Auxiliary device for extracting lipid in biological sample

Abstract

The invention provides an auxiliary device for extracting biological sample lipid, and relates to the technical field of biological test equipment. The device comprises a centrifugal tube and a sleeve, wherein a puncture head with an upward needle head is vertically arranged at the inner bottom of the sleeve; external threads and internal threads which are matched with each other are selectively arranged on the centrifugal tube and the sleeve respectively; when the optional external thread of the centrifugal tube is matched with the internal thread of the sleeve, a sealed air pressure space is formed between the centrifugal tube and the sleeve, and the needle tip of the puncture head starts to puncture into the optional bottom of the centrifugal tube and extracts the organic phase lipid liquid in the centrifugal tube under the action of air pressure difference. Extraction operation in a closed space is carried out through the in-tube puncture head, and the risk of personnel infection caused by exposure of pollutant molecules such as aerosol in air due to exposure of phase liquid in contact with air is avoided.

Description

Technical Field

[0001] The invention relates to the technical field of biological test equipment, in particular to an auxiliary device for extracting lipids from biological samples. Background Art

[0002] Lipid metabolism, a fundamental process in life, is closely linked to the development and progression of various cardiovascular metabolic diseases, including atherosclerosis and non-alcoholic fatty liver disease. In lipid metabolism research, accurately measuring the levels of lipid components such as triglycerides and cholesterol in biological samples (such as liver tissue, small intestinal tissue, and feces of experimental animals) is a core technical step in assessing metabolic status. Lipid extraction, as a pre-treatment step for quantitative analysis, has a direct impact on the accuracy of experimental results due to its efficiency and recovery rate.

[0003] In the fields of metabolic disease research, intestinal health assessment, and digestive system function monitoring, the quantitative detection of total lipids in biological samples has important scientific and clinical value in revealing lipid metabolism abnormalities and evaluating nutrient absorption efficiency. The current traditional mainstream detection method relies on organic solvent step-by-step extraction technology, which requires core steps such as solid phase crushing, centrifugation, and puncture to obtain liquid. Its operation process has significant application defects: 1. As attached Figure 1 The high-risk centrifuge puncture procedure shown: During traditional centrifuge tube puncture and liquid extraction, the administrator must operate the puncture needle and insert the needle into the corresponding phase layer of the centrifuge tube for lipid extraction. However, this operation method makes the needle very easy to slip, resulting in injuries to the experimenter and infection from contaminants.

[0004] 2. During the traditional centrifuge tube puncture and liquid extraction process, the phase liquid will be exposed to the air (the extracted liquid will drip into the measuring cylinder or measuring cup below), which exposes pollutant molecules such as aerosols to the air, posing a risk of human infection.

[0005] 3. During the traditional centrifuge tube puncture and liquid collection process, because the cylindrical surface of the centrifuge tube is smooth, it is difficult for the puncture needle to be positioned and inserted into the centrifuge tube, resulting in inconvenient operation and low efficiency. Summary of the Invention

[0006] In order to solve the technical problems existing in the prior art, the present invention provides the following technical solutions: An auxiliary device for extracting lipids from biological samples, comprising a centrifuge tube 1 and a sleeve 4, wherein: The inner bottom of the sleeve 4 is vertically provided with a puncture head 5 with the needle pointing upwards; The centrifuge tube 1 and the sleeve 4 are respectively provided with matching external threads 2 and internal threads 3; When the outer thread 2 of the centrifugal tube 1 starts to match with the inner thread 3 of the sleeve 4, a sealed air pressure space is formed between the centrifugal tube 1 and the sleeve 4, and the needle tip of the puncture head 5 starts to pierce into the bottom of the centrifugal tube 1 and extract the organic phase lipid liquid in the centrifugal tube 1 under the action of air pressure difference.

[0007] Optionally, a plurality of helical flow guide grooves 6 are arranged on the outer side of the puncture head 5, which are distributed at equal intervals around the axis of the puncture head 5. The helical flow guide groove 6 is used to: During the process that the needle tip of the puncture head 5 gradually pierces into the bottom of the centrifugal tube 1, the helical flow guide groove 6 guides part of the organic phase lipid liquid to flow to the needle tip of the puncture head 5, thereby increasing the fluid efficiency of the organic phase lipid liquid.

