Drug targeted delivery device and method

Abstract

The present invention discloses a drug targeted delivery device and method. The device includes a control light path parallel to the fluorescence excitation light path, and the control light path is perpendicular to the illumination light path; a charge-coupled device and the illumination light path are on the same light path; an illumination light path device generates illumination light to illuminate a living sample in a sample chamber; a living sample contains nanomedicine; a fluorescence excitation light path device emits a broad-spectrum white light, which excites the nanomedicine to emit fluorescence through an inverted objective lens; the control light path device emits a laser beam, which is illuminated on the living sample through an inverted objective lens; the charge-coupled device collects images in the sample chamber and converts them into electrical signals. Based on the image electrical signals, a computer adjusts the deflection angle of an acousto-optic deflector according to the speed and direction of targeted delivery to set the optical potential position. The nanomedicine is subjected to an optical gradient force exerted by the laser beam, and then moves synchronously with the irradiation position of the laser beam. The present invention can avoid invasive damage to the organism and achieve high-precision directional transmission of nanomedicines.

Description

Technical Field

[0001] The present invention relates to the field of targeted drug delivery, and in particular to a targeted drug delivery device and method. Background Art

[0002] At present, technologies based on magnetic fields and ultrasound have begun to be applied to the delivery of nanomedicines. However, the above schemes require the invasive implantation of external materials, such as transplanting magnetically controlled nanoparticles, ultrasound sources, and metal electrodes into blood vessels as control markers or excitation sources, which will cause unavoidable invasive damage to the organism. In addition, the exogenous materials used in the above schemes are usually less compatible with the organism itself, and are prone to induce immune system feedback in the body and be actively cleared, thereby reducing the accuracy and stability of the control. At the same time, due to mechanism limitations, they are usually difficult to achieve real-time control of the transport trajectory of a single nanomedicine, and face challenges in the transport direction and delivery accuracy of the drug. Summary of the Invention

[0003] The purpose of the present invention is to provide a drug targeted delivery device and method to solve the problem of invasive damage to the organism caused by the invasion of external materials to control nanomedicines and the inability to control the transport trajectory of a single nanomedicine in real time.

[0004] To achieve the above object, the present invention provides the following solutions:

[0005] A drug targeted delivery device, comprising: a control optical path device, an illumination optical path device, a fluorescence excitation optical path device, an inverted objective lens, a charge coupled device, a computer, and a sample chamber;

[0006] The computer is connected to the laser and the acousto-optic deflection plate in the control optical path device and the charge coupled device respectively;

[0007] The control light path of the control light path device is parallel to the fluorescence excitation light path of the fluorescence excitation light path device, and the control light path is perpendicular to the illumination light path of the illumination light path device; the charge coupled device and the illumination light path are on the same light path;

[0008] The illumination optical path device is used to generate illumination light to illuminate the living sample in the sample chamber; the living sample contains nanomedicine;

[0009] The fluorescence excitation optical path device is used to emit broad-spectrum white light, which is focused by the inverted objective lens to excite the nanomedicine, causing the nanomedicine to emit fluorescence;

[0010] The control optical path device is used to emit a laser beam, which is irradiated onto the living sample after passing through the inverted objective lens;

[0011] The charge coupled device is used to collect the image in the sample chamber, convert it into an image electrical signal, and upload the image electrical signal to the computer;

[0012] The computer is used to adjust the deflection angle of the acousto-optic deflection plate in real time based on the image electrical signal and the speed and direction of targeted delivery to adjust the position of the optical potential. The nanomedicine is subjected to the optical gradient force exerted by the laser beam, and the nanomedicine moves synchronously with the irradiation position of the laser beam; the irradiation position is the position of the optical potential.

[0013] Optionally, the optical path control device specifically includes: a laser, an acousto-optic deflector, a beam broadening device and a spectroscope;

[0014] The laser beam emitted by the laser sequentially passes through the acousto-optic deflection plate, the beam broadening device, the beam splitter, and the inverted objective lens and is irradiated into the living sample;

[0015] The acousto-optic deflection plate deflects according to the deflection angle set by the computer;

[0016] The beam expansion device is used to expand the diameter of the laser beam to completely cover the entrance pupil of the inverted objective lens, and the inverted objective lens focuses the laser beam and then irradiates it onto the living sample.

[0017] Optionally, the fluorescence excitation optical path device specifically includes: an LED light source, a collimator, a filter and a reflector;

[0018] The LED light source is used to generate broad-spectrum white light; the broad-spectrum white light passes through the collimator, the filter and the reflector in sequence, is reflected onto the beam splitter, and is irradiated onto the living sample through the inverted objective lens.

[0019] Optionally, the lighting optical path device specifically includes: a halogen light source and a condenser;

[0020] The halogen light source emits illumination light, which is irradiated onto the living sample through the condenser to generate an image electrical signal of the microscope field of view; the image electrical signal is collected by the inverted objective lens and sent to the charge coupled device.

[0021] Optionally, the illumination optical path device, the inverted objective lens, the beam splitter, the reflector and the charge coupled device are on the same optical path.

[0022] A targeted drug delivery method, wherein the targeted drug delivery method is applied to a targeted drug delivery device, and the targeted drug delivery method comprises:

[0023] Adjusting the deflection angle of the acousto-optic deflection plate and setting a plurality of optical potentials; the positions of the optical potentials are the irradiation positions of the laser beam; and the laser beam is irradiated on the living sample;

[0024] irradiating the laser beam to different positions of the optical potential at different times, and repeatedly scanning the laser beam between the positions of the optical potential to generate a transport trajectory;

[0025] The nanomedicine in the living sample is subjected to the optical gradient force exerted by the laser beam. According to the transport trajectory, the nanomedicine moves synchronously with the irradiation position of the laser beam until it reaches the target position.

[0026] Optionally, the nanomedicine in the living sample is subjected to the optical gradient force exerted by the laser beam, and according to the transport trajectory, the nanomedicine moves synchronously with the irradiation position of the laser beam until it reaches the target position, which specifically includes:

[0027] When the nano drug approaches the white blood cells and platelets in the living sample along the transport trajectory, the laser beam is turned off, and the nano drug is loaded on the white blood cells and platelets under endocytosis;

[0028] According to the transport trajectory, the leukocytes and platelets loaded with the nanomedicine are directed to migrate to the target location.

[0029] Optionally, the step of directionally migrating the leukocytes and platelets loaded with the nanomedicine to the target location according to the transport trajectory further comprises:

[0030] Turning on the laser beam and applying a plurality of optical potentials to deform the white blood cells and platelets loaded with the nanomedicine and actively release the nanomedicine;

[0031] Allowing the laser beam to dynamically scan a circular trajectory at the target position to form a circular trajectory;

[0032] constructing a red blood cell microfluidic pump under the action of the annular trajectory;

[0033] Driven by the red blood cell microfluidic pump, the nanomedicines are delivered to the target location in batches.

