Integrated brightfield, darkfield and fluorescence multimodal light field microscope system with stochastic confocal

Abstract

This application discloses a stochastic motion microscope integrating bright-field, dark-field, and fluorescence multimodal light field microscopy systems. The stochastic motion microscope includes a first mounting frame, a second mounting frame, a sample assembly, and a microscopy system. The second mounting frame is disposed on the first mounting frame, and the first mounting frame is adapted to drive the second mounting frame to rotate around a first axis. The second mounting frame is adapted to rotate relative to the first mounting frame around a second axis, wherein the first axis and the second axis are perpendicular. The sample assembly is disposed on the second mounting frame and is adapted to carry the sample to be tested. The microscopy system is disposed on the second mounting frame and includes an imaging component and a light source component. The light source component includes a bright-field light source, a dark-field light source, and a fluorescence light source, adapted to emit illumination light onto the sample. The imaging component is adapted to receive the reflected light from the sample for imaging. This application enables real-time observation of biological dynamics at the cellular level of biological samples while simultaneously simulating a microgravity environment.

Description

Technical Field

[0001] This application relates to the fields of optical microscopy and microgravity simulation, and in particular to a stochastic motion instrument for an integrated bright-field, dark-field and fluorescence multimodal optical field microscopy system. Background Technology

[0002] A random positioning machine is a common microgravity simulation device. It has two orthogonal, simultaneously rotating axes of rotation, which can point a sample in any direction at any given moment. By continuously changing the orientation of the sample, the sum of the gravity vectors on the sample approaches zero over a relatively long period of time, thus simulating microgravity.

[0003] In the research of microgravity biological effects based on stochastic motion instruments, biological samples usually need to be removed from the stochastic motion instrument and moved to an optical microscope for observation, making it difficult to observe the biological dynamics in a microgravity environment in real time. Summary of the Invention

[0004] This application aims to address at least one of the technical problems existing in the prior art. To this end, one objective of this application is to propose a stochastic motion microscope integrating bright-field, dark-field, and fluorescence multimodal light field microscopy systems, which aims to solve the problem that it is impossible to observe biological dynamics in real time under microgravity environment when studying the biological effects of microgravity based on a stochastic motion microscope.

[0005] This application proposes a stochastic motion microscope integrating bright-field, dark-field, and fluorescence multimodal light field microscopy systems. The stochastic motion microscope includes a first mounting frame, a second mounting frame, a sample assembly, and a microscopy system. The second mounting frame is disposed on the first mounting frame, and the first mounting frame is adapted to drive the second mounting frame to rotate around a first axis. The second mounting frame is adapted to rotate relative to the first mounting frame around a second axis, wherein the first axis is perpendicular to the second axis. The sample assembly is disposed on the second mounting frame and is adapted to carry the sample to be tested. The microscopy system is disposed on the second mounting frame and includes an imaging component and a light source component. The light source component includes a bright-field light source, a dark-field light source, and a fluorescence light source, adapted to emit illumination light onto the sample. The imaging component is adapted to receive the reflected light from the sample for imaging.

[0006] According to some embodiments of this application, the light source assembly includes an annular base, which is disposed at the object end of the imaging assembly; a plurality of dark field light sources and a plurality of fluorescence light sources are alternately disposed on the annular base along the circumferential direction.

[0007] According to some embodiments of this application, a ring-shaped base is provided with a plurality of first focusing components corresponding one-to-one with a fluorescent light source and a plurality of second focusing components corresponding one-to-one with a dark field light source; the first focusing components are arranged around the corresponding fluorescent light source; the first focusing components form two first focal points, one of which coincides with the corresponding fluorescent light source and the other of which coincides with the sample; the second focusing components are arranged around the corresponding dark field light source; the second focusing components form two second focal points, one of which coincides with the corresponding dark field light source and the other of which coincides with the sample.

[0008] According to some embodiments of this application, a first filter is provided at the emitting end of the fluorescent light source.

[0009] According to some embodiments of this application, a bright field light source is disposed on the sample assembly and located on the side of the sample away from the imaging assembly, so as to be suitable for emitting a transmitted light source onto the sample.

[0010] According to some embodiments of this application, the sample assembly includes a support base, a stage, and a sample holder. The support base is disposed on a second mounting frame; the stage is movably disposed on the support base along a first direction; and the sample holder is disposed on the stage and is adapted to hold a sample.