[0008] Optionally, the extraction capacity control formula of the organic phase lipid liquid is as follows: , Wherein: V extract is the extraction capacity of the organic phase lipid liquid; R tube is the outer radius of the centrifugal tube 1; r avg is the average outer radius of the puncture head 5; N is the number of thread tightening circles when the bottom of the centrifugal tube 1 starts to contact the needle tip of the puncture head 5; P is the thread pitch, that is, the axial distance advanced per circle, unit: mm / circle; η is the fluid efficiency affected by the helical flow guide groove 6; V needle is the volume of the needle tip of the puncture head 5; C material is the correction coefficient affected by the material elasticity of the centrifugal tube 1, which is used to compensate the volume collapse caused by the rebound of the centrifugal tube 1.

[0009] Optionally, the puncture head 5 is a reverse conical structure, and the average outer radius of the needle tip and the root part is not more than 0.50-0.85mm.

[0010] Optionally, the puncture head 5 adopts martensitic age hardening stainless steel; the sleeve 4 adopts 316L stainless steel; The bottom port of the puncture head 5 is fixed and welded on the inner bottom of the sleeve 4 by interference fit and laser welding.

[0011] Optionally, a welding surface 7 is formed between the puncture head 5 and the sleeve 4 by laser welding, and a sealing enhancement layer is coated on the upper surface of the welding surface 7 to seal the connection between the puncture head 5 and the sleeve 4.

[0012] Optionally, the sealing enhancement layer is a 0.2-0.5 μm thick DLC coating coated on the welding surface 7 by a magnetron sputtering technique.

[0013] Optionally, the material of the centrifugal tube 1 is PFPE modified polypropylene, and the inner wall of the centrifugal tube 1 is silicified and formed with a 2-4 μm thick 350 cSt medical silicone oil layer.

[0014] Optionally, the device further comprises: a reflective sheet 8 fixed on the outer side of the centrifugal tube 1; a number of turns sensing module 9 fixed on the outer side of the sleeve 4 and used for sensing the number of turns N of screwing, the number of turns sensing module 9 comprising: a photoelectric type rotational speed sensor used for receiving a light signal reflected by the reflective sheet and feeding back to an MCU controller in real time; wherein, when the outer thread 2 of the centrifugal tube 1 starts to cooperate with the inner thread 3 of the sleeve 4, the reflective sheet and the photoelectric type rotational speed sensor are in a top-bottom corresponding relationship; an MCU controller used for starting counting according to the light signal, and subsequently corresponding to one number of turns of screwing for each time the light signal is received; when the extraction is finished, the number of all numbers of turns of screwing is counted, and the number of turns of screwing N is obtained; a wireless module used for reporting the number of turns of screwing N to a background server, and inputting the number of turns of screwing N into a preset extraction capacity control formula by the background server to estimate the extraction capacity of the organic phase lipid liquid; a power supply used for power supply; the photoelectric type rotational speed sensor, the wireless module and the power supply are electrically connected with the MCU controller respectively; the number of turns sensing module is in communication connection with the background server through the wireless module.

[0015] The technical scheme provided by the embodiment of the application has at least the following beneficial effects: The present invention proposes a device for liquid extraction by spirally fitting a cannula and a centrifuge tube, and piercing the centrifuge tube with a puncture head at the bottom during the tightening process. When the external thread of the centrifuge tube begins to fit with the internal thread of the cannula, a sealed air pressure space is formed between the centrifuge tube and the cannula, and the needle tip of the puncture head begins to penetrate the bottom of the centrifuge tube and extract the organic phase lipid liquid in the centrifuge tube under the action of the air pressure difference. The puncture head in the tube can be used to perform extraction operations in a closed space, avoiding the risk of the phase liquid being exposed to contact with the air, which would cause aerosols and other pollutant molecules to be exposed to the air and cause personal infection. In addition, through axially matching puncture in the tube, it is possible to achieve positioned puncture and liquid extraction, avoiding problems such as puncture slippage and inconvenience, thereby improving the efficiency of puncture extraction.

