Heart rate regulation and control device, method and system based on light-operated otolith oscillation

Abstract

The invention discloses a heart rate regulation and control device, method and system based on light-operated otolith oscillation, and relates to the field of heart rate control. The scanning optical tweezers system comprises a laser scanning light path, an imaging light path, an illumination light path and an electric objective table; the laser scanning light path is used for generating a high-speed scanning optical potential well at the otolith of a target living body; the imaging light path comprises a high-magnification inverted objective lens imaging light path and a low-magnification bypass imaging light path which are respectively used for real-time image acquisition of a target living body and heart state monitoring of the target living body; a first illumination light path in the illumination light paths is used for illuminating an imaging view field of a target living body on the electric objective table, and a second illumination light path is used for exciting the target living body to generate a calcium ion fluorescence signal; the electric objective table is used for bearing a target living body, and the position of the target living body is moved through displacement control of the objective table. The heartbeat rhythm can be accurately regulated and controlled.

Description

Technical Field

[0001] This application relates to the field of heart rate control, and particularly to a heart rate regulation device, method and system based on optically controlled otolith oscillation. Background Art

[0002] Traditional heart rhythm regulation methods, including drug therapy and optical pacing, etc., face different challenges respectively. Drug therapy faces several challenges: on the one hand, anti-arrhythmic drugs may produce toxic side effects; optical pacing uses light-sensitive proteins to regulate the electrophysiological activities of cardiac cells. When a specific wavelength of light beam irradiates cardiac cells, the light-sensitive proteins will undergo conformational changes, thereby achieving precise regulation of the electrophysiological signals of cardiac cells. However, optical pacing technology is still in the research stage and there are some problems that need to be optimized. For example, cardiac photosensitive protein transformation needs to be carried out in the early stage, the operation is complex and the treatment cycle is long, etc. At the same time, the penetration depth of light is limited and it is difficult to reach the deep cardiac cells. Summary of the Invention

[0003] The purpose of this application is to provide a heart rate regulation device, method and system based on optically controlled otolith oscillation, which can accurately regulate the heartbeat rhythm based on the multifunctional optically controlled oscillation of otoliths by combining a scanning optical tweezer system and calcium ion imaging technology.

[0004] To achieve the above purpose, this application provides the following solutions: In the first aspect, this application provides a heart rate regulation device based on optically controlled otolith oscillation, including: a scanning optical tweezer system; the scanning optical tweezer system includes a laser scanning optical path, an imaging optical path, an illumination optical path and an electric stage; The laser scanning optical path is used to generate a high-speed scanning optical potential well at the otolith of the target living body; The imaging optical path includes an imaging optical path of a high-magnification inverted objective lens and a low-magnification side-branch imaging optical path, which are respectively used for real-time image acquisition of the target living body and for monitoring the cardiac state of the target living body; The illumination optical path includes a first illumination optical path and a second illumination optical path; the first illumination optical path is used for illuminating the imaging field of view of the target living body on the electric stage; the second illumination optical path is used for exciting the target living body to generate calcium ion fluorescence signals; The electric stage is used to carry the target living body and move the position of the target living body by controlling the displacement of the stage.

[0005] Optionally, the laser scanning optical path includes a 1064 nm laser, an acousto-optic deflector, a beam expander, a short-wave pass dichroic mirror and an inverted objective lens; the laser emitted by the 1064 nm laser is incident on the imaging optical path of the high-magnification inverted objective lens in sequence through the acousto-optic deflector, the beam expander, the short-wave pass dichroic mirror and the inverted objective lens.

[0006] Optionally, the imaging optical path of the high-magnification inverted objective lens consists of a high-magnification inverted objective lens and a first high-speed charge-coupled device; the first high-speed charge-coupled device is vertically arranged with the high-magnification inverted objective lens and is used to collect real-time image data of the target living body through the high-magnification inverted objective lens.

[0007] Optionally, the low-magnification side-branch imaging optical path consists of a low-magnification electron microscope and a second high-speed charge-coupled device; the second high-speed charge-coupled device is vertically arranged with the low-magnification electron microscope and is used to observe the heart condition of the target living body in real time.

[0008] In a second aspect, the present application provides a heart rate regulation method based on the above-mentioned heart rate regulation device based on opto-otolith oscillation, including: Generating at least two optical traps at the edge position of the otolith of the target living body through a scanning optical tweezer system; Dynamically adjusting the switching frequency, power, and spatial distribution pattern of the optical traps through the scanning optical tweezer system to drive the otolith to oscillate according to set parameters; Real-time collecting the heart beating video of the target living body and calculating the heart rate change amount through an image processing algorithm; According to the target heart rate regulation requirement, adjusting the otolith oscillation parameters to adjust the heart rate of the target living body; wherein, the otolith oscillation regulates the heart rate through the vestibular-central nervous system-heart axis neural pathway; the otolith oscillation is used to activate the hair cells connected to the bottom of the otolith to generate auditory signals; the auditory signals are transmitted to the heart through the hypothalamus and the vagus nerve motor nucleus in the dorsal medulla oblongata, causing the sympathetic nerve to release norepinephrine, which binds to the specific β-adrenergic receptor on the myocardial cell membrane, making the myocardial contraction heart rate accelerate.

[0009] Optionally, before generating at least two optical traps at the edge position of the otolith of the target living body through the scanning optical tweezer system, it further includes: a pretreatment step for the target living body, and the pretreatment step includes anesthetizing and fixing the target living body.

[0010] Optionally, generating at least two optical traps at the edge position of the otolith of the target living body through the scanning optical tweezer system specifically includes: Using a 1064 nm laser to generate a deflected light beam through an acousto-optic deflector; Based on a beam expander, expanding the diameter of the deflected light beam to be able to cover the entrance pupil of the high-magnification inverted objective lens; Reflecting the near-infrared laser in the deflected light beam to the high-magnification inverted objective lens through a short-wave pass dichroic mirror and focusing the near-infrared laser to the edge position of the otolith of the target living body to form an optical trap.

[0011] In a third aspect, the present application provides a heart rate regulation system, including: An optical potential well generation module for generating at least two optical potential wells at the edge position of the otolith of a target living body through a scanning optical tweezers system; An oscillation module for dynamically adjusting the switching frequency, power and spatial distribution mode of the optical potential well through a scanning optical tweezers system to drive the otolith to oscillate according to set parameters; A heart rate change calculation module for collecting a video of the heart beating of a target living body in real time and calculating the heart rate change through an image processing algorithm; A heart rate regulation module for adjusting the otolith oscillation parameters according to the target heart rate regulation requirement to adjust the heart rate of the target living body; wherein, the otolith oscillation regulates the heart rate through the vestibular-central nervous system-heart axis neural pathway; the otolith oscillation is used to activate the hair cells connected to the bottom of the otolith to generate auditory signals; the auditory signals are transmitted to the heart through the vagus nerve motor nucleus in the hypothalamus and the dorsal medulla oblongata, causing the sympathetic nerve to release norepinephrine, which binds to specific β-adrenergic receptors on the myocardial cell membrane, causing the myocardial contraction heart rate to increase.

[0012] Optionally, it further includes: A pretreatment module for performing pretreatment steps on the target living body, and the pretreatment steps include anesthetizing and fixing the target living body.

