Fixed tick feeding observation device

Abstract

The invention discloses a fixed tick feeding observation device. The fixed tick feeding observation device comprises an observation cylinder, a feeding seat and a fixed seat, a blood supply assembly is arranged in the feeding seat and comprises a capillary channel and a skin simulation slide glass arranged in the middle of the capillary channel, and the slide glass is of a multi-layer structure with an artificial membrane and a bionic skin matrix wrapping a capillary and can highly simulate real skin. Double temperature control is achieved through the water bath heating base and the heat preservation pad, the peristaltic pump is matched to drive blood to flow in microvessels, and dynamic blood supply is provided. The device integrates a micro pH sensor, an oxygen sensor and a microelectrode array, and can monitor the microenvironment change of a blood suction point in real time. And an adjustable observation mirror is arranged on the observation cylinder, so that multi-angle observation is facilitated. According to the invention, the problems of poor biomimetic property and insufficient monitoring capability in the prior art are solved, and refined observation and multi-parameter real-time monitoring of blood sucking behaviors of ticks under a highly bionic condition are realized.

Description

Technical Field

[0001] This invention relates to the field of tick feeding technology, specifically a fixed tick feeding observation device. Background Technology

[0002] Ticks are important disease vectors, and their blood-sucking process is a crucial step in studying tick biology, pathogen transmission mechanisms, and screening novel control methods. Controlled and observable feeding of ticks under laboratory conditions is fundamental to conducting related research. Traditional feeding methods mainly rely on live animal hosts. This method is not only costly, time-consuming, and subject to strict ethical review, but also suffers from poor data reproducibility and difficulty in controlling variables due to significant individual host variability and complex immune responses, greatly limiting the precision and standardization of research.

[0003] To overcome the limitations of live hosts, artificial feeding devices have emerged. In existing technologies, these devices typically employ a feeding chamber sealed at the bottom with an artificial membrane. Static blood or a blood meal is placed on the membrane, and the heat from the blood meal attracts ticks to penetrate the artificial membrane for blood feeding. However, these devices have significant drawbacks: First, the simulated host environment is overly simplified. The static blood meal and the single artificial membrane cannot simulate the three-dimensional structure of real skin and the physiological stimulation of dynamic blood flow in capillaries, resulting in generally low tick attachment and blood-feeding success rates, and significant deviations in behavioral data from the natural state. Second, the feeding process lacks effective real-time monitoring methods. Researchers cannot obtain information on local microenvironmental changes at the feeding point (such as pH and oxygen partial pressure) and the dynamic process of tick saliva secretion, making it difficult to quantitatively study the complex molecular interactions occurring at the tick-host interface. Finally, existing devices still have shortcomings in terms of temperature uniformity and precision, as well as in preventing blood clotting, affecting the stability and reproducibility of experiments. Summary of the Invention

[0004] (a) Technical problems to be solved To address the shortcomings of existing technologies, this invention provides a fixed tick feeding and observation device, which solves the problems mentioned in the background section.

[0005] (II) Technical Solution To achieve the above objectives, the present invention is implemented through the following technical solution: a fixed tick feeding and observation device, comprising an observation tube, a feeding seat rotatably connected to the lower side wall of the observation tube, a blood supply component installed inside the feeding seat, microvessels installed inside the blood supply component, a skin simulation slide installed in the middle of the microvessels, a water bath heating seat installed inside the feeding seat, and the blood supply component installed inside the water bath heating seat; A connecting frame is installed at the upper end of the observation tube. A mounting base is threaded inside the connecting frame. A rotating ball is rotatably connected inside the mounting base. An observation lens is fixedly connected inside the rotating ball. The observation lens is a magnifying lens, and its optical axis coincides with the center line of the observation tube. A fixing base is fixedly connected to the rear side wall of the observation tube. Two sliding plates are slidably connected to the rear side wall of the fixing base.

[0006] Preferably, the rotation range of the observation tube and the feeding seat is set to 0-30°. The bottom of the observation tube is provided with a feeding hole, which is aligned with the skin simulation slide. The observation tube is made of colorless and transparent material, which facilitates the observation of the internal condition of the observation tube 1. The rotation setting of the observation tube and the feeding seat allows the feeding hole to rotate with the skin simulation slide, thereby controlling the feeding timing.

