System and method for realizing cell regulation and control by stimulating cell nucleus based on light oscillation

Abstract

The invention discloses a system and a method for realizing cell regulation and control based on luminous oscillation stimulation of cell nucleuses, and relates to the technical field of crossing of biotechnology and optical micro-control. The method comprises the following steps: internalizing biocompatible microspheres beside a living cell nucleus, capturing the microspheres by using an optical tweezers module, and driving a light potential well to carry out double-position high-speed alternate switching by using a parameter control unit, so as to construct a programmable optical mechanical oscillator to apply oscillation stimulation to cells in a set direction, frequency and amplitude. According to the invention, the limitation of the traditional method is broken through, and direct and controllable mechanical stimulation and observation on the cell nucleus in the living cell are realized.

Description

A system and method for cell regulation based on photodynamic oscillation stimulation of the cell nucleus Technical Field

[0001] This invention relates to the field of biotechnology and optical micromanipulation, specifically to a system and method for cell regulation based on photodynamic oscillation stimulation of the cell nucleus. Background Technology

[0002] The cell nucleus is not only the center of genetic information, but also an important intracellular mechanosensor. The nuclear laminin network beneath the nuclear membrane can directly sense mechanical stimuli and convert the signals into biochemical responses, ultimately regulating gene expression.

[0003] In the fields of life sciences and regenerative medicine, precisely guiding the growth direction and connection patterns of cells (such as the specific formation of neural networks) is a crucial goal. The precise application of mechanical force, as a key physical signal influencing cell fate and function, is one of the core means to achieve this goal.

[0004] Currently, the mainstream methods for providing mechanical stimulation to cells have the following limitations: 1. Bass stiffness modulation method: This method simulates tissue stiffness by changing the elastic modulus of the culture substrate. This method provides a global, non-specific mechanical environment and cannot precisely manipulate individual target cells or specific internal structures.

[0005] 2. Atomic Force Microscopy (AFM) probe method: This method uses an AFM probe to apply localized forces to the cell surface. While it offers some spatial resolution, its target is limited to the cell membrane or surface, and it cannot simulate the natural mechanical interactions of organelles within the cell.

[0006] 3. Population mechanical loading method: This method involves stretching the cell population as a whole or applying fluid shear force. While this method has a wide range of applications, it lacks spatial specificity, affects a large number of cells simultaneously, and its signal transduction is indirect.

[0007] In summary, all existing technologies share a fundamental limitation: they cannot directly, precisely, and controllably stimulate the cell nucleus—the core organelle—within a living cell. This hinders researchers' in-depth understanding of how the cell nucleus responds to mechanical signals and how to utilize this response to guide cellular behavior.

[0008] Therefore, there is an urgent need to develop a method that can directly and dynamically manipulate the cell nucleus within living cells. Summary of the Invention

[0009] The purpose of this invention is to provide a system and method for cell regulation based on photomechanical oscillation stimulation of the cell nucleus, overcoming the limitations of existing technologies that can only apply signals from outside the cell or the whole cell. This invention utilizes optical tweezers to drive intracellular microspheres as "photomechanical oscillators" to directly stimulate the cell nucleus through programmable micro-amplitude vibrations, thereby regulating its physical state and guiding cell behavior. This technology enables direct and controllable mechanical stimulation and observation of the cell nucleus within living cells.

[0010] Based on the above objectives, the present invention provides a method for cell regulation based on photomechanical oscillation stimulation of the cell nucleus, comprising the following steps: S1, providing a live cell containing an internalized photomechanical oscillator positioned near the cell nucleus; S2, capturing the photomechanical oscillator using an optical tweezers module, bringing it into contact with or closely adjacent to the nuclear membrane of the cell nucleus; S3, according to preset oscillation parameters, controlling the optical potential trap generated by the optical tweezers system to alternately switch between a first position and a second position to drive the photomechanical oscillator to vibrate mechanically, thereby applying controllable local mechanical stimulation to the cell nucleus.

[0011] Preferably, the optomechanical oscillator is a biocompatible modified microsphere with a diameter of 0.5 μm to 2 μm. The microsphere is taken into the cell by inducing endocytosis through co-culture.

