Everted intestinal sac light-shielding experimental device

Abstract

The utility model relates to an ectropion intestinal sac shading experimental device which comprises a constant-temperature water bath kettle and gas cylinders positioned around the constant-temperature water bath kettle, the test tube rack is arranged in the constant-temperature water bath kettle and is provided with a plurality of accommodating spaces; a plurality of centrifugal tubes are correspondingly placed in the accommodating space, and a branch air supply tube, a sampling guide tube and an air pressure balance tube are inserted into a tube cover of each centrifugal tube; one end of the precise gas adjusting flow divider is connected with one end, far away from the gas cylinder, of the main gas supply pipe, is connected with a plurality of branch gas supply pipes and is correspondingly communicated with the centrifugal pipes; a light shielding layer is arranged on the outer wall of the centrifugal tube. By introducing the precise gas adjusting flow divider, uniform distribution and precise control of gas flow are achieved, air pressure balance in the device is ensured, device damage or unstable experiment conditions caused by too high air pressure are avoided, and the activity and function of the intestinal capsule are effectively maintained. The light shielding layer of the centrifugal tube effectively isolates light, solves the problem that a fluorescent marker is quenched due to illumination, and ensures the stability of a fluorescent signal and the scientific accuracy of an experimental result.

Description

TECHNICAL FIELD

[0001] The utility model relates to the technical field of everted intestinal sac model, more specifically relates to a kind of everted intestinal sac light-avoiding experimental device. BACKGROUND

[0002] In biomedical research, polysaccharide substances are of great concern due to their antioxidant, immunomodulatory and other biological activities. However, polysaccharide molecules lack characteristic UV absorption groups, making it difficult to directly track their intestinal absorption process using traditional detection methods. Currently, fluorescence labeling methods (such as FITC labeling) combined with everted intestinal sac models are mainly used for research: the intestinal segment of experimental animals is everted to form a sac-like structure, and by detecting the concentration changes of the fluorescent marker outside the sac, the intestinal absorption characteristics of polysaccharides are quantitatively analyzed.

[0003] Everted intestinal sac models have advantages such as small sample size and strong controllability, but their technical implementation faces two major challenges:

[0004] Photosensitivity problem: fluorescent markers are prone to photobleaching under visible light, leading to signal attenuation. Existing devices (such as CN217265788U) lack light-avoiding design, making it difficult to ensure the stability of fluorescent signals during the experiment.

[0005] Gas regulation defects: the maintenance of intestinal sac activity relies on the continuous supply of 95% O2 and 5% CO2 mixed gas. Traditional gas supply systems have problems such as large pressure fluctuations and uneven flow, which may cause intestinal sac inactivation or mechanical damage.

[0006] In the prior art, although there are improved everted intestinal sac experimental devices (see CN217265788U), there are still significant shortcomings:

[0007] 1. Open structure leads to full exposure of fluorescent samples to ambient light;

[0008] 2. The gas path system uses a simple shunt design, lacking precise flow control;

[0009] 3. No gas pressure compensation mechanism is set, which can easily cause pipeline pressure imbalance;

[0010] The above defects directly affect the reliability and repeatability of experimental data, restricting the application of the model in fluorescence labeling research. Therefore, it is urgent to develop an everted intestinal sac experimental device that integrates light-avoiding function and has precise gas regulation capability. Utility model content

[0011] Therefore, the purpose of the utility model is to provide an everted intestinal sac experimental device that overcomes the shortcomings of the prior art.

[0012] The utility model discloses a kind of everted intestinal sac light-avoiding experimental devices, including constant-temperature water bath, provide constant-temperature environment, and gas cylinder located around the constant-temperature water bath;Still include:

[0013] Test tube rack, arranged in the constant-temperature water bath, with multiple accommodation spaces;

[0014] Multiple centrifuge tubes, corresponding to be placed in the accommodation space, branch gas supply pipe, sampling catheter and air pressure balance pipe are inserted on the tube cover of each centrifuge tube;

[0015] Main gas supply pipe, connect the gas cylinder;

[0016] Precise gas regulating flow divider, one end is connected to the end of main gas supply pipe away from the gas cylinder, multiple branch gas supply pipes are connected thereon, and correspondingly connected to the centrifuge tube;

[0017] Among them, the centrifuge tube outer wall has light-avoiding layer.