[0034] According to the specific embodiments provided by the present invention, the present invention discloses the following technical effects: the present invention provides a drug targeted delivery device and method, which uses a laser to emit a laser beam, irradiates a living sample, sets an optical potential position, and allows the nano-drug to move synchronously with the irradiation position of the laser beam; the irradiation position is the optical potential position; through external laser irradiation, invasive damage to the organism is avoided. In addition, by setting the optical potential position to form an irradiated transport trajectory, the nano-drug can be moved synchronously with the transport trajectory to achieve directional transmission. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to more clearly illustrate the embodiments of the present invention 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 invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0036] Figure 1 This is a structural diagram of the drug targeted delivery device provided by the present invention;

[0037] Figure 2 This is a schematic diagram of the principle of targeted drug delivery provided by the present invention;

[0038] Figure 3 The present invention provides a method for achieving targeted delivery of a single nano drug in vitro and in vivo based on scanning optical tweezers; wherein, Figure 3 a is the synthesis and characterization of doxorubicin-loaded mesoporous silica particles (DOX@MSN); Figure 3 a1 is a schematic diagram of the three-dimensional structure of DOX@MSN; Figure 3 a2 is an optical microscopic image of DOX@MSN; Figure 3 a3 is a schematic diagram of the fluorescence image of DOX@MSN; Figure 3 b is the synthesis and characterization of mesoporous silica particles loaded with urokinase (URK@MSN); Figure 3 b1 is a schematic diagram of the three-dimensional structure of URK@MSN; Figure 3 b2 is a schematic diagram of the optical microscopic image of URK@MSN; Figure 3 b3 is a schematic diagram of the fluorescence image of URK@MSN; Figure 3 c is the fluorescence trajectory diagram of a single URK@MSN nanodrug manipulated in vitro; Figure 3 c1 is the fluorescence image of the nanodrug moving along the “O”-shaped trajectory; Figure 3 c2 is the fluorescence image of the nanodrug moving along the “J”-shaped trajectory; Figure 3 c3 is the fluorescence image of the nanomedicine moving along the “N”-shaped trajectory; Figure 3 c4 is the fluorescence image of the nanodrug moving along the “U”-shaped trajectory; Figure 3 d is a schematic diagram of the optical microscopy image of the reverse blood flow delivery of a single URK@MSN nanodrug in vivo; Figure 3 d1 is the initial position diagram of a single URK@MSN nanodrug in the blood vessel;

[0039] Figure 3 d2 is a schematic diagram of the spontaneous movement of URK@MSN nanomedicine along with blood flow when the laser is turned off; Figure 3 d3 is an image schematic diagram of optical manipulation of a single URK@MSN to achieve reverse blood flow delivery;

[0040] Figure 3 e is a schematic diagram showing the maximum speed that can be achieved by intravascular optical manipulation of URK@MSN nanomedicine delivery in different directions and its relationship with power; Figure 3 e1 is a schematic diagram of the maximum speed that can be achieved by optically manipulating the delivery of a single URK@MSN nanodrug in different directions; Figure 3 e2 is a schematic diagram showing the relationship between the maximum velocity achievable in reverse blood flow delivery and the laser power; Figure 3 f is a schematic diagram of optical manipulation of a single URK@MSN to achieve round-trip delivery around obstacles in blood vessels; Figure 3 f1 is a schematic diagram of the initial position of an obstacle consisting of five red blood cells and a single URK@MSN nanomedicine in a blood vessel; Figure 3 f2 shows the optical manipulation of a single URK@MSN to bypass five red blood cells and deliver them back and forth along an “S” curve; Figure 3 g is a schematic diagram of the complex trajectory delivery of a single nanomedicine based on optical manipulation; Figure 3 g1 is a schematic diagram of optical manipulation of a single URK@MSN nanomedicine to achieve “petal” trajectory delivery; Figure 3 g2 is a schematic diagram of optical manipulation of a single URK@MSN nanomedicine to achieve a "pentagram" trajectory delivery; Figure 3 h is a schematic diagram of the minimum step accuracy of optical manipulation of a single URK@MSN; Figure 3 h1 is the optical bright field image of the minimum stepping accuracy that can be achieved by optically manipulating a single URK@MSN; Figure 3 h2 is the fluorescence image of the minimum stepping accuracy that can be achieved by optically manipulating a single URK@MSN;

[0041] Figure 4 Schematic diagram of the experimental process of targeted delivery of multiple nanoparticles of drug in vivo provided by the present invention; wherein, Figure 4 a is a schematic diagram of an optical microscopic image showing the targeted delivery of three nanomedicines in a blood vessel; Figure 4 a1 is a schematic diagram of optically manipulating three nano-drugs to arrange into a triangular structure;

[0042] Figure 4a2 is a schematic diagram of delivering three nanomedicines against the blood flow and forming a triangle on the other side; Figure 4 b is a schematic diagram of an optical microscopic image for achieving overall delivery of three nanomedicines; Figure 4 b1 is a schematic diagram of an optical microscopic image showing three nanomedicines arranged into a triangular structure in a blood vessel through optical manipulation; Figure 4 b2 is a schematic diagram of an optical microscopic image of the overall reverse blood flow delivery of three nanomedicines; Figure 4 c is a schematic diagram of an optical microscopic image showing the targeted delivery of three different nanomedicines to different vascular branches;

[0043] Figure 4 c1 is a schematic diagram of an optical microscopic image of three nanodrugs captured simultaneously and arranged linearly in vivo; Figure 4 c2 is a schematic diagram of an optical microscopic image of the targeted delivery of nanodrug 1 to branch vessel Ⅰ;

[0044] Figure 4 c3 is a schematic diagram of an optical microscopic image of the targeted delivery of nanodrug 2 to branch vessel II; Figure 4 c4 is a schematic diagram of an optical microscopic image of the targeted delivery of nanodrug 3 to branch vessel III; Figure 4 d is an optical microscopic image showing five nanoparticles transported against the blood flow in zebrafish blood vessels and reconfigured into a triangle after bypassing the obstacle. Figure 4 d1 is a schematic diagram of an optical microscopic image of five nanoparticles arranged into a triangular structure within a blood vessel; Figure 4 d2 is a schematic diagram of an optical microscopic image showing four nanomedicines being manipulated sequentially to bypass an obstacle and be delivered to the other side; Figure 4 d3 is an optical microscopic image showing the fifth nanomedicine successfully bypassing the obstacle and forming a new triangular structure on the other side;

[0045] Figure 5 This is a schematic diagram of an optical microscopic image of the indirect delivery of nanomedicines using two endogenous cells in blood vessels provided by the present invention; wherein, Figure 5 a is an optical microscopic diagram showing the indirect delivery of nanoparticles using platelet phase within zebrafish blood vessels; Figure 5 a1 is an optical microscopic diagram of platelets and URK@MSN captured in blood vessels; Figure 5 a2 is a schematic diagram of an optical microscopic image of a single URK@MSN loaded onto a platelet; Figure 5 a3 is a schematic diagram of an optical microscopic image of platelets loaded with URK@MSN being delivered to the vicinity of the vascular wall; Figure 5 a4 is a schematic diagram of an optical microscopic image of the platelets induced to release nanomedicine loaded with URK@MSN; Figure 5b is a schematic diagram of an optical microscopic image of indirect delivery of nanomedicines using leukocytes in zebrafish blood vessels; Figure 5 b1 is a schematic diagram of the optical microscopic image of the distribution and capture of leukocytes and URK@MSNs in blood vessels; Figure 5 b2 is a schematic diagram of an optical microscopic image of a single URK@MSN loaded onto a leukocyte;

