CN113267389B - Lung tissue specimen fixing device and method - Google Patents

Abstract

The invention discloses a lung tissue sample fixing device and method. The lung tissue sample fixing device includes a fixed container, a sealing cover and a liquid injector. The top of the fixed container is provided with an opening, and the fixed container is also connected with a pressure regulator. The pressure regulator It communicates with the cavity of the fixed container; the sealing cover includes a cover body and a communication tube, the first end of the communication tube is located under the cover body, and the second end of the communication tube runs through the cover body or is located on the cover body. Insert the first end of the connecting tube into the trachea of the lung tissue and fix it. The cover seals the opening of the fixed container. After increasing the pressure in the fixed container through the pressure regulator, connect the liquid injector to the second end of the connecting tube, and then The pressure in the fixation container is reduced, and the lung tissue absorbs the fixation solution under the action of the pressure difference. The present invention uses bionics to simulate normal breathing state, changes the pressure of the pleural cavity through the movement of the thorax and the diaphragm to discharge the gas, and at the same time allows the fixative to be evenly injected into the lung tissue to achieve better expansion and fixation effects.

Description

A lung tissue sample fixation device and method

technical field

The invention relates to a lung tissue sample fixing device and method in the technical field of biological experiments.

Background technique

After the lung tissue is removed, it is usually shrunken due to the negative pressure in the chest cavity to positive pressure and its own elasticity. At the same time, it still contains a lot of gas inside, causing it to float in the fixative solution. The effect is poor, and the lungs are always in atrophy state during the later slicing process, resulting in unclear microstructure after slicing, which affects the later clinical and pathological judgment and the effect of medical teaching. Early efforts to solve this problem, usually by applying gravity to the lung tissue so that it is completely immersed in the fluid, still cannot solve the situation of lung tissue shrinkage. Later, some researchers used the method of directly pressurizing and perfusing the fixative solution to fill the lungs to a normal expansion state. After fixation, the tissue structure was found to be relatively clear, but this method still has some defects in practice. First of all, pressurized perfusion cannot completely discharge the air in the lungs, and some tissues are still isolated by the gas and cannot touch the fixative fluid; in addition, the pressurized perfusion liquid will flow preferentially to the larger bronchi, which will further squeeze other bronchi after dilation. tissue, resulting in partial tissue incomplete expansion, and at the same time some large bronchus-connected tissue ruptured the alveoli due to over-expansion.

Contents of the invention

The purpose of the present invention is to at least solve one of the technical problems existing in the prior art, and provide a lung tissue specimen fixation device and method to better fix the lung tissue to form a specimen.

According to the embodiment of the first aspect of the present invention, a lung tissue specimen fixing device is provided, which includes a fixing container, an opening is arranged on the top of the fixing container, and a pressure regulator is connected to the fixing container, and the pressure regulator is connected with the The cavity of the fixed container is connected; a sealing cover, the sealing cover includes a cover body and a communication pipe, the cover body is detachably connected to the opening, the first end of the communication pipe is located under the cover body, and the communication pipe The second end of the tube passes through the cover or is located on the cover, the first end of the communication tube is used to connect with the lung tissue; the injector is used to connect with the second end of the communication tube Two-terminal connection.

According to the embodiment of the first aspect of the present invention, further, the liquid injector includes a first liquid storage cylinder and a first piston push rod, and the first piston push rod is slidably installed in the first liquid storage cylinder, so The end of the first liquid storage cylinder is provided with a first protrusion, the first protrusion is provided with a first liquid hole, and the first protrusion can be interference fit with the second end of the communication pipe .

According to the embodiment of the first aspect of the present invention, further, the first liquid storage cylinder is provided with scales along the length direction.

According to the embodiment of the first aspect of the present invention, further, the fixed container is provided with a pressure regulating port, the pressure regulator includes a second liquid storage cylinder and a second piston push rod, and the second piston push rod is slidably installed In the second liquid storage cylinder, the end of the second liquid storage cylinder is provided with a second liquid through hole, and the second liquid through hole can be connected with the pressure regulating port.

According to the embodiment of the first aspect of the present invention, further, a pressure regulating tube is provided on the outside of the pressure regulating port, a second protrusion is provided at the end of the second liquid storage cylinder, and the second liquid hole is located at On the second protrusion, the second protrusion is in interference fit with the pressure regulating tube.

According to the embodiment of the first aspect of the present invention, further, an opening connecting pipe is provided on the top surface of the opening, and the cover is detachably connected to the opening connecting pipe.