[0018] According to the device of the utility model, the centrifuge tube outer surface is covered with tin foil paper to form the light-avoiding layer.

[0019] According to the device of the utility model, the sampling catheter and air pressure balance pipe are PP polypropylene transparent plastic tubes.

[0020] According to the device of the utility model, the precise gas regulating flow divider is a metal hose flow divider with independent switch, and is fixed by adjustable clamp of iron stand.

[0021] According to the device of the utility model, the main gas supply pipe is T-shaped silica gel hose.

[0022] According to the device of the utility model, the branch gas supply pipe is linear silica gel hose.

[0023] According to the device of the utility model, the end opening of the air pressure balance pipe is located above the liquid level of the centrifuge tube;The end of the branch gas supply pipe extends below the liquid level of the centrifuge tube.

[0024] According to the device of the utility model, the water level line of the constant-temperature water bath covers more than 4 / 5 of the overall height of the test tube rack.

[0025] Through the above technical scheme, compared with prior art, the utility model has the following beneficial effects:

[0026] 1, improve fluorescence signal stability: the utility model is through the light-avoiding layer (such as tin foil paper 10) of centrifuge tube outer wall full package design, effectively blocks the interference of ambient light to fluorescent marker, makes fluorescence signal decay rate reduce to 5% below (traditional device decay rate ≥30%), ensures the accuracy of experimental data.

[0027] 2. Optimizing gas regulation accuracy: The precision gas regulation shunt combines with the T-shaped main gas supply pipe and the linear branch gas supply pipe to realize the uniform distribution of mixed gas, and the flow fluctuation is controlled within ±0.05 L / min; the branch gas supply pipe extends to below the liquid surface, ensuring that the gas is continuously supplied to the intestinal sac in the form of micro-bubbles, and avoiding mechanical damage to the intestinal sac caused by sudden changes in gas pressure.

[0028] 3. Enhancing the stability of the experimental environment: The water level of the constant temperature water bath covers more than 4 / 5 of the height of the test tube rack, which cooperates with the test tube rack fixing structure to ensure the uniform immersion depth of the centrifuge tube, and the temperature fluctuation is ≤0.5℃; the opening at the end of the gas pressure balance pipe is located above the liquid surface, which dynamically adjusts the volume change of the pipe, and the internal pressure of the device is stable within ±50Pa.

[0029] 4. Operation convenience: The sampling conduit made of PP transparent plastic material has light-proof and operation visibility, which is convenient for real-time observation of the liquid flow state; the metal hose shunt fixed by the adjustable clamp supports multi-channel independent on-off control, and the experimental flux is improved by more than 3 times. BRIEF DESCRIPTION OF DRAWINGS

[0030] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiment or prior art description. Obviously, the drawings in the following description are only embodiments of the present application, and for those skilled in the art, other drawings can also be obtained without creative labor on the basis of the provided drawings.

[0031] Figure 1 The structure diagram of the light-avoiding experimental device for everted intestinal sac is provided.

[0032] Figure 2 The separation tube with light-avoiding layer is schematically shown.

[0033] Figure 3 The top view of the separation tube is schematically shown. Figure 2

[0034] The separation tube with everted intestinal sac is schematically shown. Figure 4

[0035] The schematic diagram of the test tube rack placed in the constant temperature water bath is schematically shown. Figure 5

[0036] The schematic diagram of the iron stand clamping and fixing the precision gas regulation shunt by the adjustable clamp is schematically shown. Figure 6 DETAILED DESCRIPTION

[0037] ​Embodiments of the present application are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference signs represent the same or similar elements or elements having the same or similar functions throughout. The embodiments described below by reference to the drawings are exemplary and are intended to explain the present application, and cannot be understood as a limitation of the present application.

[0038] In the description of the present application, it should be understood that the terms "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer" and the like indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and therefore cannot be understood as indicating or implying that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation of the present application.

[0039] The existing technology still has the following defects: the open structure causes the fluorescent sample to be exposed to ambient light throughout the experiment; the gas path system uses a simple shunt design, lacking precise flow control; no gas pressure compensation mechanism is provided, which can easily cause pressure imbalance in the pipeline; the above defects directly affect the reliability and repeatability of experimental data, and restrict the application of the model in fluorescence labeling research.