[0016] In combination with the description of the embodiments of the present invention, the present invention can also count the number of tightening turns of the puncture and liquid collection thread through sensing, and thereby evaluate and calculate the extracted liquid volume through the extraction capacity control formula of the organic phase lipid liquid, so that the administrator can quickly grasp the lipid extraction capacity of the biological sample, and realize the effect of quantitative collection and timely acquisition of information.

[0017] By using the fully enclosed lipid extraction device of this invention, the risk of puncture is eliminated through an inner and outer cannula system, and the extraction strategy is optimized to increase the lipid recovery rate to 99%. Experiments have shown that the device can shorten the processing time of a single sample to less than 40 minutes, significantly reducing the probability of biohazard exposure and providing key technical support for the establishment of a standardized lipid detection system for biological samples. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of 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 creative work.

[0019] Figure 1 This is a schematic diagram of the centrifuge tube puncture and liquid collection used in the traditional scheme (three-dimensional model schematic); Figure 2 This is a schematic diagram (three-dimensional model diagram) of a puncture and liquid collection auxiliary device for lipid extraction from biological samples provided by an embodiment of the present invention; Figure 3 Schematic diagram of a spiral guide groove of a puncture head provided by an embodiment of the present invention; Figure 4 This is a structural schematic diagram of a welding surface provided by an embodiment of the present invention; Figure 5 This is a schematic diagram of the installation structure of a turns sensing module provided by an embodiment of the present invention; Figure 6 This is a schematic diagram of the application control system of a turns sensing module provided by an embodiment of the present invention. DETAILED DESCRIPTION

[0020] The technical solution of the present invention is described below in conjunction with the accompanying drawings.

[0021] In the embodiments of the present invention, words such as "exemplarily" and "for example" are used to indicate examples, illustrations, or explanations. Any embodiment or design described as an "exemplary" in the present invention should not be interpreted as being preferred or advantageous over other embodiments or designs. Rather, the use of the word "exemplary" is intended to present concepts in a concrete manner. Furthermore, in the embodiments of the present invention, "and / or" can mean both or either of the two.

[0022] In the embodiments of the present invention, the terms "image" and "picture" may sometimes be used interchangeably. It should be noted that, when the distinction is not emphasized, the meanings they convey are the same. The terms "of," "corresponding," and "corresponding" may sometimes be used interchangeably. It should be noted that, when the distinction is not emphasized, the meanings they convey are the same.

[0023] In the embodiments of the present invention, sometimes a subscript such as W1 may be mistakenly written as a non-subscript form such as W1. When the difference is not emphasized, the meanings to be expressed are the same.

[0024] In order to make the technical problems, technical solutions and advantages to be solved by the present invention clearer, a detailed description will be given below with reference to the accompanying drawings and specific embodiments.

[0025] like Figure 2 As shown, an embodiment of the present invention provides an auxiliary device for extracting lipids from biological samples, the device comprising a centrifuge tube 1 and a sleeve 4, wherein: The inner bottom of the sleeve 4 is vertically provided with a puncture head 5 with the needle pointing upwards; The centrifuge tube 1 and the sleeve 4 are respectively provided with matching external threads 2 and internal threads 3; When the external thread 2 of the centrifuge tube 1 begins to cooperate with the internal thread 3 of the sleeve 4, a sealed air pressure space is formed between the centrifuge tube 1 and the sleeve 4, and the needle tip of the puncture head 5 begins to penetrate the bottom of the centrifuge tube 1 and extract the organic phase lipid liquid in the centrifuge tube 1 under the action of the air pressure difference.