According to the embodiment of the second aspect of the present invention, there is provided a method for fixing lung tissue specimens, comprising the following steps: S10. Fix the lung tissue under the sealing cap so that the lung trachea communicates with the first end of the connecting tube; S20. Stretch the tissue into the fixed container filled with fixative through the opening, so that the cover is sealed with the opening; S30. Increase the pressure in the fixed container until the lung tissue shrinks; S40. Connect the liquid injector with the second end of the connecting tube, Then reduce the pressure in the fixed container.

According to the embodiment of the second aspect of the present invention, further, in step S40, when the volume of the fixative liquid injected by the injector into the lung tissue is equal to the lung volume, the adjustment of the pressure in the fixation container is stopped.

The beneficial effect of the present invention is: insert the first end of the connecting tube into the trachea of the lung tissue and fix it, the cover body seals the opening of the fixed container, and after increasing the pressure inside the fixed container through the pressure regulator, then connect the liquid injector and the connecting tube The second end is connected, and then the pressure in the fixed container is reduced, and the lung tissue absorbs the fixative solution under the action of the pressure difference. The present invention applies bionics to simulate normal breathing state, changes the pressure of the pleural cavity through the movement of the thorax and the diaphragm to discharge the gas, and at the same time allows the fixative to be evenly injected into the lung tissue to achieve better expansion and fixation effects.

Description of drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following will briefly describe the drawings that need to be used in the description of the embodiments. Apparently, the described drawings are only some embodiments of the present invention, not all embodiments, and those skilled in the art can obtain other designs and drawings based on these drawings without creative work.

Fig. 1 is the schematic diagram of fixed container in the embodiment of the present invention;

Fig. 2 is the schematic diagram of sealing cap in the embodiment of the present invention;

Fig. 3 is the schematic diagram of the injector in the embodiment of the present invention;

Fig. 4 is a schematic diagram of a pressure regulator in an embodiment of the present invention.

Detailed ways

This part will describe the specific embodiment of the present invention in detail, and the preferred embodiment of the present invention is shown in the accompanying drawings. Each technical feature and overall technical solution of the invention, but it should not be understood as a limitation on the protection scope of the present invention.

In the description of the present invention, it should be understood that the orientation descriptions, such as up, down, front, back, left, right, etc. indicated orientations or positional relationships are based on the orientations or positional relationships shown in the drawings, and are only In order to facilitate the description of the present invention and simplify the description, it does not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as limiting the present invention.

In the description of the present invention, several means one or more, and multiple means two or more. Greater than, less than, exceeding, etc. are understood as not including the original number, and above, below, within, etc. are understood as including the original number. If the description of the first and second is only for the purpose of distinguishing the technical features, it cannot be understood as indicating or implying the relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features relation.

In the description of the present invention, unless otherwise clearly defined, words such as setting, installation, and connection should be understood in a broad sense, and those skilled in the art can reasonably determine the specific meanings of the above words in the present invention in combination with the specific content of the technical solution.

The lung tissue sample fixing device in the embodiment of the first aspect of the present invention includes a fixing container 10 , a sealing cover 20 and a liquid injector 30 , and the sealing cover 20 includes a cover body 21 and a communication tube 22 . The top of the fixed container 10 is provided with an opening, and the fixed container 10 is also connected with a pressure regulator 40. After the lung tissue is fixed in the connecting tube 22, the cover body 21 closes the opening, so that the lung tissue is put into the fixed container 10, and the liquid injector 30 It is connected with the communication tube 22, and the pressure in the fixed container 10 is changed by controlling the pressure regulator 40, and the fixed liquid in the injector 30 is filled into the lung tissue.

Referring to FIG. 1 , the fixative solution is loaded into or poured out of the fixation container 10 through the opening, and lung tissue can also be put into the fixation container 10 through the opening. Optionally, the fixing container 10 is a transparent member, so as to observe the fixing condition of the lung tissue and the fixed lung tissue. The pressure regulator 40 communicates with the cavity of the fixed container 10 to adjust the pressure in the fixed container 10 . It can be understood that an opening connecting pipe 11 is provided on the top surface of the opening, and the cover body 21 is detachably connected with the opening connecting pipe 11 . Optionally, the cover body 21 is threadedly connected to the opening connecting pipe 11 , and an elastic sealing member, such as a silicone ring, is provided between the cover body 21 and the opening connecting pipe 11 . The seal between the cover body 21 and the opening connecting pipe 11 is beneficial to control the pressure in the fixing container 10, thereby ensuring the fixing effect of the lung tissue.

In some embodiments, the fixed container 10 is provided with a pressure regulating port. Referring to FIG. In the liquid storage cylinder 41 , the end of the second liquid storage cylinder 41 is provided with a second liquid hole, and the second liquid hole can be connected with the pressure regulating port. With the movement of the second piston push rod 42 in the second liquid storage cylinder 41 , the stationary liquid flows out or flows into the second liquid storage cylinder 41 through the second liquid hole, thereby changing the cavity pressure of the stationary container 10 . Optionally, the second liquid storage cylinder 41 is provided with a scale along the length direction, so that the operator can visually observe the pressure change in the cavity of the fixed container 10 and achieve quantitative and precise control.