[0012] Preferably, the oscillation parameters include oscillation angle, oscillation frequency, and amplitude; the oscillation angle is the angle between the vibration direction of the photomechanical oscillator and the normal direction of the nuclear membrane contact point, ranging from -90° to +90°; the oscillation frequency is the number of oscillations per second of the photomechanical oscillator, which is adjusted by the switching speed of the optical potential trap, ranging from 0 to 100 Hz; the amplitude is the vibration amplitude of the photomechanical oscillator, ranging from 100 to 1000 nanometers.

[0013] Preferably, the first position and the second position are determined according to the setting of the oscillation angle and the amplitude; the switching rate of the optical potential trap between the first position and the second position is controlled according to the setting of the vibration frequency; wherein, the direction of the line connecting the first position and the second position corresponds to the oscillation angle, and the distance between them corresponds to the amplitude.

[0014] Preferably, the method is used to regulate cell behavior by observing the physical response of the cell nucleus and / or subsequent behavioral changes of the cell while or after the application of controlled local mechanical stimulation.

[0015] Preferably, the physical response of the cell nucleus and / or subsequent behavioral changes of the cell include nuclear rotation, changes in the growth direction of neurons, or the establishment of connections between the cell and neighboring cells.

[0016] The present invention also provides a system for implementing the above-mentioned method of cell regulation based on photodynamic oscillation stimulation of the cell nucleus, comprising: an optical manipulation and imaging unit for capturing and driving microspheres and observing cells in real time; an optomechanical oscillator for the microspheres captured and driven by the optical manipulation and imaging unit; a parameter control unit connected to the optical manipulation and imaging unit for receiving set oscillation parameters and controlling the optical manipulation and imaging unit to generate corresponding light field motion; and a biolabeling unit for labeling cells to observe their mechanical response.

[0017] Preferably, the optical manipulation and imaging unit includes an optical tweezers module and an integrated microscopic imaging module; the integrated microscopic imaging unit supports fluorescence microscopy and bright-field microscopy modes, and the imaging mode is switched by switching the light source and filter path; the optical tweezers module includes: a near-infrared laser source that outputs a near-infrared laser beam; an acousto-optic deflector located in the output optical path of the near-infrared laser source to control the propagation direction of the near-infrared laser beam; a beam expander located in the output optical path of the acousto-optic deflector to adjust the spot size of the near-infrared laser beam; a high numerical aperture objective lens that focuses the near-infrared laser beam to form an optical potential trap; and a dichroic mirror located in the rear optical path of the high numerical aperture objective lens to reflect the near-infrared laser beam from the beam expander to the high numerical aperture objective lens; the integrated microscopic imaging module and the optical tweezers module share the high numerical aperture objective lens. The microscope employs an aperture objective lens and a dichroic mirror. Fluorescence microscopy imaging modes include: a near-infrared laser source that outputs excitation light to excite the sample and generate fluorescence emission signals; an image detector that acquires the fluorescence emission signals generated by the sample; a dichroic mirror that reflects the excitation light output from the fluorescence excitation source to the sample and transmits the fluorescence emission signals generated by the sample to the image detector; and a high numerical aperture objective lens that collects the fluorescence emission signals generated after the sample is excited. Bright-field microscopy imaging modes include: a bright-field illumination source that outputs wide-field visible light to illuminate the sample; an image detector that acquires the visible light transmitted or scattered light from the sample; a dichroic mirror that transmits the visible light transmitted or scattered light from the sample to the image detector; and a high numerical aperture objective lens that collects the visible light transmitted or scattered light generated by the sample after being illuminated by the bright-field illumination source.

[0018] Preferably, the control logic of the parameter control unit is as follows: receiving the oscillation angle, amplitude, and vibration frequency set for the optomechanical oscillator; calculating and determining two target spatial positions, namely the first position and the second position, based on the oscillation angle and amplitude; determining a switching rate based on the vibration frequency; and outputting a drive signal to the acousto-optic deflector of the optical manipulation and imaging unit, so that it controls the optical potential trap to alternately transition between the two target spatial positions at the switching rate.

[0019] Preferably, the biomarker unit includes live cell dyes and / or specific antibodies for labeling the cell nucleus, cytoskeleton, or mechanosensitive proteins, thereby enabling the observation of stimuli-induced changes in cell nucleus morphology, localization, or protein position through an optical manipulation and imaging unit.

