Single-camera partitioned optical microscopic imaging device

Abstract

The invention discloses a single-camera partitioned optical microscopic imaging device which comprises a bottom plate, a support is mounted at the top of the rear side of the bottom plate, and a mounting plate is mounted on the front side of the support; the sample table is mounted on one side of the top of the bottom plate and is used for clamping a sample; the transmission illumination assembly is mounted on the other side of the top of the bottom plate and is used for providing uniform illumination for the sample; and the microscopic imaging assembly is mounted at the bottom of the front side of the mounting plate. According to the invention, through the arrangement of the image separation assembly, the intermediate image of the sample can be separated into microscopic images of a plurality of channels, and the microscopic images can be imaged on the target surface of the camera in real time after being transmitted through different paths, so that a single camera can carry out multi-channel imaging without considering whether a plurality of cameras are aligned or not; according to the optical microscopic imaging device, the problem of difference of time resolutions of a plurality of cameras does not exist, the optical path of the optical microscopic imaging device is relatively simple in the propagation process, some very rapid dynamic events can be synchronously captured, and the use effect is good.

Description

Technical Field

[0001] This invention specifically relates to a single-camera partitioned optical microscopy imaging device. Background Technology

[0002] Multicolor fluorescence imaging is a very common research method in the life sciences. Various fluorescent probes of different colors bind to specific proteins and are excited at different wavelengths, emitting fluorescence of different colors. This helps us distinguish different types of cells or proteins and study their interactions. It has important applications in the fields of biological microstructure imaging and high-throughput bioinformatics detection. Traditional optical microscopy imaging devices typically perform multicolor fluorescence imaging by controlling an electronic filter turntable to switch between different channels or by adding an additional camera and beam splitter to capture fluorescence images separately, and then combining them into a single multicolor fluorescence image in software. However, the above two methods have the following drawbacks: the switching of the mechanical components of the electronic filter turntable is usually slow, which will result in a lot of time loss when performing live cell imaging, and high-speed acquisition cannot be performed, making it impossible to capture some very rapid dynamic events. Adding an additional camera will increase the complexity of the optical path, and there may be differences when multiple cameras are triggered synchronously, which will affect the research.

[0003] Therefore, it is necessary to invent a single-camera partitioned optical microscopy imaging device to solve the above problems. Summary of the Invention

[0004] (a) Purpose of the invention The purpose of this invention is to provide a single-camera partitioned optical microscopy imaging device. By incorporating an image separation component, the intermediate image of the sample can be separated into multiple channels of microscopic images. After transmission through different paths, these images can be imaged in real time on the camera target surface. This allows a single camera to perform multi-channel imaging without needing to consider whether multiple cameras are aligned or not. It also eliminates the problem of differences in temporal resolution among multiple cameras. The optical path of this optical microscopy imaging device is relatively simple during propagation and can simultaneously capture some very rapid dynamic events, resulting in good performance and overcoming the aforementioned shortcomings in the technology.

[0005] (II) Technical Solution To achieve the above objectives, the present invention provides the following technical solution: a single-camera partitioned optical microscopy imaging device, comprising a base plate, a bracket mounted on the top rear side of the base plate, and a mounting plate mounted on the front side of the bracket; a sample stage, mounted on one side of the top of the base plate, for clamping samples; A transmissive illumination assembly, mounted on the other side of the top of the base plate, is used to provide uniform illumination to the sample; A microscopic imaging component is mounted on the front bottom of the mounting plate to form an intermediate image of the sample; An image separation component is installed on the top front side of the mounting plate to separate intermediate images and enable imaging at different positions on the camera target surface. An incident illumination assembly, mounted on the front side of the mounting plate, is used to provide uniform incident illumination to the sample.

[0006] Preferably, the sample stage includes a three-dimensional adjustable displacement stage and a sample holder located on top of the three-dimensional adjustable displacement stage.

[0007] Preferably, the transmissive illumination assembly includes a first cage-like frame extending along the width direction of the base plate, wherein a first white light source, a first collimating lens, and a first reflecting mirror are sequentially installed inside the first cage-like frame from front to back.