[0046] Figure 5 b3 is a schematic diagram of an optical microscopic image of leukocytes loaded with URK@MSN being delivered to the vicinity of the vascular wall; Figure 5 b4 is a schematic diagram of an optical microscopic image of the URK@MSN-loaded leukocytes inducing the release of nanomedicines; Figure 5 c is a schematic diagram of an optical microscopic image of the targeted delivery of nanomedicines to branch vessels using platelets in zebrafish blood vessels; Figure 5 c1 is a schematic diagram of optical microscopic images of platelets and URK@MSN captured in blood vessels; Figure 5 c2 is a schematic diagram of an optical microscopic image of a single URK@MSN loaded onto a platelet; Figure 5 c3 is a schematic diagram of an optical microscopic image of platelets loaded with URK@MSN being delivered to the wall of a branching vessel; Figure 5 c4 is a schematic diagram of the optical microscopic image of the platelets induced to release nanodrugs loaded with URK@MSN; Figure 5 d is an optical bright field image of intravascular targeted delivery of single nanoparticles of drug to different branch vessels using leukocytes;

[0047] Figure 5 d1 is a schematic diagram of optical microscopic images of leukocytes and URK@MSNs captured in blood vessels;

[0048] Figure 5 d2 is a schematic diagram of an optical microscopic image of a single URK@MSN loaded onto a leukocyte; Figure 5 d3 is a schematic diagram of an optical microscopic image of leukocytes loaded with URK@MSN being delivered to the vicinity of the wall of a branching vessel; Figure 5 d4 is a schematic diagram of the optical microscopy image of the URK@MSN-loaded leukocytes inducing the release of nanomedicines;

[0049] Figure 6 This is a schematic diagram of the method of assembling red blood cell micromotors in vivo to achieve batch delivery of nanomedicines provided by the present invention; wherein, Figure 6 a is a schematic diagram of assembling red blood cell micromotors in vivo to achieve bulk delivery of nanomedicines; Figure 6 b is a schematic diagram of an optical microscopic image of assembling a single red blood cell micromotor to achieve reverse blood flow delivery of two URK@MSNs; Figure 6 b1 is a schematic diagram of capturing and assembling a single red blood cell micromotor in a blood vessel; Figure 6 b2 is a schematic diagram of using a single red blood cell micromotor to deliver nanodrug 1 in the reverse direction of blood flow; Figure 6 b3 is a schematic diagram of the initial position of the nanodrug 2 in the blood vessel; Figure 6 b4 is a schematic diagram of an optical microscopic image of a single red blood cell micromotor for achieving reverse blood flow delivery of nanodrug 2; Figure 6 c is a schematic diagram showing the dependence of the red blood cell micromotor speed on the nanodrug delivery speed; Figure 6 d is a schematic diagram of the targeted delivery of four nanoparticles of drug to the blood vessel wall using a single red blood cell micromotor; Figure 6 d1 is a schematic diagram of an optical microscopic image of a single red blood cell micromotor; Figure 6 d2 is a schematic diagram of an optical microscopic image of the targeted blood vessel wall delivery of nanomedicine 1 using red blood cell micromotors; Figure 6 d3 is a schematic diagram of an optical microscopic image of the targeted blood vessel wall delivery of nanomedicine 2 using red blood cell micromotors; Figure 6 d4 is an optical microscopic diagram showing the targeted delivery of nanomedicine 3 to the blood vessel wall using red blood cell micromotors; Figure 6 d5 is an optical microscopic image showing the targeted delivery of four nanoparticles of drug to the blood vessel wall using red blood cell micromotors; Figure 6 e is a schematic diagram of using two red blood cell micromotors to achieve batch targeted delivery of nanoparticles to the blood vessel wall; Figure 6 e1 is a schematic diagram of the distribution of nanomedicine particles when assembling two red blood cell micromotors; Figure 6 e2 is a schematic diagram of the position distribution of nanomedicines after the two red blood cell micromotors rotate for 3.5s; Figure 6 e3 Schematic diagram of the position distribution of nanomedicines after the two red blood cell micromotors rotate for 6 seconds; Figure 6 e4 is a schematic diagram of the position distribution of nanomedicine particles after the two red blood cell micromotors rotated for 84.6s; Figure 6 e5 is a schematic diagram of the position distribution of the nanodrugs after the two red blood cell micromotors rotated for 118.6s;

[0050] Figure 7 Schematic diagram of the invention's targeted delivery of nanomedicine based on scanning optical tweezers to achieve precise treatment of thrombus and cancer cells; wherein, Figure 7 a is a schematic diagram of light-controlled targeted delivery of nanomedicine to a single cancer cell; Figure 7 a1 is a schematic diagram of the initial positions of cancer cells HL-60 and DOX@MSN nanomedicine; Figure 7 a2 is a schematic diagram of the optical microscopic image of DOX@MSN nanomedicine spontaneously moving in the blood; Figure 7 a3 is a schematic diagram of an optical microscopic image showing the successful delivery of DOX@MSN nanomedicine into HL-60 in the direction against blood flow; Figure 7a4 is the fluorescence image after DOX@MSN was successfully delivered to HL-60; Figure 7 b is a schematic diagram of targeted delivery of nanomedicine toward thrombus; Figure 7 b1 is a schematic diagram of the initial position of URK@MSN nanomedicine near the thrombus; Figure 7 b2 is a schematic diagram of the optical microscopic image of URK@MSN nanomedicine spontaneously moving with the blood flow; Figure 7 b3 is a schematic diagram of an optical microscopic image of a single URK@MSN nanodrug delivered to the thrombus against the blood flow; Figure 7 c is a schematic diagram of the targeted delivery of three nanomedicines toward thrombus; Figure 7 c1 is a schematic diagram of the initial position of nanodrug 1 near the thrombus;

[0051] Figure 7 c2 is a schematic diagram of an optical microscopic image showing that the nano drug 1 moves spontaneously with the blood flow and is then delivered to the thrombus in the reverse blood flow; Figure 7 c3 is a schematic diagram of an optical microscopic image of three URK@MSN nanomedicines delivered to the thrombus in the same manner; Figure 7 d is a schematic diagram of indirect delivery of nanomedicines to thrombus sites using leukocytes as drug carriers; Figure 7 d1 is a schematic diagram of an optical microscopic image showing the successful capture of leukocytes and URK@MSN nanomedicines near a thrombus; Figure 7 d2 is an optical microscopic image showing the successful loading of a single URK@MSN onto a leukocyte near a thrombus; Figure 7 d3 is a schematic diagram of an optical microscopic image of leukocytes loaded with URK@MSN being delivered to the thrombus; Figure 7 d4 is a schematic diagram of an optical microscopic image of drug release induced by URK@MSN-loaded leukocytes at the thrombus site;

[0052] Figure 7 e is a schematic diagram of the batch delivery of nanomedicines using red blood cell micromotors; Figure 7 e1 is the optical bright field image of the assembled red blood cell micromotor; Figure 7 e2 is the position diagram of the nanomedicine particles after the red blood cell micromotor worked for 7.7s; Figure 7 e3 is a schematic diagram of the position distribution of nanomedicine particles after the red blood cell micromotor has been operating for 14 seconds. DETAILED DESCRIPTION

[0053] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0054] The purpose of the present invention is to provide a drug targeted delivery device and method, which can avoid invasive damage to the organism and achieve directional transmission.

[0055] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the present invention is further described in detail below with reference to the accompanying drawings and specific embodiments.

[0056] The proper nouns involved in the present invention are as follows:

[0057] Optical tweezers: Based on the transfer of light momentum generated by the interaction of a strongly focused beam of light with matter, optical forces are generated on micro-nano particles and biological cells, trapping them at the laser focus. By changing the spatial and temporal distribution of the focus, precise alignment and rotational manipulation of multiple micro-nano particles and biological cells can be achieved.