[0011] According to some embodiments of this application, the sample holder includes at least two magnetic suction members, wherein the two magnetic suction members are respectively disposed on the stage and are arranged facing each other along the clamping direction, so as to be suitable for clamping the sample by means of the magnetic attraction between them.

[0012] According to some embodiments of this application, the sample assembly further includes a microfluidic chip having a microfluidic cavity suitable for carrying the sample, and a sample holder suitable for holding the microfluidic chip.

[0013] According to some embodiments of this application, the imaging assembly includes an objective lens, a telescopic lens, an aperture, a Fourier lens, a second filter, a microlens array, and a camera arranged sequentially along the imaging optical path.

[0014] According to some embodiments of this application, the imaging assembly further includes a sleeve and a plurality of adjustment frames. The sleeve is disposed on a second mounting frame, and each adjustment frame is movably disposed inside the sleeve along the axial direction of the sleeve. An objective lens and a camera are respectively disposed at both ends of the sleeve, and a second filter is disposed inside the sleeve. A sleeve lens, an aperture, a Fourier lens, and a microlens array are respectively disposed on the corresponding adjustment frames.

[0015] The stochastic motion instrument of the integrated bright-field, dark-field, and fluorescence multimodal light field microscopy system according to this application has the following technical advantages compared with the prior art: (1) It can observe the biological dynamics at the cell level in real time while simulating the microgravity environment with a random motion instrument; it realizes the real-time wireless transmission of imaging data in the working state of the random motion instrument, and can simultaneously record the real-time image of the microscopic system and the real-time triaxial acceleration of the random motion instrument, and can comprehensively analyze the changes in parameters and biological sample information during the microgravity simulation process. (2) The microscopic system has a multimodal light field, integrating three microscopic imaging modes: bright field, dark field, and fluorescence, which can be freely switched to observe labeled or unlabeled biological information; the miniaturized ring fluorescence / dark field dual-mode light source, in combination with the bright field backplate light source, can realize multimodal observation; among them, the fluorescence / dark field dual-mode light source uses the ellipsoidal focusing method to greatly improve the illumination power while integrating two illumination sources; at the same time, the fluorescence / dark field dual-mode light source uses the ring base to realize the object end assembly, which minimizes the size of the light source, facilitates plug-and-play replacement, and is applicable to a variety of objectives; (3) The microscopic system adopts an imaging method combining Fourier lenses and microlens arrays. It captures information of the same area of ​​the sample from multiple perspectives and then performs three-dimensional image reconstruction. With the multi-view three-dimensional imaging capability, it can observe the three-dimensional structure of cell-level samples, while increasing the depth of field, mitigating information loss caused by vibration defocusing, and reducing the impact of vibration. It can perform real-time three-dimensional observation of biological samples with high spatial resolution and multiple modes under simulated microgravity conditions. (4) The random motion instrument has a high degree of design freedom and can integrate microscopic systems with different extended functions.

[0016] Additional aspects and advantages of this application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of this application. Attached Figure Description

[0017] The above and / or additional aspects and advantages of this application will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which: Figure 1 This is a schematic diagram of the random motion instrument of an integrated bright-field, dark-field, and fluorescence multimodal optical field microscopy system according to some embodiments of this application; Figure 2 This is a schematic diagram of the structure of a microscope system according to some embodiments of this application; Figure 3 This is a schematic diagram of the structure of a light source assembly according to some embodiments of this application; Figure 4 This is a schematic diagram of the optical path of a light source assembly according to some embodiments of this application; Figure 5 This is a schematic diagram of the structure of a sample component according to some embodiments of this application.

[0018] Figure label: First mounting bracket 11; Second mounting bracket 12; First motor 13; Second motor 14; Slip ring 15; Support assembly 16; Sample assembly 20; support base 21; stage 22; sample holder 23; slide rail assembly 24; Microscopic system 30; objective lens 301; sleeve lens 302; aperture 303; Fourier lens 304; microlens array 305; second filter 306; camera 307; sleeve 308; adjustment frame 309; annular base 310; dark field light source 311; fluorescence light source 312; first filter 313; first focusing component 314; second focusing component 315. Detailed Implementation

[0019] The embodiments of this application are described in detail below. Examples of these embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain this application, and should not be construed as limiting this application.