[0020] Therefore, the present invention adopts the above-mentioned system and method for cell regulation based on photodynamic oscillation stimulation of the cell nucleus, which has the following beneficial effects: 1. It provides a new subcellular targeted mechanical stimulation approach: For the first time, it realizes direct and targeted mechanical stimulation of the cell nucleus in living cells, breaking through the limitation of traditional methods that can only apply signals from outside the cell or the whole cell, and breaking through the physical barrier of the cell membrane.

[0021] 2. Achieving a highly controllable stimulation mode for cell regulation processes: Based on an optomechanical oscillator driven by dual optical potential traps, the direction, frequency, and amplitude of stimulation can be independently and precisely set with digital parameters. This unprecedented control precision allows for the application of specific mechanical commands to cells in a programmable and quantifiable manner, thereby achieving fine-grained regulation of the cell response process, surpassing the limitations of traditional optical tweezers that can only be simply pushed and pulled.

[0022] 3. The correlation mechanism between nuclear mechanical stimulation and cell behavior was empirically demonstrated: This system revealed the complete chain that directional oscillatory stimulation can directly induce nuclear rotation, thereby activating the nuclear Lamin A / C and YAP signaling pathways, and ultimately guiding the directional growth and successful connection of nerve cells. This confirms that mechanical intervention of the cell nucleus can directly regulate cell function.

[0023] 4. High system integration and strong practicality: This system is built on a general-purpose optical microscope platform, integrating stimulation, observation, and analysis functions. It is highly practical and easy to promote. It provides a powerful and standardized research platform for studying nuclear mechanobiology and exploring new methods for neural repair and tissue engineering.

[0024] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0025] Figure 1 shows the optical manipulation and imaging unit of a system for cell regulation based on photomechanical oscillation stimulation of the cell nucleus according to the present invention; Figure 2 is a schematic diagram of the working principle of the optomechanical oscillator unit of the present invention; Figure 3 is a positioning diagram of the optomechanical oscillator around the cell nucleus, where I is a magnified view with a scale bar of 10 μm and II is a magnified view with a scale bar of 1 μm; Figure 4 shows the relationship between the position change of the optomechanical oscillator and the switching rate of the optical potential trap, where a is the change of the position of the optomechanical oscillator over time under different optical potential trap switching rates, and b is the relationship between the actual vibration frequency of the optomechanical oscillator in the cell and the optical potential trap switching rate; Figure 5 shows the vibration of the optomechanical oscillator controlled by photomechanical force; Figure 6 shows the relationship between the oscillation angle of the optomechanical oscillator and the cell nucleus. Figure 7 shows the nuclear rotation effect when the oscillation angle is 0°, 60°, and 80°; Figure 8 shows the nuclear rotation effect detected by bright-field microscopy and fluorescence microscopy when the oscillation angle is 0°; Figure 9 shows the nuclear rotation effect detected by bright-field microscopy and fluorescence microscopy when the oscillation angle is 30°; Figure 10 shows the relationship between oscillation parameters and nuclear rotation angle, where a represents the relationship between vibration frequency and nuclear rotation angle, and b represents the relationship between oscillation angle and nuclear rotation angle; Figure 11 shows the nuclear rotation effect of experimental and control group cells. Figure 12 shows a comparison of the fluorescence intensity of A / C protein on fluorescent staining, where a is a specific experimental figure and b is a statistical graph of the control and experimental groups; Figure 13 shows a comparison of the nuclear / cytoplasmic ratio of the fluorescence intensity of YAP protein on the nuclear cells of the experimental and control groups, where a is a specific experimental figure and b is a statistical graph of the control and experimental groups; Figure 14 shows a comparison of the nuclear growth direction trend of the experimental and control groups, where a is the control group and b is the experimental group; Figure 15 shows the positional relationship between the cells of the experimental and control groups and their neighboring neurons, where a is the control group and b is the experimental group. Detailed Implementation

[0026] Unless otherwise defined, the technical or scientific terms used in this invention shall have the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.

[0027] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can be appropriately combined to form other embodiments that can be understood by those skilled in the art. These other embodiments are also covered within the scope of protection of this invention.