[0058] Optical force: Specifically, it can be divided into optical gradient force and light scattering force. The light scattering force is along the direction of light propagation, and its magnitude is proportional to the laser intensity, pushing the micro-nanoparticles away along the optical axis. The optical gradient force is directed toward the laser focus, and its magnitude is proportional to the laser intensity gradient. It can be further divided into optical gradient force in the direction perpendicular to the optical axis and optical gradient force along the optical axis. When a micro-nanoparticle is near the focus, it will first be pulled toward the optical axis by the optical gradient force, and then be subjected to competing axial optical gradient force and light scattering force in the direction of the optical axis. When the axial optical gradient force dominates, stable capture and manipulation of micro-nanoparticles and biological cells can be achieved.

[0059] Optofluidic manipulation: Optofluidics studies how to control light and fluid at the micro- and nanoscale, and use the interaction between them to regulate the light field parameters. It can also stably capture and precisely manipulate micro- and nanoparticles and biological cells in microfluidics.

[0060] Scanning optical tweezers: Utilizing the interaction between an acousto-optic polarizer and a laser, rapid scanning of the laser focus between multiple positions in the focal plane (maximum scanning frequency: 100 kHz) is achieved. This allows a single laser beam to be used to create multiple optical potential wells through time-division multiplexing, enabling the stable capture and precise arrangement of multiple particles. By setting the laser focus to scan along a circular trajectory, the captured particles can be synchronously rotated, with the rotation direction and speed dynamically adjustable.

[0061] Biophotonics: Based on light or other forms of radiation energy, we understand the internal working mechanisms of living cells, enable the observation, detection, manipulation and analysis of living tissues and biological cells, and then apply them to clinical disease diagnosis and treatment.

[0062] Example 1

[0063] Figure 1 This is a structural diagram of the drug targeted delivery device provided by the present invention, such as Figure 1 As shown, a drug targeted delivery device includes: a control light path device, an illumination light path device, a fluorescence excitation light path device, an inverted objective lens, a charge coupled device, a computer and a sample chamber; the computer is respectively connected to the laser and the acousto-optic deflection plate in the control light path device and the charge coupled device; the control light path of the control light path device is parallel to the fluorescence excitation light path of the fluorescence excitation light path device, and the control light path is perpendicular to the illumination light path of the illumination light path device; the charge coupled device and the illumination light path are on the same light path; the illumination light path device is used to generate illumination light to irradiate the living sample in the sample chamber; the living sample has nanomedicine in it; the fluorescence excitation light path device is used to Broad-spectrum white light is emitted, which is focused by the inverted objective lens to excite the nanomedicine, causing the nanomedicine to emit fluorescence; the control optical path device is used to emit a laser beam, which is irradiated on the living sample after passing through the inverted objective lens; the charge-coupled device is used to collect the image in the sample chamber, and at the same time convert it into an image electrical signal, and upload the image electrical signal to the computer; the computer is used to adjust the deflection angle of the acousto-optic deflection plate in real time according to the direction and speed of targeted delivery based on the image electrical signal, so as to adjust the position of the optical potential, and the nanomedicine is subjected to the optical gradient force exerted by the laser beam, and the nanomedicine moves synchronously with the irradiation position of the laser beam; the irradiation position is the position of the optical potential.

[0064] In practical applications, acousto-optic deflection plates generate mechanical waves in a crystalline medium, causing periodic variations in the refractive index, thus 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 waves. This principle allows the focal position and intensity of a 1064nm laser to be varied.

[0065] The beam broadening device consists of two convex lenses with a distance equal to the sum of the focal lengths of the two convex lenses, which 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.

[0066] Beamsplitters are dichroic mirrors that 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. In this invention, a short-pass dichroic mirror is used, transmitting illumination light with wavelengths below 800 nm while reflecting near-infrared laser light above 800 nm.

[0067] A charge coupled device (CCD) is used to collect images in real time, and the specific collection frequency can be set through the PC.

[0068] LED light sources can generate light beams in four different wavelength bands: ultraviolet, blue, green and red.

[0069] The light beam generated by the LED light source is emitted as a parallel beam through the collimator, so that the light beam enters the lens with maximum efficiency.

[0070] By adjusting the filter, you can select any one of the four light beams: ultraviolet light, blue light, green light, and red light to reach the objective lens.

[0071] The light source and condenser provide the illumination light required to observe the sample in bright field.

[0072] Computer: The PC software, Tweez250si, controls the 1064nm laser on and off, sets the laser focus position, scanning frequency, and number of laser focal points, and controls the CCD to record images and videos. Additional software controls the LED light source module and allows for fluorescence observation and recording.

[0073] In practical applications, the wavelength of the laser is set to 1064 nanometers because biological tissue absorbs less of this wavelength, thus avoiding significant physical damage to tissues or cells due to photothermal effects.

[0074] The emitted laser interacts with the acousto-optic deflector, which can be figuratively understood as a reflector with precisely controlled deflection angles. By adjusting the deflection angle of the acousto-optic deflector, the laser beam can be directional deflected along a specific angle. The deflected laser beam first passes through a beam widening device, which consists of two convex lenses with a spacing equal to the sum of the focal lengths of the two convex lenses. This device is used to expand the output diameter of the laser so that the diameter can completely cover the entrance pupil of the inverted objective. After beam widening, the incident beam will be deflected upward by the short-wavelength dichroic mirror and focused at a preset position through the filter and the inverted objective, thereby achieving stable capture of nano-drug particles at the target position.

[0075] The specific focusing position of the laser can be set in the Tweez software interface using the mouse. The specific settings are as follows: The Tweez software interface can display the optical microscope image captured by the CCD in real time. By setting an optical potential well at a specified position on the image, the deflection angle of the acousto-optic deflector can be changed in real time through the internal feedback mechanism to focus the laser to the set position. At this time, the optical potential well set in the software interface can be changed by moving the mouse. At this time, the deflection angle of the acousto-optic deflector will also be adjusted accordingly, and the focusing position of the laser will also change in real time with the movement of the mouse. In this process, due to the existence of the optical gradient force, the captured nanomedicine will move synchronously with the laser, realizing the transportation of the nanomedicine along a specific trajectory.

[0076] In addition to capturing and manipulating individual nanotherapeutics, the acousto-optic deflector can be programmed in real time within the software interface, directing a laser beam to different locations at different times and repeatedly scanning between them. Because the acousto-optic deflector can achieve a maximum scanning frequency of 100,000 Hz, it enables simultaneous manipulation of multiple targets. For example, if 100 optical potential wells are configured at a scanning frequency of 100,000 Hz, the laser will scan back and forth in a cycle of: 1 / 2 / 3…98 / 99 / 100—1 / 2 / 3…98 / 99 / 100—1 / 2 / 3…98 / 99 / 100… In this case, within 1 second, the laser will illuminate each optical potential well 1,000 times, achieving a quasi-static distribution of the laser light across all 100 optical potential wells. Each optical potential well can then be treated as if it were continuously illuminated by a laser beam, allowing each well to stably capture a target nanotherapeutic, thereby enabling the simultaneous capture of multiple nanotherapeutics. Moreover, the position of each optical potential well can be independently controlled, thereby achieving the goal of moving a specific optical potential well while keeping the positions of other optical potential wells unchanged, thereby completing the independent control of a specific target object.

[0077] In practical applications, by using MATLAB software, the position coordinates of multiple optical potential wells can be combined into a patterned scanning sequence. At this time, the laser will move along the order set in the sequence under the action of the acousto-optic deflector, thereby realizing the multifunctional operation of nanomedicine. Taking rotation as an example, by arranging the above 100 optical potential wells into a circle with a diameter of 10μm, the laser will dynamically scan along the circular trajectory. At this time, the captured nanomedicine will also move synchronously, thereby realizing controllable rotation inside the blood vessel, such as Figure 2 shown.