[0008] Preferably, the microscopic imaging assembly includes a second cage-like frame extending along the height direction of the mounting plate, wherein a tube mirror, a first beam splitter, and a switch are sequentially installed inside the second cage-like frame from top to bottom.

[0009] Preferably, the switcher includes a rotating wheel connected to a mounting plate and a plurality of objective lenses mounted on the bottom of the rotating wheel.

[0010] Preferably, the image separation component includes two third cage-like frames arranged side by side and extending along the width direction of the mounting plate. The upper third cage-like frame is equipped with a second beam splitter and a second reflector from right to left, and the lower third cage-like frame is equipped with a third reflector and a third beam splitter from right to left. The image separation component further includes a first aperture installed inside the second cage-like frame, and a second collimating lens located above the aperture and aligned with the third beam splitter. The first aperture is vertically aligned with the tube mirror. The image separation assembly also includes a camera mounted on top of the mounting plate and a first converging lens disposed at the bottom of the camera and aligned with the second beam splitter.

[0011] Preferably, both of the third cage-type frames are equipped with a clipper.

[0012] Preferably, the incident lighting assembly includes two fourth cage-type frames arranged side by side and extending along the width direction of the mounting plate, a fifth cage-type frame extending along the height direction of the mounting plate, and a sixth cage-type frame located at the bottom of the fifth cage-type frame and extending along the width direction of the mounting plate. The fourth cage-like lens frame located at the top is equipped with a second white light source, a third collimating lens, and a fourth reflecting mirror from right to left. The fourth cage-like lens frame located at the bottom is equipped with an optical fiber interface, a fourth collimating lens, and a fourth beam splitter from right to left. The fifth cage-like lens frame is equipped with a filter module and a second aperture from top to bottom. The sixth cage-like lens frame is equipped with a fifth reflecting mirror and a second converging lens from right to left.

[0013] Preferably, the second converging lens is horizontally aligned with the first beam splitter, and the filtering module is vertically aligned with the fourth beam splitter.

[0014] Preferably, the first cage-type frame, the second cage-type frame, the third cage-type frame, the fourth cage-type frame, the fifth cage-type frame, and the sixth cage-type frame are all configured as multi-axis cage-type structure frames.

[0015] Compared with the prior art, the beneficial effects of the above-mentioned technical solution of the present invention are: 1. This invention, by setting an image separation component, enables the intermediate image of the sample to be separated into multiple channels of microscopic images. Under the action of the third beam splitter, multiple beams carrying microscopic images pass through the third mirror and the third mirror respectively, so that the microscopic images can be imaged in real time on the camera target surface after being transmitted through different paths. In this process, only one camera is needed, so that a single camera can perform multi-channel imaging, avoiding the problem of camera alignment when multiple cameras are used for imaging. That is, the optical path of this optical microscopic imaging device is relatively simple during propagation and will not cause imaging differences due to the inability of multiple cameras to be triggered simultaneously. 2. This invention enables a single camera to perform multi-channel imaging by constructing a transmission illumination component, a microscopic imaging component, an image separation component, and an incident illumination component. The entire process requires no additional operation and can acquire data in real time at high speed, thereby capturing various dynamic events. This avoids the problem of poor time resolution caused by the slow switching speed of traditional electronic filter converters. When in use, this optical microscopic imaging device has high time resolution, which is convenient for live cell imaging. Attached Figure Description

[0016] Figure 1 This is a schematic diagram of the overall structure of the present invention; Figure 2 This is a distribution diagram of the microscopic imaging component and the image separation component of the present invention; Figure 3 This is a distribution diagram of the microscopic imaging components and image separation components from another perspective of the present invention; Figure 4 This is a schematic diagram of the structure of the transmission lighting component of the present invention; Figure 5 This is an exploded view of the incident lighting assembly of the present invention; Figure 6 This is an exploded view of the image separation component of the present invention; Figure 7 This is a perspective view of the present invention.