[0013] Optionally, the optical potential well generation module specifically includes: A deflected beam generation unit for using a 1064 nm laser to generate a deflected beam through an acousto-optic deflector; An expanding beam unit for expanding the diameter of the deflected beam to be able to cover the entrance pupil of a high-magnification inverted objective lens based on an expanding lens; An optical potential well generation unit for reflecting the near-infrared laser in the deflected beam to a high-magnification inverted objective lens through a short-wave pass dichroic mirror and focusing the near-infrared laser to the edge position of the otolith of the target living body to form an optical potential well.

[0014] According to the specific embodiments provided by the present application, the present application discloses the following technical effects: The present application provides a heart rate regulation device, method and system based on opto-controlled otolith oscillation. First, the laser scanning optical path in the scanning optical tweezer system generates a high-speed scanning optical potential well at the otolith of the target living body. The optical potential well can precisely capture and manipulate the otolith to generate controllable oscillations. Then, the imaging optical path of the high-magnification inverted objective is used to collect images of the target living body in real time, ensuring that the operator can clearly observe the dynamic changes of the otolith and surrounding tissues. At the same time, the low-magnification side-branch imaging optical path is specifically used to monitor the heart state of the target living body. In terms of illumination, the first illumination optical path provides sufficient illumination for the target living body on the motorized stage to ensure clear imaging. The second illumination optical path is used to excite the target living body to generate calcium ion fluorescence signals, which is a key step in calcium ion imaging technology. The neural signal pathway of opto-controlled otolith oscillation regulating heart rate can be revealed through the intensity changes of the fluorescence signals. The motorized stage bears the target living body and can adjust its position as needed to ensure that the laser scanning optical path, imaging optical path and illumination optical path can all accurately align with the target area. Through the synergistic effect of the above system, the operator can precisely control the oscillation frequency and amplitude of the otolith, thereby achieving precise regulation of the heartbeat rhythm. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings required for use in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present application. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0016] Figure 1 Structural schematic diagram of a heart rate regulation device based on opto-controlled otolith oscillation provided by an embodiment of the present application; Figure 2 Schematic diagram of the principle of opto-controlled otolith oscillation regulating heart rate provided by an embodiment of the present application; Figure 3 Schematic diagram of realizing multi-functional opto-control oscillation of otoliths using programmable scanning optical technology in vivo provided by an embodiment of the present application; Figure 4 Schematic diagram of realizing precise heart rate regulation based on opto-controlled otolith oscillation provided by an embodiment of the present application; Figure 5 Schematic diagram of the brain nerve signal conduction pathway triggered by opto-controlled otolith oscillation provided by an embodiment of the present application; Figure 6 Analysis diagram of the neural pathway of opto-controlled otolith oscillation causing the brain to regulate heart rate provided by an embodiment of the present application; Figure 7 Biomedical application diagram of opto-controlled otolith oscillation regulating heart rate provided by an embodiment of the present application; Figure 8 Flow chart of a heart rate regulation method provided by an embodiment of the present application; Figure 9 Schematic structural diagram of a heart rate regulation system provided by an embodiment of the present application. Detailed implementation manners

[0017] Next, the technical solutions in the embodiments of the present application will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present application without creative efforts belong to the scope of protection of the present application.

[0018] To make the above objects, features, and advantages of the present application more obvious and understandable, the present application will be further described in detail below with reference to the accompanying drawings and specific implementation manners.

[0019] Embodiment 1 As Figure 1 shown, this embodiment provides a heart rate regulation device based on optically controlled otolith oscillation, including: a scanning optical tweezer system; the scanning optical tweezer system includes a laser scanning optical path, an imaging optical path, an illumination optical path, and an electric stage; The laser scanning optical path is used to generate a high-speed scanning optical potential well at the otolith of the target living body; The imaging optical path includes an imaging optical path of a high-magnification inverted objective lens and a low-magnification side-branch imaging optical path, which are respectively used for real-time image acquisition of the target living body and for monitoring the heart state of the target living body; The illumination optical path includes a first illumination optical path and a second illumination optical path; the first illumination optical path is used for illuminating the imaging field of view of the target living body on the electric stage; the second illumination optical path is used to excite the target living body to generate a calcium ion fluorescence signal; The electric stage is used to carry the target living body and move the position of the target living body by controlling the displacement of the stage.

[0020] In this embodiment, the experimental device is built around the scanning optical tweezer system (SOTs), mainly including a laser scanning optical path, an imaging optical path, an illumination optical path, and an electric stage. The laser scanning optical path is used to generate a high-speed scanning optical potential well, mainly including a 1064 nm laser, an acousto-optic deflector (AOD), a beam expander, a short-wave pass dichroic mirror, and an inverted objective lens, etc.

[0021] Selecting a laser wavelength of 1064 nm can avoid thermal damage to the manipulated tissue or cells caused by the photothermal effect of the focused beam because biological tissues absorb less in this wavelength band.

[0022] Among them, the specific working process of the device is as follows: First, the 1064 nm laser emitted by the laser is deflected after passing through the AOD, and the deflected laser beam will be expanded after passing through the beam expander, which consists of two convex lenses. The expanded laser beam is reflected by the short-wave pass dichroic mirror into the inverted objective lens, and the beam cross-sectional diameter can completely cover the entrance pupil of the objective lens. Finally, it is focused on the otolith of the zebrafish ear through a 60x water immersion objective lens (CFI Apo, NA = 1.0). At this time, due to the applied optical gradient force, the focused laser forms a trapping optical potential well, which can realize the multi-mode oscillation control of the otolith. Based on the built scanning optical tweezers system, the beam can be scanned rapidly at a maximum frequency of 100 kHz, and the position, number, power, scanning frequency, and scanning mode of the trapping optical potential well can all be independently set through software.

[0023] In addition, the optical path for microscopy is divided into two paths: (1) The imaging optical path of the high-magnification inverted objective lens, where image acquisition is performed by a high-speed charge-coupled device (CCD), and real-time monitoring, image acquisition, and video recording are carried out through a computer screen; (2) The low-magnification side-branch imaging optical path, which uses a low-magnification electron microscope and CCD and can be used to observe the general situation of the zebrafish heart in real time. The low-magnification side-branch imaging optical path is specifically used to monitor the heart state of the target living body, and the heart rate change amount is calculated through image processing algorithms.

[0024] At the same time, the system has two illumination optical paths. The first illumination optical path is emitted by a halogen light source (D-LH, 12 V / 100 W), converged by a condenser, and then irradiated on the sample stage for illuminating the imaging field of view. The second illumination optical path is a light source equipped with red, green, and blue excitation lights, and the color of the emitted excitation light can be selected through an internal filter, which is used to excite the sample to generate corresponding fluorescence.

[0025] White light source and fluorescence excitation light source: Provide the illumination light required for observing the sample in bright field or fluorescence excitation.

[0026] Acousto-optic deflector: Mechanical waves are generated in the crystal medium, which will cause periodic refractive index changes to become a phase-type diffraction grating. If a laser beam is incident on the medium, the laser beam will be diffracted, and the intensity and direction of the diffracted light change with the intensity and wavelength state of the mechanical wave. Through this principle, the focal position and intensity of the 1064 nm laser can be changed.

[0027] Beam expander: Consists of two convex lenses, and the distance between them is the sum of the focal lengths of the two convex lenses, used to expand the output diameter of the laser so that the diameter can completely cover the entrance pupil of the inverted objective lens.

[0028] Short-pass dichroic mirror: It can transmit or reflect light according to the wavelength to achieve spectral splitting. The long-pass dichroic mirror highly reflects light with a wavelength lower than the cut-off wavelength and highly transmits light with a wavelength higher than the cut-off wavelength; the short-pass dichroic mirror is the opposite, highly transmitting light with a wavelength lower than the cut-off wavelength and highly reflecting light with a wavelength higher than the cut-off wavelength. In this embodiment, a short-pass dichroic mirror is used here to allow the illumination light with a wavelength below 800 nm to pass through while reflecting the near-infrared laser with a wavelength above 800 nm.