[0020] The following is for reference. Figures 1-5 This application describes a stochastic oscillator for an integrated bright-field, dark-field, and fluorescence multimodal light field microscopy system according to embodiments of the present application.

[0021] This application proposes a stochastic motion microscope integrating bright-field, dark-field, and fluorescence multimodal light field microscopy systems. The stochastic motion microscope includes a first mounting frame 11, a second mounting frame 12, a sample assembly 20, and a microscopy system 30. The second mounting frame 12 is disposed on the first mounting frame 11, and the first mounting frame 11 is adapted to drive the second mounting frame 12 to rotate around a first axis. The second mounting frame 12 is adapted to rotate relative to the first mounting frame 11 around a second axis, wherein the first axis and the second axis are perpendicular. The sample assembly 20 is disposed on the second mounting frame 12 and is adapted to carry the sample to be tested. The microscopy system 30 is disposed on the second mounting frame 12 and includes an imaging component and a light source component. The light source component includes a bright-field light source, a dark-field light source 311, and a fluorescence light source 312, adapted to emit illumination light onto the sample. The imaging component is adapted to receive the reflected light from the sample for imaging.

[0022] In the random motion instrument of this application, by setting a first mounting bracket 11 and a second mounting bracket 12, the sample assembly 20 and the microscopic system 30 can rotate around two mutually perpendicular axes, thereby simulating a microgravity environment. Under microgravity, the microscopic system 30 and the sample assembly 20 remain relatively stationary, and the dynamic changes of the sample carried by the sample assembly 20 can be observed in real time using the microscopic system 30. The light source assembly of the microscopic system 30 can emit different illuminations to the sample through a bright field light source (not shown in the figure), a dark field light source 311, and a fluorescence light source 312. The imaging assembly receives the light reflected or excited by the sample to form an image, realizing multi-modal light field observation of bright field, dark field, and fluorescence.

[0023] The stochastic motion instrument of this application enables real-time observation of cellular dynamics in biological samples while simulating a microgravity environment. Furthermore, the microscopy system 30 integrates three illumination sources in the imaging optical path: a bright-field light source 311, a dark-field light source 311, and a fluorescence light source 312. These correspond to three microscopic imaging modes: bright-field, dark-field, and fluorescence, allowing for free switching between observing labeled and unlabeled biological information to meet diverse observation needs. This stochastic motion instrument solves the problem of real-time observation of biological dynamics in microgravity environments in studies of microgravity biological effects, providing a high-precision, multimodal solution for studying dynamic microscopic processes. It significantly expands the applicability and experimental depth of microscopy and microgravity simulation technologies.

[0024] like Figure 1 As shown, in some embodiments, the random motion instrument further includes a first motor 13 and a second motor 14. The output shaft of the first motor 13 extends along the X-axis and is circumferentially limited and connected to the first mounting frame 11, adapted to drive the first mounting frame 11 to rotate around the X-axis (i.e., the first axis). The first mounting frame 11 drives the second mounting frame 12, the sample assembly 20, and the microscopic system 30 to rotate synchronously around the X-axis. The housing of the second motor 14 is connected to the first mounting frame 11, and the output shaft of the second motor 14 extends along the Y-axis and is circumferentially limited and connected to the second mounting frame 12, adapted to drive the second mounting frame 12 to rotate relative to the first mounting frame 11 around the Y-axis (i.e., the second axis). The second mounting frame 12 drives the sample assembly 20 and the microscopic system 30 to rotate synchronously around the Y-axis.

[0025] In a specific embodiment, such as Figure 1 As shown, the first mounting bracket 11 is constructed as a rectangular outer frame, and the second mounting bracket 12 is constructed as an inner rotating plate. The inner rotating plate is located inside the rectangular outer frame and is rotatably connected to the rectangular outer frame. The inner rotating plate has a large installation area, which can realize the stable installation and fixation of the sample component 20 and the microscopic system 30.

[0026] Furthermore, the first motor 13 can be mounted on an external fixed base or other structure to achieve a rotatable connection between the first mounting frame 11 and the external fixed base. Even further, a third mounting frame is rotatably mounted on the second mounting frame 12, and the sample assembly 20 and the microscopic system 30 are specifically mounted on the third mounting frame. The third mounting frame is adapted to rotate relative to the second mounting frame 12 around the Z-axis (the Z-axis is perpendicular to the plane containing the X and Y axes), thereby achieving rotation of the third mounting frame relative to the external base around three axes, thus simulating a more complex microgravity environment.

