CN112451163A - Novel three-way valve type gravity flow experimental mouse alveolar lavage fluid acquisition device - Google Patents
Novel three-way valve type gravity flow experimental mouse alveolar lavage fluid acquisition device Download PDFInfo
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Abstract
本发明公开了一种新型三通阀式重力流实验鼠肺泡灌洗液采集装置，包括固定架、注射针筒、注射器、医用三通阀、静脉输液软管和滞留软管，其中注射针筒竖直固定在固定架上，医用三通阀的第一端连接注射针筒的注射头，第二端连接注射器的注射头，第三端连接静脉输液软管的一端，静脉输液软管的另一端连接滞留软管，滞留软管末端为倾斜面。本发明的肺泡灌洗液采集装置易于掌握操作，能够获得更高的实验成功率。
The invention discloses a novel three-way valve type gravity flow experimental rat bronchoalveolar lavage fluid collection device, comprising a fixing frame, an injection syringe, a syringe, a medical three-way valve, an intravenous infusion hose and a retention hose, wherein the injection syringe Vertically fixed on the fixing frame, the first end of the medical three-way valve is connected to the injection head of the injection syringe, the second end is connected to the injection head of the syringe, the third end is connected to one end of the intravenous infusion hose, and the other end of the intravenous infusion hose is connected. One end is connected to the retention hose, and the end of the retention hose is inclined. The bronchoalveolar lavage fluid collection device of the present invention is easy to master and operate, and can obtain a higher experiment success rate.
Description
Technical Field
The invention relates to the field of medical experimental equipment, in particular to a novel three-way valve type gravity flow experimental mouse alveolar lavage fluid collecting device.
Background
With the progress of research on respiratory system related diseases, the change of components in alveolar lavage fluid and the significance of the alveolar lavage fluid in diseases are utilized to have important values for researching lung related diseases caused by various factors. With the development of relevant experimental science, the detection of alveolar lavage fluid has been in the range from conventional cytology to cell subset labeling, enzymology, immunology, receptor function, molecular genetics and the like. In the process of animal experiment research on pathogenesis, how to successfully obtain alveolar lavage fluid specimens is a key link for evaluating airway inflammation change. Therefore, bronchoalveolar lavage is a safe and practical technique for analyzing and discussing the pathological process of lung diseases, which is an important means for fundamental research of airway diseases.
In the traditional experimental mouse alveolar lavage fluid collection process, the chest cavity is opened firstly to expose the lung, after the trachea is separated in a blunt manner, the trachea is punctured by a vein indwelling needle, the needle is inserted until the needle head reaches the bifurcation of the left main trachea and the right main trachea, the trachea and the vein indwelling catheter are ligated by cotton threads, then the syringe is used for sucking the physiological saline to be slowly injected into the mouse lung until the expansion of the mouse lung to the maximum degree is observed, then the lavage fluid is slowly sucked by the other syringe, and the process is repeated for a plurality of times. The collection method needs to adopt the vein indwelling needle, so that the problem that the trachea is easy to puncture or the lung tissue is easy to puncture exists, and the rigid needle head of the vein indwelling needle is difficult to fix at a safer position; in addition, the interface repeatedly replaces the syringe filled with the physiological saline and the syringe for suction. Therefore, the problems of complex operation, low acquisition success rate, high requirement on the proficiency of operators and the like exist.
Disclosure of Invention
In order to overcome the problems in the prior art, the invention provides a novel three-way valve type gravity flow alveolar lavage fluid collecting device.
According to one aspect of the invention, the novel three-way valve type gravity flow experimental mouse alveolar lavage fluid collection device comprises a fixing frame, an injection needle cylinder, an injector, a medical three-way valve, a venous transfusion hose and a detention hose, wherein the injection needle cylinder is vertically fixed on the fixing frame, a first end of the medical three-way valve is connected with an injection head of the injection needle cylinder, a second end of the medical three-way valve is connected with an injection head of the injector, a third end of the medical three-way valve is connected with one end of the venous transfusion hose, the other end of the venous transfusion hose is connected with the detention.
Preferably, the syringe has a capacity of 50ml and the injector has a capacity of 5 ml.