[0029] CCD camera: The abbreviation of charge coupled device, which is used to collect images in real time and can set the specific collection frequency through the PC side.

[0030] Embodiment 2 As Figure 8 shown, this embodiment provides a heart rate regulation method based on the heart rate regulation device based on optically controlled otolith oscillation, including: Step 1: Generate at least two optical potential wells at the edge position of the otolith of the target living body through the scanning optical tweezer system; Step 2: Dynamically adjust the switching frequency, power and spatial distribution mode of the optical potential wells through the scanning optical tweezer system to drive the otolith to oscillate according to the set parameters; Step 3: Collect the heart beating video of the target living body in real time and calculate the heart rate change amount through the image processing algorithm; Step 4: Adjust the otolith oscillation parameters according to the target heart rate regulation requirement to adjust the heart rate of the target living body; wherein, the otolith oscillation regulates the heart rate through the vestibular-central nervous system-heart axis neural pathway; the otolith oscillation is used to activate the hair cells connected to the bottom of the otolith to generate auditory signals; the auditory signals are transmitted to the heart through the vagus nerve motor nucleus in the hypothalamus and the dorsal medulla oblongata, causing the sympathetic nerve to release norepinephrine, which binds to the specific β-adrenergic receptor on the myocardial cell membrane, making the myocardial contraction heart rate increase.

[0031] In this embodiment, zebrafish is selected as the target living body.

[0032] In this embodiment, a technology for regulating the heart rate by optically controlling otolith oscillation is developed. By using programmable optical scanning technology, the otolith oscillation is precisely manipulated to induce auditory signals in auditory hair cells and applied to non-invasive heart rate regulation of living bodies. Specifically, the optically controlled otolith oscillation technology uses a programmable scanning optical tweezer system to generate scanning optical potential wells with specific wavelengths and intensities, and then completes the precise, real-time and multi-functional regulation of the otolith oscillation mode.

[0033] In this embodiment, through quantitative characterization, the effects of different otolith oscillation modes on heart rate were determined, and the differences in the effects of otolith oscillation on the heart rates of zebrafish larvae at different developmental stages were verified. Meanwhile, with the help of neuron calcium labeling technology and specific drugs, the internal neural circuit of otolith oscillation regulating heart rate was preliminarily verified. Finally, the treatment and improvement of various heart disease models were achieved by regulating heart rate with otolith oscillation. Compared with existing research methods, optical manipulation has unique advantages of non-contact, non-damage, and programmability, and can accurately trigger and change the motion state of otoliths at the microscopic level, thereby providing an unprecedented technical solution for in-depth and accurate analysis of the change mechanisms of physiological indicators such as auditory signal conduction, emotion regulation, and heart rate regulation.

[0034] Specifically, the otoliths of zebrafish larvae have a high refractive index. When a focused laser beam irradiates the otoliths, the generated optical potential well can stably capture the otoliths. Moreover, when the optical potential well is located at the edge of the otolith, the capture ability is the strongest under the same laser power. In the experiment, this embodiment used a scanning optical tweezer system to generate two optical potential wells, which were respectively set at the edge positions of the otoliths, and then relied on the scanning optical tweezer system to adjust the switching frequency of the two optical potential wells to drive the otoliths to oscillate at a set frequency at high speed. At the same time, according to the experimental requirements, a variety of complex oscillation modes were set, and the oscillation frequency, oscillation amplitude, and oscillation direction were adjusted in real time, so as to complete the precise control of otolith oscillation and simulate a variety of auditory input modes. As shown in a of Figure 2 , since the otoliths are connected to the underlying hair cells through ciliary bundles, oscillating the otoliths can deflect the ciliary bundles, thereby activating the hair cells to form auditory signal input. At the same time, the auditory nerve signal is transmitted through the eighth cranial nerve fibers to the vagal motor nucleus in the medulla oblongata (for preliminary processing and integration of auditory information), and then upward to the semicircular ring in the midbrain (for more complex processing and analysis of auditory information). The auditory information processed by the midbrain is then transmitted to the thalamus and hypothalamus. As an important relay station for sensory conduction, the thalamus can screen, integrate, and modulate auditory information (including regulating the sympathetic and parasympathetic nerves). During this process, the nerve signal can be transmitted from the hypothalamus and the vagal motor nucleus in the dorsal part of the medulla oblongata to the heart, causing the sympathetic nerve to release norepinephrine, which binds to specific β-adrenergic receptors on the myocardial cell membrane and acts on the sinoatrial node (SAP) of the heart to accelerate myocardial contraction and heart rate. Therefore, in this embodiment, through the multifunctional oscillation of the otoliths, precise regulation of the heart beating rhythm was successfully achieved, and its specific state was mainly quantitatively evaluated and measured through changes in heart rate data ( Figure 2 b of

[0035] In some embodiments, before generating at least two optical potential wells at the edge position of the otoliths of the target living body through the scanning optical tweezer system, it further includes: a pretreatment step for the target living body, and the pretreatment step includes anesthetizing and fixing the target living body.

[0036] Specifically, when caring for and treating zebrafish, zebrafish larvae (3 days after fertilization) were purchased from a commercial supplier (Company A). They were raised in a culture dish with a volume of 100 ml, at (28 ± 0.5 °C, 14 h of light / 10 h of darkness), fed with first-feeding diet 3 times a day, 50% of the water body was changed every 24 hours, and a 100-μm pore size nylon mesh was used to separate excreta and residual bait. All experiments were conducted in accordance with the ethical standards of the Animal Ethics Committee.

[0037] In the experiment, to ensure a good operation posture, 2% agarose was used to fix them on a glass slide with a thickness of 1 mm, facilitating subsequent experimental operations and imaging records. When verifying neural pathways with drugs, zebrafish were treated with drug incubation: the same batch of 5 dpf zebrafish embryos were incubated in the culture solutions of MS222, propranolol, and atropine at the same concentration (within the drug concentration tolerance range). After 24 hours, they developed to 6 dpf, and then the otolith oscillation experiment was immediately carried out and the corresponding heart rate was recorded.

[0038] In this example, three drugs, terfenadine, sunitinib, and tolterodine, were used to establish atrioventricular block, cardiac escape, and bradycardia heart disease models. Before the experiment, a stock solution of terfenadine (Terfenadine) was prepared in advance, and an appropriate amount of the terfenadine stock solution was added to the culture solution and sonicated for 10 minutes to adjust the terfenadine concentration in the culture solution to 5 μM. Subsequently, in this example, 5 dpf zebrafish embryos were placed in the culture solution containing the drug for co-incubation. After 24 hours of cultivation, zebrafish embryos with a successfully constructed atrioventricular block disease model were screened out through a microscope; similarly, 6 dpf zebrafish embryos were co-incubated with the embryo culture solution containing 0.5 μM sunitinib (Sunitinib) for 5 hours, and then the cardiac escape disease larvae were screened out; 5 dpf zebrafish embryos were co-incubated with the culture solution containing 50 μM tolterodine (Tolterodine) for 24 hours, and then the bradycardia disease larvae were screened out.