[0040] Therefore, the technical scheme provided by the present application is a light-avoiding experimental device for everted gut sac, which comprises a constant temperature water bath pot 1, which provides a constant temperature environment (37℃±0.5℃ water bath temperature), and a gas cylinder 2 (containing 95% oxygen and 5% carbon dioxide mixed gas) located around the constant temperature water bath pot 1; see Figures 1-5 It also comprises a test tube rack 13, a plurality of centrifuge tubes 7, a main gas supply pipe 5, and a precise gas regulating shunt 4.

[0041] The test tube rack 13 is arranged in the constant temperature water bath pot 1 and has a plurality of accommodation spaces (more than 6 cylindrical accommodation cavities).

[0042] The plurality of centrifuge tubes 7 are placed in the accommodation spaces, and each centrifuge tube 7 (which can be a standard centrifuge tube of 15ml) is provided with three through holes on the tube cover, with hole diameters of: a branch gas supply pipe 6 insertion hole: φ4.0±0.1mm; a sampling catheter 8 insertion hole: φ5.0±0.1mm, and a gas pressure balance pipe insertion hole: φ4.5±0.1mm; the branch gas supply pipe 6, the sampling catheter 8 and the gas pressure balance pipe 9 are respectively inserted.

[0043] The main gas supply pipe 5 is connected to the gas cylinder 2.

[0044] The precise gas regulating shunt 4 is connected with one end of the main gas supply pipe 5 far away from the gas cylinder 2, a plurality of branch gas supply pipes 6 are connected with the precise gas regulating shunt 4, and the centrifugal tubes 7 are correspondingly connected, so that the uniform distribution and precise control of the gas are realized.

[0045] The centrifugal tube 7 is provided with a light-proof layer on an outer wall.

[0046] In the utility model, the precise gas regulating shunt is introduced, the uniform distribution and precise control of the gas flow are realized, the internal pressure balance of the device is ensured, the device damage or unstable experimental conditions caused by excessively high gas pressure are avoided, and the activity and function of the intestinal capsule are effectively maintained. The light-proof layer on the outer wall of the centrifugal tube effectively isolates light, solves the problem that the fluorescent marker is quenched by light, and ensures the stability of the fluorescent signal and the scientific accuracy of the experimental results.

[0047] In the embodiment of the utility model, the outer surface of the centrifugal tube 7 is tightly wrapped by tin foil paper 10 (0.02mm thick, the joint is sealed by aluminum foil tape) to form the light-proof layer. The sampling conduit 8 and the gas pressure balance pipe 9 can be PP polypropylene transparent plastic pipes with an outer diameter of 4-6mm and an inner diameter of 3-5mm.

[0048] In the embodiment of the utility model, the precise gas regulating shunt 4 is a metal hose shunt with an independent switch, and is fixed through the adjustable clamp 14 of the iron stand 3, as shown in the accompanying drawings. Figure 6 The stability and safety of the device in the experimental process are ensured.

[0049] In the utility model everted intestinal capsule light-proof experimental device, the precise gas regulating shunt 4 comprises:

[0050] Metal hose: made of flexible metal bellows (such as stainless steel 304 / 316), with pressure resistance (usually >=1MPa), corrosion resistance (acid and alkali and organic solvent resistance) characteristics, and a bending radius of 3-5 times of the pipe diameter.

[0051] Shunt body: multi-channel metal manifold (commonly 4 / 6 / 8 channels), each channel is provided with an independent valve (knob type or needle valve type).

[0052] Switch mechanism 15: the valve of each channel is provided with a precise adjusting knob, and the gas flow can be independently controlled (the adjusting precision can reach ±2% of the full scale).

[0053] The precise gas regulating shunt 4 is reflected in:

[0054] Intestinal capsule activity maintenance: the gas supply flow of each centrifugal tube 7 (such as 0.5±0.05L / min) is independently adjusted, the oxygen supply of the intestinal capsule 11 is ensured to be uniform, and local hypoxia or overoxidation damage is avoided.

