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

Abstract

The present application discloses a heart rate control device, method, and system based on optically controlled otolith oscillations, relating to the field of heart rate control. The method includes 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 a 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 first illumination optical path in the illumination optical path is used to illuminate the imaging field of the target living body on the motorized stage, and the second illumination optical path is used to excite the target living body to generate a calcium ion fluorescence signal; the motorized stage is used to carry the target living body and to move the position of the target living body through stage displacement control. The present application can precisely control the heartbeat rhythm.

Description

Technical Field

[0001] The present application relates to the field of heart rate control, and in particular to a heart rate control device, method and system based on light-controlled otolith oscillation. Background Art

[0002] Traditional methods of heart rhythm regulation, including drug therapy and optical pacing, each face different challenges. Drug therapy faces several challenges: on the one hand, antiarrhythmic drugs may have toxic side effects; optical pacing uses light-sensitive proteins to regulate the electrophysiological activity of heart cells. When a light beam of a specific wavelength is used to irradiate heart cells, the light-sensitive protein undergoes conformational changes, thereby achieving precise regulation of the electrophysiological signals of heart cells. However, optical pacing technology is still in the research stage, and there are some problems that need to be optimized, such as the need for cardiac photosensitive protein conversion in the early stage, complex operations and long treatment cycles, etc. At the same time, the penetration depth of light is limited, making it difficult to reach cells deep in the heart. Summary of the Invention

[0003] The purpose of this application is to provide a heart rate control device, method and system based on light-controlled otolith oscillation. By combining a scanning optical tweezers system with calcium ion imaging technology, precise control of heart rhythm is achieved based on the multifunctional light-controlled oscillation of otoliths.

[0004] To achieve the above objectives, this application provides the following solutions:

[0005] In a first aspect, the present application provides a heart rate control device based on light-controlled otolith oscillation, comprising: a scanning optical tweezers system; the scanning optical tweezers system comprises a laser scanning optical path, an imaging optical path, an illumination optical path, and a motorized stage;

[0006] The laser scanning optical path is used to generate a high-speed scanning optical potential well at the otolith of the target living body;

[0007] The imaging optical path includes a high-magnification inverted objective lens imaging optical path 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 cardiac state monitoring of the target living body;

[0008] The illumination light path includes a first illumination light path and a second illumination light path; the first illumination light path is used to illuminate the imaging field of the target living body on the electric stage; the second illumination light path is used to excite the target living body to generate a calcium ion fluorescence signal;

[0009] The electric stage is used to carry the target living body, and the position of the target living body is moved by displacement control of the stage.

[0010] Optionally, the laser scanning optical path includes a 1064 nm laser, an acousto-optic deflector, a beam expander, a short-wavelength dichroic mirror and an inverted objective lens; the laser light emitted by the 1064 nm laser passes through the acousto-optic deflector, the beam expander, the short-wavelength dichroic mirror and the inverted objective lens in sequence and is incident on the imaging optical path of the high-magnification inverted objective lens.

[0011] Optionally, the imaging light path of the high-magnification inverted objective lens is composed of the high-magnification inverted objective lens and a first high-speed charge-coupled device; the first high-speed charge-coupled device is arranged vertically to 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.

[0012] Optionally, 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 arranged vertically to the low-magnification electron microscope for real-time observation of the heart condition of the target living body.

[0013] In a second aspect, the present application provides a heart rate control method based on the aforementioned heart rate control device based on light-controlled otolith oscillation, comprising:

[0014] At least two optical potential wells are generated at the edge of the otolith of the target living body by a scanning optical tweezers system;

[0015] The switching frequency, power and spatial distribution pattern of the optical potential well are dynamically adjusted by the scanning optical tweezers system to drive the otoliths to oscillate according to the set parameters.

[0016] Real-time capture of the target living body's heartbeat video, and calculation of heart rate changes through image processing algorithms;

[0017] According to the target heart rate control requirements, the otolith oscillation parameters are adjusted 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 via the vagus motor nucleus in the hypothalamus and the dorsal medulla oblongata, causing the sympathetic nerves to release norepinephrine, which binds to specific β-adrenergic receptors on the myocardial cell membrane, thereby accelerating the myocardial contraction heart rate.

[0018] Optionally, before generating at least two optical potential wells at the edge of the otolith of the target living body by the scanning optical tweezers system, the method further includes: performing a pretreatment step on the target living body, wherein the pretreatment step includes anesthetizing and fixing the target living body.

[0019] Optionally, generating at least two optical potential wells at the edge of the otolith of the target living body by a scanning optical tweezers system specifically includes:

[0020] A 1064 nm laser is used to generate a deflected beam through an acousto-optic deflector;

[0021] Using a beam expander, the diameter of the deflected light beam is expanded to cover the entrance pupil of a high-magnification inverted objective lens;

[0022] The near-infrared laser in the deflected beam is reflected by a short-wavelength dichroic mirror to a high-magnification inverted objective lens, and the near-infrared laser is focused to the edge of the otolith of the target living organism to form an optical potential well.

[0023] In a third aspect, the present application provides a heart rate control system, comprising:

[0024] An optical potential well generation module, used for generating at least two optical potential wells at the edge of the otolith of the target living body through a scanning optical tweezers system;

[0025] An oscillation module is used to dynamically adjust the switching frequency, power, and spatial distribution pattern of the optical potential well through a scanning optical tweezers system, driving the otolith to oscillate according to the set parameters;

[0026] The heart rate variation calculation module is used to collect the heartbeat video of the target living body in real time and calculate the heart rate variation through image processing algorithm;

[0027] The heart rate control module is used to adjust the otolith oscillation parameters and the heart rate of the target living body according to the target heart rate control requirements; 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 base of the otoliths to generate auditory signals; the auditory signals are transmitted to the heart via the vagus motor nucleus in the hypothalamus and the dorsal medulla oblongata, causing the sympathetic nerves to release norepinephrine, which binds to specific β-adrenergic receptors on the myocardial cell membrane, thereby accelerating the myocardial contraction heart rate.

[0028] Optionally, it also includes:

[0029] The pre-processing module is used to perform a pre-processing step on the target living body, wherein the pre-processing step includes anesthetizing and fixing the target living body.

[0030] Optionally, the optical potential well generation module specifically includes:

[0031] a deflected beam generating unit, configured to generate a deflected beam using a 1064 nm laser via an acousto-optic deflector;

[0032] a beam expansion unit, configured to expand the diameter of the deflected light beam to a diameter capable of covering an entrance pupil of a high-magnification inverted objective lens using a beam expander;

[0033] The optical potential well generating unit is used to reflect the near-infrared laser in the deflected light beam to the high-magnification inverted objective lens through a short-wavelength dichroic mirror, and focus the near-infrared laser to the edge of the otolith of the target living body to form an optical potential well.