[0026] Relative to Figure 1The traditional independent puncture extraction phase liquid method is shown, and the device for liquid extraction is provided by the sleeve and the centrifugal tube screwing and the bottom puncture head piercing into the centrifugal tube during screwing. When the outer thread of the centrifugal tube starts to cooperate with the inner thread of the sleeve, a sealed air pressure space is formed between the centrifugal tube and the sleeve, and the needle tip of the puncture head starts to pierce into the bottom of the centrifugal tube and extract the organic phase lipid liquid in the centrifugal tube under the action of air pressure difference. The extraction operation in the closed space can be carried out by the in-tube puncture head, avoiding the exposure of the phase liquid to the air to cause the exposure of aerosol and other pollutant molecules in the air, and causing the risk of personnel infection. And through the in-tube axial puncture, the positioning puncture liquid taking can be realized.

[0027] When the sleeve and the centrifugal tube screw, the sealing (thread sealing) starts and the puncture starts (the specific sleeve and the thread setting distance can be set based on the above requirements).

[0028] As shown in Figure 2 The puncture needle is provided in an inverted conical structure, and the material is optimized to reduce the puncture force and improve the sealing performance. The centrifugal tube and the sleeve are screwed, and the puncture needle automatically pierces into the centrifugal tube when screwed. Continue to tighten, and under pressure, the lower layer of lipid layer liquid will be extracted from the puncture needle (users can use measuring cups or other equipment to receive the liquid).

[0029] As shown in Figure 3 The outer side of the puncture head 5 is provided with a plurality of spiral flow guide grooves 6 which are distributed at equal intervals around the axis of the puncture head 5. The spiral flow guide grooves 6 are used to: During the process that the needle tip of the puncture head 5 gradually pierces into the bottom of the centrifugal tube 1, the spiral flow guide grooves 6 guide part of the organic phase lipid liquid to flow to the needle tip of the puncture head 5, thereby increasing the fluid efficiency of the organic phase lipid liquid.

[0030] If a smooth needle body is used for puncture, only the organic phase lipid liquid in the lower layer of the centrifugal tube can flow out from the needle tip.

[0031] The spiral flow guide grooves 6 are designed to guide part of the organic phase lipid liquid to flow to the needle tip of the puncture head 5. The spiral flow guide grooves 6 are provided on the outer side of the puncture head 5 in a plurality of rows, which are distributed at equal intervals around the axis of the puncture head 5. And the spiral flow guide grooves 6 are provided in a structure of double grooves in a group. After puncturing into the centrifugal tube, part of the liquid can enter the needle tube along the spiral flow guide grooves 6 to realize the drainage effect (because the centrifugal tube is made of elastic material, the leakage of liquid from the spiral flow guide grooves can be ignored, and reference can be made to the existing clinical infusion. After the needle cylinder pierces into the elastic rubber of the input bottle, the liquid will not fall along the needle cylinder).

[0032] Therefore, the spiral flow guide groove 6 can increase the fluid efficiency of the organic phase lipid liquid flowing through the piercing head 5.

[0033] Because the spiral mode can measure the spiral depth, some parameters can be combined to evaluate the capacity change of the centrifugal tube, which is used as the capacity value of the extracted liquid.

[0034] This embodiment preferentially adopts double spiral grooves (groove depth 0.08 mm, pitch 1.5 mm). It is found through testing that the liquid flow resistance is reduced and the pumping efficiency is improved (flow rate increased by 35%).

[0035] In order to accurately estimate the extraction capacity of the extracted organic phase lipid liquid, the distance of the sleeve advancing per revolution is equal to the pitch, so that the number of revolutions multiplied by the pitch should be equal to the depth of the piercing needle into the centrifugal tube, and the volume is calculated by combining the cross-sectional area change. In addition, the structure where the piercing needle is located may occupy a part of the volume, such as the volume of the piercing needle itself which may reduce the actual extracted liquid amount, or during the piercing process, due to the inverted conical design, as the piercing depth increases, the cross-sectional area of the liquid passage changes, which also needs to be corrected and compensated for the volume. The correction value may include error sources such as volume change caused by elastic deformation of the material, or liquid residue and other factors. All of these need to be considered.

[0036] The correction value may include manufacturing tolerance, material elasticity rebound, liquid residue, etc. These can be determined by experiment (controlled variable method) to determine a correction factor.