[0055] Experimental stability guarantee, multi-channel independent switch design can quickly isolate the fault unit (such as a centrifugal tube leakage), prevent the whole system from stopping;

[0056] Metal material resistant to water bath environment hot and humid corrosion (compared with plastic shunt life more than 3 times).

[0057] Flexible operation, experimenters can quickly set different gas flow through the knob scale (such as 0-10 gears) to adapt to the gas demand of different intestinal segments (jejunum / ileum).

[0058] In specific embodiments, the main gas supply pipe 5 can be a T-shaped silicone hose with an outer diameter of 5-8 mm and an inner diameter of 4-7 mm. The manufacturing method is as follows: select a silicone hose with an outer diameter of 5-8 mm, an inner diameter of 4-7 mm, and a length of 10-15 cm, accurately drill a hole with a diameter of 5-8 mm in the middle section, insert another silicone hose with a length of 30-40 cm and the same specifications into the hole, and use special silicone adhesive to firmly bond the interface. The design ensures the stability of gas flow and the convenience of connection.

[0059] The branch gas supply pipe 6 can be a linear silicone hose with an outer diameter of 3-6 mm and an inner diameter of 2-5 mm to meet the more precise gas distribution requirements.

[0060] In the embodiment of the utility model, the end opening of the gas pressure balance pipe 9 is located above the liquid surface of the centrifugal tube 7; the water level line of the constant temperature water bath kettle 1 covers more than 4 / 5 of the overall height of the test tube rack 13, maintaining a constant experimental temperature environment. The branch gas supply pipe 6 transports mixed gas to the centrifugal tube 7 through the precise regulation of the gas regulating shunt 4. To ensure the activity of the intestinal capsule, the branch gas supply pipe 6 should extend below the liquid surface.

[0061] Referring to the accompanying drawings Figure 4 The end of the sampling catheter 8 is connected to the ligation end of the everted intestinal capsule 11 (see Figure 4), for timing extraction transport medium; air pressure balance pipe 9 is open to the end 1cm from the liquid surface, the pipe orifice is installed 0.22 μm hydrophobic filter membrane, prevents liquid backflow. The specific everted intestinal sac 11 one end is tightly sleeved in sampling catheter 8, and is firmly fastened with fine wire 12, the other end is also ligated with fine wire 12, so that intestinal sac 11 is naturally vertically hung in centrifugal tube. In the process of intestinal sac balance, incubation and experiment, the intestinal sac needs to be totally immersed in nutrient solution containing glucose, amino acid and other nutrients to keep the wet state, prevent dry inactivation, and provide energy for the intestinal sac to maintain biological activity. Sampling catheter 8 is used to extract transport medium using a point sampling head at a preset transport time point, and to supplement blank liquid in time to ensure the continuity of the experiment. Air pressure balance pipe 9 should be above the liquid level in centrifugal tube 7, for discharging excess gas, ensuring the air pressure balance inside the device, avoiding the damage of the device or the fluctuation of experimental conditions caused by excessive air pressure. In the embodiment, the cover of centrifugal tube 7 is precisely drilled, and branch gas supply pipe 6, sampling catheter 8 and air pressure balance pipe 9 are connected in turn, to ensure efficient and pollution-free sampling. During the experiment, the water surface of water bath 1 is always covered with test tube rack 13 to maintain a constant experimental temperature environment. Iron stand 3 fixes gas regulating flow divider 4 through adjustable clamp 14 to ensure the stability and safety of the device during the experiment.

[0062] In the utility model, unless there is definite stipulation and limitation, the terms "mount", "link", "connect", "fix" and so on should be understood in broad sense, for example, can be fixed connection, also can be detachable connection, or integral; can be mechanical connection, also can be electrical connection; can be direct connection, also can be indirect connection through intermediate medium, can be the communication of two elements or the interaction of two elements. For ordinary skilled in the art, the specific meaning of the above terms in the utility model can be understood according to specific circumstances.

[0063] Experimental operation flow:

[0064] Intestinal sac preparation: the jejunum segment of SD rat is everted after being washed by pre-cooled Krebs-Ringer buffer solution, the proximal end is fixed to sampling catheter 8 and is double-ligated by surgical suture, and the distal end is closed to form a sac-like structure.

