Puncture needle for molding and virus injection of cavernous blood vessel deformity of newborn mouse brain

Abstract

The invention relates to a puncture needle for injection virus in molding of cavernous blood vessel deformity of newborn mouse brain. The puncture needle comprises an operation handle, a limiting lug, a needle tube, a needle tube connector, a fiber grating recognition structure and a depth limiting structure. By arranging a fiber bragg grating recognition structure, in the penetrating process of a puncture needle, according to the FBG wavelength offset, a microprocessor receives an electric signal of a fiber FBG sensor, the microprocessor transmits the electric signal to a fiber bragg grating demodulator, the fiber bragg grating demodulator receives FBG reflection spectrum information, and a neural network model for deformation reconstruction of the puncture needle is constructed; micro deformation of the puncture needle is monitored, so that an experimenter is assisted in judging tissue resistance mutation in the puncture process, and accidental injury of blood vessels is prevented; by arranging the depth limiting structure and rotating the fine adjustment screw clockwise, the fine adjustment screw is in threaded fit with the inner side of the fixed seat, so that the connecting seat and the jacking mechanism are pushed to move towards the front end, the position of the jacking mechanism is changed, and the depth of the needle tube is changed.

Description

Technical Field

[0001] The invention relates to the technical field of biomedical instruments, in particular to a puncture needle for injecting viruses into a newborn mouse cerebral cavernous malformation model. Background Art

[0002] Establishing a cerebral cavernous malformation (CCMs) model by injecting viruses into the brain of newborn mice is a highly sophisticated and challenging technique, which is mainly used to study the pathogenesis, gene function and potential therapeutic strategies of CCMs.

[0003] When performing virus injection, a puncture needle is often required. However, since the skull of newborn mice is thin and not ossified, fixation must be extremely gentle. Traditional puncture needles cannot sense sudden changes in tissue resistance during the puncture process, resulting in accidental injury to blood vessels. At the same time, they cannot target the depth-limiting structure of the newborn mouse skull. Needle placement can only rely on the operator's experience, which makes the operation difficult. Summary of the Invention

[0004] (1) Technical problems solved

[0005] In order to solve the above technical problems, the present invention provides a puncture needle for injecting virus for modeling cerebral cavernous malformations in newborn mice.

[0006] (2) Technical solution

[0007] Based on this, the present invention provides the following technical solution: a puncture needle for injecting virus into a newborn mouse cerebral cavernous malformation model, comprising an operating handle, a limiting ear, a needle tube, a needle tube connector, a fiber Bragg grating recognition structure, and a depth limiting structure;

[0008] The rear end of the operating handle is fixed with a limiting ear, the middle part of the operating handle is embedded with a needle tube, the rear end of the needle tube is provided with a needle tube joint, the front end of the needle tube is provided with a fiber Bragg grating recognition structure, and the front end of the operating handle is installed with a limiting depth structure;

[0009] The fiber Bragg grating identification structure includes a microgroove and an optical fiber FBG sensor. The outer diameter of the needle tube is ≤100μm, and a microgroove is provided 150-250μm away from the needle tip. The optical fiber FBG sensor is embedded in the inner side of the microgroove and fixed with epoxy resin glue. The optical fiber FBG sensors are distributed in three groups along the circumference of the needle tube to form a strain sensing array.

[0010] Preferably, the three groups of optical fiber FBG sensors are symmetrically distributed at 120° along the radial direction of the needle tube 3 .

[0011] Preferably, the depth limiting structure includes a fixed tube, a fixed seat, a fine-tuning screw, a connecting seat, a supporting mechanism, and a guide rod. The fixed tube is fixed to the front end of the operating handle, the fixed tube is fixedly connected to the bottom of the fixed seat, the fine-tuning screw is matched with the inner thread of the fixed seat, the fine-tuning screw is movably connected to the rear end of the connecting seat, and the connecting seat is fixed to the top of the supporting mechanism.