[0007] Preferably, each of the sliding pieces is provided with a fixing hole inside for easy fixation to the outside. A connecting side piece is fixedly connected to the outer side wall of the sliding piece for easy pulling out of the sliding piece. The connecting side piece is provided with anti-slip texture. A nano-adhesive is installed on the rear side wall of the mounting base for further fixation to the outside.

[0008] Preferably, an injection tube is installed on one side wall of the blood supply component, a sealing cap is installed at the outer end of the injection tube, a one-way valve is provided at the connection between the injection tube and the blood supply component to prevent blood backflow, and the blood supply component is made of transparent biocompatible material with a hollow internal structure to store blood.

[0009] Preferably, a heat-insulating pad is installed on the upper sidewall of the blood supply component, and the microvessels are partially disposed inside the heat-insulating pad from the blood supply component. The heat-insulating pad wraps the skin simulation carrier. The heat-insulating pad is a flexible heating pad with a controllable operating temperature maintained at 35-38°C. A peristaltic pump is disposed inside the heat-insulating pad, and a portion of the microvessels is disposed inside the peristaltic pump. The flow rate of the peristaltic pump is adjustable, ranging from 1-100µL / min.

[0010] Preferably, a miniature pH sensor is placed upstream and downstream of the microvessel, and an oxygen sensor is integrated at both the inlet and outlet of the microvessel. The signal lines of the miniature pH sensor and the oxygen sensor are integrated into the wall of the blood supply component and led out to the control component. Several pairs of microelectrodes are disposed on both sides of the microvessel and inside the skin simulation substrate. The microelectrodes are interdigitated electrodes with their wires embedded in the substrate of the substrate for measuring tissue impedance changes.

[0011] Preferably, the bottom of the skin-simulating slide is set as a slide base, the upper sidewall of the slide base is provided with a biomimetic skin matrix, the outer sidewall of the biomimetic skin matrix is ​​wrapped with an artificial membrane, the artificial membrane is an ultrathin silicone membrane, the biomimetic skin matrix is ​​a three-dimensional hydrogel containing type I collagen, the microvessels in the part of the skin-simulating slide are wrapped with the biomimetic skin matrix, and the wall of the microvessels in this area is a semi-permeable membrane, allowing the exchange of small molecules.

[0012] Preferably, the front sidewall of the fixing base is provided with an illumination panel, which faces the observation tube. The illumination panel is an LED ring light that provides a cold light source and has adjustable brightness. The fixing base is provided with a control component, which integrates a microprocessor for receiving sensor data and controlling the operation of the peristaltic pump, water bath heating base, heat preservation pad and illumination panel. The feeding base is provided with a power supply component, which is a rechargeable lithium battery or an external power interface.

[0013] (III) Beneficial Effects This invention provides a fixed tick feeding and observation device, which has the following beneficial effects: 1. This invention creates a highly biomimetic three-dimensional structure of host skin by constructing a multilayered skin-simulating slide composed of an artificial membrane, a biomimetic skin matrix, and embedded microvessels. Combined with dynamic blood flow driven by a peristaltic pump, it provides ticks with crucial physiological stimulation, effectively solving the problems of low tick attachment and blood-feeding success rates in traditional static blood-feeding devices. This design significantly improves the standardization of experiments, making the observed tick behavioral data closer to the natural state, and providing a reliable platform for tick blood-feeding biology research.

[0014] 2. This device integrates a miniature pH sensor, an oxygen sensor, and a microelectrode array, enabling in-situ, real-time monitoring of the chemical parameters (pH value, oxygen partial pressure) and physical parameters (tissue impedance) of the microenvironment at the tick feeding site during tick feeding. This overcomes the limitations of traditional devices that can only perform endpoint observations, achieving quantitative capture of microscopic dynamic processes such as tick saliva secretion and local tissue reactions, providing unprecedented data support for a deeper understanding of the molecular mechanisms of tick-host interface interactions.