[0039] Among them, when generating at least two optical traps at the edge position of the otolith of the target living body through a scanning optical tweezer system, it specifically includes: Using a 1064 nm laser to generate a deflected beam through an acousto-optic deflector; Based on an expander, expanding the diameter of the deflected beam to be able to cover the entrance pupil of a high-magnification inverted objective lens; Reflecting the near-infrared laser in the deflected beam through a short-wave pass dichroic mirror to achieve opto-controlled oscillation of the otolith; According to the high-magnification inverted objective lens, focusing the near-infrared laser to the edge position of the otolith of the target living body to form an optical trap.

[0040] Specifically, a programmable scanning optical technique is used in zebrafish to achieve multifunctional optical control of otolith oscillation: The prerequisite for precisely controlling otolith oscillation is that the optical trap can stably capture the otolith and drive its directional movement. Therefore, in this embodiment, the performance of the optical trap for capturing the otolith is first experimentally characterized. As Figure 3 shown in a1 of , the otolith of zebrafish larvae is connected to the ciliary bundles of hair cells. When there is no external vibration, the otolith remains stationary. When a single optical trap is set at the left edge of the otolith, it can be found that the otolith moves about 1.37 Figure 3 to the left compared to the original position ( a2 in Figure 3 ). Similarly, when the optical trap is located at the right edge of the otolith, the otolith moves 1.28 to the right ( Figure 3 a3 in ). In addition to moving left and right, this embodiment can also control the up and down movement of the otolith, driving the otolith to displace 1.06 and 0.97 vertically ( Figure 3 a4 - a5 in

[0041] ). The above experimental phenomena preliminarily prove that the otolith can be stably captured based on the scanning optical tweezer system, and then the multifunctional oscillation of the otolith can be precisely controlled based on two optical traps. Figure 3 As shown in b of

[0042] , it can be found from the waveform diagram that the oscillation frequency of the otolith always remains synchronized with the set OT scanning frequency. Figure 3 Furthermore, in this embodiment, the scanning frequency is gradually increased to 40 Hz, and the otolith can also synchronously respond to the oscillation ( Figure 3 c in

[0043] ). After proving that the otolith can respond well to the scanning frequency of the two optical traps and oscillate, this embodiment explores the oscillation amplitude and the maximum oscillation displacement in different directions of the otolith. As Figure 3 shown in d of

[0043] , as the scanning frequency of the optical trap continuously increases (the power remains unchanged), the oscillation amplitude of the otolith decreases. This is because the ciliary bundles of hair cells connected to the bottom of the otolith have a certain rigidity. When the scanning frequency of the optical trap increases, the acting time of the optical force applied to the left and right edges of the otolith decreases, resulting in a decrease in the oscillation displacement of the otolith with the increase of frequency, that is, the oscillation amplitude decreases.

[0043] Furthermore, in this embodiment, by changing the power of the two optical potential wells (while keeping the frequency unchanged), it can be seen that as the trapping force increases, the oscillation amplitudes of the utricular otolith (Ut) and the saccular otolith (Sac) increase accordingly. As can be seen from Figure 3 e in, under the same trapping force of the optical potential well, Ut has a larger displacement than Sac, which is due to the different masses of the two and the different lengths of the cilia connected at the bottom.

[0044] On this basis, this embodiment further explores the oscillation flexibility of the otoliths in different directions. As shown in Figure 3 f, along the head-tail direction of the zebrafish (0-180° horizontal direction), the otoliths can generate the largest displacement, and both types of otoliths show the same pattern. The above experiments confirm that flexible oscillation of the otoliths can be achieved based on the scanning optical tweezers, laying a foundation for the next experimental research.

[0045] On the basis of stably trapping and precisely oscillating the otoliths using the optical potential well (Trap), this embodiment realizes the real-time programming of the optical potential well by means of an acousto-optic deflector, and then develops a variety of complex otolith oscillation modes, which can perform multi-mode switching in terms of the number of optical traps, oscillation direction, oscillation frequency, and oscillation amplitude, providing a variety of means for exploring auditory input and auditory-induced physiological activities. As shown in Figure 3 g1, two optical potential wells can achieve unidirectional oscillation of the otoliths, and the scanning sequence switches back and forth between Trap1 and Trap2 in turn to achieve controllable oscillation of the otoliths along the horizontal direction. When controlling three optical potential wells to scan in the order of Trap1, Trap2 to Trap3, multi-directional oscillation of the otoliths around the center to the outside can be achieved ( Figure 3 g2). If four optical potential wells are introduced and scanned in the order shown in Figure 3 g3-g4, the otoliths can be made to oscillate in an "8" shape and a "cross" shape around the center respectively.

[0046] It should be noted that different oscillation modes cause different deflection modes and intensities of the ciliary bundles, and at this time, the mechanical force stimuli perceived by the hair cells will also be different, so different intensities of auditory input signals are generated. Based on this, this embodiment can apply different otolith oscillation modes by setting different oscillation frequencies and amplitudes, and then complete complex and variable auditory input manipulation to further simulate the complex state of real auditory input, so as to more accurately study the influence law of otolith oscillation on heart rate.

[0047] Among them, when performing step 4, precise regulation of the heart rate is realized based on optically controlled otolith oscillation, and the specific process is as follows: In the above experiment, this embodiment successfully realized the multi-mode oscillation of otoliths based on a scanning optical tweezers system. It should be noted that in the experiment, this embodiment observed that the heart rate changed correspondingly with the oscillation of the otoliths. To explore the influence law of optically controlled otolith oscillation on the heart rate, this embodiment quantitatively characterized the corresponding heart rate change amount (HRV) by changing the oscillation amplitude, frequency, and number of otoliths.

[0048] Figure 4 a in [reference] shows the position distribution of otoliths and the heart in zebrafish larvae: there is an otolith cavity on each side of the zebrafish head, and there are two types of otoliths, Sac and Ut, in the otolith cavity (the corresponding fluorescence images are as shown in Figure 4 b in [reference]). The red fluorescence is the blood vessels in the brain, and the green fluorescence is the heart. In the experiment, this embodiment first performed optical oscillation of the otoliths at 50 Hz. After the stimulation lasted for about 5 seconds, the laser was turned off, and the oscillation of the otoliths stopped immediately. At the same time, a bypass low-magnification electron microscope was used to collect images and record videos of the heart beating process. By analyzing the heartbeat video, a zebrafish heartbeat waveform diagram ( Figure 4 c in [reference]) can be obtained. Through quantitative analysis of it, this embodiment found that the normal heartbeat frequency of zebrafish before otolith oscillation was about 2.1 Hz; when an optical potential well was applied to oscillate the otoliths, the heartbeat frequency gradually increased to 2.8 Hz. As the oscillation of the otoliths stopped, the heartbeat frequency returned to 2.1 Hz again. To reduce errors and more accurately evaluate the change in the zebrafish heartbeat frequency, this embodiment introduced the heart rate change amount (Heart Rate Variability, HRV) for quantitative analysis. It is defined as the time variance between successive heartbeat cycles and can be used to reflect the heartbeat frequency fluctuations of the heart at different time points. The magnitude represents the speed of heart rate change.