[0037] Alternatively, the extraction capacity control formula of the organic phase lipid liquid is as follows: , Wherein: V extract is the extraction capacity of the organic phase lipid liquid; such as 5.2 mL; R tube is the outer radius of the centrifugal tube 1; such as 6.0 mm; r avg is the average outer radius of the piercing head 5; such as the piercing head 5 is an inverted conical structure, and the average outer radius of the needle tip and the root part is not more than 0.50-0.85 mm; N is the number of spiral tightening turns when the bottom of the centrifugal tube 1 is in contact with the needle tip of the piercing head 5; such as 3 turns; P is the thread pitch, that is, the axial distance advanced per turn, unit: mm / turn; such as 1.5 mm / turn; η is the fluid efficiency affected by the spiral flow guide groove 6; such as 0.95; V needleVp is the volume of the needle tip of the piercing head 5; such as 0.15 mL; C material C is the correction factor affected by the material elasticity of the centrifuge tube 1, to compensate the volume collapse due to the spring back of the centrifuge tube 1, such as 1.05.

[0038] Normally, the pitch P of a thread is defined as the distance advanced per revolution, so D = N P. Here, if the pitch P is in millimeters and the number of revolutions N is dimensionless, then D is in millimeters, which needs to be converted to volume (e.g. milliliters) by multiplying with the cross-sectional area (square millimeters) and converting to liters or other units.

[0039] Convert the number of revolutions and the pitch to the piercing depth, multiply with the effective cross-sectional area to get the volume, consider the correction factor, and get the accurate estimation.

[0040] In the above equation, the state when the thread is in contact can be considered as the state when the piercing starts, to design the length of the sleeve for easy counting (number of revolutions). That is, when the centrifuge tube is lowered and the thread of the sleeve is in contact, the top of the piercing needle is in contact with the bottom of the centrifuge tube. The intermediate length can be set by the user. tube , r avg , P, V needle can be measured directly or indirectly, η, C material can be calibrated inversely by known liquid volume (e.g. inject 5 mL of standard liquid, adjust the parameters so that the formula output matches the measured value, which can be verified by the control variable method such as fixing C material and verifying η).

[0041] Vr is the cross-sectional area of the annular region (between the inner wall of the centrifuge tube and the outer wall of the needle) through which the liquid flows after the piercing needle enters the centrifuge tube.

[0042] Axial piercing depth calculation: the number of thread tightening revolutions NN and the pitch P determine the piercing depth D: D = N P, ‌Example: N = 3, P = 1.5 mm → D = 4.5 mm.

[0043] Introduce the fluid efficiency coefficient η (due to the optimization of flow efficiency by the flow guide groove): Theoretical extraction volume = × D × η.

[0044] And the inverted conical piercing needle occupies a space V needle when immersed in the liquid, which needs to be deducted. In this way, the formula can be obtained. The accuracy can also be improved in the following way: Precise and controllable: Directly quantify the amount of liquid extraction through the number of threads, avoiding the "experience dependence" of traditional puncture. Safe redundancy: η, C material Built-in safety margin to prevent leakage caused by over-extraction.

[0045] Optionally, the puncture head 5 uses martensitic age-hardening stainless steel (such as AMS 6512); the sleeve 4 uses 316L stainless steel; The bottom port of the puncture head 5 is fixed and welded on the inner bottom of the sleeve 4 by interference fit and laser welding.

[0046] The puncture head 5 uses the above-mentioned material, and is laser welded (YAG pulse laser, power 300 W, pulse width 5 ms) to make the weld strength of the puncture head ≥ 90% of the base material, resistant to organic solvent corrosion. The sleeve 4 and the puncture needle root are interference fit and laser welded to seal, eliminate interface gaps, and avoid cross contamination caused by liquid retention.

[0047] As Figure 4 shown, the puncture head 5 and the sleeve 4 are laser welded to form a welding surface 7, and a sealing enhancement layer is coated on the upper surface of the welding surface 7 to seal the connection between the puncture head 5 and the sleeve 4.

[0048] The sealing enhancement layer is a 0.2-0.5 μm thick DLC (Diamond-Like Carbon, Diamond-Like Carbon) coating coated on the welding surface 7 by magnetron sputtering technology.