[0028] Example 1 This example provides a system for cell regulation based on photodynamic oscillation stimulation of the cell nucleus, including: an optical manipulation and imaging unit, an optomechanical oscillator, a parameter control unit, and a biomarker unit.

[0029] In this embodiment, the optical manipulation and imaging unit is based on a modified inverted fluorescence microscope, which integrates an optical tweezers module and an integrated microscopic imaging module.

[0030] The optical tweezers module is the core of the mechanical manipulation. Its workflow is shown in Figure 1. The laser beam emitted from a near-infrared laser source first passes through an acousto-optic deflector. This device can control the propagation direction of the laser beam at high speed and with precision according to electronic control signals. Subsequently, the laser beam is expanded by a beam expander to ideally match the spot size with the aperture of subsequent optical elements. The expanded laser beam is guided by a dichroic mirror, which has high reflectivity for near-infrared lasers. After reflection, the laser beam enters vertically upwards into a high numerical aperture objective lens and is highly focused by this lens, forming a micrometer-scale optical potential trap on the sample plane, capable of stably capturing the target microsphere.

[0031] Its integrated microscopic imaging module and optical tweezers module share the core optical path, and the switching between fluorescence microscopic imaging mode and bright-field microscopic imaging mode can be achieved by switching the light source and the filter path.

[0032] In bright-field microscopy, a bright-field illumination source (such as an LED) provides wide-field visible light illumination. After the light passes through the sample, its transmitted or scattered light signal is collected by a high numerical aperture objective lens. This visible light imaging signal is also transmitted through a dichroic mirror and is ultimately received by the same image detector, generating a bright-field image that reflects the overall morphology, position, and dynamics of the cells.

[0033] In fluorescence microscopy, a specific wavelength of excitation light emitted from a fluorescence excitation source (in this embodiment, the LED light source) illuminates the sample, exciting a fluorescent label. The fluorescence emission signal generated by the sample is collected by the same objective lens, transmitted through a dichroic mirror, and finally acquired by an image detector to generate a high-contrast image.

[0034] The photomechanical oscillator in this embodiment is essentially a microsphere made of biocompatible materials (such as silica, polystyrene, etc.). This microsphere can be captured and driven by an external light field, thus acting as a controlled mechanical vibration source within the cell, precisely converting the modulation of the light field into local mechanical signals acting on the cell nucleus. The oscillator is delivered non-invasively through co-culturing with cells. During this process, the cells actively take the microsphere into their cells through endocytosis.

[0035] The diameter range is preferably 0.5 μm to 2 μm. This range ensures that the microspheres can be stably and efficiently captured and driven by the optical potential trap provided by the optical tweezers system, avoiding insufficient capture force due to excessive size, and can be effectively internalized by cells through endocytosis, avoiding excessive burden on cell metabolism or difficulty in transporting to the perinuclear region due to excessive size.

[0036] The surface of microspheres can be functionalized according to experimental requirements. For example, specific chemical groups can be introduced through amination or carboxylation, or biomolecules such as cell-penetrating peptides can be modified to regulate their interaction with cells, thereby optimizing their endocytosis efficiency, intracellular stability, and perinuclear localization.

[0037] In this embodiment, the parameter control unit is communicatively connected to the optical manipulation and imaging unit, converting mechanical parameters into control commands to drive the optical tweezers module to generate specific light field motion modes. The direct input and adjustment of the following three key parameters define the specific vibration mode of the photomechanical oscillator: Oscillation angle (α): used to set the angle of the microsphere's vibration direction relative to the normal direction of the nuclear membrane contact point, typically ranging from -90° to +90°. This parameter directly determines the directionality of the stimulating force applied to the cell nucleus.

[0038] Vibration frequency (f): Used to set the number of times the microsphere reciprocates per second. It can be directly controlled by adjusting the switching speed of the optical potential trap, and the range is within 100 Hz.

[0039] Amplitude (A): Used to set the spatial amplitude of each vibration of the microsphere, typically in the hundreds of nanometers range. This parameter is coordinated with the vibration frequency.

[0040] The parameter control unit executes the following core control logic: 1. Parameter analysis and position calculation: Receive the set oscillation angle (α) and amplitude (A), and calculate and determine two target spatial positions (i.e., the first position and the second position) based on the relative geometric relationship between the microsphere and the nuclear membrane. These two positions are symmetrical about the contact point, the direction of the line connecting them is defined by α, and the distance between them is defined by A.