[0049] To rule out the possibility that the change in heart rhythm might be due to the heart rate stress acceleration caused by the body perturbation or stimulation of zebrafish, this embodiment set up multiple control experiments for verification. In the experiment, this embodiment uniformly fixed the larvae on the sample stage with 2% agarose. At the same time, two scanning optical tweezers with fixed power and frequency were set up. The two optical tweezers were applied outside the larval body, on the back of the fish body, in the ear cavity (non-hair cell area), and on the otoliths respectively, while the Control group did not apply an optical potential well. As Figure 4 shown in d in [reference], the optical potential wells applied on the back of the fish body and in the ear cavity (non-hair cell area) would cause slight changes in the heart rate of the larvae, while acting on the otoliths could increase the heart rate by up to about 50%. Further, this embodiment set the optical potential well parameters respectively for directly capturing the non-oscillating otoliths and the dynamically oscillating otoliths after capture. As Figure 4As shown by e in [the figure], the photothermal stimulation that only captures the otolith without oscillation is not sufficient to effectively increase the heart rate. In contrast, optically controlled otolith oscillation can cause a significant increase in the heart rate. Based on the above experiments, in this embodiment, it can be determined that the main factor affecting the heart rate change by optically controlled otolith oscillation is not photothermal stimulation, but the auditory signal conduction of hair cells caused by otolith oscillation, which in turn triggers the heart rate change.

[0050] After determining the main factors affecting the heart rate change, this embodiment further controls the amplitude and frequency of otolith oscillation to quantitatively explore its influence law on the heart rate change. As Figure 4 shown by f in [the figure], in this embodiment, otolith oscillation is performed on 6-dpf larvae, and at the same time, the oscillation amplitude is adjusted from the initial 1 gradually increased to 4 . It can be seen that as the otolith oscillation amplitude increases, the value of HRV also increases linearly. This may be because the magnitude of the otolith oscillation amplitude affects the signal conduction intensity of hair cells. When the otolith oscillation amplitude is small, the mechanical deflection of the bottom-connected ciliary bundle is reduced, resulting in a relatively weak mechanical conduction signal of hair cells. Therefore, the signal conduction intensity affecting the heart rate change is also relatively weak, leading to a relatively small change in HRV. On the contrary, when the otolith oscillation amplitude is the largest, the mechanical deflection of the ciliary bundle is the largest, and the nerve signal intensity is also the strongest. At the same time, the signal transmitted to the next neuron region is stronger, which in turn acts on the heart to accelerate its contraction frequency, and finally significantly increases the change in HRV. At the same time, this embodiment also characterized the change law of HRV with the oscillation frequency when the oscillation amplitude is the same. As Figure 4 shown by g in [the figure], when the otolith oscillates at 10 Hz, 50 Hz, and 100 Hz, the magnitude of HRV shows a certain growth trend, but the difference is relatively small. This may be because the types of ciliary bundles connected to the oscillating otolith are relatively few, different from the whole ear which contains a variety of hair cells that can detect mechanical oscillations of different frequencies respectively. At this time, the ability of the ciliary bundle connected to the otolith to distinguish the oscillation frequency is relatively weak, so the difference in the influence on the heart rate is relatively small.

[0051] Since the types of ciliary bundles connected to different types of otoliths are different, the influence law on the heart beat will also be different. In response to this, in this embodiment, the same optical potential well oscillation parameters are respectively applied to the Sac and Ut otoliths on the same side of zebrafish larvae, and their respective influence laws on the heart rate are compared in real time. As Figure 4As shown by h in [reference], at the same oscillation amplitude and oscillation frequency, the oscillating Sac causes a greater change in heart rate compared to the oscillating Ut. From this example, it can be known that for the utricular otoliths (Ut) responsible for maintaining the body's vestibular balance, their response to heart rate is smaller than that of the sacculus otoliths (Sac) responsible for hearing. In subsequent experiments, this example also uniformly uses the experimental data of the Sac otoliths for illustration. On this basis, this example also compares the differences in heart rate changes when oscillating Ut and Sac separately and when oscillating both otoliths simultaneously. As Figure 4 As shown by i in [reference], the sum of the HRV values when oscillating the utricular otoliths (Ut) and the sacculus otoliths (Sac) separately is relatively close to the HRV value when oscillating both otoliths simultaneously. From this, it can be known that the effect of oscillating the otoliths on heart rate has an additive effect.

[0052] In addition, in some embodiments, the analysis and verification of the neural pathway for heart rate regulation based on optically controlled otolith oscillation can be as follows: Based on the above experiments, this example further explores the internal signal pathway of optically controlled otolith oscillation regulating heart rate. Based on the previous literature research, this example preliminarily proposes a hypothesis for the neural pathway of otolith oscillation regulating heart rate: the vestibular - central nervous system - heart axis. Specifically, the vestibular auditory signals generated by otolith oscillation will enter the relevant regions of the brain for processing, and then the brain makes corresponding decisions, and then adjusts the myocardial contraction of the heart to regulate the heart rate. To verify the above signal pathway mechanism, this example quantitatively explores the signal response of the central nervous system caused by optically controlled otolith oscillation based on calcium ion fluorescence labeling. Specifically, calcium imaging is used to image the neurons in the zebrafish brain to characterize the neural signal circuit for regulating heart rhythm after optically controlled otolith oscillation.

[0053] First, this example characterized the response of otolith oscillation to relevant neural regions of the brain. The commonly used methods for detecting the signal transmission activities of brain neurons mainly include electrophysiological measurement and neural calcium ion fluorescence detection. The former has a certain degree of invasiveness and is difficult to perform non - destructive detection on zebrafish larvae. Therefore, this example selects a transgenic zebrafish strain (huc:GCaMP6f&nacre) with whole - brain neuron calcium ion fluorescence labeling for experimental exploration. As Figure 5 As shown by a in [reference], this example uses 2% agarose to fix the transgenic zebrafish and at the same time adjusts its body angle to a suitable field of view for brain fluorescence imaging. At the same time, two scanning optical potential traps are set to act on the Sac otoliths, and the specific parameters of the action position, laser power, and scanning frequency are optimized to achieve controllable oscillation of the otoliths at a specific frequency and power. The illumination light is turned off, and at the same time, the blue excitation light is turned on to continuously excite the calcium ion fluorescence signal in the brain. On this basis, based on the programmable optical potential trap to control otolith oscillation, the video data of fluorescence changes is synchronously recorded. As Figure 5As shown by b in [reference], it can be observed that the fluorescence signal of calcium neurons in the whole brain of transgenic zebrafish is successfully excited, where the white box and the yellow box correspond to the eye and the otic cavity respectively.

[0054] Next, the picture sequence of the calcium fluorescence signal in the zebrafish brain during the otolith oscillation process was processed to obtain the neuron calcium fluorescence signal response map of the otolith oscillation at different time stages. The specific operations are as follows: The picture with fluorescence change response after the otolith oscillation starts is recorded as the initial frame (at the 2.8 s moment), and one picture is taken every 0.2 s and the gray value subtraction is performed on the previous picture. Since the fluorescence response time is short once, a total of 6 fluorescence pictures at different moments are intercepted for subtraction. At the same time, a threshold screening is set in this embodiment. The fluorescence changes below the sub-threshold belong to the spontaneous fluorescence response. By analogy, 5 fluorescence pictures with regional fluorescence changes can be obtained. In order to distinguish the fluorescence response regions at each time stage, this embodiment assigns them the colors of red, yellow, green, blue, and purple respectively ( Figure 5 as shown in c1 - c5 in [reference]). Finally, they are aligned and superimposed to obtain the integrated map of the fluorescence response region. As Figure 5 shown by c6 in [reference], the fluorescence response gradually spreads from the initial red region to the final purple region, that is, the signal generated by the otolith oscillation in the brain is transmitted from the hindbrain to the midbrain and then to the forebrain. To clarify the brain regions and their functional roles of the otolith oscillation response, this embodiment matches the obtained fluorescence response region with the zebrafish brain atlas. The three dotted boxes of different colors indicate that the brain signal response regions caused by the otolith oscillation can be divided into three categories. After comparing them with the zebrafish brain atlas, it can be seen that, in the order of signal transmission from bottom to top, they are the vagus motor nucleus region (VMN), the semicircular region (TS), and the thalamus region (Thalamus).

