then reflected by the DM505 filter 402, and further passes through the microscope objective lens 2, and further light is emitted to the photographed sample 3.

[0023] Preferably, the reflector 9 is a total reflection reflector.

[0024] Preferably, the second fluorescence filter block 6 comprises: a second filter block holder, a BA510IF filter, a BA578-633 filter and a BA510-550 filter; The BA510IF filter, the BA578-633 filter and the BA510-550 filter are arranged in the second filter block holder, and the BA510IF filter is arranged at a preset angle with the BA578-633 filter and the BA510-550 filter respectively; The emitted light 11 reflected by the reflector 9 is filtered by the BA510IF filter and then filtered by the BA578-633 filter to be emitted to the shooting port of the first camera 7; The emitted light 11 reflected by the reflector 9 is also reflected by the BA510IF filter and filtered by the BA510-550 filter before being emitted to the shooting port of the second camera 8 .

[0025] Understandably, Figure 3FIG. 1 is a schematic diagram of the structure of the second fluorescent filter block 6 provided in an embodiment of a multi-modal synchronous imaging myocardial cell function detection optical path structure of the present invention, as shown in FIG. Figure 3 As shown, 601 is the second filter block holder, 602 is the BA510IF filter, 603 is the BA578-633 filter, and 604 is the BA510-550 filter.

[0026] Preferably, the first camera 7 is a high quantum efficiency CMOS or SCMOS camera, which is used to photograph fluorescent microbeads.

[0027] It can be understood that the emission light 11 excited by the exciting and photographing sample 3 passes through the microscope objective lens 2, the DM505 filter 402, the BA510-550 filter 403, the reflector 9, the BA510IF filter 602, and the BA578-633 filter 603 in sequence, and the further light signal is finally received by the first camera 7 to complete the mechanical data acquisition.

[0028] Preferably, the second camera 8 is a high quantum efficiency CMOS or SCMOS camera, which is used for calcium imaging.

[0029] It can be understood that the emission light 11 excited by the exciting shooting sample 3 passes through the microscope objective lens 2DM505 filter 402, the BA510-550 filter 403, and the reflector 9 in sequence, is further reflected by the BA510IF filter 602, and further passes through the BA510-550 filter 604. The light signal is finally received by the second camera 8 to complete calcium imaging.

[0030] It can be understood that the same or similar parts of the above embodiments can be referenced to each other, and the contents not described in detail in some embodiments can refer to the same or similar contents in other embodiments.

[0031] It should be noted that, in the description of the present invention, the terms "first", "second", etc. are only used for descriptive purposes and cannot be understood as indicating or implying relative importance. In addition, in the description of the present invention, unless otherwise specified, the meaning of "plurality" refers to at least two.

[0032] Any process or method description in a flowchart or otherwise described herein may be understood to represent a module, segment or portion of code that includes one or more executable instructions for implementing the steps of a specific logical function or process, and the scope of the preferred embodiments of the present invention includes alternative implementations in which functions may not be performed in the order shown or discussed, including performing functions in a substantially simultaneous manner or in the reverse order depending on the functions involved, which should be understood by those skilled in the art to which the embodiments of the present invention belong.

[0033] It should be understood that the various parts of the present invention can be implemented by hardware, software, firmware or a combination thereof. In the above-mentioned embodiments, multiple steps or methods can be implemented by software or firmware stored in a memory and executed by a suitable instruction execution system. For example, if implemented by hardware, as in another embodiment, it can be implemented by any one of the following technologies known in the art or their combination: a discrete logic circuit having a logic gate circuit for implementing a logic function for a data signal, a dedicated integrated circuit having a suitable combination of logic gate circuits, a programmable gate array (PGA), a field programmable gate array (FPGA), etc.

[0034] A person skilled in the art may understand that all or part of the steps in the method for implementing the above-mentioned embodiment may be completed by instructing related hardware through a program, and the program may be stored in a computer-readable storage medium, which, when executed, includes one or a combination of the steps of the method embodiment.