[0041] 2. Timing control signal generation: Based on the set vibration frequency (f), determine the precise rate at which the optical potential trap alternates between the two target positions.

[0042] 3. Execution: The calculated position coordinates and switching rate are converted into a specific radio frequency control signal to drive the acousto-optic deflector (AOD). This signal drives the AOD to operate, causing the single optical potential well generated by the optical tweezers unit to alternately transition between the first and second positions at a set switching rate.

[0043] In this embodiment, the biomarker unit provides "stains" for observing cellular responses, including live-cell dyes that label the cell nucleus and cytoskeleton, and antibodies that label specific mechanosensitive proteins (such as Lamin A / C, YAP). This allows for the observation of stimulus-induced nuclear rotation, changes in neuronal growth direction, or the establishment of connections with neighboring cells through optical manipulation and imaging units.

[0044] Based on the above system, this embodiment can achieve the following effects: a. It can achieve non-invasive and precise intervention in living cells, directly apply controllable mechanical stimulation to the cell nucleus, and break through the cell membrane barrier.

[0045] b. It provides highly parameterized and programmable stimulation modes, precisely controlling the direction, frequency, and amplitude of stimulation to achieve micro-amplitude directional oscillations. As shown in Figure 2, this method of oscillating the cell nucleus causes it to rotate, and the change in the nucleus's position affects the cell's growth direction. Furthermore, it activates the Lamin A / C protein, which influences nuclear structure, and promotes the nuclear translocation of the mechanotransduction factor YAP protein, which is related to cell growth and proliferation. These effects enable the oscillating nucleus to regulate the growth direction of nerve cells.

[0046] c. Real-time observation and revelation of changes in nuclear physical state (nuclear rotation), responses to key nuclear signaling pathways (lamin A / C and mechanotransducer YAP) and ultimate changes in cell behavior (changes in cell growth direction) triggered by direct nuclear stimulation, establishing a complete stimulus-response causal chain.

[0047] Example 2 This example uses the system shown in Example 1 to realize a method for cell regulation based on photodynamic oscillation stimulation of the cell nucleus.

[0048] In this embodiment, a Nikon Eclipse Ti-U inverted microscope was used, integrating a Tweez250si scanning optical tweezers system (1064 nm laser). A 60x water immersion objective was employed. HT22 neurons were co-cultured with 1-micron silica microspheres for 12 hours to internalize the microspheres into the neurons, as shown in Figure 3.

[0049] In the parameter control unit, dual optical potential traps are set. The oscillation angle (α) is set by adjusting the direction of the line connecting the two optical potential trap points. The frequency (f) is set by changing the scanning rate of AOD (the switching speed of the two optical potential traps). Under a fixed optical potential trap switching rate, the amplitude (A) is controlled by setting the distance between the two optical potential trap points.

[0050] During this process, by setting different switching rates of the optical potential trap (20, 40, 60, 80, 100 Hz), the actual oscillation of the optomechanical oscillator is shown in Figure 4. As the switching rate of the optical potential trap increases from 20 Hz to 100 Hz, the oscillation amplitude decreases, while the actual oscillation frequency of the optomechanical oscillator increases linearly with the switching rate of the optical potential trap. When the switching rate of the optical potential trap is about 80 Hz, the peak response is reached. After exceeding this frequency, the optomechanical oscillator begins to decrease as the switching rate of the optical potential trap continues to increase.

[0051] Figure 5 shows a schematic diagram of the vibration of the microspheres after being controlled by photomechanical forces. By switching the optical potential trap at a frequency of 80 Hz, the photomechanical oscillator generates oscillatory motion on the nuclear membrane. Through repeated collisions with the nuclear membrane, the photomechanical oscillator induces localized periodic micro-deformations of the nuclear membrane.

[0052] In this embodiment, as shown in Figure 6, the angle between the long axis of the cell nucleus before and after rotation is defined as the nuclear rotation angle, with the long axis direction of the cell nucleus as the reference.