[0055] Based on the above experiments and data analysis, the brain nerve signal conduction pathway triggered by the otolith oscillation can be obtained. However, the signal conduction path of the part of the brain regulating the heart rate still needs to be explored. Since the nerve pathway regulating the heart cannot be directly imaged by calcium fluorescence labeling temporarily, this embodiment uses propranolol and atropine drugs to act on the specific receptors on the heart, and then explores whether the path of the specific receptor action is the key to controlling the heart rate change. Figure 6 a in [reference] shows the flow chart of the drug incubation treatment for zebrafish: Select the same batch of zebrafish embryos at 5 dpf and incubate them in the culture solutions of the same concentration of MS222, propranolol, and atropine (within the drug concentration tolerance range). After 24 hours, they develop to 6 dpf, and then the otolith oscillation experiment is carried out and the corresponding heart rate is recorded.

[0056] Previous studies and pharmacological experiments have demonstrated that propranolol is a β-adrenergic receptor blocker that mainly acts on the sympathetic nervous system. Under normal circumstances, when the sympathetic nerve is in an excited state, its terminals release norepinephrine, which binds to β-adrenergic receptors on the heart, thereby achieving the effects of increased heart rate and enhanced myocardial contractility. As Figure 6 shown in b of

[0057] , after incubation with propranolol, when the otolith is optically oscillated, it can be found that the heart HRV value decreases with the increase in drug concentration. This is because propranolol can competitively block β-adrenergic receptors and prevent norepinephrine from binding to the receptors. Therefore, when zebrafish are in the state of otolith oscillation, if propranolol is administered, even if the sympathetic nervous system is excited by otolith oscillation stimulation, the norepinephrine it releases cannot effectively bind to β-receptors and thus cannot exert its effect. As a result, the heart rate acceleration effect originally caused by sympathetic nerve excitation is weakened, and the heart rate increase amplitude decreases.

[0057] On the contrary, after incubation with atropine, the corresponding heart HRV value continuously increases with the increase in drug concentration ( Figure 6 shown in c of Figure 6 ). This is because atropine is an M-type acetylcholine receptor antagonist that mainly acts on the parasympathetic nervous system. In normal physiological conditions, when the parasympathetic nerve is excited, its terminals release acetylcholine, which binds to M-type acetylcholine receptors on the heart, increasing the permeability of the myocardial cell membrane to potassium ions, and potassium ions thus flow outwards. Such a change leads to hyperpolarization of myocardial cells, thereby slowing down the heart rate and weakening myocardial contractility at the same time. When atropine is administered to zebrafish, it blocks the binding process of acetylcholine to M-type receptors. At this time, when the otolith is oscillated with a laser in this embodiment, even if the parasympathetic nervous system releases acetylcholine after activation, due to the drug effect of atropine, acetylcholine cannot bind to the receptor normally and thus cannot exert its effect of slowing down the heart rate. At this time, the heart rate slowing effect caused by parasympathetic nerve excitation is inhibited, so that the heart rate will not decrease as it does under normal circumstances during otolith oscillation stimulation, and may even increase under the relative action of the sympathetic nerve. At the same time, as Figure 6 shown in d of Figure 6 , this embodiment compared the effect of the anesthetic MS222 on the heart rate and found that although MS222 can reduce the heart rate value of zebrafish, compared with propranolol and atropine, which can reduce and increase the heart HRV of larvae respectively, MS222 has almost no effect on the change in heart HRV value ( Figure 6 shown in e of

[0058] The above experimental results confirm that propranolol and atropine can respectively block the regulatory pathways of the sympathetic nerve and the parasympathetic nerve on the heart rate, thus proving that otolith oscillation needs to utilize this neural pathway to regulate the heart's beating rhythm. Based on the above experiments and data analysis, this embodiment successfully obtained a neural pathway model for precise regulation of heart rate by optogenetic oscillation of zebrafish otoliths. As Figure 6 shown in f of Figure 6 , the otolith is connected to the basal hair cells through the ciliary bundle. Oscillating the otolith deflects the ciliary bundle, which in turn activates the hair cells to form an auditory signal input. At the same time, the auditory nerve signal is transmitted through the fibers of the eighth cranial nerve to the vagal motor nucleus in the medulla oblongata (for preliminary processing and integration of auditory information), and then upward to the semicircular ring in the midbrain (for more complex processing and analysis of auditory information). The auditory information processed by the midbrain is then transmitted to the thalamus and hypothalamus. As an important relay station for sensory conduction, the thalamus can screen, integrate, and modulate auditory information (including regulating the sympathetic nerve and the parasympathetic nerve). During this process, the nerve signal can be transmitted from the hypothalamus and the vagal motor nucleus in the dorsal part of the medulla oblongata to the heart, causing the sympathetic nerve to release norepinephrine, which binds to specific β-adrenergic receptors on the myocardial cell membrane and acts on the sinoatrial node (SAP) of the heart to make the myocardium contract and the heart rate increase. The above experimental system reveals the "vestibular-central nervous system-heart axis" neural pathway for regulating heart rate based on otolith oscillation.

[0059] In addition, in some embodiments, the biomedical applications of optogenetic otolith oscillation for regulating heart rate can be as follows: This embodiment first verified the therapeutic effect of optogenetic otolith oscillation on the atrioventricular block disease model. Before the experiment, this embodiment prepared a stock solution containing terfenadine in advance, and took an appropriate amount of the terfenadine stock solution and added it to the culture medium and ultrasonicated for 10 minutes to adjust the terfenadine concentration in the culture medium to 5 μM. Subsequently, this embodiment placed 5 dpf zebrafish embryos in the culture medium containing the drug for co-incubation. After 24 hours of cultivation, zebrafish embryos with successfully constructed atrioventricular block disease models were screened out through a microscope. Further, this embodiment placed the modeled zebrafish embryos on the sample stage, and used the constructed optical tweezer system to perform programmable optogenetic oscillation on their otoliths, while observing and recording the beating conditions of the zebrafish hearts before and after otolith oscillation. Through experimental data, it was found that under normal conditions (Control group), the atrial and ventricular beating frequencies of the zebrafish heart were consistent, that is, 1:1. However, after drug incubation, atrioventricular block occurred in the zebrafish embryos, and the beating of the atria and ventricles was significantly inconsistent. Specifically, when the atria beat twice, the ventricles beat only once, and the atrioventricular ratio became 2:1. This situation is as Figure 7In phase I of a in [description]. In response to this phenomenon, in this embodiment, the scanning optical trap is accurately applied to the otolith to generate a specific oscillation mode. At the same time, in this embodiment, it is observed that during the oscillation of the otolith, the atrioventricular ratio of the zebrafish heart has been significantly improved, and the two gradually tend to synchronize. On this basis, in this embodiment, the optically controlled otolith oscillation is continuously carried out for 20 minutes, and then the laser is turned off. As Figure 7 shown in phase III of a in [description], the atrioventricular beating ratio returns to the normal 1:1. At the same time, it can be seen from the heartbeat waveform diagram that within a period of time after the otolith stops oscillating, the atrial and ventricular beating ratio of the zebrafish heart is still 1:1, that is, the phenomenon of atrioventricular block no longer occurs, confirming that optically controlled oscillating otoliths can achieve targeted treatment of atrioventricular block. Further, in this embodiment, the influence law of the otolith oscillation amplitude on the treatment of atrioventricular block disease is explored. As Figure 7 shown in b in [description], when the otolith oscillation amplitude increases from 0.5 μm to 2 μm, the otolith oscillation treatment time required for the atrioventricular block to return to normal heartbeat is significantly shortened, from the original 30 minutes to only 10 minutes. This experiment shows that the greater the oscillation amplitude of the otolith controlled by the scanning optical trap, the stronger its effect on correcting the heartbeat, so the required treatment time is shorter, and the treatment effect is more obvious.