[0027] In addition, accelerometers can be installed on the first mounting bracket 11 and / or the second mounting bracket 12 and / or the third mounting part to detect the acceleration of the sample rotating around each axis in real time, providing microgravity environment data for observation and analysis.

[0028] In some embodiments, both the first mounting bracket 11 and the second mounting bracket 12 are provided with slip rings 15, which can supply power to the first motor 13 and the second motor 14.

[0029] According to some embodiments of this application, the imaging assembly includes an objective lens 301, a sleeve lens 302, an aperture 303, a Fourier lens 304, a second filter 306, a microlens array 305, and a camera 307 arranged sequentially along the imaging optical path. In this embodiment, the objective lens 301 is used to receive reflected or emitted light from the sample and magnify the image; the sleeve lens 302 works in conjunction with the objective lens 301 to focus the near-parallel light emitted from the objective lens 301; the aperture 303 is located at the primary image plane and is specifically configured as an adjustable aperture, which can adjust the field of view according to imaging requirements; the combination of the Fourier lens 304 and the microlens array 305 can acquire the spatial spectrum information of the light field to achieve multi-view three-dimensional imaging; the second filter 306 can filter out light in non-target wavelength bands, making the image clearer; the camera 307, as the terminal element of the imaging assembly, is responsible for converting optical signals into electrical signals and generating digital images. In some embodiments, the microlens array 305 consists of three microlenses, which can capture multi-view information to achieve three-dimensional imaging. The distance between the back focal plane of the telescopic lens 302 and the objective lens 301 is equal to the focal length of the telescopic lens 302, and the distance between the Fourier lens 304 and the telescopic lens 302 is the sum of their focal lengths. The camera 307 is connected to the microcontroller and can transmit digital images in real time to observation terminals such as computers via a wireless transmitter.

[0030] According to some embodiments of this application, the imaging assembly further includes a sleeve 308 and a plurality of adjustment brackets 309. The sleeve 308 is disposed on the second mounting bracket 12, and each adjustment bracket 309 is movably disposed inside the sleeve 308 along the axial direction of the sleeve 308. The objective lens 301 and the camera 307 are respectively disposed at both ends of the sleeve 308, and the second filter is disposed inside the sleeve 308. The sleeve lens 302, the aperture 303, the Fourier lens 304, and the microlens array 305 are respectively disposed on the corresponding adjustment brackets 309. In this embodiment, the sleeve 308 can provide a mounting base for the other components of the imaging assembly. The objective lens 301 and the camera 307 are detachably disposed at both ends of the sleeve 308, specifically by means of threaded connection for assembly; the second filter 306 is disposed inside the sleeve 308. The sleeve 308 is equipped with multiple movable adjustment brackets 309. The sleeve lens 302, aperture 303, Fourier lens 304, and microlens array 305 are respectively mounted on the corresponding adjustment brackets 309. Each adjustment bracket 309 can adjust the axial position of the corresponding element within the sleeve 308 to optimize the imaging effect. This embodiment can improve the flexibility of the imaging assembly, realize flexible adjustment of the optical path, and improve imaging quality.

[0031] Furthermore, a set of fixing components is provided between the sleeve 308 and each adjusting bracket 309. The fixing components are used to restrict the axial relative movement between the adjusting bracket 309 and the sleeve 308, thereby fixing the adjusting bracket 309.

[0032] In some embodiments, the sleeve 308 is constructed as a cylindrical structure with a perforated surface and a hollow interior, facilitating observation and adjustment of the optical path. The sleeve 308 has a slot in which the second filter 306 is detachably installed. Further, the sleeve 308 has an axially extending connecting groove, and the adjusting frame 309 has a limiting hole on its outer periphery. When the adjusting frame 309 moves axially relative to the sleeve 308 for adjustment, the limiting hole and the connecting groove are always radially aligned, and fasteners can pass through the connecting groove and engage with the limiting hole. Through the tight engagement of the fasteners and the limiting hole, the inner peripheral wall of the sleeve 308 and the outer peripheral wall of the adjusting frame 309 can be tightly fitted, thereby achieving a fixed connection between the sleeve 308 and the adjusting frame 309, locking the relative position of the adjusting frame 309 and the sleeve 308, and ensuring stable assembly under microgravity conditions. Furthermore, the adjustment frame 309 is equipped with springs and set screws. When imaging elements such as the sleeve lens 302, aperture 303, Fourier lens 304, and microlens array 305 are set on the corresponding adjustment frame 309, the set screws and springs abut against the corresponding imaging elements and are suitable for adjusting the relative position of the imaging elements in the radial plane of the adjustment frame 309.