[0035] In addition, each functional unit in each embodiment of the present invention may be integrated into a processing module, or each unit may exist physically separately, or two or more units may be integrated into one module. The above-mentioned integrated module may be implemented in the form of hardware or in the form of a software functional module. If the integrated module is implemented in the form of a software functional module and sold or used as an independent product, it may also be stored in a computer-readable storage medium.

[0036] The storage medium mentioned above can be a read-only memory, a magnetic disk or an optical disk, etc.

[0037] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "examples", "specific examples", or "some examples" means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner.

[0038] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A multi-modal synchronous imaging myocardial cell function detection optical path structure, characterized in that: include: A fluorescent light source, a microscope objective lens, a first fluorescent filter block, a reflector, a second fluorescent filter block, a first camera, a second camera and a supporting structure; The microscope objective lens is arranged at the sample focusing port of the support structure, and is used to focus and photograph the sample; wherein the photographed sample is a cell sample spread on a hydrogel containing nano fluorescent microbeads, and the cell sample is treated with fluorescent dyeing; The fluorescent light source is aligned with the light source entrance of the supporting structure to provide excitation light; The first fluorescent filter block is arranged in the supporting structure and at the entrance of the light source. The excitation light is filtered and reflected by the first fluorescent filter block and passes through the objective lens of the microscope to be emitted toward the photographed sample. The reflector is arranged at a corner of the support structure, and is used to reflect the emission light excited by the photographed sample after being irradiated by the excitation light; wherein the emission light is filtered by the first fluorescence filter block; The support structure has a first camera shooting port and a second camera shooting port; The second fluorescent filter block is arranged at the intersection of the first camera shooting port and the second camera shooting port, and is used to receive the emission light reflected by the reflector, and filter and reflect the emission light so that the emission light is respectively emitted into the first camera shooting port and the second camera shooting port; The first camera is aligned with the first camera shooting port to receive the emission light filtered by the second fluorescent filter block to complete the collection of mechanical data; The second camera is aligned with the second camera shooting port to receive the emission light reflected and filtered by the second fluorescent filter block to complete calcium imaging.

2. The optical path structure according to claim 1, characterized in that: The fluorescent light source is a mercury lamp or a multi-color LED light source.

3. The optical path structure according to claim 2, characterized in that: The first fluorescent filter block includes: a first filter block bracket, a DM505 filter, a BA510-550 filter and a BP460-495 filter; The DM505 filter, the BA510-550 filter and the BP460-495 filter are arranged in the first filter block holder, and the DM505 filter is arranged at a preset angle with the BA510-550 filter and the BP460-495 filter respectively; The excitation light is filtered by the BP460-495 filter, and then reflected by the DM505 filter, passes through the microscope objective lens and is emitted to the photographed sample; The emitted light is filtered by the DM505 filter, and then filtered by the BA510-550 filter to be emitted to the reflector.

4. The optical path structure according to claim 3, characterized in that: The reflector is a total reflection reflector.

5. The optical path structure according to claim 4, characterized in that: The second fluorescence filter block includes: a second filter block holder, a BA510IF filter, a BA578-633 filter and a BA510-550 filter; The BA510IF filter, the BA578-633 filter and the BA510-550 filter are arranged in the second filter block holder, and the BA510IF filter is arranged at a preset angle with the BA578-633 filter and the BA510-550 filter respectively; The emitted light reflected by the reflector is filtered by the BA510IF filter and then filtered by the BA578-633 filter to be emitted to the shooting port of the first camera; The emitted light reflected by the reflector is also reflected by the BA510IF filter and filtered by the BA510-550 filter before being emitted to the shooting port of the second camera.

6. The optical path structure according to claim 5, characterized in that: The first camera is a high quantum efficiency CMOS or SCMOS camera, which is used to photograph fluorescent microbeads.

7. The optical path structure according to claim 6, characterized in that: The second camera is a high quantum efficiency CMOS or SCMOS camera, which is used for calcium imaging.

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