[0078] The experimental system consists of three optical paths: the control optical path on the left, the illumination optical path in the middle, and the fluorescence excitation optical path on the right:

[0079] Controlled Optical Path: A 1064nm laser serves as the laser source, emitting near-infrared laser light. The laser light first passes through an acousto-optic deflector (AOD), whose deflection angle can be set using the Tweez250si software interface. The beam then passes through a beam expander, expanding the laser's output diameter to completely cover the entrance pupil of the inverted objective. The expanded laser light then passes through a shortwave beam splitter, reflected into the inverted objective, and focused by the inverted objective before irradiating the sample within the sample chamber.

[0080] Illumination Optical Path: A halogen light source above generates visible light as illumination, which is focused by a condenser and then illuminates the sample in the sample chamber. The condenser's height can be flexibly adjusted in the z-direction. During operation, the image of the microscope's field of view is collected by the inverted objective lens below, passes through a short-wavelength dichroic beamsplitter and a reflector, and is ultimately captured by a charge-coupled device (CCD). The CCD converts the image into an electrical signal, which is transmitted to a connected computer for real-time observation on the computer screen.

[0081] Fluorescence excitation light path: The right side is the excitation module for fluorescence imaging. The LED light source generates broad-spectrum white light, which is then calibrated by a collimator and then hits the filter module. In the experiment, a filter module with three transmission bands was selected, allowing the passing wavelength ranges of 361-398nm, 465-495nm, and 540-580nm, corresponding to ultraviolet light, blue light, and green light, respectively. By selecting the appropriate filter, you can select the appropriate excitation laser: Figure 3 In a3, green excitation light is required to excite DOX@MSN nanomedicine, so a filter with a wavelength of 540-580 nm is selected to allow green light to illuminate the surface of the particles and stimulate red fluorescence. Figure 3 In b3, blue excitation light is required to excite the URK@MSN nanomedicine. Therefore, a filter with a wavelength range of 465-495 nm is selected to allow blue light to illuminate the particle surface, stimulating green fluorescence. The excitation light is reflected upward by a reflector, passes through a short-wavelength beam splitter, and enters the inverted objective lens, where it illuminates the sample in the sample chamber.

[0082] The present invention operates as follows: After entering a blood vessel, nanoparticles typically diffuse randomly under the influence of blood flow. At this point, the laser focus position and specific laser intensity are set using a mouse on the real-time optical image captured by the CCD. The laser can then be precisely focused on the target nanoparticle by adjusting the deflection angle of the acousto-optic deflector. The nanoparticle is then subjected to the optical gradient force exerted by the focused laser beam, allowing it to be stably captured. By shifting the position of the focused laser beam, the captured nanoparticle moves synchronously with the focused laser beam, enabling it to be transported against the blood flow, enter a specific branch, and be targeted near the HL-60 blood cancer cell. Furthermore, with the assistance of optical manipulation, the nanoparticle can reach the interior of the cell.

[0083] In addition, by taking advantage of the rapid scanning of the acousto-optic deflector, two optical potential wells can be set up at the same time to capture a single nano-drug particle and white blood cells or platelets in the blood vessels, respectively, and precisely move the nano-drug particles close to the captured cells; at this time, the laser is turned off, and the released nano-drug particles will be loaded onto white blood cells and platelets under endocytosis. Furthermore, by changing the deflection angle of the acousto-optic deflector, the laser focus irradiation position can be changed in real time, and the white blood cells (or platelets) loaded with nano-drugs can be directed to migrate near the thrombus. After reaching the target position, multiple optical potential wells are applied simultaneously to deform the cells, thereby realizing the active release of the drug, and acting on the thrombus to achieve controllable dissolution of the thrombus.

[0084] Finally, the focused beam is set to dynamically scan along a circular trajectory. The captured red blood cells are then controllably rotated by the optical force, creating a red blood cell microfluidic pump. This pump then generates a driving flow field that allows a large number of drug particles to move, completing the bulk delivery of the nanoparticles toward the clot.

[0085] Example 2

[0086] A targeted drug delivery method is provided, wherein the targeted drug delivery method is applied to the targeted drug delivery device of embodiment 1, and the targeted drug delivery method comprises:

[0087] The deflection angle of the acousto-optic deflection plate is adjusted, and a plurality of optical potential points are set; the positions of the optical potential points are the irradiation positions of the laser beam; and the laser beam is irradiated on the living sample.

[0088] The laser beam is irradiated to different positions of the optical potential at different times, and the laser beam is repeatedly scanned between the positions of the optical potential to generate a transport trajectory.

[0089] The nanomedicine in the living sample is subjected to the optical gradient force exerted by the laser beam. According to the transport trajectory, the nanomedicine moves synchronously with the irradiation position of the laser beam until it reaches the target position.

[0090] In actual applications, the nanomedicine in the living sample is subjected to the optical gradient force exerted by the laser beam. According to the transport trajectory, the nanomedicine moves synchronously with the irradiation position of the laser beam until it reaches the target position. Specifically, the process includes: when the nanomedicine approaches the white blood cells and platelets in the living sample according to the transport trajectory, the laser beam is turned off, and the nanomedicine is loaded on the white blood cells and platelets under endocytosis; and according to the transport trajectory, the white blood cells and platelets loaded with the nanomedicine are directed to migrate to the target position.

[0091] In actual application, the white blood cells and platelets loaded with the nanodrug are directed to migrate to the target position according to the transport trajectory, and then the method further includes: turning on the laser beam, applying multiple optical potentials, deforming the white blood cells and platelets loaded with the nanodrug, and actively releasing the nanodrug; making the laser beam dynamically scan a circular trajectory at the target position to form a circular trajectory; constructing a red blood cell microfluidic pump under the action of the circular trajectory; and delivering the nanodrug in batches to the target position under the drive of the red blood cell microfluidic pump.

[0092] In practical applications, the basic manipulation of a single nanoparticle of drug is as follows:

[0093] In order to characterize the feasibility of using optical manipulation technology to achieve targeted delivery of nanodrugs, the dynamic delivery of single nanodrug particles in vivo and in vitro was first quantitatively characterized.

[0094] First, two nanoparticles of drug ( Figure 3 a and Figure 3 b): DOX@MSN particles are mesoporous silica nanoparticles (MSN) loaded with doxorubicin (DOX) with a diameter of 620nm; while URK@MSN particles are mesoporous silica nanoparticles (MSN) loaded with urokinase (URK) with a diameter of 700nm. The above two drugs were chosen because current research has confirmed that DOX can be used to kill and eliminate cancer cells, while urokinase (URK) can be used to effectively dissolve blood clots. By observing with a fluorescence microscope, it can be seen that the two particles exhibit red fluorescence ( Figure 3 a3) and green fluorescence ( Figure 3 b3), indicating that DOX and URK have been successfully encapsulated on the surface of MSNs.