[0034] According to the specific embodiments provided in this application, this application discloses the following technical effects:

[0035] The present application provides a heart rate control device, method and system based on light-controlled otolith oscillation. First, the laser scanning optical path in the scanning optical tweezers system generates a high-speed scanning optical potential well at the otolith of the target living organism. The optical potential well can accurately capture and manipulate the otoliths, causing them to produce controllable oscillations. Next, the imaging optical path of the high-magnification inverted objective lens is used to capture images of the target living organism in real time, ensuring that the operator can clearly observe the dynamic changes of the otoliths 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 organism. In terms of lighting, the first lighting optical path provides sufficient lighting for the target living organism on the electric stage to ensure clear imaging. The second lighting optical path is used to excite the target living organism to produce a calcium ion fluorescence signal, which is a key step in calcium ion imaging technology. The changes in the intensity of the fluorescence signal can reveal the neural signal pathway that controls the heart rate through light-controlled otolith oscillation. The electric stage carries the target living organism and can adjust its position as needed to ensure that the laser scanning optical path, imaging optical path and lighting optical path can all be accurately aligned with the target area. Through the synergistic effect of the above systems, the operator can precisely control the oscillation frequency and amplitude of the otoliths, thereby achieving precise regulation of the heart rhythm. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0037] Figure 1 A schematic structural diagram of a heart rate control device based on light-controlled otolith oscillation provided in one embodiment of the present application;

[0038] Figure 2 A schematic diagram illustrating the principle of light-controlled otolith oscillation to regulate heart rate according to an embodiment of the present application;

[0039] Figure 3 A schematic diagram of achieving multifunctional optically controlled oscillation of otoliths using programmable scanning optical technology in vivo according to one embodiment of the present application;

[0040] Figure 4 A schematic diagram of achieving precise heart rate control based on light-controlled otolith oscillations according to an embodiment of the present application;

[0041] Figure 5 A schematic diagram of a brain nerve signal conduction pathway triggered by light-controlled otolith oscillations provided in one embodiment of the present application;

[0042] Figure 6 This is an analysis diagram of the neural pathway by which the brain regulates heart rate by light-controlled otolith oscillations provided in one embodiment of the present application;

[0043] Figure 7 A diagram showing a biomedical application of light-controlled otolith oscillation to regulate heart rate according to an embodiment of the present application;

[0044] Figure 8 A flow chart of a heart rate control method provided in one embodiment of the present application;

[0045] Figure 9 A schematic diagram of the structure of a heart rate control system provided in one embodiment of the present application. DETAILED DESCRIPTION

[0046] The following will be combined with the drawings in the embodiments of this application to clearly and completely describe the technical solutions in the embodiments of this application. Obviously, the embodiments described are only part of the embodiments of this application, not all of the embodiments. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0047] In order to make the above-mentioned purposes, features and advantages of the present application more obvious and easy to understand, the present application is further described in detail below with reference to the accompanying drawings and specific implementation methods.

[0048] Example 1

[0049] like Figure 1 As shown, this embodiment provides a heart rate control device based on light-controlled otolith oscillation, including: 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;

[0050] The laser scanning optical path is used to generate a high-speed scanning optical potential well at the otolith of the target living body;

[0051] The imaging optical path includes a high-magnification inverted objective lens imaging optical path 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 cardiac state monitoring of the target living body;

[0052] The illumination light path includes a first illumination light path and a second illumination light path; the first illumination light path is used to illuminate the imaging field of the target living body on the electric stage; the second illumination light path is used to excite the target living body to generate a calcium ion fluorescence signal;

[0053] The electric stage is used to carry the target living body, and the position of the target living body is moved by displacement control of the stage.

[0054] In this example, the experimental setup is built around a scanning optical tweezers system (SOTs), primarily comprising a laser scanning optical path, an imaging optical path, an illumination optical path, and a motorized stage. The laser scanning optical path, used to generate a high-speed scanning optical potential well, primarily includes a 1064 nm laser, an acousto-optic deflector (AOD), a beam expander, a short-wavelength dichroic mirror, and an inverted objective lens.

[0055] Selecting a laser wavelength of 1064 nm can avoid thermal damage to manipulated tissues or cells caused by the photothermal effect of the focused beam, because biological tissues have less absorption in this band.

[0056] 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 widened after passing through the beam expander, which is composed of two convex lenses. The widened laser beam is reflected by the short-wave dichroic mirror into the inverted objective lens, and the beam cross-sectional diameter can completely cover the incident light 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 capture optical potential well, which can realize the multi-mode oscillation manipulation of the otolith. Based on the constructed scanning optical tweezers system, the light beam can be rapidly scanned at a maximum frequency of 100 kHz. At the same time, the position, number, power, scanning frequency and scanning mode of the capture optical potential well can be independently set by software.

[0057] In addition, the optical path for microscopic imaging is divided into two paths: (1) the imaging optical path of the high-magnification inverted objective lens, which uses a high-speed charge-coupled device (CCD) for image acquisition and real-time monitoring, image acquisition, and video recording on a computer screen; (2) the low-magnification side-branch imaging optical path, which uses a low-magnification electron microscope and CCD to observe the general state 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 calculate the heart rate change through image processing algorithms.

[0058] The system features two illumination paths. The first, a halogen light source (D-LH, 12 V / 100 W), is focused by a condenser and illuminates the sample stage, illuminating the imaging field. The second, a light source with red, green, and blue excitation light, uses built-in filters to select the color of the emitted excitation light to stimulate the sample and produce the corresponding fluorescence.

[0059] White light source and fluorescence excitation light source: Provides the illumination light required to observe samples in bright field or fluorescence excitation.

[0060] Acousto-optic deflectors: Generating mechanical waves in a crystalline medium causes periodic refractive index variations, forming a phase-type diffraction grating. When a laser beam is incident on the medium, it diffracts, and the intensity and direction of the diffracted light vary with the intensity and wavelength of the mechanical wave. This principle allows the focal position and intensity of a 1064 nm laser to be varied.

[0061] Beam broadening device (beam expander): consists of two convex lenses with a distance equal to the sum of the focal lengths of the two convex lenses. It is used to expand the output diameter of the laser so that the diameter can completely cover the incident light pupil of the inverted objective lens.

[0062] Short-pass dichroic mirrors transmit or reflect light based on wavelength, achieving spectral separation. Long-pass dichroic mirrors highly reflect light below the cutoff wavelength and transmit light above it. Short-pass dichroic mirrors, on the other hand, transmit light below the cutoff wavelength and reflect light above it. This example uses a short-pass dichroic mirror, which transmits illumination light with wavelengths below 800 nm while reflecting near-infrared laser light above 800 nm.