[0033] like Figure 1As shown, in some embodiments, a support assembly 16 is provided on the second mounting bracket 12, and the sleeve 308 is connected to the support assembly 16 to fix the imaging assembly, allowing it to rotate stably and synchronously with the second mounting bracket 12. The support assembly 16 and the second mounting bracket 12 can be connected by fasteners. Further, the support assembly 16 includes a clamping member adapted to clamp the sleeve 308; the clamping member can specifically be constructed as an annular clamping plate.

[0034] According to some embodiments of this application, the light source assembly includes an annular base 310, which is disposed at the object end of the imaging assembly; a plurality of dark field light sources 311 and a plurality of fluorescence light sources 312 are alternately spaced along the circumference on the annular base 310. In this embodiment, as... Figure 3 As shown, by setting up the annular base 310, the dark-field light source 311 and the fluorescence light source 312 can be installed on the objective lens, realizing spatial coupling between the light source and the imaging component. This avoids the redundant structure of traditional external light sources, maintains illumination stability in microgravity environments, and reduces interference from external ambient light. Furthermore, the dark-field light source 311 and the fluorescence light source 312 are integrated on the annular base 310, forming a dark-field / fluorescence dual-mode light source. This allows for independent or combined operation of the dark-field light source 311 and the fluorescence light source 312, without the need for mechanical movement of the light sources. Rapid mode switching can be achieved through electronic control, improving observation efficiency. In addition, the dark-field light source 311 on the annular base 310 can generate oblique light on the sample, allowing light to enter the objective lens 301 only through scattering from the sample edge or surface, while direct light is blocked, significantly improving the observation effect. Furthermore, multiple dark-field light sources 311 and fluorescence light sources 312 are constructed and alternately arranged along the circumference of the annular base 310 to improve the uniformity of sample illumination; setting multiple fluorescence light sources 312 can improve the uniformity of fluorescence illumination on the sample, reduce differences in fluorescence signal intensity, and improve the consistency of imaging. Figure 3 As shown, both the dark field light source 311 and the fluorescent light source 312 are constructed in triplicate.

[0035] In some embodiments, the dark field light source 311 is constructed as a green LED that can emit green light; the fluorescent light source 312 is constructed as a blue LED that can emit blue light; after the biological sample cells are labeled with green fluorescent protein (GFP), they can emit green light after being excited by the blue light emitted by the fluorescent light source 312.

[0036] According to some embodiments of this application, a ring-shaped base 310 is provided with a plurality of first focusing members 314 corresponding one-to-one with the fluorescent light source 312 and a plurality of second focusing members 315 corresponding one-to-one with the dark field light source 311. The first focusing members 314 are arranged around the corresponding fluorescent light source 312. The first focusing members 314 form two first focal points, one of which coincides with the corresponding fluorescent light source 312, and the other of which coincides with the sample. The second focusing members 315 are arranged around the corresponding dark field light source 311. The second focusing members 315 form two second focal points, one of which coincides with the corresponding dark field light source 311, and the other of which coincides with the sample. In this embodiment, by setting the first focusing members 314, the light emitted by the fluorescent light source 312 can be focused on the sample surface; by setting the second focusing members 315, the light emitted by the dark field light source 311 can be focused on the sample surface, thereby maximizing the illumination power. In some embodiments, the first focusing member 314 is formed with an ellipsoidal surface surrounding the fluorescent light source 312, and the second focusing member 315 is formed with an ellipsoidal surface surrounding the dark field light source 311. The ellipsoidal surface forms two focal points that coincide with the corresponding light source and the sample, respectively, so as to utilize light reflection and focusing to improve the fluorescent illumination power.