[0095] On this basis, the prepared URK@MSN solution was placed on the stage of the scanning optical tweezers (SOT) system. By manipulating the position of the laser focus, the target URK@MSN particles were successfully captured stably (a mark point was set in the Tweez250si software, and the focus of the laser beam could be hit at the corresponding position to capture the particles. Specifically, the computer's Tweez software interface can display the optical microscope image captured by the CCD in real time. By moving the stage, the URK@MSN in the solution was found in the image, and an optical potential well was set at the position of a single URK@MSN. The deflection angle of the acousto-optic deflector can be changed in real time through the inherent feedback mechanism, and the laser can be focused to the set position (that is, the location of the particle). Due to the existence of the optical gradient force, the particle is captured in the optical potential well) and manipulated to move dynamically along the trajectory formed by the four letters "O", "J", "N" and "U" ( Figure 3c) confirmed that it is feasible to achieve stable capture and controllable delivery of single nanoparticles using optical manipulation technology. Based on this, DOX@MSN particles were injected into the posterior (tail) cardinal vein of 5-day-old zebrafish larvae (5dpf). Then, single URK@MSN nanoparticles were observed to diffuse freely with the blood flow in vivo, and their movement trajectory is shown in Figure 2. Figure 3 As shown by the arrowed curve in d2. At this time, by controlling the acousto-optic modulator to focus the laser on the target nanoparticles, it is successfully captured stably and can be driven to migrate in the direction opposite to the blood flow and perpendicular to the blood vessels ( Figure 3 d).

[0096] Furthermore, the ability to deliver nanoparticles in different directions in flowing blood using optical manipulation technology was quantitatively characterized. Figure 3 As shown in e, the blood in the blood vessel flows from right to left at a speed of 5 μm / s.

[0097] At this time, an optical potential trap was set up to stably capture a URK@MSN nanoparticle (laser power was 85 mW), and it was dynamically migrated in 8 directions, and the maximum migration speed that could be achieved while ensuring stable capture of the particle was recorded. Figure 3 As can be seen from e1, the maximum migration speed that can be achieved by migrating along the blood flow is 10.7 μm / s, while the maximum migration speed that can be achieved by migrating against the blood flow is the smallest, which is 8 μm / s. The migration speeds in other directions are between the two.

[0098] On this basis, we also quantitatively characterized the maximum velocity of a single URK@MSN particle that can be achieved in reverse blood flow migration as a function of laser power (set the marker point in the same way as above, drag the marker point to the particle with the mouse to capture the particle, and drag the marker point along the reverse blood flow direction with the mouse to achieve reverse blood flow movement of the particle). Figure 3 As shown in e2, the maximum migration speed increases with the increase of laser power. This is because as the laser power increases, the capture force exerted on the nanoparticles also increases, thereby achieving a greater transport speed.

[0099] In addition to transporting nanoparticles along linear trajectories, the ability to control the transport of nanoparticles along complex trajectories using optical manipulation techniques has also been explored. Figure 3 As shown in Figure f, a single URK@MSN particle was successfully manipulated to precisely bypass an obstacle consisting of five red blood cells in the blood vessel and be repeatedly transported along an "S"-shaped trajectory, and was manipulated to return to its initial position in 40.9 seconds.

[0100] In practical applications, similar to setting up a circular trajectory, create an S-shaped coordinate distribution and import it into the Tweez250si. Set an appropriate scanning frequency, such as 10kHz, and turn on the laser. The laser beam will rapidly scan along the coordinate points, and the CCD camera will record the image. The video will show the particle moving along the S-shaped trajectory. Specifically, the Tweez software interface displays the optical microscope image captured by the CCD in real time. By setting an optical potential well at a specific location on the image, the inherent feedback mechanism can be used to change the deflection angle of the acousto-optic deflector in real time, focusing the laser at the set position. The optical potential well set in the software interface can be changed by moving the mouse, which in turn adjusts the deflection angle of the acousto-optic deflector, and the laser focus position also changes in real time with the mouse movement. During this process, due to the optical gradient force, the trapped nanoparticle moves synchronously with the laser, achieving nanoparticle transport along a specific trajectory. In this scenario, after trapping the nanoparticle, dragging the mouse back and forth along the S-shaped trajectory can achieve this operation.

[0101] Furthermore, it was confirmed that scanning optical tweezers can be used to manipulate single nanoparticles of drug to move along more complex trajectories, such as along petal-shaped trajectories ( Figure 3 g1) and the pentagram ( Figure 3 g2) trajectory for controllable transport, and the speed of the particles during transport can be controlled in real time, so that they can be delivered at different speeds in stages. Finally, the step accuracy of transporting a single nanoparticle is quantitatively characterized, such as Figure 3 As shown in h, optical manipulation technology was successfully used to achieve the controllable delivery of single nanoparticles of drug with a minimum size of 500nm.

[0102] In practical applications, the delivery of multiple nanoparticles of drug in vivo is as follows:

[0103] Based on the targeted delivery of a single nanoparticle, the ability to simultaneously capture and target multiple nanoparticles within a living blood vessel using optical manipulation technology was demonstrated. Leveraging the rapid scanning and switching capabilities of the acousto-optic modulator (maximum switching frequency: 100kHz), multiple optical potential wells can be set up in specific areas within the blood vessel, thereby enabling the simultaneous capture of multiple target particles. Figure 4 As shown in a, three optical potential wells were used to capture three nanoparticles of drug at the same time and arranged them into a triangular array ( Figure 4 a1). On this basis, by independently controlling the position of each optical potential well, the nanoparticles are controlled to migrate dynamically in the direction opposite to the blood flow, and then reassemble into a new triangular array at the new position ( Figure 4 a2). In addition to achieving independent migration of particles in the triangular array, the synchronous movement of the triangular array can also be achieved. Figure 4As shown in b, the spatial positions of the three optical potential wells are moved simultaneously, and the three captured nanomedicines will also move synchronously, thereby realizing the synchronous transport of multiple particles against the direction of blood flow.

[0104] In addition to achieving targeted delivery of nanoparticles in a single blood vessel, optical manipulation technology can also be used to achieve controllable sorting of nanomedicines at vascular branches.

[0105] like Figure 4 As shown in Figure c, three optical potential wells are set up simultaneously in the blood vessel to capture three nano-drug particles and arrange them in a linear pattern. At this time, by independently manipulating the positions of the three optical potential wells, the three nano-particles were successfully delivered to three different vascular branches (three calibration points were set on the Tweez250si software, and the appropriate scanning frequency was set. The laser was turned on to search for particles, and the calibration points were placed at the location of the particles to capture them. The mouse was moved at different times to achieve the directional movement of the three particles). This confirms the potential application of optical manipulation technology in achieving targeted delivery of nano-drugs along different branches.

[0106] In addition to capturing and manipulating individual nanoparticles, the acousto-optic deflector can be programmed in real time within the software interface, directing a laser beam to different locations at different times and repeatedly scanning between them. Because the acousto-optic deflector has a maximum scanning frequency of 100,000 Hz, it enables simultaneous manipulation of multiple targets. For example, with a scanning frequency of 30,000 Hz, if three optical wells are simultaneously positioned at the locations of three nanoparticles, the laser will scan back and forth in a cycle of 1 / 2 / 3—1 / 2 / 3—1 / 2 / 3… In this case, within 1 second, the laser will illuminate each optical well 10,000 times, achieving a quasi-static distribution of the nanoparticles at the three optical wells. Each optical well can then be treated as if it were continuously illuminated by a laser beam, allowing each to stably capture a single target nanoparticle, thereby achieving simultaneous capture of all three nanoparticles. Furthermore, the position of each optical well can be independently manipulated, enabling independent manipulation of specific targets by moving a specific well while maintaining the positions of the others. By independently dragging the mouse at different times, dragging the three optical potential wells separately, you can complete the separate capture and movement of the three particles.