[0063] CCD camera: short for charge coupled device, used to capture images in real time. The specific acquisition frequency can be set through the PC.

[0064] Example 2

[0065] like Figure 8 As shown, this embodiment provides a heart rate control method based on the heart rate control device based on light-controlled otolith oscillation, including:

[0066] Step 1: Generate at least two optical potential wells at the edge of the otolith of the target living organism using a scanning optical tweezers system;

[0067] Step 2: Dynamically adjust 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;

[0068] Step 3: Real-time capture of the target living person's heartbeat video and calculation of the heart rate variation using image processing algorithms;

[0069] Step 4: According to the target heart rate control requirements, the otolith oscillation parameters are adjusted 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 motor nucleus in the hypothalamus and the dorsal medulla oblongata, causing the sympathetic nerves to release norepinephrine, which binds to specific β-adrenergic receptors on the myocardial cell membrane, thereby accelerating the myocardial contraction heart rate.

[0070] In this embodiment, zebrafish is selected as the target living organism.

[0071] In this embodiment, a technology for light-controlled otolith oscillation to regulate heart rate has been developed. By precisely manipulating otolith oscillations using programmable optical scanning technology, auditory hair cells are induced to produce auditory signals, which are then applied to non-invasive heart rate regulation in vivo. Specifically, this light-controlled otolith oscillation technology utilizes a programmable scanning optical tweezers system, using a focused beam of light of specific wavelength and intensity to generate a scanning optical potential well, thereby achieving precise, real-time, and multifunctional control of the otolith oscillation pattern.

[0072] This embodiment further determines the effects of different otolith oscillation modes on heart rate through quantitative characterization, and verifies the differences in the effects of otolith oscillation on the heart rate of zebrafish larvae at different developmental stages. At the same time, with the help of neuronal calcium ion labeling technology and specific drugs, the intrinsic neural circuit of otolith oscillation on heart rate regulation was preliminarily verified. Finally, the treatment and improvement of various heart disease models were achieved by using otolith oscillation to regulate heart rate. Compared with existing research methods, optical manipulation has the unique advantages of being non-contact, non-destructive and programmable. It can accurately trigger and change the movement state of otoliths at the microscopic level, thereby providing an unprecedented technical solution for in-depth and accurate analysis of the changing mechanisms of physiological indicators such as auditory signal conduction, emotion regulation and heart rate regulation.

[0073] Specifically, the otoliths of zebrafish larvae have a relatively high refractive index. When a focused laser is irradiated onto the otoliths, the optical potential well generated can stably capture the otoliths. Moreover, when the optical potential well is located at the edge of the otoliths, the capture capability is strongest under the same laser power. In the experiment, this embodiment uses a scanning optical tweezers system to generate two optical potential wells, which are respectively set at the edge of the otoliths, and then the switching frequency of the two optical potential wells is adjusted by the scanning optical tweezers system to drive the otoliths to oscillate at a high speed at the set frequency. At the same time, a variety of complex oscillation modes are set according to the experimental requirements, and the oscillation frequency, oscillation amplitude and oscillation direction are adjusted in real time to achieve precise control of the otolith oscillations to simulate a variety of auditory input modes. Such as Figure 2As shown in figure a, because the otoliths are connected to the base of the hair cells via ciliary bundles, oscillating otoliths can deflect the ciliary bundles, activating the hair cells and generating auditory signal input. Simultaneously, auditory nerve signals travel via the eighth pair of cranial nerve fibers to the vagus motor nucleus in the medulla oblongata (for initial processing and integration of auditory information), and then upward to the semicircular rings in the midbrain (for more complex processing and analysis of auditory information). This processed auditory information 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, nerve signals can be transmitted from the hypothalamus and the vagus motor nucleus in the dorsal medulla oblongata to the heart, causing the sympathetic nerves to release norepinephrine, which binds to specific β-adrenergic receptors on the myocardial cell membrane and acts on the sinoatrial node (SAP) to accelerate myocardial contraction and heart rate. Therefore, this embodiment successfully achieves precise regulation of heart rhythm through the multifunctional oscillation of the otoliths. Its specific status is mainly quantitatively evaluated and measured through changes in heart rate data ( Figure 2 b) in the above example.

[0074] In some embodiments, before generating at least two optical potential wells at the edge of the otolith of the target living body by the scanning optical tweezers system, the method further includes: performing a pretreatment step on the target living body, wherein the pretreatment step includes anesthetizing and fixing the target living body.

[0075] Specifically, in the care and treatment of zebrafish, juvenile zebrafish (3 days post-fertilization) were purchased from a commercial supplier (Company A). They were maintained in 100 ml culture dishes at 28 ± 0.5°C, 14 h light / 10 h dark, and fed with a starter diet three times daily. 50% of the water was replaced every 24 hours, and 100 The nylon mesh with large aperture separates excrement and leftover bait.

[0076] To ensure a good operating posture during the experiment, the fish were fixed to a 1 mm thick glass slide using 2% agarose, facilitating subsequent experimental manipulation and imaging. When using drugs to validate neural pathways, zebrafish were treated with the drugs: 5 dpf zebrafish embryos were selected and incubated in culture medium containing the same concentrations of MS222, propranolol, and atropine (within the drug tolerance range). After 24 hours, the fish developed to 6 dpf, at which time they were subjected to otolith oscillation experiments and their corresponding heart rates were recorded.

[0077] In this example, three drugs, terfenadine, sunitinib, and tolterodine, were used to establish heart disease models of atrioventricular block, cardiac escape, and bradycardia. Before the experiment, a stock solution containing terfenadine was prepared in advance, and an appropriate amount of the stock solution was added to the culture medium and ultrasonicated for 10 minutes. The terfenadine concentration in the culture medium was adjusted to 5 Subsequently, in this example, 5 dpf zebrafish embryos were placed in a culture medium containing drugs for co-incubation. After 24 hours of incubation, zebrafish embryos with successfully established atrioventricular block disease models were screened under a microscope. Similarly, 6 dpf zebrafish embryos were co-incubated with a 0.5% concentration of dpf. After 5 hours of incubation with Sunitinib embryo culture medium, juvenile fish with cardiac escape disease were screened out; 5 dpf zebrafish embryos and 50 After incubation with the culture medium containing tolterodine at a high concentration for 24 hours, juvenile fish with bradycardia disease were screened out.