[0037] According to some embodiments of this application, the emitting end of the fluorescent light source 312 is provided with a first filter 313. In this embodiment, the first filter 313 can filter out light of non-target wavelengths. Specifically, when the light emitted by the fluorescent light source 312 passes through the first filter 313, the first filter 313 selectively transmits the fluorescent excitation wavelength light while blocking stray light of other wavelengths, which can ensure the monochromaticity of the excitation light and avoid excitation of unintended fluorescent substances by light of non-target wavelengths, thereby improving the accuracy of imaging.

[0038] In one specific embodiment, at the emitting end of the blue LED, the first filter 313 is configured as a blue light filter, which only allows light of the corresponding wavelength band of blue light to pass through.

[0039] In some embodiments, such as Figure 3As shown, an annular base 310 is sleeved on the end of the objective lens 301 of the imaging assembly. Multiple first mounting portions and multiple second mounting portions are formed on the annular base 310. The multiple first mounting portions and multiple second mounting portions are alternately spaced along the circumference. Multiple fluorescent light sources 312 are connected one-to-one with the multiple first mounting portions, and the fluorescent light sources 312 and the first mounting portions can be assembled using detachable connection methods such as snap-fit ​​or sleeve connection. Multiple dark-field light sources 311 are connected one-to-one with the multiple second mounting portions, and the dark-field light sources 311 and the second mounting portions can be assembled using detachable connection methods such as snap-fit ​​or sleeve connection. A third mounting portion is correspondingly provided on the outer periphery of each first mounting portion and each second mounting portion. Each first focusing member 314 and each second focusing member 315 are respectively connected to the corresponding third mounting portion, specifically assembled using methods such as snap-fit, sleeve connection, or adhesive bonding.

[0040] According to some embodiments of this application, a bright-field light source is disposed on the sample assembly 20 and located on the side of the sample away from the imaging assembly, suitable for emitting a transmitted light source onto the sample. In this embodiment, the bright-field light source is disposed on the side of the sample away from the imaging assembly, which can emit a uniform transmitted light source onto the sample. Light that is not absorbed or scattered by the sample passes through the sample and is collected by the objective lens 301 and focused onto the imaging assembly, forming a bright background; areas in the sample that absorb or scatter light appear dark, thus constructing an image with contrast between light and dark; furthermore, different components of the sample absorb and scatter light differently, therefore a structurally clear image can be obtained in the imaging assembly. In this embodiment, the bright-field light source is integrated into the sample assembly 20, which can improve the stability and uniformity of bright-field illumination, while making the structure more compact and facilitating stable operation in a microgravity environment.

[0041] In some embodiments, the bright field light source is constructed as an LED array backplate, on which a diffuser is provided to provide uniform illumination. After the LED array backplate is fixed to the sample, the sample and the LED array backplate can be simultaneously held by the sample holder 23, and the LED array backplate provides transmissive illumination to the sample.

[0042] According to some embodiments of this application, the sample assembly 20 includes a support base 21, a stage 22, and a sample holder 23. The support base 21 is disposed on a second mounting frame 12; the stage 22 is movably disposed on the support base 21 along a first direction; the sample holder 23 is disposed on the stage 22 and is adapted to hold a sample. Figure 5As shown, in this embodiment, the support base 21 is disposed on the second mounting frame 12, providing the mounting base for the stage 22 and the sample holder 23. The sample holder 23 holds the sample to be tested and is disposed on the stage 22. The stage 22 is movably disposed on the support base 21, which can drive the sample to move relative to the support base 21 along a first direction, making the distance between the sample and the imaging component adjustable, thereby achieving precise focusing and helping to improve the clarity of the image. This embodiment can achieve flexible movement and stable holding of the sample, effectively reducing the vibration of the sample in a microgravity environment and improving the imaging quality. The first direction coincides with the axis of the imaging component. It should be noted that when the sample holder 23 holds the sample, it does not directly contact the biological sample, but rather holds the container or support structure such as the glass slide that carries the biological sample.

[0043] In some embodiments, such as Figure 5 As shown, a slide rail assembly 24 is provided between the support base 21 and the platform 22. Specifically, the support base 21 is provided with a guide rail, and the platform 22 is provided with a sliding member that slides with the guide rail; when the sliding member slides relative to the guide rail, it drives the platform 22 to move relative to the guide rail along a first direction.

[0044] Furthermore, the sample assembly 20 includes a drive assembly for driving the slider to slide relative to the guide rail, and a locking assembly for locking the relative position of the slider and the guide rail. When the drive assembly drives the slider to move the stage 22 to the target sample position, the locking assembly can restrict the slider from continuing to move relative to the guide rail, keeping the sample stable during observation and reducing vibration.