[0065] System assembly: the everted intestinal sac 11 is vertically hung in centrifugal tube 7, and Krebs nutrient solution containing 10mm glucose is injected, and the liquid level is 1cm from the pipe orifice.

[0066] Gas activation: open gas cylinder 2, adjust the flow divider to make the flow rate of each branch gas supply pipe stable at 0.5L / min, and activate the intestinal sac activity for 10min.

[0067] Light-avoiding experiment: the centrifuge tube 7 wrapped with tin foil 10 is placed in the constant temperature water bath 1, and the transport experiment is carried out under light-avoiding condition, 200 mu L of the external liquid is collected through the sampling pipe 8 every 30 min, and an equal amount of blank nutrient solution is supplemented.

[0068] In one embodiment, the inner diameter of the branch gas supply pipe 6 is adjusted to 2mm, the gas flow rate is increased to 0.8L / min, and the bubble diameter is reduced to less than 0.5mm, further reducing the mechanical stimulation to the intestinal capsule;

[0069] In the utility model, the circulating water pump can be additionally arranged in the constant temperature water bath 1, so that the temperature gradient difference is less than or equal to 0.2 DEG C;

[0070] The digital pressure sensor can also be connected to the end of the air pressure balance pipe 9 to monitor the air pressure change in the pipe in real time.

[0071] The everted intestinal capsule light-avoiding experiment device is convenient and efficient, can effectively prevent the fluorescence sample from being quenched by light, ensures signal stability and result accuracy, realizes uniform distribution and accurate control of air flow by introducing the precise gas regulating shunt 4, and maintains air pressure balance and intestinal capsule activity. The device has low cost, simple operation and is suitable for promotion.

[0072] In the description of the present specification, the description of the terms "one embodiment", "some embodiments", "example", "specific example" or "some examples" means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the utility model. In the present specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in the present specification.

[0073] Although the embodiments of the utility model have been shown and described above, it can be understood that the above-mentioned embodiments are exemplary and cannot be understood as limiting the utility model, and those skilled in the art can change, modify, replace and modify the above-mentioned embodiments within the scope of the utility model.

Claims

1. An everted intestinal sac photophobotometer device, comprising a constant temperature water bath (1) for providing a constant temperature environment, and a gas cylinder (2) located around the constant temperature water bath (1); characterized in that, Also include: Test tube rack (13), arranged in the constant temperature water bath (1), with a plurality of accommodation space; A plurality of centrifuge tubes (7), corresponding to the placement of the accommodation space, each of the centrifuge tube (7) cover inserted with branch gas pipe (6), sampling catheter (8) and gas pressure balance tube (9); Main gas pipe (5), connecting the gas cylinder (2); Precise gas regulating shunt (4), one end connected to the main gas pipe (5) away from the gas cylinder (2) end, on which a plurality of branch gas pipe (6) is connected, and corresponding to the communication of the centrifuge tube (7); Wherein, the centrifuge tube (7) outer wall has a light barrier layer.

2. The everted gut sac photophobic assay device of claim 1, wherein, The centrifuge tube (7) outer surface covered with tin foil (10) to form the light barrier layer.

3. The everted gut sac photophobic assay device of claim 1, wherein, The sampling catheter (8) and gas pressure balance tube (9) are PP polypropylene transparent plastic tube.

4. The everted gut sac photophobic assay device of claim 1, wherein, The precise gas regulating shunt (4) is a metal hose shunt with independent switch, and is fixed by the adjustable clamp (14) of the iron stand (3).

5. The everted gut sac photophobic assay device of claim 1, wherein, The main gas pipe (5) is T type silica gel hose.

6. The everted gut sac photophobic assay device of claim 5, wherein, The branch gas pipe (6) is a linear silica gel hose.

7. The everted gut sac photophobia test device according to any one of claims 1-6, wherein, The end of the gas pressure balance tube (9) is open above the liquid level of the centrifuge tube (7); the end of the branch gas pipe (6) extends below the liquid level of the centrifuge tube (7).

8. The everted gut sac photophobia test device according to any one of claims 1-6, wherein, The water level line of the constant temperature water bath (1) covers more than 4 / 5 of the overall height of the test tube rack (13).

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