[0053] First, cell nuclei were stimulated and photographed at a fixed frequency f = 80 Hz with different oscillation angles α. The results are shown in Figures 7-9. The induced nuclear rotation angle θ was largest at α = 30°, averaging approximately 32°, while almost no rotation was observed at α = 0°. This demonstrates that the oscillation angle is a key parameter controlling the direction and efficiency of nuclear rotation, and that an optimal angle exists. Through systematic analysis of the relationship between θ and the optical trap oscillation angle and switching rate, the measured rotation angle θ first increased and then decreased with α (Figure 10a). The rotation angle was smallest at α ≈ 0°, then increased, reaching a peak near α ≈ 36° (maximum average θ approximately 28°). After exceeding this peak, θ gradually decreased. Quantifying the relationship between nuclear rotation and the optical trap switching rate, as shown in Figure 10b, revealed that increasing the optical trap switching rate significantly increased the nuclear rotation angle θ. The rotation angle was minimal at low frequencies, but increased with higher frequencies, reaching a plateau at approximately 80 Hz with an average rotation angle of approximately 26.4°.

[0054] Under the conditions of α=30° and f=80 Hz, an experimental group and an unstimulated control group were established. After stimulation, cells were fixed with 4% paraformaldehyde and subjected to immunofluorescence staining: first treated with rabbit anti-Lamin A / C primary antibody and rabbit anti-YAP primary antibody, then visualized with green fluorescent secondary antibody. Fluorescence microscopy revealed (Figures 11-12) that the Lamin A / C fluorescence signal at the edge of the cell nucleus in the experimental group was significantly stronger than that in the control group; at the same time, the YAP fluorescence signal in the cell nucleus of the experimental group was also significantly stronger. This reveals that the mechanical stimulation in this embodiment can directly and specifically enhance nuclear laminin and promote YAP entry into the cell nucleus, that is, activate the key intranuclear mechanotransmission pathway.

[0055] Two HT22 cells that are close to each other but not connected were selected. One cell was randomly chosen as the target cell, and its nucleus was stimulated with the optimal parameters described above (α=30°, f=80 Hz). As shown in Figures 13-14, the nucleus of the stimulated cell rotated, and the overall growth direction of the cell followed the nucleus's deflection. Furthermore, the growth tip (growth cone) of the stimulated cell gradually turned towards the neighboring cell, eventually successfully contacting and forming a connection; while the control group cells did not exhibit this directional behavior. This confirms that directional oscillatory stimulation of a single cell nucleus can achieve active guidance of intercellular connection behavior.

[0056] The specific embodiments described above further illustrate the purpose, technical solution, and beneficial effects of the present invention. It should be understood that the above description is only a specific embodiment of the present invention and does not limit the scope of protection of the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for cell regulation based on photodynamic oscillation stimulation of the cell nucleus, characterized in that, Includes the following steps: S1. Provide live cells containing an internalized photomechanical oscillator positioned near the cell nucleus; S2. Capture the photomechanical oscillator using an optical tweezers module, bringing it into contact with or close proximity to the nuclear membrane of the cell nucleus; S3. According to preset oscillation parameters, control the optical potential trap generated by the optical tweezers system to alternate between a first and a second position to drive the photomechanical oscillator to vibrate mechanically, thereby applying controllable local mechanical stimulation to the cell nucleus.

2. The method for cell regulation based on photodynamic oscillation stimulation of the cell nucleus according to claim 1, characterized in that, The optomechanical oscillator is a biocompatible modified microsphere with a diameter of 0.5 μm to 2 μm. It is induced by co-culture to induce endocytosis in cells, thereby taking the microsphere into the cell.

3. The method for cell regulation based on photodynamic oscillation stimulation of the cell nucleus according to claim 2, characterized in that, The oscillation parameters include oscillation angle, vibration frequency, and amplitude; the oscillation angle is the angle between the vibration direction of the photomechanical oscillator and the normal direction of the nuclear membrane contact point, ranging from -90° to +90°; the vibration frequency is the number of oscillations per second of the photomechanical oscillator, which is adjusted by the switching speed of the optical potential trap, ranging from 0 to 100 Hz; the amplitude is the vibration amplitude of the photomechanical oscillator, ranging from 100 to 1000 nanometers.