[0078] Wherein, when generating at least two optical potential wells at the edge of the otolith of the target living body by using a scanning optical tweezers system, the method specifically includes:

[0079] A 1064 nm laser is used to generate a deflected beam through an acousto-optic deflector;

[0080] Using a beam expander, the diameter of the deflected light beam is expanded to cover the entrance pupil of a high-magnification inverted objective lens;

[0081] The near-infrared laser in the deflected beam is reflected by a short-wavelength dichroic mirror to achieve optically controlled oscillation of the otoliths;

[0082] Using a high-magnification inverted objective lens, the near-infrared laser is focused onto the edge of the otolith of the target living organism to form an optical potential well.

[0083] Specifically, programmable scanning optics technology was used to achieve multifunctional light-controlled oscillation of otoliths in zebrafish:

[0084] The premise of precise control of otolith oscillation is that the optical potential well can stably capture the otolith and drive its directional motion. Therefore, this embodiment first experimentally characterizes the performance of the optical potential well in capturing the otolith. Figure 3 As shown in a1 in the figure, the otolith of zebrafish larvae is connected to the ciliary bundles of hair cells. When there is no external vibration, the otolith remains motionless. When a single optical potential well is set at the left edge of the otolith, it is found that the otolith shifts to the left by about 1.37 μm compared to its original position ( Figure 3 Similarly, when the optical potential well is located at the right edge of the otolith, the otolith moves to the right by 1.28 μm ( Figure 3 In addition to left-right movement, this embodiment can also control the otolith to move up and down, driving the otolith to move 1.06μm and 0.97μm in the vertical direction respectively ( Figure 3 The above experimental phenomena preliminarily prove that the otoliths can be stably captured by the scanning optical tweezers system, and then the otoliths can be precisely manipulated based on two optical potential wells to achieve multifunctional oscillations.

[0085] Furthermore, this embodiment quantitatively characterizes the phenomenon of oscillation of otoliths captured by two optical potential wells. The scanning frequencies of the scanning optical tweezers between the two optical potential wells are set to 5Hz, 10Hz and 15Hz respectively, and then the oscillation frequencies of the otoliths under different optical tweezers scanning frequencies are obtained by high-speed CCD imaging and analysis. Figure 3 As shown in b in FIG, it can be found from the waveform diagram that the oscillation frequency of the otolith is always synchronized with the set OT scanning frequency.

[0086] Furthermore, this embodiment gradually increases the scanning frequency to 40 Hz, which can also make the otoliths respond to oscillation synchronously ( Figure 3 c). After proving that the otolith can respond well to the scanning frequency of the two optical potential wells and oscillate, this embodiment investigates the oscillation amplitude of the otolith and the maximum oscillation displacement along different directions. Figure 3 As shown in Figure d, as the optical potential well scanning frequency increases (while the power remains constant), the otolith oscillation amplitude decreases. This is because the hair cell ciliary bundles connected to the base of the otolith have a certain degree of rigidity. When the optical potential well scanning frequency increases, the optical force applied to the left and right edges of the otolith decreases in duration, resulting in a decrease in the otolith oscillation displacement with increasing frequency, and thus a decrease in the oscillation amplitude.

[0087] Furthermore, in this embodiment, the power of the two optical potential traps is changed (the frequency remains unchanged), and it can be seen that as the trapping force increases, the oscillation amplitude of the utricle otolith (Ut) and the saccule otolith (Sac) increases accordingly. Figure 3 As can be seen from e in the figure, under the same light potential trap capture force, Ut produces a larger displacement than Sac. This is because the mass size and the length of the cilia connected to the bottom are different.

[0088] On this basis, this example further explores the oscillation flexibility of the otoliths in different directions. Figure 3 As shown in Figure 5, along the zebrafish's head-tail direction (0-180° horizontally), the otoliths can produce the largest displacement, and both otoliths exhibit the same pattern. These experiments confirm that scanning optical tweezers can achieve flexible otolith oscillation, laying the foundation for further experimental research.

[0089] Based on the use of optical potential traps (Trap) to stably capture and accurately oscillate otoliths, this embodiment uses an acousto-optic deflector to achieve real-time programming of the optical potential trap, and then develops a variety of complex otolith oscillation modes. It can perform multi-mode switching in terms of the number of optical traps, oscillation direction, oscillation frequency, and oscillation amplitude, providing multiple means for exploring auditory input and physiological activities triggered by hearing. Figure 3As shown in g1 in the figure, two optical potential traps can realize unidirectional oscillation of the otolith. The scanning order switches back and forth between Trap1 and Trap2 in turn, realizing controllable oscillation of the otolith in the horizontal direction. When the three optical potential traps are controlled to scan in the order of Trap1, Trap2 to Trap3, the otolith can be realized to oscillate in multiple directions around the center to the outside ( Figure 3 If four optical potential wells are introduced and the Figure 3 The sequential scanning shown by g3-g4 in the figure can respectively make the otolith oscillate in the shape of "8" and "cross" around the center.

[0090] It's worth noting that different oscillation modes cause different deflection patterns and intensities of the ciliary bundles, resulting in different mechanical force stimuli perceived by hair cells, thus generating auditory input signals of varying intensities. Based on this, this embodiment can apply different otolith oscillation modes by setting different oscillation frequencies and amplitudes, thereby achieving complex and varied auditory input manipulation, further simulating the complex state of real auditory input and enabling more accurate research on the influence of otolith oscillations on heart rate.

[0091] Among them, when executing step 4, accurate heart rate control is achieved based on light-controlled otolith oscillation. The specific process is as follows:

[0092] In the above experiments, this example successfully achieved multi-mode oscillations in otoliths using a scanning optical tweezers system. It should be noted that heart rate changes were observed in response to otolith oscillations. To investigate the influence of light-controlled otolith oscillations on heart rate, this example quantitatively characterized the corresponding heart rate variation (HRV) by varying the amplitude, frequency, and number of otolith oscillations.

[0093] Figure 4 Figure a shows the distribution of otoliths and heart in zebrafish larvae: There is an ear cavity on each side of the zebrafish head, and the ear cavity contains two types of otoliths, Sac and Ut (the corresponding fluorescence image is shown in Figure 4). Figure 4 (As shown in b in the figure), red fluorescence indicates brain blood vessels, and green fluorescence indicates the heart. In this experiment, the otoliths were first subjected to 50 Hz optical oscillation. After the stimulation lasted for about 5 seconds, the laser was turned off, and the otolith oscillations immediately ceased. Simultaneously, a bypass low-magnification electron microscope was used to capture images and record videos of the heartbeat process. By analyzing the heartbeat video, a zebrafish heartbeat waveform can be obtained ( Figure 4c in the figure). Through quantitative analysis, this example found that the normal heart rate of the zebrafish before otolith oscillation was approximately 2.1 Hz; after the otolith oscillation was applied with an optical potential well, the heart rate gradually increased to 2.8 Hz. As the otolith oscillation ceased, the heart rate returned to 2.1 Hz. To reduce errors and more accurately assess zebrafish heart rate changes, this example introduced heart rate variability (HRV) for quantitative analysis. HRV is defined as the time variation between successive heartbeat cycles. It can be used to reflect heart rate fluctuations at different time points, with the magnitude indicating the speed of heart rate changes.