[0017] Explanation of reference numerals in the attached figures: 1. Base plate, 2. Support bracket, 3. Mounting plate, 4. Sample stage, 41. Three-dimensional adjustment displacement stage, 42. Sample rack; 5. Transmitted illumination assembly; 51. First cage-type frame; 52. First white light source; 53. First collimating lens; 54. First reflecting mirror; 6. Microscopic imaging assembly, 61. Second cage frame, 62. Tube lens, 63. First beam splitter, 64. Switcher, 65. Rotary wheel, 66. Objective lens; 7 Image separation component, 71 Third cage frame, 72 Second beam splitter, 73 Second reflecting mirror, 74 Third reflecting mirror, 75 Third beam splitter, 76 First aperture, 77 Second collimating lens, 78 Camera, 79 First converging lens; 8. Radiated illumination assembly; 81. Fourth cage-type frame; 82. Fifth cage-type frame; 83. Sixth cage-type frame; 84. Second white light source; 85. Third collimating lens; 86. Fourth reflecting mirror; 87. Fiber optic interface; 88. Fourth collimating lens; 89. Fourth beam splitter; 810. Filter module; 811. Second aperture; 812. Fifth reflecting mirror; 813. Second converging lens; 9. Card holder. Detailed Implementation

[0018] To enable those skilled in the art to better understand the technical solution of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings.

[0019] This invention provides, for example Figure 1-7 The single-camera 78-segment optical microscopy imaging device shown includes a base plate 1, a bracket 2 mounted on the top rear side of the base plate 1, and a mounting plate 3 mounted on the front side of the bracket 2. Sample stage 4 is installed on one side of the top of the base plate 1 and is used to clamp samples; A transmissive illumination assembly 5 is installed on the other side of the top of the base plate 1 to provide uniform illumination for the sample; The microscopic imaging component 6 is installed at the bottom front side of the mounting plate 3 and is used to form an intermediate image of the sample; The image separation component 7 is installed on the top front side of the mounting plate 3 and is used to separate the intermediate image so that it can be imaged at different positions on the target surface of the camera 78. The incident illumination assembly 8 is installed on the front side of the mounting plate 3 to provide uniform incident illumination to the sample.

[0020] In one embodiment, the sample stage 4 includes a three-dimensional adjustable displacement stage 41 and a sample holder 42 located on top of the three-dimensional adjustable displacement stage 41. The sample holder 42 can hold samples of various sizes and can be horizontally driven by the three-dimensional adjustable displacement stage 41, thereby making the position of the sample adjustable and facilitating the use of the optical microscopic imaging device. In one embodiment, the transmission illumination assembly 5 includes a first cage-like frame 51 extending along the width direction of the base plate 1. Inside the first cage-like frame 51, a first white light source 52, a first collimating lens 53, and a first reflecting mirror 54 are installed sequentially from front to back to provide illumination to the bottom of the sample, so that the sample can obtain uniform illumination conditions, thereby removing all external interference, such as uneven light, and allowing the sample's own characteristics, such as absorption, refraction, and thickness, to be accurately measured and recorded.

[0021] In one embodiment, the microscopic imaging assembly 6 includes a second cage-like frame 61 extending along the height direction of the mounting plate 3. Inside the second cage-like frame 61, a tube mirror 62, a first beam splitter 63, and a switcher 66 are installed sequentially from top to bottom. The switcher 66 includes a rotating wheel 65 connected to the mounting plate 3 and a plurality of objective lenses 66 installed at the bottom of the rotating wheel 65. Different magnification objective lenses 66 can be switched according to observation needs, which is convenient and quick.