[0012] Preferably, there are two guide rods in total, and the guide rods are arranged opposite to each other on the left and right sides of the rear end of the supporting mechanism, and the guide rods are both slidably matched with the inner side of the fixed tube.

[0013] By adopting the above technical solution, the supporting mechanism can be easily moved horizontally.

[0014] Preferably, a microprocessor is provided at the bottom of the fixing tube, and the microprocessor is electrically connected to the optical fiber FBG sensor.

[0015] By adopting the above technical solution, the electrical signal of the optical fiber FBG sensor can be received by the microprocessor, and then the microprocessor is connected to the optical fiber Bragg grating demodulator to receive the FBG reflection spectrum information.

[0016] Preferably, the supporting mechanism includes a connecting seat, a damping turntable, a receiving seat, a supporting claw 1, a supporting claw 2, a movable rod, and a support pad. The connecting seat is fixed to the front end of the guide rod, the connecting seat is movably connected to the rear end of the damping turntable, the front end of the damping turntable is fixed to the receiving seat, the right end of the supporting claw 1 is hinged to the receiving seat, the left end of the supporting claw 2 is hinged to the receiving seat, the supporting claw 1 is movably connected to the upper and lower sides of the movable rod, and the movable rod is fixed to the rear end of the support pad.

[0017] Preferably, the structure of the supporting claw 1 is consistent with the structure of the supporting claw 2, and the supporting claw 1 and the supporting claw 2 are symmetrically arranged with respect to each other.

[0018] Preferably, the right end of the supporting claw 1 is provided with a tooth angle, and the left end of the supporting claw 2 is provided with a tooth angle, and the tooth angle of the supporting claw 1 and the tooth angle of the supporting claw 2 are meshed with each other.

[0019] By adopting the above technical solution, when one of the support claws, support claw 1 or support claw 2, is moved, the tooth angles that mesh with each other can cause one support claw to swing, so that support claw 1 and support claw 2 can be opened or closed synchronously, thereby adjusting the angle between support claw 1 and support claw 2.

[0020] Preferably, the support claw 1 and the support claw 2 are both arranged in an arc shape, and a movable rod and a support pad are provided at the right end of the support claw 2. A through hole is provided in the middle of the connecting seat and the damping turntable, and the front end of the operating handle passes through the through hole.

[0021] By adopting the above technical solution, the supporting mechanism can be easily moved toward the front end of the operating handle.

[0022] (3) Beneficial effects

[0023] Compared with the prior art, the present invention provides a puncture needle for virus injection for modeling cavernous malformations in newborn mice, which has the following beneficial effects:

[0024] 1. The puncture needle used for virus injection in the modeling of cavernous malformations in the brain of newborn mice is equipped with a fiber grating recognition structure. During the penetration of the puncture needle, the microprocessor receives the electrical signal from the optical fiber FBG sensor based on the FBG wavelength offset. The microprocessor transmits the electrical signal to the fiber grating demodulator, which receives the FBG reflection spectrum information and constructs a neural network model for puncture needle deformation reconstruction to monitor the slight deformation of the puncture needle, thereby assisting experimenters in judging sudden changes in tissue resistance during the puncture process and preventing accidental injury to blood vessels.

[0025] 2. The puncture needle for virus injection for modeling cavernous malformations in the brain of newborn mice is equipped with a depth-limiting structure. By turning the fine-tuning screw clockwise, the fine-tuning screw is threadedly engaged with the inner side of the fixing seat, thereby pushing the connecting seat and the supporting mechanism to move toward the front end, changing the position of the supporting mechanism, and thus changing the penetration depth of the needle tube. In addition, the support angles of the first and second supporting claws can be adjusted according to the size of different newborn mice to facilitate fit with the head and achieve depth-limiting operation. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is a schematic diagram of the overall structure of the present invention;

