A method for taking out a brain of a laboratory rat

Abstract

The application relates to a method for taking brain of experimental rats, which comprises the following steps: using tissue scissors to cut the occipital bone to the inion along the direction of the sagittal suture from the foramen magnum of the rat; using a curved hemostat to vertically pierce the frontal bone until the skull base at 0.5-0.8 cm from the anterior fontanelle to the frontal bone of the rat along the extension direction of the sagittal suture; using a curved hemostat to separate the parietal bone, and then taking out the complete brain tissue. Since the frontal bone of the rat is pierced until the skull base at the selected position by using the curved hemostat, there is no brain tissue under the frontal bone at the selected position, the frontal bone and the parietal bone of the rat are connected, and there is a suture between the left and right parietal bones, that is, the sagittal suture, so after the frontal bone of the rat is pierced, the left and right parietal bones are separated into two parts along the sagittal suture by the curved hemostat, at this time, the parietal bone above the brain tissue of the rat does not produce broken bone pieces, so that the sampled brain tissue does not contain other tissues, and therefore, the integrity of the brain tissue of the rat can be ensured.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of life science, and particularly relates to a method for taking brain of experimental rat. BACKGROUND

[0002] The use of animal models for the study of neuropsychiatric diseases is essential. This is because neuropsychiatric diseases often involve complex neural circuits and molecular mechanisms, which are difficult to study directly in humans. Using animal models can better study the pathogenesis and pathophysiology of neuropsychiatric diseases, as well as evaluate the effectiveness and safety of treatment methods. These animal models can provide a systematic, controllable and repeatable method to help scientists study the occurrence and development of the disease. Using animal models for the study of neuropsychiatric diseases also helps to develop new drugs and treatment methods. In the animal models for the study of neuropsychiatric diseases, brain tissue is an important material for the study of neuropsychiatric diseases, which can provide important information on neurons, neural circuits and neurotransmitters, etc. Using brain tissue can obtain a lot of useful data.

[0003] The current method for taking brain tissue of experimental rat is to use a drill to make a large enough skull window on the rat's head, use a brain scoop or other sampling tool to take out the rat's brain tissue; or directly use bone forceps to cut open the skull bit by bit to expose the brain tissue, and then take out the brain tissue.

[0004] However, the above method of obtaining brain tissue inevitably affects or even destroys the integrity of the brain tissue, and the sampled brain tissue may contain other tissues, thereby affecting the interpretation and analysis of data. SUMMARY

[0005] Embodiments of the present application provide a method for taking brain of experimental rat to solve the problem that in the related art, a skull window is made on the rat's head using a drill, and the rat's brain tissue is taken out; or the skull is directly cut open using bone forceps, and then the brain tissue is taken out, and the sampled brain tissue may contain other tissues, which affects the integrity of the brain tissue.

[0006] In a first aspect, a method for taking brain of experimental rat is provided, which includes the following steps: using tissue scissors to cut the occipital bone from the foramen magnum of the rat along the sagittal suture direction to the inion; using a curved hemostat to vertically pierce the frontal bone at a position 0.5-0.8 cm from the anterior fontanelle to the frontal bone direction along the extension direction of the sagittal suture until the skull base; using the curved hemostat to pry open the parietal bone to take out the complete brain tissue.

[0007] In some embodiments, the step of using a curved hemostat to vertically pierce the frontal bone at a position 0.5-0.8 cm from the anterior fontanelle to the frontal bone direction along the extension direction of the sagittal suture further includes locking the curved hemostat.

[0008] In some embodiments, the jaw of the curved hemostatic forceps is provided with a spike head.

[0009] In some embodiments, the method of using the curved hemostatic forceps to pry open the parietal bone and remove the intact brain tissue comprises: unlocking the curved hemostatic forceps, prying open the curved hemostatic forceps in a direction perpendicular to the sagittal suture, and evenly dividing the parietal bone above the brain tissue of the rat along the sagittal suture; and cutting off the connection between the brain tissue and the nerves of the skull base, and removing the brain tissue completely.

[0010] In some embodiments, the method of cutting off the connection between the brain tissue and the nerves of the skull base, and removing the brain tissue completely comprises: using the spike head detachably connected to the curved hemostatic forceps to cut off the connection between the brain tissue and the nerves of the skull base; and detaching the spike head from the curved hemostatic forceps, and using the curved hemostatic forceps to remove the brain tissue, so that the intact brain tissue is separated from the skull base.

[0011] In some embodiments, the curved hemostatic forceps comprises a main body and an arc-shaped portion arranged at the front end of the main body; and the method of using the curved hemostatic forceps to remove the brain tissue, so that the intact brain tissue is separated from the skull base comprises: using the front end of the arc-shaped portion to touch the skull base, and using the inner side of the arc-shaped portion to contact the brain tissue, so that the intact brain tissue is pried out of the skull base.