[0094] In order to exclude the possibility that the heart rhythm changes may be due to the stress-induced acceleration of the heart rate caused by disturbance or stimulation of the zebrafish body, this example set up multiple groups of control experiments for verification. In the experiment, this example fixed the larvae on the sample stage with 2% agarose, and set up two scanning optical traps with fixed power and frequency. The two optical traps were applied to the outside of the larvae body, the back of the fish body, the ear cavity (non-hair cell area) and the otolith respectively. The control group did not apply an optical potential trap. Figure 4 As shown in d, the optical potential well applied to the fish's back and the ear cavity (non-hair cell area) can slightly change the heart rate of the juvenile fish, while acting on the otolith can increase the heart rate by up to about 50%. Furthermore, this embodiment sets the optical potential well parameters to directly capture the non-oscillation of the otolith and to capture the dynamic oscillation of the otolith. Figure 4 As shown in Figure e, photothermal stimulation that only captures non-oscillating otoliths is insufficient to effectively increase heart rate variation. In contrast, light-controlled otolith oscillations can significantly increase heart rate. Based on the above experiments, this embodiment can determine that the primary factor affecting heart rate variation through light-controlled otolith oscillations is not photothermal stimulation, but rather the conduction of auditory signals from hair cells caused by otolith oscillations, which in turn triggers heart rate variation.

[0095] After determining the main factors affecting heart rate changes, this embodiment further manipulates the amplitude and frequency of otolith oscillations to quantitatively explore their influence on heart rate changes. Figure 4As shown in f, in this embodiment, otolith oscillations were performed on juvenile fish of 6 dpf, and the oscillation amplitude was adjusted to gradually increase from the initial 1 μm to 4 μm. It can be seen that as the otolith oscillation amplitude increases, the HRV value also increases linearly, which may be because the magnitude of the otolith oscillation amplitude affects the conduction signal strength of the hair cells. The otolith oscillation amplitude is small, and its mechanical deflection of the bottom connecting ciliary bundle is reduced, causing the mechanical conduction signal of the hair cells to be weak, so the signal conduction strength that affects the heart rate change is also weak, resulting in 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 level of neuronal area is strong, which in turn acts on the heart to speed up its contraction frequency, and ultimately significantly increases the change in HRV. At the same time, this embodiment also characterizes the change pattern of HRV with oscillation frequency when the oscillation amplitude is the same. As shown Figure 4 As shown in Figure g, the magnitude of HRV increases at 10 Hz, 50 Hz, and 100 Hz oscillation frequencies, but the differences are relatively small. This may be because the oscillating otoliths are connected to relatively few types of ciliary bundles, unlike the entire ear, which contains a variety of hair cells that can detect mechanical oscillations of different frequencies. At this time, the ciliary bundles connected to the otoliths are less able to distinguish oscillation frequencies, resulting in relatively small differences in their impact on heart rate.

[0096] Since different types of otoliths are connected to different types of ciliary bundles, their influence on heartbeat will also be different. To address this, this embodiment performs optical potential trap oscillation with the same parameters on the Sac and Ut otoliths on the same side of the zebrafish larvae, and compares their respective influences on heart rate in real time. Figure 4 As shown in h, under the same oscillation amplitude and oscillation frequency, oscillating Sac causes greater changes in heart rate than oscillating Ut. Therefore, this embodiment knows that the Ut otolith responsible for maintaining the body's vestibular balance has a smaller response to heart rate than the Sac otolith responsible for hearing. In subsequent experiments, this embodiment also uniformly uses the experimental data of the Sac otolith for illustration. On this basis, this embodiment also compares the difference in heart rate changes when oscillating Ut and Sac separately and oscillating the two otoliths simultaneously. As shown in h, under the same oscillation amplitude and oscillation frequency, oscillating Sac causes greater changes in heart rate than oscillating Ut and Sac separately and oscillating the two otoliths simultaneously. Figure 4 As shown in (i), the sum of the HRV values ​​of oscillating the Ut otolith and the Sac otolith separately is close to the HRV value of oscillating the two otoliths simultaneously. This shows that the influence of oscillating otoliths on heart rate has an additive effect.

[0097] Furthermore, in some embodiments, the neural pathway analysis and verification of heart rate regulation based on light-controlled otolith oscillations can be specifically as follows:

[0098] On the basis of the above experiments, this embodiment further explores the intrinsic signal pathway of light-controlled otolith oscillations in regulating heart rate. Based on previous literature research, this embodiment preliminarily proposes a hypothesis of the neural pathway of otolith oscillations in regulating heart rate: vestibular-central nervous system-heart axis. Specifically, the vestibular auditory signals generated by otolith oscillations will enter the relevant areas of the brain for processing, and then the brain will make corresponding decisions, thereby regulating the myocardial contraction of the heart to regulate heart rate. In order to verify the above-mentioned signal pathway mechanism, this embodiment quantitatively explores the central nervous system signal response caused by light-controlled otolith oscillations based on calcium ion fluorescence labeling. Specifically, calcium ion imaging is performed on zebrafish brain neurons to characterize the neural signal circuit that regulates heart rhythm after light-controlled otolith oscillations.

[0099] First, this example characterizes the response of otolith oscillations to relevant neural areas of the brain. Commonly used methods for detecting brain neuron signal transmission activity mainly include electrophysiological measurement and neural calcium ion fluorescence detection. The former is somewhat invasive and difficult to perform non-destructive detection on zebrafish larvae. Therefore, this example selects a transgenic zebrafish strain (huc:GCaMP6f&nacre) with whole-brain neuronal calcium ion fluorescence labeling for experimental research. Figure 5 As shown in a in the figure, this embodiment uses 2% agarose to fix the transgenic zebrafish, and adjusts its body angle to a suitable brain fluorescence imaging field of view. At the same time, two scanning light potentials are set to be applied to 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 according to a specific frequency and power. Turn off the illumination light and turn on the blue excitation light at the same time to stimulate the brain calcium ion fluorescence signal in real time. On this basis, the otolith oscillation is manipulated based on the programmable optical potential well, and the video data of the fluorescence changes are synchronously recorded. As shown in FIG. Figure 5 As shown in b, the fluorescent signal of calcium ion neurons in the whole brain of the transgenic zebrafish can be observed to be successfully excited, where the white box and yellow box correspond to the eyes and ear cavity, respectively.