4. The method for cell regulation based on photodynamic oscillation stimulation of the cell nucleus according to claim 3, characterized in that, Based on the oscillation angle and amplitude settings, the first position and the second position are determined; based on the vibration frequency settings, the switching rate of the optical potential trap between the first position and the second position is controlled; wherein, the direction of the line connecting the first position and the second position corresponds to the oscillation angle, and the distance between them corresponds to the amplitude.

5. The method for cell regulation based on photodynamic oscillation stimulation of the cell nucleus according to claim 1, characterized in that, The method is used to regulate cell behavior by observing the physical response of the cell nucleus and / or subsequent behavioral changes of the cell while or after the application of controlled local mechanical stimulation.

6. The method for cell regulation based on photodynamic oscillation stimulation of the cell nucleus according to claim 5, characterized in that, Physical responses of the cell nucleus and / or subsequent behavioral changes of the cell include nuclear rotation, altered growth direction of neurons, or the establishment of connections between cells and neighboring cells.

7. A system for cell regulation based on photodynamic oscillation stimulation of the cell nucleus, characterized in that, A method for implementing cell regulation based on photodynamic oscillation stimulation of the cell nucleus as described in any one of claims 1-6, comprising: an optical manipulation and imaging unit for capturing and driving microspheres and observing cells in real time; an optomechanical oscillator for the microspheres captured and driven by the optical manipulation and imaging unit; a parameter control unit connected to the optical manipulation and imaging unit for receiving set oscillation parameters and controlling the optical manipulation and imaging unit to generate corresponding light field motion; and a biolabeling unit for labeling cells to observe their mechanical response.

8. A system for cell regulation based on photodynamic oscillation stimulation of the cell nucleus according to claim 7, characterized in that, The optical manipulation and imaging unit includes an optical tweezers module and an integrated microscopy imaging module. The integrated microscopy imaging unit supports fluorescence microscopy and bright-field microscopy modes, switching between imaging modes by changing the light source and filter path. The optical tweezers module includes: a near-infrared laser source that outputs a near-infrared laser beam; an acousto-optic deflector located in the output optical path of the near-infrared laser source to control the propagation direction of the near-infrared laser beam; a beam expander located in the output optical path of the acousto-optic deflector to adjust the spot size of the near-infrared laser beam; a high numerical aperture (HFA) objective lens that focuses the near-infrared laser beam to form an optical potential trap; and a dichroic mirror located in the rear optical path of the HFA objective lens to reflect the near-infrared laser beam from the beam expander back to the HFA objective lens. The integrated microscopy imaging module and the optical tweezers module share the same HFA. The microscope employs a high numerical aperture objective lens and a dichroic mirror. Fluorescence microscopy imaging modes include: a fluorescence excitation source that outputs excitation light to excite the sample and generate a fluorescence emission signal; an image detector that acquires the fluorescence emission signal generated by the sample; a dichroic mirror that reflects the excitation light output from the fluorescence excitation source to the sample and transmits the fluorescence emission signal generated by the sample to the image detector; and a high numerical aperture objective lens that collects the fluorescence emission signal generated after the sample is excited. Bright-field microscopy imaging modes include: a bright-field illumination source that outputs wide-field visible light to illuminate the sample; an image detector that acquires the visible light transmitted or scattered light from the sample; a dichroic mirror that transmits the visible light transmitted or scattered light from the sample to the image detector; and a high numerical aperture objective lens that collects the visible light transmitted or scattered light generated after the sample is illuminated by the bright-field illumination source.

9. A system for cell regulation based on photodynamic oscillation stimulation of the cell nucleus according to claim 7, characterized in that, The control logic of the parameter control unit is as follows: receiving the oscillation angle, amplitude, and vibration frequency set for the optomechanical oscillator; calculating and determining two target spatial positions, namely the first position and the second position, based on the oscillation angle and amplitude; determining a switching rate based on the vibration frequency; and outputting a drive signal to the acousto-optic deflector of the optical manipulation and imaging unit, so that it controls the optical potential trap to alternately transition between the two target spatial positions at the switching rate.

10. A system for cell regulation based on photodynamic oscillation stimulation of the cell nucleus according to claim 7, characterized in that, The biomarker unit includes live cell dyes and / or specific antibodies for labeling the cell nucleus, cytoskeleton, or mechanosensitive proteins, thereby enabling the observation of stimuli-induced changes in cell nucleus morphology, localization, or protein position through optical manipulation and imaging units.

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