[0022] In one embodiment, the image separation component 7 includes two vertically arranged third cage-like frames 71 extending along the width of the mounting plate 3. The upper third cage-like frame 71 is equipped with a second beam splitter 72 and a second reflecting mirror 73 sequentially from right to left, while the lower third cage-like frame 71 is equipped with a third reflecting mirror 74 and a third beam splitter 75 sequentially from right to left. The image separation component 7 also includes a first aperture 76 installed inside the second cage-like frame 71, and a third beam splitter 75 located above the aperture and adjacent to the third beam splitter 75. The second collimating lens 77 is aligned with the mirror 75, and the first aperture 76 is vertically aligned with the tube mirror 62. The image separation assembly 7 also includes a camera 78 mounted on the top of the mounting plate 3, and a first converging lens 79 located at the bottom of the camera 78 and aligned with the second beam splitter 72. It can separate the intermediate image and make the separated image image real-time at different positions on the target surface of the camera 78 along different paths, so that a single camera 78 can perform multi-channel imaging, avoiding the problem of needing to align the cameras 78 when imaging with multiple cameras 78.

[0023] In one embodiment, each of the two third cage-type frames 71 is equipped with a mounting device 9, which can be used to install different types of optical devices, facilitating component replacement. This allows the optical microscopy imaging device to not only perform multicolor fluorescence imaging, but also to be used for the study of material anisotropy, thereby improving the applicability of the optical microscopy imaging device.

[0024] In one embodiment, the incident illumination assembly 8 includes two fourth cage-like frames 81 arranged side-by-side and extending along the width direction of the mounting plate 3, a fifth cage-like frame 82 extending along the height direction of the mounting plate 3, and a sixth cage-like frame 83 located at the bottom of the fifth cage-like frame 82 and extending along the width direction of the mounting plate 3; wherein, the upper fourth cage-like frame 81 is equipped with a second white light source 84, a third collimating lens 85, and a fourth reflector 86 sequentially from right to left, and the lower fourth cage-like frame... From right to left, the optical fiber interface 87, the fourth collimating lens 88, and the fourth beam splitter 89 are installed in sequence on the fifth cage-like frame 82. From top to bottom, the filter module 810 and the second aperture 811 are installed in sequence on the sixth cage-like frame 83. From right to left, the fifth reflecting mirror 812 and the second converging lens 813 are installed in sequence on the sixth cage-like frame 83. The second converging lens 813 is horizontally aligned with the first beam splitter 63. The filter module 810 and the fourth beam splitter 89 are vertically aligned to provide illumination above the sample, so that the sample can obtain uniform illumination conditions.

[0025] The filter module 810 consists of a lens and a pinhole filter. Specifically, from top to bottom, it consists of an input lens, a pinhole filter, and an output lens. Its function is to selectively filter out impurities by adjusting the spatial frequency distribution of the beam, thereby optimizing the beam uniformity and collimation. In one embodiment, the first cage frame 51, the second cage frame 61, the third cage frame 71, the fourth cage frame 81, the fifth cage frame 82, and the sixth cage frame 83 are all configured as multi-axis cage structure frames, which can make the complex and fragile optical alignment process stable, efficient, and scalable. It is a common design in modern optical experimental setups and is relatively easy to use.