[0045] According to some embodiments of this application, the sample holder 23 includes at least two magnetic suction elements, which are respectively disposed on the stage 22 and face each other along the clamping direction to facilitate clamping the sample using their magnetic attraction. In this embodiment, the sample is clamped and fixed by using magnetic suction elements, resulting in high stability and reliability, and adaptability to dynamic microgravity environments. Moreover, the magnetic clamping structure enables rapid clamping and release, improving operational efficiency. Furthermore, by adjusting the magnitude of the magnetic attraction, the sample holder 23 can adapt to biological samples of different shapes and sizes, exhibiting wide applicability and flexibility, and meeting the requirements for stable clamping.

[0046] In some embodiments, such as Figure 5 As shown, the sample holder 23 is provided in two sets, which are respectively set on both sides of the sample to further improve the stability of sample holding.

[0047] According to some embodiments of this application, the sample assembly 20 further includes a microfluidic chip with a microfluidic cavity suitable for carrying the sample, and a sample holder 23 suitable for holding the microfluidic chip. In this embodiment, the biological sample is placed in the microfluidic cavity and cultured using adherent or suspension culture; the microfluidic chip can carry a small number of samples; the sample holder 23 holds the microfluidic chip to achieve sample positioning. This embodiment can utilize the characteristics of the microfluidic chip to dynamically adjust the sample culture environment, improving the accuracy and flexibility of sample observation.

[0048] Furthermore, the sample component 20 also includes a circulating micro water pump and a small water tank. The microfluidic chip has an input port and an output port that are connected to the microfluidic cavity. The input port and the output port are connected in series with the circulating micro water pump and the small water tank through pipelines, which can realize continuous liquid exchange in the microfluidic cavity and meet the survival needs of biological samples during the observation period.

[0049] When applied, this application allows for real-time viewing of multimodal cell culture results via a wireless transmission module; observation of cell morphology and geometric features via bright-field mode; and free switching to fluorescent mode during cell culture to observe fluorescently labeled proteins or autofluorescence in the cells, or switching to dark-field mode to observe the backscattering signal of the cells.

[0050] The random motion instrument of the integrated optical microscopy system 30 according to this application has the following technical advantages compared with the prior art: (1) It can observe the biological dynamics at the cell level in real time while simulating the microgravity environment with a random motion instrument; it realizes the real-time wireless transmission of imaging data in the working state of the random motion instrument, and can simultaneously record the real-time image of the microscopic system 30 and the real-time triaxial acceleration of the random motion instrument, and can comprehensively analyze the changes in parameters and biological sample information during the microgravity simulation process. (2) The microscopic system 30 has a multimodal light field, integrating three microscopic imaging modes: bright field, dark field, and fluorescence, and can freely switch to observe labeled or unlabeled biological information; the miniaturized annular fluorescence / dark field dual-mode light source, in conjunction with the bright field backplate light source, can realize multimodal observation; among them, the fluorescence / dark field dual-mode light source, by using the ellipsoidal focusing method, greatly improves the illumination power while integrating two illumination sources; at the same time, the fluorescence / dark field dual-mode light source uses the annular base 310 to realize the object end assembly, which minimizes the size of the light source to the greatest extent and is easy to plug and replace, and can be used for various objectives 301; (3) The microscopic system 30 adopts an imaging method combining Fourier lens 304 and microlens array 305. It uses multiple perspectives to capture information of the same area of ​​the sample and then performs three-dimensional image reconstruction. With the multi-view three-dimensional imaging capability, it can observe the three-dimensional structure of cell-level samples, while increasing the depth of field, mitigating information loss caused by vibration defocusing, and reducing the impact of vibration. It can perform real-time three-dimensional observation of biological samples with high spatial resolution and multiple modes under simulated microgravity conditions. (4) The random motion instrument has a high degree of design freedom and can integrate microscopic systems 30 with different extended functions.

[0051] In the description of this application, it should be understood that the terms "center," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," and "circumferential" indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this application.

[0052] In the description of this application, "first feature" and "second feature" may include one or more of the features.

[0053] In the description of this application, "multiple" means two or more.

[0054] In the description of this application, the first feature being "above" or "below" the second feature may include the first and second features being in direct contact, or the first and second features being in contact through another feature between them.