Furthermore, the inner diameter of the joint of the medical three-way valve is 5.0mm, the inner diameter of the retention hose is 1.0mm, and the outer diameter is 1.1 mm; the venous transfusion hose is a disposable venous transfusion needle, the steel needle end of the disposable venous transfusion needle is cut off, the upper end of the disposable venous transfusion needle is an expanding end, the disposable venous transfusion needle is just in sealing sleeve connection with a connector of a medical three-way valve, and the detention hose is just in sealing insertion connection with the lower end of the venous transfusion hose without extra sealing.
According to another aspect of the present invention, there is provided a method for collecting alveolar lavage fluid of a laboratory rat, comprising the steps of:
(1) the invention provides a novel three-way valve type gravity flow experimental mouse alveolar lavage fluid collecting device;
(2) carrying out intraperitoneal injection on a mouse, disinfecting the skin of a neck and a chest by using alcohol conventionally, opening the chest to expose the lung, carrying out blunt trachea separation, cutting a transverse small opening below the cricoid cartilage of the trachea by using an ophthalmic scissors, inserting a retention hose into an incision, and pushing the retention hose inwards until the tip of the retention hose reaches the bifurcation of a left main trachea and a right main trachea;
(3) the injection syringe at the upper end is filled with normal saline, and the knob is rotated to enable the three-way valve to be communicated up and down, so that the normal saline naturally flows into the lung through gravity;
(4) when the lung expands to the maximum degree, the three-way valve is rotated to connect the injector on the left side and the retention hose on the lower side, lavage liquid is slowly sucked through the injector, and the suction is stopped when resistance is met or the suction is not performed, and the lavage liquid is collected.
The experimental mouse comprises a rat and a mouse, when the experimental mouse is suitable for the rat, a retention hose can be omitted from the device, and the tail end of an intravenous infusion hose can be cut into an inclined surface and directly inserted into the trachea of the rat.
The novel three-way valve type gravity flow alveolar lavage fluid collection device has the following advantages:
(1) the device fully utilizes the core structure of the three-way valve, a 50ml syringe is connected above the device, large-capacity physiological saline can be contained for filling the lung once, the side surface of the device is connected with an injector for sucking alveolar lavage fluid, the lower part of the device is connected with a detention hose and is inserted into a trachea, lavage can be continuously carried out for many times by simply rotating the three-way valve, the injector filled with the physiological saline and the injector for suction are saved from being repeatedly replaced at the interface of the venous indwelling needle, and the step of ligation can be saved because the hose is inserted instead of the rigid needle head of the venous indwelling needle which is difficult to fix at a safe position;
(2) the success rate is high, because the retention tube inserted into the organ is a soft tube, the problem that the rigid needle head of the venous retention needle punctures the trachea or the lung tissue when inserted into the trachea is basically avoided, and in addition, the normal saline makes the lung reach the maximum filling by utilizing the natural gravity, and the problems that the lung tissue is damaged and the normal saline leaks outwards because the excessive normal saline is injected into the lung by the traditional method are also avoided;
(3) the lavage is sufficient, enough physiological saline can be infused in one time, the lung tissues of the improved group can be observed to be paler than the lung tissues of the traditional group by the same times of lavage, and the amount of the lavage fluid collected and the number of white blood cells finally obtained are also more.
Drawings
FIG. 1 is a schematic structural view of the novel three-way valve type gravity flow experimental mouse alveolar lavage fluid collection device of the present invention.
Fig. 2 is a working principle diagram of the medical three-way valve adopted by the invention.
FIG. 3 is a photograph of an experiment in which alveolar lavage fluid was collected using a conventional intravenous indwelling needle method.
FIG. 4 is a photograph of an experiment using the novel three-way valve device of the present invention to collect alveolar lavage fluid.
Detailed Description
The following describes in detail a specific embodiment of the novel three-way valve type gravity flow experimental mouse alveolar lavage fluid collection device of the present invention with reference to the accompanying drawings. It should be understood by those skilled in the art that the following described embodiments are only illustrative of the present invention and are not intended to limit the same in any way.