[0026] Implementation: In use, the external optical fiber is connected to the incident illumination assembly 8 through the optical fiber interface 87. The sample is placed on the sample holder 42. The position of the sample holder 42 is adjusted by the three-dimensional adjustment displacement stage 41 so that it is located between the transmission illumination assembly 5 and the microscopic imaging assembly 6, and directly above the first reflecting mirror 54. At this time, the operator can rotate the rotating wheel 65 to switch between different magnification objective lenses 66 for use, thereby selecting the objective lens 66 that is suitable for the current sample observation needs, which is convenient for observing the sample. Furthermore, the diverging light emitted by the second white light source 84 is collimated into parallel light when it passes through the third collimating lens 85. The parallel light then continues to propagate to the left, and after being reflected by the fourth reflecting mirror 86, its propagation path changes to vertically downwards, subsequently reaching the fourth beam splitter 89. Simultaneously, the diverging light emitted by the external optical fiber is collimated by the fourth collimating lens 88, and then continues to propagate to the left into the fourth beam splitter 89. At this point, the fourth beam splitter 89 combines the parallel light reflected by the fourth reflecting mirror 86 with the parallel light collimated by the fourth collimating lens 88. The beams are combined to form a combined beam, which continues to propagate downwards into the filter module 810. At this time, the filter module filters the combined beam and continues to propagate downwards. The filtered beam first passes through the second aperture 811 and is reflected by the fifth reflecting mirror 812. It then continues to the left through the second converging lens 813 to reach the first beam splitter 63. After being processed by the first beam splitter 63, it is focused on the back focal plane of the objective lens 66. Finally, the beam emitted by the incident illumination assembly 8 is collimated by the objective lens 66 and directly illuminates the sample, thereby providing uniform incident illumination for the sample in the field of view. At the same time, the divergent light emitted by the first white light source 52 is collimated into parallel light when it passes through the first collimating lens 53. The parallel light then continues to propagate backward and is reflected by the first reflecting mirror 54. The propagation path changes to vertical upward, so as to illuminate the sample on the sample holder 42 and provide uniform illumination for the sample above it. Furthermore, objective lens 66 collects scattered light carrying sample information and collimates it into parallel light, which then propagates upward and is converged by tube lens 62 after passing through first beam splitter 63 to form an intermediate image. Furthermore, the beam carrying the intermediate image propagates vertically upwards to the second collimating lens 77, where it is collimated into parallel light and continues to propagate upwards to the third beam splitter 75. At this point, the intermediate image beam is separated into multiple beams carrying images by the third beam splitter 75, meaning the intermediate image is separated into several microscopic images. One portion of the beam continues to propagate upwards to the second reflecting mirror 73, and is reflected by the second reflecting mirror 73 to the second beam splitter 72, and then propagated upwards to the target surface of the camera 78. The other portion of the beam is separated by the third beam splitter 75, propagated horizontally to the right to the third reflecting mirror 74, and reflected by the third reflecting mirror 74 to the second beam splitter 72, and then also propagated upwards to the target surface of the camera 78. This process separates the intermediate image and allows the separated beams carrying images to perform real-time imaging at different positions on the target surface of the camera 78 along different paths. This implementation specifically addresses the problems in the prior art where traditional optical microscopy imaging devices lose a lot of time when performing multicolor fluorescence imaging, i.e., live cell imaging, and cannot perform high-speed acquisition, thus failing to capture some very rapid dynamic events. Furthermore, adding an additional camera 78 increases the complexity of the optical path, and there may be differences when multiple cameras 78 are triggered synchronously, affecting the research. Furthermore, the potential sources of differences in the synchronous triggering of multiple cameras 78 include the delay and processing of the trigger signal, differences in the processing speed of the cameras 78 themselves, and minute errors in internal mechanical or electronic components. Although these differences are very small in most cases and imperceptible to the naked eye, they can still cause problems in research for scenarios requiring extremely precise synchronization of multicolor fluorescence imaging.

[0027] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used above are only some embodiments described in this invention. Obviously, those skilled in the art can obtain other drawings based on these drawings.

[0028] The foregoing has only described certain exemplary embodiments of the present invention by way of illustration. Undoubtedly, those skilled in the art can modify the described embodiments in various ways without departing from the spirit and scope of the present invention. Therefore, the foregoing drawings and descriptions are illustrative in nature and should not be construed as limiting the scope of protection of the claims of the present invention.

Claims

1. A single-camera partitioned optical microscopic imaging apparatus, characterized by, It includes: The bottom plate (1) is provided with a support (2) on the top of the back side, and the front side of the support (2) is provided with a mounting plate (3); The sample table (4) is installed on one side of the top of the bottom plate (1) and used for clamping the sample; The transmission illumination assembly (5) is installed on the other side of the top of the bottom plate (1) and used for providing uniform illumination for the sample; The microscopic imaging assembly (6) is installed on the front side of the bottom of the mounting plate (3) and used for forming the intermediate image of the sample; The image separation assembly (7) is installed on the front side of the top of the mounting plate (3) and used for separating the intermediate image and imaging on the target surface of the camera (78) at different positions; The fall illumination assembly (8) is installed on the front side of the mounting plate (3) and used for providing uniform fall illumination for the sample.