[0100] Next, the sequence of calcium ion fluorescence signal images of the zebrafish brain during otolith oscillation was processed to obtain neuronal calcium ion fluorescence signal response images of otolith oscillation at different time stages. The specific operation is as follows: the image at which the fluorescence change response begins after the otolith oscillation is recorded as the initial frame (2.8 s moment), a frame of image is taken every 0.2 s and the grayscale value of the previous frame of image is subtracted. Since the fluorescence response time is short, a total of 6 fluorescence images at different moments are intercepted for subtraction. At the same time, this embodiment sets a threshold screening. Fluorescence changes below the subthreshold value belong to spontaneous fluorescence responses. Similarly, 5 fluorescence images of regional fluorescence changes can be obtained. In order to distinguish the fluorescence response areas at each time stage, this embodiment assigns red, yellow, green, blue and purple colors to them respectively ( Figure 5Finally, they are aligned and superimposed to obtain the integrated fluorescence response area map. Figure 5 As shown in c6 in the figure, the fluorescence response gradually shifts from the red region at the beginning to the purple region at the end. This indicates that the brain signal generated by otolith oscillations is transmitted from the hindbrain to the midbrain and then to the forebrain. To clarify the brain regions affected by otolith oscillations and their functional roles, this example matched the obtained fluorescence response regions with a zebrafish brain atlas. The three different colored dashed boxes indicate that the brain signal response regions induced by otolith oscillations can be divided into three categories. Comparison with the zebrafish brain atlas reveals that, in order of signal transmission, they are, from bottom to top, the vagal motor nucleus (VMN), the semicircular tract (TS), and the thalamus.

[0101] Based on the above experiments and data analysis, we can obtain the brain neural signal transmission pathways triggered by otolith oscillations. However, the signal transmission pathways in which the brain controls heart rate remain to be explored. Because the neural pathways that regulate the heart cannot currently be directly imaged using calcium ion fluorescent labeling, this example uses propranolol and atropine to act on specific receptors on the heart to explore whether the pathways of specific receptor action are the key to controlling heart rate changes. Figure 6 Figure a shows a flowchart of drug incubation treatment of zebrafish: the same batch of 5 dpf zebrafish embryos were selected and incubated in the same concentrations of MS222, propranolol and atropine culture medium (within the tolerance range of drug concentration). After 24 hours, they developed to 6 dpf, and then the otolith oscillation experiment was performed and the corresponding heart rate was recorded.

[0102] Existing studies and pharmacological experiments have shown that propranolol is a β-adrenergic receptor blocker that mainly acts on the sympathetic nervous system. Under normal circumstances, when the sympathetic nerves are in an excited state, their terminals will release norepinephrine, which will bind to the β-adrenergic receptors on the heart, thereby achieving the effects of accelerating heart rate and enhancing myocardial contractility. Figure 6 As shown in Figure b, after incubation with propranolol, when the otoliths were optically oscillated, it was found that the heart HRV value decreased with increasing drug concentration. This is because propranolol can competitively block β-adrenergic receptors, preventing norepinephrine from binding to the receptors. Therefore, when the zebrafish is in the otolith oscillation state, if propranolol is administered, even if the sympathetic nervous system is excited by the otolith oscillation stimulation, the norepinephrine released by it cannot effectively bind to the β-receptor and exert its effect, thereby weakening the heart rate acceleration effect originally caused by sympathetic nerve excitement and reducing the increase in heart rate.

[0103] In contrast, after incubation with atropine, the corresponding cardiac HRV value continued to increase with increasing drug concentration ( Figure 6 c in the figure). This is because atropine is an M-type acetylcholine receptor antagonist that mainly acts on the parasympathetic nervous system. Under normal physiological conditions, when the parasympathetic nerves are excited, their terminals will release acetylcholine, and these acetylcholines will bind to the M-type acetylcholine receptors on the heart, increasing the permeability of the myocardial cell membrane to potassium ions, and potassium ions will flow out. Such changes lead to hyperpolarization of myocardial cells, which in turn slows the heart rate and weakens the myocardial contractility. When atropine is administered to zebrafish, it blocks the binding process of acetylcholine to the M-type receptors. At this time, when the present embodiment uses laser oscillation otoliths, even if the parasympathetic nervous system is activated and acetylcholine is released, the drug effect of atropine causes acetylcholine to be unable to bind normally to the receptors, and thus cannot play its role in slowing down the heart rate. At this time, the heart rate slowing effect caused by parasympathetic nerve excitement will be suppressed, so that the heart rate will not drop as normally when stimulated by otolith oscillation, and may even rise under the relative action of the sympathetic nerves. At the same time, if Figure 6 As shown in d, this example compares the effects of the anesthetic MS222 on heart rate and finds 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 juvenile fish respectively, MS222 has almost no effect on the change of heart HRV value ( Figure 6 e in the above example).

[0104] The above experimental results confirm that propranolol and atropine can respectively block the sympathetic and parasympathetic nerves that regulate heart rate, thus proving that otolith oscillations need to utilize this neural pathway to regulate the heart's rhythm. Based on the above experiments and data analysis, this example successfully obtained a neural pathway model for zebrafish otolith light-controlled oscillations to achieve precise heart rate regulation. Figure 6 As shown in figure f, the otoliths are connected to the base of the hair cells via ciliary bundles. Oscillations of the otoliths deflect these bundles, activating the hair cells and generating auditory input. Simultaneously, auditory nerve signals travel via the eighth pair of cranial nerve fibers to the vagal motor nucleus in the medulla oblongata (for preliminary processing and integration of auditory information), and then ascend to the semicircular rings in the midbrain (for more complex processing and analysis of auditory information). This processed auditory information is then transmitted to the thalamus and hypothalamus. As a crucial relay station for sensory transmission, the thalamus filters, integrates, and modulates auditory information (including regulating both sympathetic and parasympathetic nerves). During this process, neural signals are transmitted from the hypothalamus and vagal motor nucleus in the dorsal medulla oblongata to the heart, triggering the release of norepinephrine from sympathetic nerves. This sympathetic release binds to specific β-adrenergic receptors on the myocardial cell membrane, acting on the sinoatrial node (SAP) to induce myocardial contraction and increase the heart rate. This experimental system reveals a neural pathway in the vestibular-central nervous system-heart axis that regulates heart rate based on otolith oscillations.