[0012] In some embodiments, the method of using the tissue scissors to cut the occipital bone from the foramen magnum of the rat to the inion along the sagittal suture comprises: using the tissue scissors to cut the occipital bone from the foramen magnum of the rat to a position 0.5-0.8 cm away from the inion along the sagittal suture.

[0013] In some embodiments, the method of using the tissue scissors to cut the occipital bone from the foramen magnum of the rat to the inion along the sagittal suture further comprises: using the tissue scissors to cut the skin of the skull along the sagittal suture, and cleaning the fascia on the skull.

[0014] In some embodiments, the method of using the tissue scissors to cut the skin of the skull along the sagittal suture, and cleaning the fascia on the skull further comprises: sacrificing the rat after the rat is anesthetized, and cutting off the cervical spinal cord of the rat.

[0015] The technical solutions provided by the present application have the following beneficial effects:

[0016] The embodiment of the present application provides a method for taking brain tissue of a laboratory rat, wherein a curved hemostatic forceps is used to pierce the frontal bone of the rat at a selected position until the base of the skull, no brain tissue is under the frontal bone at the selected position, the frontal bone of the rat is connected with the parietal bone, and there is a suture between the left and right parietal bones, that is, a sagittal suture, so that after the frontal bone of the rat is pierced, the left and right parietal bones are separated into two parts along the sagittal suture by the curved hemostatic forceps, at this moment, the parietal bone above the brain tissue of the rat does not produce broken bone pieces, so that the sampled brain tissue does not contain other tissues, and therefore, the integrity of the brain tissue of the rat is ensured. BRIEF DESCRIPTION OF DRAWINGS

[0017] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed in the following embodiment description will be briefly introduced, and obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without any creative effort on the basis of these drawings.

[0018] Figure 1 A flow chart of the method for taking brain tissue of a laboratory rat is provided in the embodiment of the present application.

[0019] Figure 2 A schematic diagram of the skull dissection and brain tissue positioning structure of a rat is provided in the embodiment of the present application.

[0020] Figure 3 A top view of the skull structure of a rat is provided in the embodiment of the present application.

[0021] Figure 4 A side view of the skull structure of a rat is provided in the embodiment of the present application.

[0022] Figure 5 A top view of the position relationship between the brain tissue and the skull of a rat is provided in the embodiment of the present application.

[0023] Figure 6 A side view of the position relationship between the brain tissue and the skull of a rat is provided in the embodiment of the present application.

[0024] Figure 7 A schematic diagram of the steps for taking brain tissue of a rat is provided in the embodiment of the present application.

[0025] Figure 8 A schematic diagram of the three-dimensional structure of a curved hemostatic forceps is provided in the embodiment of the present application.

[0026] 21, sagittal suture; 22, inion; 23, frontal bone; 24, parietal bone; 25, anterior fontanel; 26, occipital bone;

[0027] 3, brain tissue;

[0028] 5, curved hemostatic forceps; 51, sharp head; 52, arcuate portion. DETAILED DESCRIPTION

[0029] In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the following will clearly and completely describe the technical solutions in the embodiments of the present application with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by a person of ordinary skill in the art without creative work belong to the protection scope of the present application.

[0030] The embodiments of the present application provide a method for taking brain tissue of a test rat, which can solve the problems in the prior art that a drill is used to make a skull window in a head of a rat to take out brain tissue of the rat, or a bone forceps is directly used to cut a skull to then take out brain tissue, and the sampled brain tissue may contain other tissues, which affects the integrity of the brain tissue.

[0031] Referring to Figures 1 to 7 As shown in the figure, the method for taking brain tissue of a test rat provided by the embodiments of the present application comprises the following steps:

[0032] S1: using tissue scissors to cut occipital bone 26 from a foramen magnum of a rat along a sagittal suture 21 direction to a vertex point 22, so as to facilitate separation of the entire parietal bone 24 of the rat in subsequent operations, and the tissue scissors are used because they are blunt and will not damage the brain tissue 3.