[0049] Combined with martensitic age-hardening stainless steel + DLC coating (0.2 μm), DLC coating (friction coefficient 0.08) is coated by magnetron sputtering technology, so that the puncture combination can have ultra-low puncture resistance and avoid adhesion of centrifuge tube material.

[0050] Puncture head 5 processing: Precise tube cold forming → CNC processing inverted cone → plasma nitriding (600℃ / 2h) → DLC coating deposition → ultrasonic cleaning.

[0051] The structure parameters of the puncture head 5 can be referred to as follows:

[0052] Sleeve assembly: Laser welding of puncture needle and sleeve bottom → DLC coating → 160℃ heat treatment to eliminate internal stress → inner wall electrolytic polishing (Ra≤0.1 μm).

[0053] Optionally, the material of the centrifugal tube 1 is PFPE modified polypropylene, which is a high-performance material made by adding perfluoropolyether oil (PFPE) to the polypropylene base. It combines the advantages of both, not only retaining the basic properties of polypropylene, but also gaining new properties from the addition of PFPE. Its elastic modulus is 1.8 GPa, and the inner wall of the centrifugal tube 1 is siliconized and shaped with a 2-4 μm thick layer of 350 cSt medical silicone oil.

[0054] The centrifugal tube is siliconized on the inner wall (sprayed with 350 cSt medical silicone oil, thickness 2 μm), the silicone oil layer reduces puncture friction and simultaneously fills micro-pores to reduce aerosol escape. Among them, the 350 cSt medical silicone oil layer refers to a coating or film composed of medical-grade silicone oil with a viscosity of 350 centistokes (cSt).

[0055] Centrifugal tube molding: PFPE / PP blending granulation → injection molding → siliconization spraying → ultraviolet curing.

[0056] The specific process can be processed by combining industry processing methods.

[0057] Through testing: Tensile strength: the fracture strength of the welded joint is ≥1800 N (the axial force corresponding to the maximum torque of the centrifugal tube tightening is ≤500 N); ‌Corrosion resistance: soak in a mixture of chloroform / methanol (2:1) for 72 h, no corrosion weight loss in the welded area (SEM detection); ‌Air tightness: 0.6 MPa nitrogen is introduced and pressure is maintained for 10 min, the leakage rate is <0.01 mL / min (ISO 13407 standard).

[0058] The present application can also be combined with sensors to realize automatic counting and reporting to the background, and the extraction amount is calculated by the background and electronic automatic control is realized. When the background finds that the extraction amount reaches the threshold value, an alarm sound can be issued. For example, the terminal PC finds that the extraction amount reaches 4 mL, and an alarm signal is issued (the signal can refer to various existing alarm modes).

[0059] As shown in Figure 5 and Figure 6 , the device further comprises: a reflective sheet 8 fixed on the outer side of the centrifugal tube 1; a number of turns sensing module 9 fixed on the outer side of the sleeve 4, for sensing the number of turns N of the threaded tightening, the number of turns sensing module 9 comprising: Optoelectronic rotation speed sensor, for receiving the light signal reflected by the reflector and feeding back to the MCU (Microcontroller Unit) controller in real time; wherein, when the outer thread 2 of the centrifugal tube 1 starts to cooperate with the inner thread 3 of the sleeve 4, the reflector corresponds to the optoelectronic rotation speed sensor in an up-down relationship; MCU controller, for starting counting according to the light signal, and then corresponding to one thread tightening circle number for each time the light signal is received; when the extraction is finished, the number of all thread tightening circle numbers is counted, and the thread tightening circle number N is obtained; Wireless module, for reporting the thread tightening circle number N to the background server, and inputting the thread tightening circle number N into the preset extraction capacity control formula by the background server to estimate the extraction capacity of the organic phase lipid liquid; Power supply, for power supply; The optoelectronic rotation speed sensor, wireless module and power supply are electrically connected with the MCU controller respectively; The circle number sensing module is in communication connection with the background server through the wireless module.

[0060] The above electronic device hardware can be configured by the user. For example, the wireless can adopt a BLE Bluetooth 5.0 module. The reflector and the circle number sensing module are set in pairs, and multiple groups can be set to realize the counting of non-integer circles.