2. The single-camera partitioned optical microscopic imaging apparatus of claim 1, wherein: The sample table (4) includes a three-dimensional adjustment displacement table (41) and a sample holder (42) located on the top of the three-dimensional adjustment displacement table (41).

3. The single-camera partitioned optical microscopic imaging apparatus of claim 1, wherein: The transmission illumination assembly (5) includes a first cage frame (51) extending along the width direction of the bottom plate (1), and the inside of the first cage frame (51) is sequentially provided with a first white light source (52), a first collimating lens (53) and a first mirror (54) from front to back.

4. A single camera partitioned optical microscopic imaging device according to claim 3, wherein: The microscopic imaging assembly (6) includes a second cage frame (61) extending along the height direction of the mounting plate (3), and the inside of the second cage frame (61) is sequentially provided with a tube mirror (62), a first beam splitter (63) and a switch (66) from top to bottom.

5. A single camera partitioned optical microscopic imaging device according to claim 4, wherein: The switch (66) includes a rotating wheel (65) connected with the mounting plate (3) and a plurality of objective lenses (66) installed on the bottom of the rotating wheel (65).

6. A single camera partitioned optical microscopic imaging device according to claim 4, wherein: The image separation assembly (7) includes two third cage frames (71) arranged in parallel from top to bottom and extending along the width direction of the mounting plate (3), wherein the third cage frame (71) located on the top is sequentially provided with a second beam splitter (72) and a second mirror (73) from right to left, and the third cage frame (71) located on the bottom is sequentially provided with a third mirror (74) and a third beam splitter (75) from right to left. The image separation assembly (7) further includes a first diaphragm (76) installed in the inside of the second cage frame (61), a second collimating lens (77) located above the diaphragm and aligned with the third beam splitter (75), and the first diaphragm (76) is aligned with the tube mirror (62) from top to bottom. The image separation assembly (7) further includes a camera (78) installed on the top of the mounting plate (3) and a first converging lens (79) arranged on the bottom of the camera (78) and aligned with the second beam splitter (72).

7. A single camera partitioned optical microscopic imaging device according to claim 6, wherein: The clamping device (9) is installed on the two third cage frames (71).

8. A single camera partitioned optical microscopic imaging device according to claim 6, wherein: The said downlight illuminating assembly (8) comprises two fourth cage-shaped mirror holders (81) arranged in parallel from top to bottom and extending along the width direction of the mounting plate (3), a fifth cage-shaped mirror holder (82) extending along the height direction of the mounting plate (3), and a sixth cage-shaped mirror holder (83) located at the bottom of the fifth cage-shaped mirror holder (82) and extending along the width direction of the mounting plate (3); Wherein, the fourth cage-shaped mirror holder (81) located at the top is sequentially provided with a second white light source (84), a third collimating lens (85), and a fourth reflecting mirror (86) from right to left, the fourth cage-shaped mirror holder (81) located at the bottom is sequentially provided with a fiber interface (87), a fourth collimating lens (88), and a fourth beam splitter (89) from right to left, the fifth cage-shaped mirror holder (82) is sequentially provided with a filter module (810), and a second diaphragm (811) from top to bottom, and the sixth cage-shaped mirror holder (83) is sequentially provided with a fifth reflecting mirror (812) and a second converging lens (813) from right to left.

9. A single camera partitioned optical microscopic imaging device according to claim 8, wherein: The second converging lens (813) is horizontally aligned with the first beam splitter (63), and the filter module (810) is vertically aligned with the fourth beam splitter (89).

10. The single-camera partitioned optical microscopic imaging apparatus of claim 8, wherein: The first cage-shaped mirror holder (51), the second cage-shaped mirror holder (61), the third cage-shaped mirror holder (71), the fourth cage-shaped mirror holder (81), the fifth cage-shaped mirror holder (82), and the sixth cage-shaped mirror holder (83) are all provided as multi-axis cage-shaped structure holders.

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