[0027] Figure 2 It is a schematic diagram of a partial three-dimensional structure of the present invention;

[0028] Figure 3 This is a schematic diagram of the planar structure of the needle tube of the present invention;

[0029] Figure 4 This is a schematic diagram of the three-dimensional structure of the depth limiting structure of the present invention;

[0030] Figure 5 This is a schematic diagram of the cross-sectional structure of the fixed pipe of the present invention;

[0031] Figure 6 This is a schematic diagram of the three-dimensional structure of the supporting mechanism of the present invention;

[0032] Figure 7 It is a schematic diagram of the planar structure of the supporting mechanism of the present invention.

[0033] In the figure: operating handle-1, limiting ear-2, needle tube-3, needle tube connector-4, fiber Bragg grating recognition structure-5, depth limiting structure-6;

[0034] Micro-groove 51, optical fiber FBG sensor 52, fixing tube 61, fixing base 62, fine-tuning screw 63, connecting base 64, supporting mechanism 65, guide rod 66, microprocessor 67;

[0035] Connecting seat 651, damping turntable 652, receiving seat 653, supporting claw 1 654, supporting claw 2 655, movable rod 656, supporting pad 657. DETAILED DESCRIPTION

[0036] 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.

[0037] See also Figure 1-Figure 3 The invention discloses a puncture needle for virus injection for modeling cavernous malformations in the brain of newborn mice, comprising an operating handle 1, a limiting ear 2, a needle tube 3, a needle tube connector 4, a fiber grating identification structure 5, and a depth limiting structure 6; the limiting ear 2 is fixed at the rear end of the operating handle 1, the needle tube 3 is embedded in the middle part of the operating handle 1, the rear end of the needle tube 3 is provided with a needle tube connector 4, the front end of the needle tube 3 is provided with a fiber grating identification structure 5, and the front end of the operating handle 1 is installed with a depth limiting structure 6; the fiber grating identification structure 5 includes a microgroove 51 and an optical fiber FBG sensor 52, the outer diameter of the needle tube 3 is ≤100μm, and the microgroove 51 is provided at 150-250μm from the needle tip, the optical fiber FBG sensor 52 is embedded in the inner side of the microgroove 51 and is fixed with epoxy resin glue with a thickness of 1-5μm, and the optical fiber FBG sensor 52 is distributed in three groups along the circumference of the needle tube 3 to form a strain sensing array.

[0038] In some embodiments, three groups of optical fiber FBG sensors 52 are symmetrically distributed at 120° along the radial direction of the needle tube 3. The optical fiber FBG sensor 52 is a bare optical fiber FBG etched to a diameter of ≤5 μm and embedded in the microgroove 51. The fiber core is directly exposed to the medium to be measured. The temperature-strain decoupling algorithm module can be integrated inside the optical fiber FBG sensor 52. According to the FBG wavelength offset, the tissue strain, temperature and chemical concentration are synchronously calculated. A scale ruler is marked on the periphery of the needle tube 3 to facilitate observation of the penetration depth during puncture positioning. The scale ruler can be fixed on the needle tube 3 by laser marking, ink, coating, etc.

[0039] See also Figure 4-Figure 5, a puncture needle for injecting viruses for modeling cavernous malformations in the brain of newborn mice, the depth limiting structure 6 includes a fixed tube 61, a fixed seat 62, a fine-tuning screw 63, a connecting seat 64, a supporting mechanism 65, and a guide rod 66. The fixed tube 61 is fixed to the front end of the operating handle 1, the fixed tube 61 is fixedly connected to the bottom of the fixed seat 62, the fine-tuning screw 63 is engaged with the inner thread of the fixed seat 62, the fine-tuning screw 63 is movably connected to the rear end of the connecting seat 64, the connecting seat 64 is fixed to the top of the supporting mechanism 65, and there are two guide rods 66, and the guide rods 66 are arranged opposite to each other on the left and right sides of the rear end of the supporting mechanism 65, and the guide rods 66 are both slidably engaged with the inner side of the fixed tube 61.