Further, the outside of the pressure regulating port is provided with a pressure regulating tube 12, and the end of the second liquid storage cylinder 41 is provided with a second protrusion 43, and the second liquid hole is located on the second protrusion 43, and the second protrusion 43 Interference fit with the pressure regulating tube 12. Due to the interference fit between the second protrusion 43 and the pressure regulating tube 12 , the sealed connection between the second liquid storage cylinder 41 and the fixed container 10 can be ensured, and the pressure in the cavity of the fixed container 10 can be controlled more accurately.

Referring to FIG. 2 , the cover body 21 is detachably connected to the opening, and the sealing cover 20 can also fix the lung tissue while covering the opening. Specifically, the first end of the communication tube 22 is located below the cover 21 , and the second end of the communication tube 22 passes through the cover 21 or is located on the cover 21 , so that the communication tube 22 can communicate with the space inside and outside the cover 21 . The first end of the communication tube 22 is used to connect with the lung tissue, and the second end of the communication tube 22 is connected to the injector 30 . In this embodiment, the first end of the connecting tube 22 penetrates into the trachea of the lung tissue and is connected. The fixative in the liquid injector 30 flows into the lung tissue through the connecting tube 22 to fully expand and fix the lung tissue.

In some embodiments, referring to FIG. 3 , the liquid injector 30 includes a first liquid storage cylinder 31 and a first piston push rod 32 , the first piston push rod 32 is slidably installed in the first liquid storage cylinder 31 , and the first liquid storage cylinder 31 The end of the barrel 31 is provided with a first protrusion 33 , the first protrusion 33 is provided with a first liquid hole, and the first protrusion 33 can be interference fit with the second end of the communication pipe 22 . By pushing and pulling the first piston push rod 32, the first piston push rod 32 moves in the first liquid storage cylinder 31, and the fixative fluid flows into the communication tube 22 through the first liquid hole, and then the fixative fluid enters the lung tissue. Due to the interference fit between the first protrusion 33 and the communication pipe 22, it can ensure that the stationary liquid will not leak from the gap between the liquid injector 30 and the communication pipe 22, and also ensure that the first liquid storage cylinder 31 will not be filled with gas. into the lung tissue.

Further, the first liquid storage cylinder 31 is provided with a scale along the length direction, and the volume of the fixed liquid filled in the lung tissue can be judged according to the scale, so as to judge whether to continue perfusion or stop the perfusion, so as to ensure that the lung tissue is fully expanded and avoid Part of the bronchi is over-inflated and squeezes other tissues, and even causes alveolar rupture, which affects the production effect of lung tissue specimens.

Taking SD rats of 200g as an example, the lung tissue sample fixing method in the second aspect embodiment of the present invention comprises the following steps:

S10 . Fix the lung tissue under the sealing cap 20 , specifically, bind the lung tissue to the first end of the connecting tube 22 so that the lung trachea communicates with the first end of the connecting tube 22 .

S20. Extending the lung tissue through the opening into the fixation container 10 filled with the fixation solution, so that the cover 21 is sealed and connected with the opening.

S30. Increase the pressure in the fixed container 10 through the air pressure regulator until the lung tissue shrinks. Specifically, before filling the stationary container 10 with the stationary liquid, the second protrusion 43 is first connected to the pressure regulating pipe 12 . When the pressure in the fixed container 10 is to be increased, the second piston push rod 42 is pushed to make the fixed liquid in the second liquid storage cylinder 41 flow into the fixed container 10 through the second liquid hole, thereby increasing the pressure in the fixed container 10 . As the pressure in the fixed container 10 rises, the lung tissue is discharged out of the fixed container 10 through the connecting tube 22 under the action of pressure, so that the lung tissue gradually shrinks until it shows atrophy.

S40. After the lung tissue shrinks, connect the liquid injector 30 with the second end of the communication tube 22, and then reduce the pressure in the fixed container 10. Specifically, the second piston push rod 42 is pulled, so that the stationary liquid in the stationary container 10 flows into the second liquid storage cylinder 41 through the second liquid hole, and the pressure in the stationary container 10 decreases. The lung tissue can be evenly expanded by negative pressure. At the same time, the liquid in the first liquid storage cylinder 31 flows into the lung tissue through the connecting tube 22 under the action of pressure difference, and the fixative fluid flows from the trachea to each bronchi, simulating The circulation of gas in the lungs when animals breathe keeps various structures in the lung tissue in a normal and good shape. Since the lung tissue actively "absorbs" the fixative in the first liquid storage cylinder 31 under the action of the pressure difference, it is avoided that the lung tissue passively receives the fixative when the liquid injector 30 injects liquid into the lung tissue. The structure of the lung tissue may be destroyed under the action; in addition, the inside and outside of the lung tissue are soaked in the fixative solution, so that a better fixation effect can be achieved, and the microstructure after sectioning is closer to the physiological state.