[0107] Finally, the obstacle avoidance performance during multi-particle delivery was verified. Figure 4As shown in Figure d, optical manipulation simultaneously captures five drug nanoparticles, directs their migration against the bloodstream, and sequentially circumvents an intravascular obstacle (a damaged cell adhered to the vessel wall) to reassemble into a new triangular array. This experiment demonstrates that optical manipulation can achieve simultaneous delivery of multiple drug nanoparticles (same as above, with five calibration points), and that the trajectory of each drug particle can be independently controlled in real time, which is crucial for achieving targeted delivery of nanoparticles and effectively improving transport efficiency.

[0108] In practical applications, indirect delivery of nanomedicines in vivo using leukocytes and platelets as carriers is shown below:

[0109] It is well known that after nano drug particles enter the body, they are easily actively cleared by the mononuclear phagocytic system (MPS) in the body, which greatly reduces the efficiency of drug delivery. In order to circumvent the phagocytic effect of MPS and further improve the efficiency of drug delivery, researchers have proposed that endogenous cells in blood vessels (such as white blood cells and platelets) can be used to achieve indirect delivery of nano drugs. However, this requires precise control of the spatial position of cells and nano drugs to achieve dynamic contact between the two. With the help of optical manipulation technology, the manipulation target can be precisely controlled, and white blood cells (or platelets) can be remotely directed to actively load nano drug particles, thereby transporting nano drugs to the target location for controlled release.

[0110] The experimental process is as follows Figure 5 As shown in a, at t=0s, two optical potential wells are applied to capture single URK@MSN particles and platelets that diffuse freely with the blood flow, respectively, and then the platelets are manipulated to approach the nanodrug particles. Under endocytosis, the platelets actively swallow the nanodrugs into the interior, realizing the dynamic loading of the drug. At this time, by adjusting the spatial position of the optical potential well, the platelets can be manipulated to migrate dynamically along the set trajectory and reach the target position of the blood vessel wall in 33s. After reaching the blood vessel wall, multiple optical potential wells are applied to the drug-loaded platelets at the same time to cause them to deform, ultimately achieving the controlled release of the nanodrug. In addition to platelets, white blood cells were also used as experimental subjects, and the light-controlled indirect delivery of nanodrugs was also achieved ( Figure 5 b) (Same as above, set the calibration point, move to the platelet to capture, and drag the mouse to perform displacement manipulation).

[0111] On this basis, we try to see whether we can selectively transport nanomedicines to different vascular branches through indirect delivery of cells. Figure 5As shown in Figure c, the drug was successfully loaded onto the platelets again. By precisely controlling the spatial position of the optical potential well, the platelets loaded with nanoparticles were transported from branch vessel I to branch vessel II. At the same time, by applying optical stretching to the platelets, the dynamic release of nanoparticles was successfully completed in branch II, achieving targeted delivery of nanoparticles to different branch vessels ( Figure 5 c). Furthermore, leukocytes were successfully used to deliver nanoparticles to specific branches, achieving controlled release of nanoparticles ( Figure 5 d).

[0112] In practical applications, the red blood cell microfluidic pump for bulk drug delivery is shown below:

[0113] In order to further improve the transmission speed and delivery efficiency of nanomedicines, a red blood cell microfluidic pump was constructed using optical manipulation technology to achieve batch delivery of nanomedicines. Figure 6 As shown in a, a stationary optical potential well is set to fix the center of the red blood cell, and a circular scanning optical potential well is applied at the same time, and the diameter of the circular optical potential well is equal to the red blood cell. At this time, under the action of optical force, the captured red blood cells will begin to rotate (set a calibration point to the center of the red blood cell in Tweez250si, import a circular scanning trajectory to the position shown by the dotted line, set the scanning frequency, turn on the laser, and the rotation of the red blood cell can be achieved), and its rotation speed and direction can be flexibly adjusted by optimizing the optical tweezers system (the higher the set laser scanning frequency, the faster the rotation speed, and the change of direction is achieved by editing the coordinate point text of the scan, that is: by using MATLAB software, the position coordinates of multiple optical potential wells can be combined into a patterned scanning sequence (sequence), and the laser will be moved along the set point in the sequence under the action of the acousto-optic deflector). Move in a certain order, thereby realizing the multifunctional operation of nanomedicine and cells. In order to complete the rotational manipulation, when the scanning frequency of the acousto-optic deflector is set to 100000 Hz, 1000 optical potential wells are arranged into a circle with a diameter of 10μm, and the laser will scan back and forth according to the cycle of: 1 / 2 / 3…998 / 999 / 1000——1 / 2 / 3…998 / 999 / 1000——1 / 2 / 3…998 / 999 / 1000…. At this time, within 1s, the laser will irradiate the position of each optical potential well 100 times, completing 100 cycles. At this time, a single optical potential well is set to capture and fix the red blood cells ( Figure 6 b1 solid dots), around the red blood cells Figure 6The circular sequence is placed at the position indicated by the dotted circle b1. The laser will dynamically scan along the circular trajectory, and one end of the captured red blood cell will also move synchronously, thereby achieving a rotation rate of 100 revolutions per second within the blood vessel. After setting the scanning frequency of the acousto-optic deflector to 50,000 Hz, the red blood cell rotation rate becomes 50 revolutions per second. Similarly, by varying the scanning frequency of the acousto-optic deflector, the red blood cell rotation rate can be flexibly controlled. At this point, the rotating red blood cell acts like a microfluidic pump, and a driving flow field is synchronously generated around it. Particles in the flow field will undergo directional motion along with the nanomedicine, thereby achieving batch delivery of nanomedicine particles.

[0114] To verify the above ideas, a series of delivery experiments were conducted. Figure 6 As shown in Figure b, by regulating the spatial and temporal distribution of the optical potential well, a red blood cell was stably captured and driven to rotate counterclockwise, thus successfully assembling a red blood cell microfluidic pump inside a living blood vessel. With the help of the microfluidic field generated by the rotation of the microfluidic pump, two URK@MSNs moving along the blood flow were successfully decelerated and achieved reverse migration ( Figure 5 b). Furthermore, by regulating the scanning speed between the optical potential wells, the rotation speed of the red blood cell microfluidic pump can be changed in real time, thereby achieving controllable adjustment of the delivery speed of nanoparticles ( Figure 5 c). Based on the single particle trajectory manipulation, the team successfully used a counterclockwise rotating red blood cell microfluidic pump to complete the controlled delivery of four nanoparticles of drug toward the blood vessel wall, ultimately successfully transporting them to the endothelial cells of the blood vessel wall ( Figure 5 d). Finally, two relatively rotating red blood cell microfluidic pump arrays were constructed. Under the action of the microfluidic field generated by them, the batch delivery of nanoparticles of drug was achieved and they were successfully delivered to the vicinity of the thrombus on the blood vessel wall ( Figure 5 e).