[0040] In some embodiments, in order to facilitate the horizontal movement of the supporting mechanism 65, a microprocessor 67 is provided at the bottom of the fixed tube 61, and the microprocessor 67 is electrically connected to the optical fiber FBG sensor 52. The electrical signal of the optical fiber FBG sensor 52 can be received through the microprocessor 67, and then the microprocessor 67 is connected to the optical fiber Bragg grating demodulator to receive the FBG reflection spectrum information, and a neural network model for the deformation reconstruction of the puncture needle is constructed to monitor the micro-deformation of the puncture needle. The fine-tuning screw 63 is a component for fine distance adjustment, and the thread accuracy is ±0.01mm.

[0041] See also Figure 6-Figure 7 , a puncture needle for injecting virus for modeling cavernous malformation of the brain of newborn mice, the supporting mechanism 65 includes a connecting seat 651, a damping turntable 652, a receiving seat 653, a supporting claw 1 654, a supporting claw 2 655, a movable rod 656, and a supporting pad 657. The connecting seat 651 is fixed to the front end of the guide rod 66, the connecting seat 651 is movably connected to the rear end of the damping turntable 652, the front end of the damping turntable 652 is fixed to the receiving seat 653, the right end of the supporting claw 1 654 is hinged to the receiving seat 653, the left end of the supporting claw 2 655 is hinged to the receiving seat 653, the supporting claw 1 654 is movably connected to the upper and lower sides of the movable rod 656, and the movable rod 656 is fixed to the rear end of the support pad 657.

[0042] In some embodiments, the structure of the support claw 1 654 is consistent with the structure of the support claw 2 655, and the support claw 1 654 and the support claw 2 655 are symmetrically arranged. The right end of the support claw 1 654 is provided with a tooth angle, and the left end of the support claw 2 655 is provided with a tooth angle. The tooth angle of the support claw 1 654 and the tooth angle of the support claw 2 655 are meshed with each other. When one of the support claws 1 654 or the support claw 2 655 is dialed, the tooth angles that mesh with each other can make one support claw move. Swing so that the support claw 1 654 and the support claw 2 655 can be opened or closed synchronously, and the angle between the support claw 1 654 and the support claw 2 655 is adjusted. The support claw 1 654 and the support claw 2 655 are both arranged in an arc shape, and the right end of the support claw 2 655 is provided with a movable rod 656 and a support pad 657, and a through hole is provided in the middle of the connecting seat 651 and the damping turntable 652, and the front end of the operating handle 1 passes through the through hole, so that the supporting mechanism 65 can move toward the front end of the operating handle 1.

[0043] In summary, when in use, first adjust the depth of the needle tube 3 inserted into the operating handle 1 as needed, and rotate the fine-tuning screw 63 clockwise so that the fine-tuning screw 63 is threadedly engaged with the inner side of the fixing seat 62, thereby pushing the connecting seat 64 and the supporting mechanism 65 to move forward. When the supporting mechanism 65 moves, the guide rod 66 moves inside the fixing tube 61 to guide the supporting mechanism 65, and connects the needle tube connector 4 to the micro-liquid delivery pump for delivering the virus;

[0044] Then, experimental animals were prepared: wild-type mice or conditional knockout mice, pregnant mice or newly born mother mice and newborn mice were used;

[0045] Environmental requirements: strictly sterile, warm and quiet.

[0046] Virus selection and preparation:

[0047] Commonly used viruses: recombinant adeno-associated virus.