Optionally, when the volume of the fixative fluid injected into the lung tissue by the injector 30 is equal to the lung volume, stop adjusting the pressure in the fixation container 10 to avoid tissue damage caused by excessive expansion of the lung tissue. In this embodiment, the lung volume is calculated according to the body weight of the experimental animal. The formula is: lung volume (ml)=0.981+0.038×body weight (g), and the fixed liquid volume injected into the lung tissue by the injector 30 can be obtained.

In this fixation method, positive pressure is used to discharge the residual gas in the lung tissue first, and then negative pressure is used to inhale the fixative solution. Bionics is used to simulate the movement of the thorax and diaphragm under normal breathing conditions, and the pressure of the pleural cavity is changed to make the gas discharge. At the same time, the fixative is evenly injected into the lung tissue to achieve a better expansion and fixation effect. Compared with the control group that only underwent conventional perfusion, the structure of the lung tissue fixed by this fixation method was normal, the loose vesicular structure of the lung tissue was clear, the alveolar septum was thinner, and all structures maintained normal and good shape without obvious collapse , Shrinkage or distortion of lung tissue structure destruction.

The above is a specific description of the preferred embodiments of the present invention, but the invention is not limited to the embodiments, and those skilled in the art can also make various equivalent modifications or replacements without violating the spirit of the present invention. Equivalent modifications or replacements are all included within the scope defined by the claims of the present application.

Claims (6)

1. A method for fixing a lung tissue sample, adopting a lung tissue sample fixing device with a fixed container, a sealing cover and a liquid injector, is characterized in that the top of the fixed container is provided with an opening, and the fixed container is also connected with a pressure A regulator, the pressure regulator communicates with the cavity of the fixed container, the pressure regulator includes a second liquid storage cylinder and a second piston push rod, and the second liquid storage cylinder is provided with a scale along the length direction , the end of the second liquid storage cylinder is provided with a second liquid hole; the sealing cover includes a cover and a communication tube, the cover can be detachably connected to the opening, and the first end of the communication tube is located at Below the cover, the second end of the communication tube runs through the cover or is located on the cover, the first end of the communication tube is used to connect with the lung tissue; the injector is used to communicate with the The second end of the tube is connected, the liquid injector includes a first liquid storage cylinder and a first piston push rod, and the first liquid storage cylinder is provided with a scale along the length direction; The lung tissue specimen fixing method comprises the following steps: S10. Fixing the lung tissue under the sealing cap, so that the lung trachea communicates with the first end of the connecting tube; S20. Stretch the lung tissue through the opening into the fixed container that has been equipped with fixative, so that the lid is sealed with the opening; S30. Pushing the second piston push rod, so that the fixative in the second liquid storage cylinder flows into the fixation container through the second liquid hole, increasing the pressure in the fixation container until the lung tissue shrinks; S40. Connect the liquid injector to the second end of the connecting pipe, pull the second piston push rod, so that the fixed liquid in the fixed container flows into the second liquid storage cylinder, reduce the pressure in the fixed container, and the fixed liquid in the first liquid storage cylinder Under the action of pressure difference, the liquid flows into the lung tissue through the connecting tube. 2. The lung tissue specimen fixation method according to claim 1, characterized in that: in step S40, when the volume of the fixative fluid injected into the lung tissue by the injector is equal to the lung volume, stop adjusting the pressure in the fixation container. 3. The lung tissue specimen fixing method according to claim 1, characterized in that: the first piston push rod is slidably installed in the first liquid storage cylinder, and the end of the first liquid storage cylinder is provided with A first protrusion, the first protrusion is provided with a first liquid hole, and the first protrusion can be interference-fitted with the second end of the communication pipe. 4. The lung tissue specimen fixing method according to claim 1, characterized in that: said fixing container is provided with a pressure regulating port, said second piston push rod is slidably installed in said second liquid storage cylinder, and said The second liquid hole can be connected with the pressure regulating port. 5. The lung tissue specimen fixing method according to claim 4, characterized in that: a pressure regulating tube is provided on the outside of the pressure regulating port, and a second protrusion is provided at the end of the second liquid storage cylinder, so that The second liquid hole is located on the second protrusion, and the second protrusion is in interference fit with the pressure regulating tube. 6 . The method for fixing lung tissue samples according to claim 1 , wherein an open connecting tube is provided on the top surface of the opening, and the cover is detachably connected to the open connecting tube. 7 .

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