[0105] Furthermore, in some embodiments, the biomedical application of light-controlled otolith oscillation to regulate heart rate can be specifically as follows:

[0106] This example first verified the therapeutic effect of light-controlled otolith oscillation on the atrioventricular block disease model. Before the experiment began, this example prepared a stock solution containing terfenadine, and added an appropriate amount of terfenadine stock solution to the culture medium and ultrasonicated it for 10 minutes. The terfenadine concentration in the culture medium was adjusted to 5 . Subsequently, in this embodiment, 5 dpf zebrafish embryos were placed in a culture medium containing drugs for co-incubation. After 24 hours of incubation, zebrafish embryos with a successful atrioventricular block disease model were screened out under a microscope. Furthermore, in this embodiment, the modeled zebrafish embryos were placed on a sample stage, and the constructed optical tweezers system was used to perform programmable light-controlled oscillations on the otoliths. At the same time, the beating of the zebrafish heart before and after the otolith oscillations was observed and recorded. Experimental data showed that under normal circumstances (Control group), the beating frequencies of the atria and ventricles of the zebrafish heart remained consistent, that is, 1:1. However, after drug incubation, the zebrafish embryos underwent atrioventricular block, and the beating of the atria and ventricles became obviously inconsistent. Specifically, when the atria beat twice, the ventricles beat only once, and the atrioventricular ratio became 2:1. This situation is like Figure 7 In response to this phenomenon, the present embodiment applies a scanning light trap precisely to the otolith to generate a specific oscillation pattern. At the same time, the present embodiment observed that during the otolith oscillation, the atrioventricular ratio of the zebrafish heart improved significantly, and the two gradually became synchronized. On this basis, the present embodiment continued the light-controlled otolith oscillation for 20 minutes, and then turned off the laser. Figure 7 As shown in stage III in a, the atrioventricular beat ratio returns to the normal 1:1. At the same time, it can be seen from the heartbeat waveform that for a period of time after the otoliths stop oscillating, the atrial and ventricular beat ratio of the zebrafish heart is still 1:1, that is, the phenomenon of atrioventricular block no longer occurs, which confirms that light-controlled oscillating otoliths can achieve targeted treatment of atrioventricular block. Further, this embodiment explores the influence of otolith oscillation amplitude on the treatment of atrioventricular block disease. Figure 7 As shown in b, when the otolith oscillation amplitude changes from 0.5 Increase to 2 The time required for otolith oscillation therapy to restore atrioventricular block to normal heartbeat was significantly shortened from 30 minutes to just 10 minutes. This experiment shows that the greater the amplitude of otolith oscillation manipulated by the scanning optical trap, the stronger the effect on heartbeat correction, thus shortening the treatment time and achieving a more pronounced therapeutic effect.

[0107] In addition to atrioventricular block, this example further explored the therapeutic effect of light-controlled otolith oscillation on cardiac escape disease. In this experimental study, this example used 6 dpf zebrafish embryos and 0.5 Sunitinib was co-incubated with embryo culture medium. After 5 hours, the zebrafish embryos incubated with the drug were observed under a microscope. It was found that the zebrafish heart beat would briefly pause, also known as cardiac arrest. Moreover, different concentrations of the drug caused different pause times, and the longer the pause time, the more serious the deterioration of heart function. Figure 7 As shown in c in , at t = 3 s, this embodiment observed a pause time of about 2 s in the zebrafish embryonic heart, confirming that this embodiment successfully constructed a zebrafish model with cardiac esophageal disease, laying the foundation for subsequent therapeutic experiments. Next, this embodiment uses optical tweezers to perform continuous controllable oscillations on the zebrafish otoliths for 30 minutes. At this time, the optical tweezers apply a regular mechanical stimulation signal to the otoliths, which will be transmitted to the heart through the neural regulation mechanism, thereby affecting and correcting its heart rhythm. After turning off the laser, this embodiment continues to observe and record the heartbeat. After the light-controlled otolith oscillation, the intermittent heartbeats that originally appeared disappeared, the heartbeat rhythm returned to normal, and it continued to beat normally after the laser was turned off. Furthermore, the treatment time will increase with the increase in the heartbeat pause time ( Figure 7 (d) This means that the more severe the deterioration of heart function, the longer the required otolith oscillation treatment time. This finding further confirms the effectiveness of light-controlled otolith oscillation in improving abnormal heart rhythms and provides new research ideas for the development of non-invasive treatments for abnormal heart rhythms.

[0108] As a common heart disease, bradycardia can have many adverse effects on the patient's daily life and physical health. For example, patients may experience symptoms such as fatigue, dizziness, and syncope. In severe cases, they may also develop more serious secondary heart diseases such as heart failure. Therefore, the prevention and treatment of bradycardia is particularly important. It can not only help patients improve their quality of life, but also effectively reduce the risk of other complications caused by bradycardia. In view of this, this example further verifies the therapeutic effect of light-controlled otolith oscillation on bradycardia. First, this example combines 5 dpf zebrafish embryos and 50 The zebrafish embryos were co-incubated with a culture medium containing a high concentration of tolterodine, and then used to construct a bradycardia disease model. During the experiment, the zebrafish embryos were placed under a high-power microscope for observation after 24 hours of drug incubation. The results showed that compared with the resting heart rate of 2.3 Hz of normal zebrafish, the heart rate of the incubated zebrafish was significantly lower (only 1.3Hz), indicating that the bradycardia model of zebrafish was successfully constructed in this embodiment. Subsequently, the laser was manipulated to controllably oscillate the otoliths. After ten minutes of otolith oscillation, the heart rate of the zebrafish was successfully observed to increase to 1.51Hz, confirming the improvement effect of light-controlled otolith oscillation on bradycardia ( Figure 7 Furthermore, when the laser was removed 20 minutes after the otolith oscillation, the zebrafish's heart rate eventually stabilized at 1.75 Hz, an increase of 0.45 Hz compared to before treatment, confirming the potential of otolith oscillation to improve bradycardia.

[0109] In addition to achieving optimized treatment of diseases, this embodiment also quantitatively explores the regulatory effects of different music on heart rhythm based on light-controlled otolith oscillation technology. The specific principle is as follows: By real-time programming of the spatial scanning sequence of the laser, this embodiment loads different musical rhythms into the oscillation mode of the otolith, thereby simulating the auditory input of different types of music, and quantitatively characterizing its regulatory effect on heart rhythm. Among them, this embodiment defines the mode with large sudden changes in the light trap oscillation amplitude and frequency as exciting music, and conversely, the oscillation mode with small changes in the light trap oscillation amplitude and frequency is defined as soothing music ( Figure 7 g in the figure). By loading two different types of music onto the otoliths, this example obtained heartbeat waveforms for different music types. When the resting heart rate was 2.32 Hz, the heart rate fluctuated slightly, increasing to 2.55 Hz when soothing music was added to the otoliths. However, when exciting music was added, the heart rate increased sharply and rose to 2.98 Hz. Furthermore, this example verified the heart rate response patterns of these two conditions on 20 zebrafish embryos. Figure 7 As shown in h, compared to soothing music, exciting music increased heart rate by approximately 20% on average. These experiments validate the ability of light-controlled otolith oscillations to characterize heart rhythm regulation under different music therapy modalities, and are expected to provide new technologies and approaches for further unraveling the physical mechanisms and physiological properties behind music therapy.