[0015] 3. This invention employs a dual temperature control system combining water bath heating and a flexible insulation pad, ensuring temperature uniformity and stability throughout the entire path from blood storage to the skin simulation interface. Simultaneously, the dynamically circulating blood flow path design effectively prevents blood coagulation. This precise environmental control greatly improves the repeatability and success rate of the experiment, ensuring smooth long-term feeding observation and overcoming the technical difficulties of large temperature fluctuations and easy blood deterioration in existing technologies. Attached Figure Description

[0016] Figure 1 This is a schematic diagram of the structure of the present invention; Figure 2 This is a schematic diagram of the rear side of the present invention; Figure 3 This is a cross-sectional schematic diagram of the present invention; Figure 4 This is a top view of the present invention; Figure 5 This is a schematic diagram of the blood supply component in this invention; Figure 6 This is a schematic diagram of the structure of the skin simulation substrate in this invention.

[0017] The components include: 1. Observation tube; 101. Feeding hole; 102. Connecting frame; 2. Feeding seat; 201. Blood supply component; 2011. Microvessel; 2012. Miniature pH sensor; 2013. Oxygen sensor; 2014. Injection tube; 2015. Peristaltic pump; 2016. Insulation pad; 202. Power supply component; 203. Skin simulation slide; 2031. Artificial membrane; 2032. Bionic skin matrix; 2033. Slide substrate; 2034. Microelectrode; 204. Water bath heating seat; 3. Fixing seat; 301. Sliding piece; 3011. Fixing hole; 3012. Connecting side piece; 302. Control component; 303. Illumination panel; 4. Mounting seat; 5. Observation mirror. Detailed Implementation

[0018] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0019] Example 1: like Figure 1-6 As shown, this embodiment of the invention provides a fixed tick feeding and observation device, including an observation tube 1. The observation tube 1 is made of transparent polycarbonate and has a feeding hole 101 with a diameter of 5 mm at the bottom. The observation tube 1 is connected to the feeding seat 2 via a rotating shaft with adjustable damping, allowing the observation tube to be suspended at any angle within the range of 0-30 degrees.

[0020] The main body of the feeding seat 2 is made of ABS engineering plastic, and a blood supply component 201 is embedded inside. The blood supply component 201 is made of biocompatible PDMS (polydimethylsiloxane), and has microvascular channels 2011 with a cross-section of 100µm×50µm processed inside. The skin simulation slide 203 is fixed to the feeding seat 2 by a slot and is located above the center of the microvascular channel 2011. The slide substrate 2033 of the skin simulation slide 203 is a glass slide, on which a biomimetic skin matrix 2032 containing type I collagen (concentration of 3mg / mL) is laid, and the outermost layer is covered with an ultra-thin silicone artificial membrane 2031 with a thickness of about 20µm.

[0021] The blood supply component 201 is immersed in a water bath heating seat 204 within the feeding seat 2. The water bath heating seat 204 maintains the water temperature at 37±0.5℃ via a built-in heating rod and temperature sensor (not shown in the figure). Blood is injected through an injection tube 2014, the inlet of which is equipped with a sealing cap.

[0022] The upper end of the observation tube 1 is connected to a mounting base 4 by a threaded connection. The mounting base 4 is equipped with a rotating ball, which contains an observation lens 5 with a magnification of 5x. Its optical axis coincides with the center line of the observation tube 1, so that the user can observe ticks from different angles without losing focus.

[0023] The mounting base 3 is fixed to the rear side of the observation tube 1 by screws. The sliding plates 301 on both sides can be pulled out through the connecting side plates 3012, and the entire device is fixed to the experimental rack with bolts through the fixing holes 3011 on the sliding plates 301. The mounting base 4 has nano-adhesive attached to its rear side, which can help it adhere to smooth surfaces and enhance stability.

[0024] Example 2: Based on Example 1, this embodiment adds environmental maintenance, sensing and monitoring, and intelligent control functions, providing a highly integrated and automated research platform.

[0025] In this embodiment, a flexible heat-insulating pad 2016 is attached tightly to the upper surface of the blood supply component 201. The heat-insulating pad 2016 is a silicone heating pad with temperature feedback, and its operating temperature is controllable and maintained at 37°C. A miniature peristaltic pump 2015 is encapsulated inside the heat-insulating pad 2016. The pump tube section of the peristaltic pump 2015 is part of the microvessel 2011, and its flow rate can be precisely adjusted within the range of 1-100µL / min by the control system to simulate different blood flow velocities.