[0048] Virus-carrying element: Cre recombinase: injected into the brain of a conditional knockout mouse carrying a specific CCM gene flanked by loxP sites, resulting in local excision of the target gene at the injection site;

[0049] shRNA or CRISPR-Cas9: Directly injected into the brain of wild-type mice to locally knock down or knock out the target CCM gene;

[0050] Reporter genes: Usually co-expressed with Cre or gene editing tools to mark transfected cells and locate the injection area;

[0051] Viral titer: High titer is required for efficient transduction;

[0052] Injection volume: This is extremely critical. The brain volume of newborn mice is extremely small (about 50 μL). Usually, the single-point injection volume is controlled between 0.1-0.5 μL. A larger volume can easily lead to acute injury, brain edema, and death.

[0053] Intracerebral injection procedure:

[0054] Anesthesia: Neonatal mice are usually anesthetized with brief deep hypothermia or low-concentration isoflurane inhalation anesthesia;

[0055] Fixation: Carefully fix the head of the anesthetized newborn mouse on a stereotaxic apparatus suitable for newborn mice; the skull is thin and not ossified, so fixation should be extremely gentle;

[0056] Positioning: Calculate three-dimensional coordinates based on the target brain area; the brain atlas of newborn mice is different from that of adult mice, and a special atlas is required. In addition, according to the size of the skull of the newborn mouse, one of the support claws 1 654 and 2 655 is moved to expand the support claws 1 654 and 2 655 to a suitable arc for the head of the newborn mouse. At the same time, the movable rod 656 on the support claws 1 654 and 2 655 can be rotated to change the support position of the support pad 657. During puncture, the puncture needle is supported and limited by the support pad 657. At the same time, the damping turntable 652 can be rotated to rotate on the inner side of the connecting seat 651 to change the support position of the support claws 1 654 and 2 655, so as to cope with different scenarios.

[0057] Craniotomy: After making a tiny incision in the scalp, drilling is usually not necessary because the skull is very thin and soft. A fine needle tip is used to carefully puncture the skull and dura mater at the designated point. The operation must be performed very gently to avoid damaging the underlying brain tissue.

[0058] Injection: Use a puncture needle for virus injection in neonatal mouse cerebral cavernous malformation models; carefully insert the needle tube 3 of the puncture needle vertically to the predetermined depth;

[0059] During the penetration process, the microprocessor 67 receives the electrical signal from the optical fiber FBG sensor 52 according to the FBG wavelength offset. The microprocessor 67 transmits the electrical signal to the fiber optic Bragg grating demodulator, which receives the FBG reflection spectrum information and constructs a neural network model for puncture needle deformation reconstruction to monitor the slight deformation of the puncture needle, thereby assisting the experimenter in judging the sudden change of tissue resistance during the puncture process. By observing the sudden change of tissue resistance during the puncture process, medical staff can adjust the puncture force and puncture angle in time to prevent accidental injury to blood vessels.

[0060] When the needle tube 3 reaches the predetermined position, inject very slowly, leave the needle in place for at least 1-2 minutes after injection, and then slowly withdraw the needle.

[0061] Suturing / closing: The incision is usually small and only requires gentle pressure or a small amount of tissue glue to complete the virus injection operation for the newborn mouse cerebral cavernous malformation model.

[0062] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A puncture needle for virus injection in neonatal mouse cerebral cavernous malformation modeling, characterized by: It comprises an operating handle (1), a limiting ear (2), a needle tube (3), a needle tube joint (4), a fiber optic Bragg grating identification structure (5), and a depth limiting structure (6); A limiting ear (2) is fixed at the rear end of the operating handle (1), a needle tube (3) is embedded in the middle of the operating handle (1), a needle tube connector (4) is provided at the rear end of the needle tube (3), a fiber grating identification structure (5) is provided at the front end of the needle tube (3), and a limiting depth structure (6) is installed at the front end of the operating handle (1); The fiber grating identification structure (5) comprises a microgroove (51) and an optical fiber FBG sensor (52). The outer diameter of the needle tube (3) is ≤100 μm, and the microgroove (51) is provided at a distance of 150-250 μm from the needle tip. The optical fiber FBG sensor (52) is embedded in the inner side of the microgroove (51) and is fixed with epoxy resin glue. The optical fiber FBG sensor (52) is distributed in three groups along the circumference of the needle tube (3) to form a strain sensing array.