Referring to fig. 1, the novel three-way valve type gravity flow experimental mouse alveolar lavage fluid collection device comprises a fixing frame 1, an injection syringe 2, an injector 3, a medical three-way valve 4, a venous transfusion hose 5 and a retention hose 6. Wherein, the fixing frame 1 can adopt an iron stand commonly used in a laboratory. The injection syringe 2 adopts the existing injector on the market to remove the push rod and the injection needle, only the syringe part with the specification of 50ml is reserved, and the syringe part is vertically fixed on the fixed frame 1 through the fixture 7. The injector 3 adopts the existing injector on the market to remove the injection needle, and the needle cylinder and the push rod are reserved, and the specification is 5 ml. The lower ends of the injection syringe 2 and the injector 3 are provided with injection heads with the outer diameter of 5 mm. The upper end joint of the medical three-way valve 4 is connected with the injection head of the injection syringe 2, the side end joint is connected with the injection head of the injector 3, the lower end joint is connected with one end of the venous transfusion hose 5, and the other end of the venous transfusion hose 5 is connected with the detention hose 6. The inner diameter of the connector of the medical three-way valve 4 is 5.0mm, the inner diameter of the retention hose 6 is 1.0mm, and the outer diameter is 1.1 mm; the venous transfusion hose 5 adopts a common disposable venous transfusion needle on the market, the steel needle end is cut off, the upper end is an expanding end, the third end of the medical three-way valve 4 is just in sealing sleeve joint, one end of the detention hose 6 is just in sealing insertion in the lower end of the venous transfusion hose 5, and extra sealing is not needed. In order to facilitate the insertion of the cut laboratory mouse trachea, the other end of the retention hose 6 is cut into an inclined plane. The intravenous transfusion hose 5 and the retention hose 6 are made of polyethylene.
Referring to fig. 2, the medical three-way valve 4 is composed of a three-way pipe, a valve core in the pipe, and a T-shaped knob connected with the valve core. Three communicated pipelines are arranged in the valve core, and the direction of the pipelines is consistent with the three directions of outward pointing of the T-shaped knob. When the T-shaped knob is rotated to the position shown in fig. 2A, the three-way pipeline is communicated in the up-and-down direction, and the physiological saline can naturally flow out downwards under the action of gravity; turning the "T" knob as shown in FIG. 2B closes the upper channel, stops the saline infusion, and communicates the sides and bottom, allowing for aspiration of alveolar lavage fluid via syringe.
The following describes the implementation effect of the novel three-way valve type gravity flow experimental mouse alveolar lavage fluid collection device according to the present invention by using specific examples.
12 Kunming mice were divided into 2 groups of 6 mice each. One group collected alveolar lavage fluid using conventional intravenous indwelling needle (conventional group) and the other group collected alveolar lavage fluid using the novel three-way valve device of the present invention (modified group). The operator is the experimenter who implements 2 kinds of devices for the first time, and the specific conditions are as follows:
conventional group: referring to fig. 3, mice were injected with an intraperitoneal anesthetic, the neck and chest skin was disinfected with 75% alcohol, the chest was opened to expose the lungs, the trachea was bluntly isolated, a venous indwelling needle was inserted into the trachea (start timing), the needle was inserted until the needle reached the bifurcation of the left and right main bronchi, the trachea and venous indwelling catheter were ligated with cotton thread, 5ml syringe was used to aspirate physiological saline and slowly inject into the mice lungs until the mice lungs were observed to swell to the maximum, another 5ml syringe was used to aspirate lavage fluid slowly, aspiration was stopped when resistance or aspiration failed, 15ml centrifuge tube was used to hold lavage fluid, repeat 6 times, stop timing, and record the time taken by each mouse and the amount of lavage fluid collected.