[0060] In addition to atrioventricular block, in this embodiment, the treatment effect of optically controlled otolith oscillation on cardiac escape disease is further explored. In this experimental study, zebrafish embryos at 6 dpf are co-incubated with embryo culture medium containing 0.5 μM sunitinib. After 5 hours, the zebrafish embryos after drug incubation are placed under a microscope for observation, and it can be found that the heart beating of the zebrafish will experience a short pause at this time, that is, cardiac escape. Moreover, the pause time caused by different concentrations of the drug is different, and the longer the pause time, the more serious the cardiac function degradation. As Figure 7 shown in c in [description], at t = 3 s, in this embodiment, it is observed that the zebrafish embryo heart has a pause time of about 2 s, confirming that in this embodiment, a zebrafish model with cardiac escape disease is successfully constructed, laying a foundation for subsequent treatment experiments. Next, in this embodiment, the optical tweezer is used to continuously and controllably oscillate the zebrafish otolith for 30 minutes. At this time, the optical tweezer applies a regular mechanical stimulation signal to the otolith, which will be transmitted to the heart through the neuromodulation mechanism, thereby affecting and correcting its heart rhythm. After turning off the laser, in this embodiment, the heart beating condition is continuously observed and recorded. After optically controlled otolith oscillation, the originally occurring intermittent heart pauses disappear, the heart rhythm returns to normal, and it remains normal after turning off the laser. Further, the treatment time increases with the increase of the heart pause time ( Figure 7In d), that is, the otolith oscillation treatment time required increases as the heart function degradation becomes more severe. This finding once again verifies the effectiveness of light-controlled otolith oscillation in improving abnormal heart rhythms, providing a new research idea for developing non-invasive means to treat cardiac rhythm abnormalities.

[0061] As a common heart disease, bradycardia can bring many adverse effects to patients' daily life and physical health. For example, patients may experience symptoms such as fatigue, dizziness, and syncope. In severe cases, it may even trigger more serious secondary heart diseases such as heart failure. Therefore, the prevention and treatment of bradycardia are particularly important, as it can not only help patients improve their quality of life but also effectively reduce the risk of other complications caused by bradycardia. In response to this, this embodiment further verifies the therapeutic effect of light-controlled otolith oscillation on bradycardia. First, this embodiment co-incubates 5 dpf zebrafish embryos with a 50 μM concentration of tolterodine culture solution and then uses it to construct a bradycardia disease model. During the experiment, this embodiment places the zebrafish embryos after 24 hours of drug incubation under a high-power microscope and observes that the heart rate of the incubated zebrafish is significantly lower (only 1.3 Hz) compared to the resting heart rate of 2.3 Hz of normal zebrafish, indicating that this embodiment has successfully constructed a bradycardia model of zebrafish. Subsequently, this embodiment controls the laser to perform controllable oscillation on its otoliths. After ten minutes of otolith oscillation, this embodiment successfully observes that the heart rate of the zebrafish increases to 1.51 Hz, confirming the improvement effect of light-controlled otolith oscillation on bradycardia ( Figure 7 In e-f). Further, after removing the laser 20 minutes after light-controlled otolith oscillation, the heart rhythm of the zebrafish finally stabilizes at 1.75 Hz, which is 0.45 Hz higher than before treatment, confirming the potential of light-controlled otolith oscillation in improving bradycardia.

[0062] In addition to achieving optimized treatment of diseases, this embodiment also quantitatively explores the regulatory effect of different music on heart rhythms based on the light-controlled otolith oscillation technology. The specific principle is as follows: By real-time programming the spatial scanning timing of the laser, this embodiment loads different music rhythms into the oscillation mode of the otoliths, thereby simulating the auditory input of different types of music and quantitatively characterizing its regulatory effect on the heart rhythm. Among them, this embodiment defines the mode with large mutations in the amplitude and frequency of the optical trap oscillation as exciting music, and on the contrary, defines the oscillation mode with small changes in the amplitude and frequency of the optical trap oscillation as soothing music ( Figure 7in g). By loading two different pieces of music onto the otoliths, this embodiment obtained electrocardiogram waveforms of different music types. When the resting heart rate was 2.32 Hz, loading soothing music to oscillate the otoliths resulted in relatively small fluctuations in heart rate, which slightly increased to 2.55 Hz. When exciting music was loaded, the heart rate increased sharply and rose to 2.98 Hz. Further, this embodiment verified the heart rate response laws in these two situations for 20 zebrafish embryos. As Figure 7 shown in h, compared with soothing music, loading exciting music increased the heart rate by about 20% on average. The above experiments verified the ability of optically controlled otolith oscillation to characterize heart rhythm regulation under different music mode treatments, and are expected to provide new technologies and means for further revealing the physical mechanisms and physiological characteristics behind music therapy.

[0063] Embodiment 3 As Figure 9 shown, this embodiment provides a heart rate regulation system, including: An optical potential well generation module for generating at least two optical potential wells at the edge position of the otoliths of a target living body through a scanned optical tweezer system; An oscillation module for dynamically adjusting the switching frequency, power, and spatial distribution pattern of the optical potential wells through a scanned optical tweezer system to drive the otoliths to oscillate according to set parameters; A heart rate change calculation module for real-time collecting a video of the heart beating of the target living body and calculating the heart rate change through an image processing algorithm; A heart rate regulation module for adjusting the otolith oscillation parameters according to the target heart rate regulation requirement to adjust the heart rate of the target living body; wherein, the otolith oscillation regulates the heart rate through the vestibular - central nervous system - heart axis neural pathway; the otolith oscillation is used to activate the hair cells connected to the bottom of the otoliths to generate auditory signals; the auditory signals are transmitted to the heart through the vagus nerve motor nucleus in the hypothalamus and the dorsal medulla oblongata, causing the sympathetic nerve to release norepinephrine, which binds to specific β - adrenergic receptors on the myocardial cell membrane, accelerating myocardial contraction and heart rate.

[0064] Among them, the heart rate regulation system further includes: A preprocessing module for performing preprocessing steps on the target living body, and the preprocessing steps include anesthetizing and fixing the target living body.

[0065] Among them, the optical potential well generation module in the heart rate regulation system specifically includes: A deflected beam generation unit for using a 1064 nm laser to generate a deflected beam through an acousto - optic deflector; A beam expansion unit for expanding the diameter of the deflected beam to be able to cover the entrance pupil of a high - magnification inverted objective lens based on a beam expander; An optical potential well generating unit is configured to reflect the near-infrared laser in the deflected light beam to a high-magnification inverted objective lens through a short-wave pass dichroic mirror, and focus the near-infrared laser to the otolith edge position of the target living body to form an optical potential well.

[0066] In summary, the present application has the following technical effects: 1. Based on the scanning optical tweezer system, the programmed regulation of the optical potential well is realized, and then it is applied to the stable capture and multi-mode oscillation manipulation of zebrafish larvae otoliths, providing a powerful tool for simulating complex auditory inputs.