[0026] Upstream and downstream of the microvessel 2011, a miniature pH sensor 2012 (such as a commercially available needle-tip pH microelectrode) and a miniature oxygen sensor 2013 (such as a Clarke-type oxygen microelectrode) are integrated, respectively. The signal lines of these sensors are embedded in the wall of the blood supply component 201 made of PDMS material and are finally connected to the control component 302 inside the fixture 3.

[0027] The structure of the skin-simulating slide 203 is more complex. Within its biomimetic skin matrix 2032, a pair of interdigitated microelectrodes 2034 made of gold are embedded, with electrode leads also encapsulated within the slide substrate 2033 and extended outwards. These microelectrodes 2034 are connected to an external impedance analyzer (via the interface of the control component 302) for real-time monitoring of impedance changes around the tick's mouthparts caused by salivary sheath formation and tissue reactions.

[0028] An LED ring light panel 303 is mounted on the front of the mounting base 3. Its color temperature is 5000K, and its brightness is infinitely adjustable, providing a uniform cold light source for observation. The control component 302 inside the mounting base 3 is the core, integrating an STM32 series microprocessor, an analog-to-digital converter module, and a power management module. The microprocessor is responsible for receiving signals from the pH sensor 2012 and the oxygen sensor 2013, and simultaneously controlling the rotation speed of the peristaltic pump 2015, the temperature of the water bath heating base 204 and the insulation pad 2016, as well as the on / off state and brightness of the lighting panel 303.

[0029] The power supply component 202 is a rechargeable lithium battery pack integrated inside the feeder 2, which can provide more than 8 hours of continuous power for the entire device (excluding the external impedance analyzer). It also has an external power interface for long-term experimental use.

[0030] Working Principle: In use, animal blood containing an anticoagulant is first injected into the blood supply component 201 through the injection tube 2014. A hungry adult or nymph tick is placed on the artificial membrane 2031 of the skin simulation slide 203 through the feeding hole 101. By rotating the observation tube 1, the feeding hole 101 is aligned with or offset from the skin simulation slide 203, thus controlling the timing of the tick's initial contact with the feed. After sensing the stimulation from the 37°C warm "skin" below, the tick attempts to pierce the artificial membrane 2031 and the biomimetic skin matrix 2032 with its mouthparts, eventually penetrating the microvessels 2011 to begin sucking blood. Throughout the process, researchers can observe through the observation mirror 5 and an external microscope. Simultaneously, the control component 302 records and saves real-time data from various sensors, enabling refined and quantitative research on tick-feeding behavior.

[0031] Example 3: This embodiment further optimizes the device fixing method and observation system based on Embodiment 1.

[0032] A positioning spring is added between the sliding plate 301 and the fixed base 3. When the sliding plate is pulled out, the spring can provide appropriate damping force, so that the sliding plate can be stably stopped at any extended position, which is convenient for fixing at the edge of the experimental platform at different depths. A copper threaded sleeve is embedded in the fixing hole 3011, which improves the durability of the thread.

[0033] The space between the rotating ball of the observation mirror 5 and the mounting base 4 is filled with high-viscosity silicone grease, which ensures both rotational flexibility and stability of the observation mirror at any angle, preventing changes in the viewing angle due to slight touches. The observation mirror 5 uses an achromatic composite lens, which effectively reduces image chromatic aberration and improves the clarity of observation.

[0034] A flexible silicone sealing ring is added to the edge of the feeding hole 101. When the observation tube 1 is rotated to fit tightly with the skin simulation slide 203, the sealing ring can effectively prevent ticks from escaping from the gaps, while avoiding interference from the external environment to the feeding process.

[0035] Example 4: This embodiment expands the functionality of the sensing system and control system based on Embodiment 2.

[0036] The control component 302 incorporates a wireless communication module (such as Wi-Fi or Bluetooth) to transmit data with the laboratory's central computer or researchers' mobile terminals, enabling remote monitoring and data recording. Researchers can view changes in various physiological parameters during tick feeding in real time from their offices.