2. The puncture needle for virus injection for neonatal mouse cerebral cavernous malformation modeling according to claim 1, characterized in that: The three groups of optical fiber FBG sensors (52) are symmetrically distributed at 120 degrees along the radial direction of the needle tube (3).

3. The puncture needle for virus injection for neonatal mouse cerebral cavernous malformation modeling according to claim 1, characterized in that: The depth limiting structure (6) comprises a fixed tube (61), a fixed seat (62), a fine-adjusting screw (63), a connecting seat (64), a supporting mechanism (65), and a guide rod (66); the fixed tube (61) is fixed to the front end of the operating handle (1); the fixed tube (61) is fixedly connected to the bottom of the fixed seat (62); the fine-adjusting screw (63) is engaged with the inner thread of the fixed seat (62); the fine-adjusting screw (63) is movably connected to the rear end of the connecting seat (64); and the connecting seat (64) is fixed to the top of the supporting mechanism (65).

4. The puncture needle for virus injection for neonatal mouse cerebral cavernous malformation modeling according to claim 3, characterized in that: There are two guide rods (66) in total, and the guide rods (66) are arranged opposite to each other along the left and right sides of the rear end of the supporting mechanism (65). The guide rods (66) are slidably matched with the inner side of the fixed tube (61).

5. The puncture needle for virus injection for neonatal mouse cerebral cavernous malformation modeling according to claim 3, characterized in that: A microprocessor (67) is provided at the bottom of the fixing tube (61), and the microprocessor (67) is electrically connected to the optical fiber FBG sensor (52).

6. The puncture needle for virus injection for neonatal mouse cerebral cavernous malformation modeling according to claim 3, characterized in that: The supporting mechanism (65) includes a connecting seat (651), a damping turntable (652), a receiving seat (653), a supporting claw 1 (654), a supporting claw 2 (655), a movable rod (656), and a supporting pad (657). The connecting seat (651) is fixed to the front end of the guide rod (66), the connecting seat (651) is movably connected to the rear end of the damping turntable (652), the front end of the damping turntable (652) is fixed to the receiving seat (653), the right end of the supporting claw 1 (654) is hinged to the receiving seat (653), the left end of the supporting claw 2 (655) is hinged to the receiving seat (653), the supporting claw 1 (654) is movably connected to the upper and lower sides of the movable rod (656), and the movable rod (656) is fixed to the rear end of the supporting pad (657).

7. The puncture needle for virus injection for neonatal mouse cerebral cavernous malformation modeling according to claim 6, characterized in that: The structure of the supporting claw 1 (654) is consistent with the structure of the supporting claw 2 (655), and the supporting claw 1 (654) and the supporting claw 2 (655) are symmetrically arranged with respect to each other.

8. The puncture needle for virus injection for neonatal mouse cerebral cavernous malformation modeling according to claim 6, characterized in that: The right end of the supporting claw 1 (654) is provided with a tooth angle, and the left end of the supporting claw 2 (655) is provided with a tooth angle, and the tooth angle of the supporting claw 1 (654) and the tooth angle of the supporting claw 2 (655) are meshed with each other.

9. The puncture needle for virus injection for neonatal mouse cerebral cavernous malformation modeling according to claim 6, characterized in that: The supporting claw 1 (654) and the supporting claw 2 (655) are both arranged in an arc shape, and a movable rod (656) and a supporting pad (657) are arranged at the right end of the supporting claw 2 (655).

10. The puncture needle for virus injection for neonatal mouse cerebral cavernous malformation modeling according to claim 6, characterized in that: A through hole is provided in the middle of the connecting seat (651) and the damping rotary disk (652), and the front end of the operating handle (1) passes through the through hole.

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