[0067] 2. Compared with the existing research methods, the optically controlled otolith oscillation has unique advantages of non-contact, non-damage, and programmability, and can accurately trigger and change the motion state of the otolith at the microscopic level.

[0068] 3. Precise manipulation of the otolith provides an unprecedented technical solution for in-depth and accurate analysis of the change mechanisms of physiological indicators such as auditory signal conduction, emotion regulation, and heart rate regulation.

[0069] 4. Indirect heart rate regulation is achieved through optically controlled otolith oscillation, avoiding the deficiencies of traditional heart rate regulation methods, including drug therapy, electrical pacing, and optical pacing, which respectively face challenges such as potential toxicological damage, invasive implantation, and complex transgenic processing.

[0070] 5. By characterizing the ability of optically controlled otolith oscillation in regulating heart rhythm under different music mode treatments, it is expected to provide new technologies and means for further revealing the physical mechanisms and physiological characteristics behind music therapy.

[0071] The technical features of the above embodiments can be combined arbitrarily. For the sake of brevity of description, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, it should be considered as the scope recorded in this specification.

[0072] In this article, specific examples are used to elaborate on the principle and implementation manner of the present application. The description of the above embodiments is only used to help understand the method and its core idea of the present application; at the same time, for those of ordinary skill in the art, according to the idea of the present application, there will be changes in the specific implementation manner and application scope. In summary, the content of this specification should not be construed as a limitation to the present application.

Claims

1. A heart rate regulation device based on optically controlled otolith oscillation, characterized in that, Comprising: A scanning optical tweezers system; the scanning optical tweezers system includes a laser scanning optical path, an imaging optical path, an illumination optical path, and a motorized stage; The laser scanning optical path is used to generate a high-speed scanning optical potential well at the otolith of the target living body; The imaging optical path includes an imaging optical path of a high-magnification inverted objective lens and a low-magnification side-branch imaging optical path, which are respectively used for real-time image acquisition of the target living body and for monitoring the cardiac state of the target living body; The illumination optical path includes a first illumination optical path and a second illumination optical path; the first illumination optical path is used for illuminating the imaging field of view of the target living body on the motorized stage; the second illumination optical path is used for exciting the target living body to generate a calcium ion fluorescence signal; The motorized stage is used to carry the target living body and move the position of the target living body by controlling the displacement of the stage.

2. The heart rate regulation device based on opto-controlled otolith oscillation according to claim 1, wherein The laser scanning optical path includes a 1064 nm laser, an acousto-optic deflector, a beam expander, a short-wave pass dichroic mirror, and an inverted objective lens; the laser emitted by the 1064 nm laser sequentially passes through the acousto-optic deflector, the beam expander, the short-wave pass dichroic mirror, and the inverted objective lens and is incident on the imaging optical path of the high-magnification inverted objective lens.

3. The heart rate regulation device based on opto-controlled otolith oscillation according to claim 1, characterized in that, The imaging optical path of the high-magnification inverted objective lens is composed of a high-magnification inverted objective lens and a first high-speed charge-coupled device; the first high-speed charge-coupled device is perpendicularly arranged with the high-magnification inverted objective lens and is used to collect real-time image data of the target living body through the high-magnification inverted objective lens.

4. The heart rate regulation device based on opto-controlled otolith oscillation according to claim 1, wherein The low-magnification side-branch imaging optical path is composed of a low-magnification electron microscope and a second high-speed charge-coupled device; the second high-speed charge-coupled device is perpendicularly arranged with the low-magnification electron microscope and is used to observe the cardiac condition of the target living body in real time.

5. A heart rate regulation method for a heart rate regulation device based on opto-controlled otolith oscillation according to any one of claims 1-4, characterized in that, Comprising: Generating at least two optical potential wells at the edge position of the otolith of the target living body through the scanning optical tweezers system; Dynamically adjusting the switching frequency, power, and spatial distribution pattern of the optical potential well through the scanning optical tweezers system to drive the otolith to oscillate according to the set parameters; Real-time collecting the cardiac beating video of the target living body and calculating the heart rate change amount through an image processing algorithm; According to the target heart rate regulation requirement, adjusting the otolith oscillation parameters to adjust the heart rate of the target living body; wherein, the otolith oscillation regulates the heart rate through the vestibular-central nervous system-heart axis neural pathway; the otolith oscillation is used to activate the hair cells connected to the bottom of the otolith to generate an auditory signal; the auditory signal is transmitted to the heart through the hypothalamus and the vagus nerve motor nucleus in the dorsal medulla oblongata, causing the sympathetic nerve to release norepinephrine, which binds to the specific β-adrenergic receptor on the myocardial cell membrane, making the myocardial contraction heart rate increase.

6. The heart rate regulation method according to claim 5, wherein Before generating at least two optical potential wells at the edge position of the otolith of the target living body through the scanning optical tweezers system, it further includes: a pretreatment step for the target living body, and the pretreatment step includes anesthetizing and fixing the target living body.

7. The heart rate regulation method according to claim 5, characterized in that, Generating at least two optical potential wells at the edge position of the otolith of the target living body through the scanning optical tweezers system, specifically including: Using a 1064 nm laser to generate a deflected beam through an acousto-optic deflector; Based on the beam expander, expanding the diameter of the deflected beam to be able to cover the entrance pupil of the high-magnification inverted objective lens; The near-infrared laser in the deflected light beam is reflected by a short-wave pass dichroic mirror to a high-magnification inverted objective lens, and the near-infrared laser is focused on the edge position of the otolith of the target living body to form an optical potential well.

8. A heart rate regulation system, characterized in that, It includes: An optical potential well generation module for generating at least two optical potential wells at the edge position of the otolith of the target living body through a scanning optical tweezers system; An oscillation module for dynamically adjusting the switching frequency, power and spatial distribution mode of the optical potential well through a scanning optical tweezers system to drive the otolith to oscillate according to set parameters; A heart rate change calculation module for real-time collecting the heart beating video of the target living body and calculating the heart rate change through an image processing algorithm; A heart rate regulation module for adjusting the otolith oscillation parameters according to the target heart rate regulation requirement to adjust the heart rate of the target living body; wherein, the otolith oscillation regulates the heart rate through the vestibular-central nervous system-heart axis neural pathway; the otolith oscillation is used to activate the hair cells connected to the bottom of the otolith to generate auditory signals; the auditory signals are transmitted to the heart through the vagus nerve motor nucleus in the hypothalamus and the dorsal medulla oblongata, causing the sympathetic nerve to release norepinephrine, which binds to the specific β-adrenergic receptor on the myocardial cell membrane, making the myocardial contraction heart rate increase.

9. The heart rate regulation system according to claim 8, characterized in that, It also includes: A pretreatment module for performing pretreatment steps on the target living body, and the pretreatment steps include anesthetizing and fixing the target living body.

10. The heart rate regulation system according to claim 8, characterized in that, The optical potential well generation module specifically includes: A deflected light beam generation unit for using a 1064 nm laser to generate a deflected light beam through an acousto-optic deflector; An expanding unit for expanding the diameter of the deflected light beam to be able to cover the entrance pupil of the high-magnification inverted objective lens based on an expanding lens; An optical potential well generation unit for reflecting the near-infrared laser in the deflected light beam by a short-wave pass dichroic mirror to a high-magnification inverted objective lens, and focusing the near-infrared laser on the edge position of the otolith of the target living body to form an optical potential well.

Images