The improvement group is as follows: referring to fig. 4, the mouse is injected with an anesthetic in the abdominal cavity, the skin of the neck and chest is disinfected with 75% alcohol conventionally, the chest cavity is opened to expose the lung, after the trachea is bluntly separated, a transverse small opening (timing is started) is cut slightly below the cricoid cartilage of the trachea with an ophthalmic scissors, the retention hose is inserted into the incision and pushed inwards to the tip of the retention hose to the bifurcation of the left and right main bronchi, and the outer diameter of the retention hose is consistent with the inner diameter of the trachea of the mouse, so that the trachea and the retention hose do not need to be ligated by using cotton threads. 50ml of normal saline is contained in a 50ml syringe at the upper end, a knob is rotated to enable a three-way valve to be communicated up and down, the normal saline is naturally injected into the lung, when the lung expands to the maximum degree, a 5ml syringe connected to the left side is rotated to be communicated with a detention hose at the lower part, lavage fluid is slowly sucked through the syringe, the suction is stopped when resistance or suction is unavailable, a 15ml centrifugal tube is used for containing the lavage fluid, the process is repeated for 6 times, the timing is stopped, and the time used by each mouse and the collected lavage fluid volume are recorded.
Leukocyte extraction and reading in lavage:
placing the alveolar lavage fluid on ice, then centrifuging the alveolar lavage fluid in a centrifuge at 4 ℃ and 1500r/s for 10 minutes, discarding the supernatant, adding 1500 mu l of erythrocyte lysate, uniformly mixing the alveolar lavage fluid and the erythrocyte lysate, placing the alveolar lavage fluid on the ice for 10 minutes, then centrifuging the alveolar lavage fluid in the centrifuge at 4 ℃ and 1500r/s for 10 minutes again, discarding the supernatant, adding 200 mu l of normal saline, uniformly mixing the alveolar lavage fluid and the erythrocyte lysate, and finally counting the number by adopting a cell counting plate.
The experimental results are as follows:
as can be seen from tables 1 and 2, the conventional group failed twice, while the improved group failed, with the success rates of 66.7% and 100%, respectively. The reasons for the failure of the conventional group are: firstly, a vein remaining needle is inserted into a trachea; ② excessive physiological saline is injected into the lung, and the physiological saline of the lung tissue leaks out. The retention hose used by the improved group can avoid puncturing the trachea to a great extent, and the normal saline naturally and downwards fills the lung by utilizing gravity until the whole lung is full, so that the normal saline can be prevented from being injected into the lung as much as possible, but the normal saline of the lung tissue leaks out due to the bad control. Therefore, the lung irrigation device with the improved group is easier to master and operate, and higher experiment success rate can be obtained.
TABLE 1 results of the conventional group experiments
Table 2 modified set of experimental results
In contrast to the two sets of lavage times, although one was saline injected by syringe and the other was naturally saline injected by gravity, the lavage fluid was aspirated by syringe, so the difference was not statistically significant, although the two methods used different times (Table 3).
Comparing the two groups of lavage fluid amounts, finding that the difference of the two groups of total lavage fluid amounts has statistical difference (table 3), naturally flowing the normal saline into the lung by the improved group by using gravity, naturally stopping when the whole lung is full, and averagely filling 2-2.5ml of normal saline into the lung each time; in the traditional group, the air pipe is punctured through the syringe needle, then liquid is injected by artificially giving pressure, and the problem of side leakage of the liquid caused by damaging lung tissues due to puncturing the air pipe, or over-violent too fast, excessive liquid injection amount and the like exists, so that 1-1.5ml of the liquid is usually injected at most at one time; and both groups use syringes to draw lavage fluid, the improved group has more lavage fluid than the conventional group.
Comparing the number of leukocytes in the lavage fluids in the two groups, the improved group was significantly more than the conventional group, and the differences were statistically significant (Table 3). In the case of the same lavage times of 6 times, the number of leukocytes collected by the improved group was significantly greater than that of the conventional group because the improved group was filled with a significantly larger amount of physiological saline than the conventional group and the improved group was also filled with a significantly larger amount of lavage fluid than the conventional group in the same manner of aspirating the lavage fluid with the syringe.
TABLE 3 comparison of the results of the conventional and improved groups
Note: p <0.05
The above description is only an embodiment of the present invention, and is not intended to limit the present invention, and all equivalent changes and modifications according to the spirit of the present invention are within the scope of the technical protection scheme of the present invention.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116343566A (en) * | 2023-03-21 | 2023-06-27 | 挪度医疗器械（苏州）有限公司 | A reusable multifunctional airway management trainer |
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Application publication date: 20210309 |
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| RJ01 | Rejection of invention patent application after publication |