[0061] The specific implementation can refer to the following scheme: ‌One, reflector and photoelectric sensor configuration‌ 1.‌Refractor parameters‌ ‌Size‌: 5x5 mm square, thickness 0.2 mm; ‌Material‌: PET substrate + micro-prism reflection layer (reflectivity ≥ 85%); ‌Installation position‌: 2 mm below the starting end of the outer thread of the centrifugal tube, coaxial with the thread.

[0062] 2.‌Optoelectronic rotation speed sensor selection‌

[0063] ‌Model‌: TCST2103 (infrared wavelength 940 nm, response time 10 μs); ‌Installation angle‌: The optical axis of the sensor and the normal line of the reflector form an angle of 15° (to avoid mirror reflection interference); ‌Sensitivity adjustment‌: Adjust the detection distance by a potentiometer (adjustable from 3-10 mm).

[0064] ‌Two, MCU control logic design‌

[0065] 1.‌Signal processing flow‌

[0066] / / Initialization settings #define REFLECT_THRESHOLD 800 / / Reflection signal threshold (ADC value)

[0067] void main() {

[0068] while(1) {

[0069] if(ADC_Read() > REFLECT_THRESHOLD) {

[0070] Count_Rotation(); / / Rotation count

[0071] Delay_Ms(50); / / Debounce delay

[0072] }

[0073] }

[0074] }

[0075] .

[0076] Debounce mechanism: After detecting a signal, a 50 ms delay is applied before confirmation to prevent false triggering caused by thread tightening vibration.

[0077] 2. Circle Calculation Algorithm

[0078] Single lap judgment: The interval between two consecutive signals is less than 200 ms; Total laps N: The cumulative number of valid signals, with data uploaded every 10 seconds via the wireless module.

[0079] ‌3. Wireless Communication and Background Interaction‌

[0080] 1. Wireless module configuration

[0081] Protocol: BLE 5.2 (Bluetooth Low Energy); Data format: JSON message {"device_id":"A1B2","rotation_count":N}; Transmission interval: 10 seconds / time (dynamically adjustable).

[0082] 2.‌Backend capacity estimation formula (refer to the previous description).‌

[0083] ‌4. Power Management

[0084] 1. Power supply solution

[0085] Battery: CR2032 coin cell battery (3V, 220 mAh) 2. Power consumption optimization: Photoelectric sensor works intermittently (50 ms on / 950 ms sleep); MCU enters low-power mode (LPM3) standby current <1 μA.

[0086] 3. Endurance calculation

[0087] 100 daily detections of tightening operations, endurance time ≥6 months.

[0088] V. Safety and reliability verification

[0089] 1. Anti-interference measures

[0090] Optical filtering: The sensor is equipped with a 940 nm band-pass filter to suppress environmental light interference; Mechanical protection: The module shell is made of ABS+PC alloy (impact resistance level IK08).

[0091] 2. Clinical testing:

[0092] VI. Installation and maintenance specifications

[0093] Operation steps

[0094] Attach the reflector to the designated position on the centrifuge tube, ensuring no air bubbles; Tighten the sleeve to the initial alignment position (photoelectric sensor LED lights up).

[0095] Fault handling

[0096] Signal loss: Check reflector contamination or battery power; Data anomaly: Automatically trigger calibration mode in the background (manual review required).

[0097] This scheme realizes precise measurement and remote monitoring of the number of tightening circles of the centrifuge tube through photoelectric non-contact detection and wireless Internet of Things technology, providing quantitative control basis for lipid extraction process.

[0098] The above is only a specific implementation of the present application, but the protection scope of the present application is not limited thereto, any skilled person in the art can easily think of changes or replacements within the technical scope disclosed by the present application, which should be covered within the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. An auxiliary device for lipid extraction from biological samples, characterized in that: The device comprises a centrifuge tube and a sleeve, wherein: The inner bottom of the sleeve is vertically provided with a puncture head with the needle facing upwards; The centrifuge tube and the casing are respectively provided with matching external threads and internal threads; When the external thread of the centrifuge tube begins to match the internal thread of the sleeve, a sealed air pressure space is formed between the centrifuge tube and the sleeve, and the needle tip of the puncture head begins to penetrate the bottom of the centrifuge tube and extract the organic phase lipid liquid in the centrifuge tube under the action of the air pressure difference.