[0115] In practical applications, the targeted delivery of nanomedicine to single cancer cells and thrombi in vivo is shown below:

[0116] Based on the above results, we further explored how to use scanning optical tweezers technology to achieve targeted drug delivery to single cancer cells and thrombus locations. It is well known that achieving single-cell targeted delivery of nanomedicines is of great significance in the fields of treatment, biomarker detection and disease diagnosis. In the experiment, based on the microinjection technology, the human promyelocytic leukemia cell line HL-60 and DOX@MSN solution were injected into the blood vessels of zebrafish embryos through their veins. At t=0s, a free HL-60 near the blood vessel wall was found in the blood vessel, and then a DOX@MSN particle appeared near it, which passively diffused with the blood flow. With the help of optical manipulation technology of acousto-optic modulation, at t=25.5s, the DOX@MSN particles were stably captured and dynamically delivered to the vicinity of a single HL-60 cell, achieving single-cell precision targeted delivery of nanomedicines ( Figure 7 a) In addition to single cancer cells, targeted drug delivery to thrombus is also achieved. Figure 7 As shown in Figure b, with the help of scanning optical tweezers technology, a single nanoparticle was first accurately delivered to the thrombus location by direct delivery. Furthermore, using multi-potential well manipulation technology, the motion state of three nanoparticles, which originally passively diffused with the blood flow, was successfully changed, and they were simultaneously and precisely delivered to the thrombus ( Figure 7 c). Next, thrombosis treatment was performed using indirect delivery with the help of white blood cells and platelets. At time t = 0s, a free white blood cell and URK@MSN were captured, and the URK@MSN was successfully loaded onto the white blood cell and delivered to the vicinity of the thrombus with the help of optical force. At this time, by applying multiple optical potential wells on the white blood cell, the controlled release of the drug at the thrombus was successfully achieved ( Figure 7 d). Finally, the assembled red blood cell microfluidic pump was used to deliver batches of nanoparticles of drug to the thrombus location by means of the microfluidic field generated by its rotation ( Figure 7 e).

[0117] The present invention realizes the targeted delivery of a single nano drug particle toward a single cancer cell in a living blood vessel based on scanning optical tweezers technology using acousto-optic modulation.

[0118] The present invention uses this system to achieve simultaneous capture, controllable arrangement and precise delivery of multiple nanoparticles of drug.

[0119] The present invention uses optical manipulation technology to load nano-drugs on leukocytes or platelets, thereby achieving indirect transport of nano-drugs.

[0120] The present invention utilizes optical manipulation technology to assemble a red blood cell microfluidic pump to achieve batch and precise delivery of nanomedicines toward thrombi.

[0121] In summary, the advantages of the present invention include:

[0122] 1) Leveraging the non-contact and non-destructive advantages of optical tweezers, scanning optical tweezers technology with acousto-optic modulation is used to achieve stable capture and multifunctional delivery of nanoparticles with high spatiotemporal precision, and can achieve directional delivery with single-cell precision.

[0123] 2) By utilizing the rapid scanning of the laser focus, multiple nanoparticles can be manipulated simultaneously and precisely arranged without the need for additional optical control components. The system has high integration and is easy to operate.

[0124] 3) Using naturally existing leukocytes and platelets in blood vessels to construct a light-controlled nano-drug delivery system can avoid the complex particle surface modification process and potential immune feedback, and has good biocompatibility.

[0125] 4) Using dynamically programmed scanning optical tweezers technology, multi-modal nanodrug delivery can be achieved for different lesion locations, thereby solving the challenges of low spatiotemporal precision, poor targeting effect, easy clearance by the immune system, and low transmission efficiency in traditional drug delivery.

[0126] The present invention proposes to use non-contact, non-destructive, and high-temporal and spatial precision optical manipulation to achieve targeted transport of nanomedicines in vivo, while the transport can achieve the precision of a single nanoparticle.

[0127] Specifically, the present invention utilizes time-division multiplexing scanning optical tweezers technology and dynamically programs an acousto-optic modulator to precisely control the spatial distribution of the laser focus, stably capturing nanoparticles flowing spontaneously within zebrafish blood vessels and enabling their reverse blood flow transport. Furthermore, the nanoparticles can be manipulated to actively avoid intravascular obstacles and immune cell engulfment, with flexible and controllable transport speed and direction. Furthermore, the present invention can simultaneously capture multiple nanoparticles and independently manipulate their trajectories, allowing them to circumvent intravascular obstacles and enter different vascular branches. To further optimize the biocompatibility of nanoparticles, the present invention attaches nanoparticles to the interior of platelets and red blood cells within blood vessels, enabling indirect delivery of nanoparticles. To address the transport efficiency shortcomings of optical manipulation, the present invention captures and rotates naturally occurring red blood cells in the body, constructing an endogenous red blood cell microfluidic pump and utilizing the generated driving flow field to achieve bulk nanoparticle transport. Unlike traditional manipulation schemes, this optical manipulation-based nanoparticle delivery method does not require the implantation of exogenous materials, avoiding potential physiological damage to the organism and providing a biocompatible solution for further research into targeted drug delivery at disease sites in vivo.

[0128] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on the differences from other embodiments. Reference can be made to the common and similar parts between the various embodiments. For the systems disclosed in the embodiments, since they correspond to the methods disclosed in the embodiments, the description is relatively simple, and the relevant parts can be referred to the method description.

[0129] This document uses specific examples to illustrate the principles and implementation methods of the present invention. The above examples are only intended to help understand the method and core concept of the present invention. At the same time, those skilled in the art will find that the specific implementation methods and application scopes may vary based on the concept of the present invention. In summary, the contents of this specification should not be construed as limiting the present invention.

Claims

1. A drug targeted delivery device, characterized in that: include: Control optical components, illumination optical components, fluorescence excitation optical components, inverted objective lens, charge-coupled device, computer and sample chamber; The computer is connected to the laser and the acousto-optic deflection plate in the control optical path device and the charge coupled device respectively; The control light path of the control light path device is parallel to the fluorescence excitation light path of the fluorescence excitation light path device, and the control light path is perpendicular to the illumination light path of the illumination light path device; the charge coupled device and the illumination light path are on the same light path; The illumination optical path device is used to generate illumination light to illuminate the living sample in the sample chamber; the living sample contains nanomedicine; The fluorescence excitation optical path device is used to emit broad-spectrum white light, which is focused by the inverted objective lens to excite the nanomedicine, causing the nanomedicine to emit fluorescence; The fluorescence excitation optical path device specifically includes: an LED light source, a collimator, a filter and a reflector; The LED light source is used to generate broad-spectrum white light; the broad-spectrum white light passes through the collimator, the filter, and the reflector in sequence, is reflected onto the beam splitter, and is irradiated onto the living sample through the inverted objective lens; The control optical path device is used to emit a laser beam, which is irradiated onto the living sample after passing through the inverted objective lens; The charge coupled device is used to collect the image in the sample chamber, convert it into an image electrical signal, and upload the image electrical signal to the computer; The computer is used to adjust the deflection angle of the acousto-optic deflection plate in real time based on the image electrical signal and the direction and speed of targeted delivery to adjust the position of the optical potential. The nanomedicine is subjected to the optical gradient force exerted by the laser beam, and the nanomedicine moves synchronously with the irradiation position of the laser beam; the irradiation position is the position of the optical potential.

2. The drug targeted delivery device according to claim 1, characterized in that The control optical path components specifically include: a laser, an acousto-optic deflector, a beam broadening device and a spectroscope; The laser beam emitted by the laser sequentially passes through the acousto-optic deflection plate, the beam broadening device, the beam splitter, and the inverted objective lens and is irradiated into the living sample; The acousto-optic deflection plate deflects according to the deflection angle set by the computer; The beam expansion device is used to expand the diameter of the laser beam to completely cover the entrance pupil of the inverted objective lens, and the inverted objective lens focuses the laser beam and then irradiates it onto the living sample.

3. The drug targeted delivery device according to claim 2, characterized in that The lighting optical path device specifically includes: a halogen light source and a concentrator; The halogen light source emits illumination light, which is irradiated onto the living sample through the condenser to generate an image electrical signal of the microscope field of view; the image electrical signal is collected by the inverted objective lens and sent to the charge coupled device.

4. The drug targeted delivery device according to claim 3, characterized in that The illumination optical path device, the inverted objective lens, the beam splitter, the reflector and the charge coupled device are on the same optical path.

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