[0037] The 303 lighting panel has been upgraded to an RGB LED light assembly, which not only offers adjustable brightness but also provides light sources of different wavelengths. For example, specific wavelengths of red light can be used for observation, reducing behavioral interference with light-sensitive ticks; or ultraviolet light can be used to excite specific fluorescent markers for special research needs.

[0038] In the microvessels 2011 of the blood supply component 201, an additional miniature ultrasonic flow sensor is integrated, which can accurately measure the actual blood flow velocity in real time and form a closed-loop control with the peristaltic pump 2015 to ensure the accurate and stable blood flow velocity with an error controlled within ±0.5µL / min.

[0039] The power supply component 202 has added a power display function and a low power alarm function. When the power is below 20%, the control component 302 will remind researchers to charge in time or switch to an external power source through audible and visual signals to avoid data loss due to power outages during the experiment.

[0040] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A fixed tick feeding and observation device, comprising an observation tube (1), characterized in that: The lower side wall of the observation tube (1) is rotatably connected to a feeding seat (2). A blood supply component (201) is installed inside the feeding seat (2). A microvessel (2011) is installed inside the blood supply component (201). A skin simulation slide (203) is installed in the middle of the microvessel (2011). A water bath heating seat (204) is installed inside the feeding seat (2). The blood supply component (201) is installed inside the water bath heating seat (204). The upper end of the observation tube (1) is equipped with a connecting frame (102), the connecting frame (102) is internally threaded with a mounting base (4), the mounting base (4) is internally rotatably connected with a rotating ball, the rotating ball is internally fixedly connected with an observation mirror (5), the rear side wall of the observation tube (1) is fixedly connected with a fixing base (3), and the rear side wall of the fixing base (3) is slidably connected with two sliding pieces (301).

2. The fixed tick feeding and observation device according to claim 1, characterized in that: The rotation range of the observation tube (1) and the feeding seat (2) is set to 0-30°. The bottom of the observation tube (1) is provided with a feeding hole (101). The feeding hole (101) is aligned with the skin simulation slide (203). The observation tube (1) is made of colorless and transparent material.

3. The fixed tick feeding and observation device according to claim 1, characterized in that: Each of the sliding pieces (301) has a fixing hole (3011) inside, and a connecting side piece (3012) is fixedly connected to the outer side wall of the sliding piece (301).

4. A fixed tick feeding and observation device according to claim 1, characterized in that: A liquid injection tube (2014) is installed on one side wall of the blood supply assembly (201), and a sealing cap is installed on the outer end of the liquid injection tube (2014).

5. A fixed tick feeding and observation device according to claim 1, characterized in that: A heat-insulating pad (2016) is installed on the upper side wall of the blood supply component (201). The microvessels (2011) are partially disposed inside the heat-insulating pad (2016) from the blood supply component (201). The heat-insulating pad (2016) wraps the skin simulation slide (203). A peristaltic pump (2015) is disposed inside the heat-insulating pad (2016). A portion of the microvessels (2011) is disposed inside the peristaltic pump (2015).

6. A fixed tick feeding and observation device according to claim 1, characterized in that: A micro pH sensor (2012) is placed upstream and downstream of the microvessel (2011). An oxygen sensor (2013) is integrated at both the inlet and outlet of the microvessel (2011). Several pairs of microelectrodes (2034) are arranged on both sides of the microvessel (2011) and inside the skin simulation substrate (203).

7. A fixed tick feeding and observation device according to claim 1, characterized in that: The bottom of the skin simulation slide (203) is set as a slide base (2033), and the upper sidewall of the slide base (2033) is provided with a biomimetic skin matrix (2032). The outer sidewall of the biomimetic skin matrix (2032) is wrapped with an artificial membrane (2031), and the microvessels (2011) in the skin simulation slide (203) are partially wrapped by the biomimetic skin matrix (2032).

8. A fixed tick feeding and observation device according to claim 1, characterized in that: The front side wall of the fixed seat (3) is provided with an illumination lamp plate (303), which is directly opposite the observation tube (1). The fixed seat (3) is provided with a control component (302), and the feeding seat (2) is provided with a power supply component (202).

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