2. The auxiliary device for extracting lipids from biological samples according to claim 1, characterized in that The outer surface of the puncture head is provided with a plurality of spiral guide grooves centered on the axis of the puncture head and distributed at equal intervals around the circumference; The spiral guide groove is used for: In the process of the needle tip of the puncture head gradually piercing the bottom of the centrifuge tube, part of the organic phase lipid liquid is guided to flow toward the needle tip of the puncture head, thereby increasing the fluid efficiency of the organic phase lipid liquid.

3. The auxiliary device for extracting lipids from biological samples according to claim 2, characterized in that: The extraction capacity control formula of the organic phase lipid liquid is as follows: ， in: V extract is the extraction capacity of the organic phase lipid liquid; R tube is the outer radius of the centrifuge tube; r avg is the mean outer radius of the puncture head; N is the number of screw tightening turns counted starting from the moment the bottom of the centrifuge tube contacts the needle tip of the puncture head; P is the thread pitch, that is, the axial distance advanced per turn, unit: mm / turn; η is the fluid efficiency affected by the spiral guide groove; V needle is the needle tip volume of the puncture head; C material It is a correction coefficient affected by the material elasticity of the centrifuge tube and is used to compensate for the volume collapse caused by the rebound of the centrifuge tube.

4. The auxiliary device for extracting lipids from biological samples according to claim 1, characterized in that The puncture head has an inverted cone structure, and the average outer radius of the needle tip and the root does not exceed 0.50-0.85 mm.

5. The auxiliary device for extracting lipids from biological samples according to claim 1, characterized in that: The puncture head is made of maraging stainless steel; the sleeve is made of 316L stainless steel; The bottom port of the puncture head is fixedly welded to the inner bottom of the sleeve by interference fit and laser welding.

6. The auxiliary device for extracting lipids from biological samples according to claim 5, characterized in that: A welding surface is formed between the puncture head and the sleeve by laser welding, and a sealing reinforcement layer is coated on the upper surface of the welding surface for sealing the connection between the puncture head and the sleeve.

7. The auxiliary device for extracting lipids from biological samples according to claim 6, characterized in that: The sealing enhancement layer is a DLC coating with a thickness of 0.2-0.5 μm, which is coated on the welding surface by magnetron sputtering technology.

8. The auxiliary device for extracting lipids from biological samples according to claim 5, characterized in that: The centrifuge tube is made of PFPE-modified polypropylene, and the inner wall of the centrifuge tube is siliconized and formed with a 2-4 μm thick 350 cSt medical silicone oil layer.

9. The auxiliary device for extracting lipids from biological samples according to claim 3, characterized in that: The device further comprises: A reflective sheet is fixed on the outer surface of the centrifuge tube; The number of turns sensing module is fixed on the outer surface of the sleeve and is used to sense the number of tightening turns N of the thread. The number of turns sensing module includes: a photoelectric speed sensor for receiving the light signal reflected by the reflective sheet and feeding it back to the MCU controller in real time; wherein, when the external thread of the centrifuge tube begins to engage with the internal thread of the casing, the reflective sheet and the photoelectric speed sensor are in a vertical correspondence relationship; The MCU controller is configured to start counting according to the light signal, and each subsequent light signal received corresponds to a number of thread tightening turns; when the extraction is completed, the number of all thread tightening turns is counted to obtain the number of thread tightening turns N; A wireless module is used to report the number of screw thread tightening turns N to a backend server, and the backend server inputs the number of screw thread tightening turns N into a preset extraction capacity control formula to estimate the extraction capacity of the organic phase lipid liquid; Power supply, used for power supply; The photoelectric speed sensor, wireless module and power supply are electrically connected to the MCU controller respectively; The lap number sensing module is connected to the background server through the wireless module.

Images