[0110] Example 3

[0111] like Figure 9 As shown, this embodiment provides a heart rate control system, including:

[0112] An optical potential well generation module, used for generating at least two optical potential wells at the edge of the otolith of the target living body through a scanning optical tweezers system;

[0113] An oscillation module is used to dynamically adjust the switching frequency, power, and spatial distribution pattern of the optical potential well through a scanning optical tweezers system, driving the otolith to oscillate according to the set parameters;

[0114] The heart rate variation calculation module is used to collect the heartbeat video of the target living body in real time and calculate the heart rate variation through image processing algorithm;

[0115] The heart rate control module is used to adjust the otolith oscillation parameters and the heart rate of the target living body according to the target heart rate control requirements; 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 base of the otoliths to generate auditory signals; the auditory signals are transmitted to the heart via the vagus motor nucleus in the hypothalamus and the dorsal medulla oblongata, causing the sympathetic nerves to release norepinephrine, which binds to specific β-adrenergic receptors on the myocardial cell membrane, thereby accelerating the myocardial contraction heart rate.

[0116] The heart rate control system also includes:

[0117] The pre-processing module is used to perform a pre-processing step on the target living body, wherein the pre-processing step includes anesthetizing and fixing the target living body.

[0118] Among them, the optical potential well generation module in the heart rate control system specifically includes:

[0119] a deflected beam generating unit, configured to generate a deflected beam using a 1064 nm laser via an acousto-optic deflector;

[0120] a beam expansion unit, configured to expand the diameter of the deflected light beam to a diameter capable of covering an entrance pupil of a high-magnification inverted objective lens using a beam expander;

[0121] The optical potential well generating unit is used to reflect the near-infrared laser in the deflected light beam to the high-magnification inverted objective lens through a short-wavelength dichroic mirror, and focus the near-infrared laser to the edge of the otolith of the target living body to form an optical potential well.

[0122] In summary, this application has the following technical effects:

[0123] 1. Based on a scanning optical tweezers system, we achieved programmable control of an optical potential well, which was then applied to the stable capture and multi-mode oscillation manipulation of the otoliths of zebrafish larvae, providing a powerful tool for simulating complex auditory input.

[0124] 2. Compared with existing research methods, light-controlled otolith oscillation has the unique advantages of being non-contact, non-destructive, and programmable, and can accurately trigger and change the movement state of otoliths at the microscopic level.

[0125] 3. Precise manipulation of otoliths provides an unprecedented technical solution for in-depth and accurate analysis of the changing mechanisms of physiological indicators such as auditory signal transmission, emotion regulation, and heart rate control.

[0126] 4. Indirect heart rate regulation through light-controlled otolith oscillation avoids the shortcomings of traditional heart rate regulation methods, including drug therapy, electrical pacing and optical pacing, which face challenges such as potential toxicological damage, invasive implantation and complex transgenic processing.

[0127] 5. The ability of light-controlled otolith oscillations to characterize heart rhythm regulation under different music therapy patterns is expected to provide new technologies and means to further reveal the physical mechanisms and physiological characteristics behind music therapy.

[0128] The technical features of the above embodiments can be combined arbitrarily. To make the description concise, 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, they should be considered to be within the scope of this specification.

[0129] This document uses specific examples to illustrate the principles and implementation methods of this application. The description of the above examples is only intended to help understand the method and core concept of this application. At the same time, for those skilled in the art, based on the concept of this application, there may be changes in the specific implementation methods and application scope. In summary, the content of this specification should not be understood as limiting this application.

Claims

1. A heart rate control device based on light-controlled otolith oscillation, characterized in that: include: 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 a high-magnification inverted objective lens imaging optical path 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 cardiac status monitoring of the target living body; The illumination light path includes a first illumination light path and a second illumination light path; the first illumination light path is used to illuminate the imaging field of the target living body on the electric stage; the second illumination light 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 the position of the target living body is moved by displacement control of the stage.

2. A heart rate control device based on light-controlled otolith oscillation according to claim 1, characterized in that: The laser scanning optical path includes a 1064 nm laser, an acousto-optic deflector, a beam expander, a short-wavelength dichroic mirror and an inverted objective lens; the laser light emitted by the 1064 nm laser passes through the acousto-optic deflector, the beam expander, the short-wavelength dichroic mirror and the inverted objective lens in sequence and is incident on the imaging optical path of the high-magnification inverted objective lens.

3. The heart rate control device based on light-controlled otolith oscillation according to claim 1, characterized in that: The imaging optical path of the high-magnification inverted objective lens is composed of the high-magnification inverted objective lens and a first high-speed charge-coupled device; the first high-speed charge-coupled device is arranged vertically to 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 control device based on light-controlled otolith oscillation according to claim 1, characterized in that: 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 vertically arranged with the low-magnification electron microscope for real-time observation of the heart condition of the target living body.

5. A heart rate control system, characterized in that: include: An optical potential well generation module, used for generating at least two optical potential wells at the edge of the otolith of the target living body through a scanning optical tweezers system; An oscillation module is used to dynamically adjust the switching frequency, power, and spatial distribution pattern of the optical potential well through a scanning optical tweezers system, driving the otolith to oscillate according to the set parameters; The heart rate variation calculation module is used to collect the heartbeat video of the target living body in real time and calculate the heart rate variation through image processing algorithm; The heart rate control module is used to adjust the otolith oscillation parameters and the heart rate of the target living body according to the target heart rate control requirements; 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 base of the otoliths to generate auditory signals; the auditory signals are transmitted to the heart via the vagus motor nucleus in the hypothalamus and the dorsal medulla oblongata, causing the sympathetic nerves to release norepinephrine, which binds to specific β-adrenergic receptors on the myocardial cell membrane, thereby accelerating the myocardial contraction heart rate.

6. The heart rate control system according to claim 5, characterized in that: Also includes: The pre-processing module is used to perform a pre-processing step on the target living body, wherein the pre-processing step includes anesthetizing and fixing the target living body.

7. The heart rate control system according to claim 5, characterized in that: The optical potential well generation module specifically includes: a deflected beam generating unit, configured to generate a deflected beam using a 1064 nm laser via an acousto-optic deflector; a beam expansion unit, configured to expand the diameter of the deflected light beam to a diameter capable of covering an entrance pupil of a high-magnification inverted objective lens using a beam expander; The optical potential well generating unit is used to reflect the near-infrared laser in the deflected light beam to the high-magnification inverted objective lens through a short-wavelength dichroic mirror, and focus the near-infrared laser to the edge of the otolith of the target living body to form an optical potential well.

Images