[0055] In the description of this application, the terms "above," "over," and "on top" for the first feature and the second feature include the first feature being directly above or diagonally above the second feature, or simply indicate that the first feature is at a higher horizontal level than the second feature.

[0056] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "illustrative embodiment," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0057] Although embodiments of this application have been shown and described, those skilled in the art will understand that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of this application, the scope of which is defined by the claims and their equivalents.

Claims

1. A stochastic confocal microscope system integrating brightfield, darkfield and fluorescence multi-modal light field microscopy, characterized in that, include: A first mounting bracket and a second mounting bracket, the second mounting bracket being disposed on the first mounting bracket, the first mounting bracket being adapted to drive the second mounting bracket to rotate around a first axis, and the second mounting bracket being adapted to rotate relative to the first mounting bracket around a second axis; wherein, the first axis is perpendicular to the second axis; A sample assembly, which is disposed on the second mounting bracket and is adapted to carry the sample to be tested; A microscopy system is disposed on the second mounting bracket; the microscopy system includes an imaging component and a light source component, the light source component including a bright field light source, a dark field light source and a fluorescence light source, adapted to emit illumination light onto the sample; the imaging component is adapted to receive reflected light from the sample for imaging.

2. The integrated brightfield, darkfield and fluorescence multimodal light field microscope system of claim 1, wherein, The light source assembly includes: A ring-shaped base is disposed on the object end of the imaging assembly; a plurality of dark field light sources and a plurality of fluorescence light sources are alternately disposed on the ring-shaped base along the circumferential direction.

3. The integrated brightfield, darkfield and fluorescence multimodal light field microscope system of claim 2, wherein, The annular base frame is provided with a plurality of first focusing components corresponding to the fluorescent light source and a plurality of second focusing components corresponding to the dark field light source. The first focusing component is arranged around the corresponding fluorescent light source; the first focusing component forms two first focal points, one of which coincides with the corresponding fluorescent light source, and the other of the first focal points coincides with the sample; The second focusing member is arranged around the corresponding dark field light source; the second focusing member forms two second focal points, one of which coincides with the corresponding dark field light source, and the other of which coincides with the sample.

4. The integrated brightfield, darkfield and fluorescence multi-modal light field microscope system of claim 1, wherein, The emitting end of the fluorescent light source is provided with a first filter.

5. The integrated brightfield, darkfield and fluorescence multimodal light field microscope system of claim 1, wherein, The bright-field light source is disposed on the sample assembly and located on the side of the sample away from the imaging assembly, so as to be suitable for emitting a transmitted light source onto the sample.

6. The integrated brightfield, darkfield and fluorescence multimodal light field microscope system of claim 1, wherein, The sample components include: A support base, wherein the support base is disposed on the second mounting bracket; A platform, which is movably disposed on the support base along a first direction; A sample holder is disposed on the stage and is adapted to hold the sample.

7. The integrated brightfield, darkfield and fluorescence multimodal light field microscope system of claim 6, wherein, The sample holder includes at least two magnetic suction elements, wherein the two magnetic suction elements are respectively disposed on the stage and are arranged facing each other along the clamping direction, so as to be adapted to clamp the sample by means of the magnetic attraction between them.

8. The integrated brightfield, darkfield and fluorescence multimodal light field microscope system of claim 6, wherein, The sample component also includes: A microfluidic chip having a microfluidic cavity adapted to hold the sample, and a sample holder adapted to hold the microfluidic chip.

9. The integrated brightfield, darkfield and fluorescence multimodal light field microscope system of claim 1, wherein, The imaging assembly includes an objective lens, a telescopic lens, an aperture, a Fourier lens, a second filter, a microlens array, and a camera arranged sequentially along the imaging optical path.

10. The stochastic motion instrument of the integrated bright-field, dark-field, and fluorescence multimodal light field microscopy system according to claim 9, characterized in that, The imaging assembly further includes a sleeve and a plurality of adjustment brackets. The sleeve is disposed on the second mounting bracket, and each adjustment bracket is movably disposed inside the sleeve along the axial direction of the sleeve. The objective lens and the camera are respectively disposed at both ends of the sleeve, and the second filter is disposed inside the sleeve; the sleeve lens, the aperture, the Fourier lens and the microlens array are respectively disposed on the corresponding adjustment frame.

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