[0033] S2: using a curved hemostatic forceps 5 to vertically pierce a frontal bone 23 of the rat until a skull base at a position of 0.5-0.8 cm in a direction from a bregma 25 to the frontal bone 23 along an extension direction of the sagittal suture 21, the curved hemostatic forceps 5 has a certain arc at a front end compared with a straight hemostatic forceps, which can make an operator more convenient and labor-saving when vertically piercing the frontal bone 23 of the rat fixed on a table until the skull base, and the frontal bone 23 is pierced at the position of 0.5-0.8 cm in the direction from the bregma 25 to the frontal bone 23 along the extension direction of the sagittal suture 21 because there is no brain tissue 3 below the position, and piercing from this position will not produce bone fragments to affect the integrity of the brain tissue 3, if the position selected is less than 0.5 cm in the direction from the bregma 25 to the frontal bone 23, brain tissue 3 may exist below the frontal bone 23, and piercing the frontal bone 23 may produce bone fragments to affect the integrity of the brain tissue 3, and if the position selected is greater than 0.8 cm in the direction from the bregma 25 to the frontal bone 23, the curved hemostatic forceps 5 may pierce the frontal bone 23 until the skull base, and because the piercing position is too far from the sagittal suture 21, the effect of separating the left and right parietal bones 24 cannot be achieved.

[0034] S3: using the curved hemostatic forceps 5 to pry open the parietal bone 24, and take out the complete brain tissue 3, the head of the rat is generally small, and the frontal bone 23 is pierced until the skull base, at this time the body of the curved hemostatic forceps 5 enters the inside of the skull of the rat, because the body has a certain thickness, the parietal bone 24 of the rat is directly stretched and divided into left and right parts along the sagittal suture 21, and the brain tissue 3 of the rat is completely exposed, and the operation of taking out the complete brain tissue 3 can be performed.

[0035] The embodiment of the present application provides a method for taking out brain tissue of a test rat. Since the sagittal suture 21 is a gap between the left and right parietal bones 24, the parietal bones 24 located at the connection of the left and right sides of the skull are connected. In the process of obtaining the brain tissue 3, the occipital bone is cut from the foramen magnum to the vertex point 22 using tissue scissors, and the vertex point 22 is located at the junction of the occipital bone 26 and the sagittal suture 21. At this time, the occipital bone is divided into left and right parts along the extension direction of the sagittal suture. Then, the curved hemostatic forceps 5 is used to vertically pierce the frontal bone 23 until the skull base. There is no brain tissue 3 below the positioned frontal bone 23, and the frontal bone 23 of the rat is connected with the parietal bone 24. At this time, the parietal bone 24 of the rat head is subjected to a force perpendicular to the extension direction of the sagittal suture 21 and vertically downward. Then, the parietal bone 24 of the rat is separated along the sagittal suture 21 under the action of the external force, and the brain tissue 3 located at the skull base is exposed. The complete brain tissue 3 can be taken out. In the process of obtaining the brain tissue 3, the parietal bone 24 located above the brain tissue 3 is directly split along the sagittal suture 21, and basically no skull fragments are generated. Therefore, there is no other tissue falling onto the brain tissue 3 to contaminate the brain tissue 3.

[0036] Referring to Figure 5 and Figure 6 As shown in the figure, the curved hemostatic forceps 5 is used to vertically pierce the frontal bone 23 until the skull base in the direction of 0.5-0.8 cm of the extension direction of the sagittal suture 21 of the rat from the anterior fontanel 25 to the frontal bone 23. In addition, the curved hemostatic forceps 5 can also include a locking structure. The structure of the curved hemostatic forceps 5 includes a beak, a joint and a handle. Locking the curved hemostatic forceps 5 means locking the locking structure on the handle. After the curved hemostatic forceps 5 is locked, it is then pierced into the frontal bone 23. Compared with the thickness of only one side of the curved hemostatic forceps 5, the thickness is increased, which can better separate the left and right parietal bones 24 along the sagittal suture 21. After the curved hemostatic forceps 5 is locked, the operator can hold the curved hemostatic forceps 5 more easily. At this time, the operator fixes the head of the rat with one hand and holds the curved hemostatic forceps 5 with the other hand. The front end of the curved hemostatic forceps 5 has a curvature, which can adjust the angle of holding the curved hemostatic forceps 5, and can save more labor when piercing.

[0037] Referring to Figure 8 As shown in the figure, the beak of the curved hemostatic forceps 5 is provided with a sharp head 51. The beak of the curved hemostatic forceps 5 is provided with the sharp head 51, which can realize more accurate positioning. The operator can save more labor when piercing the frontal bone 23.

[0038] Referring to Figure 7 As shown, the use of the curved hemostatic forceps 5 to pry open the parietal bone 24, take out the complete brain tissue 3, including: unlocking the curved hemostatic forceps 5, that is, unlocking the buckle structure in the curved hemostatic forceps 5 at the handle position, in some alternative embodiments, after the curved hemostatic forceps 5 is inserted into the skull of the rat, the parietal bone 24 of the rat is not directly separated, at this time, the curved hemostatic forceps 5 is prised open in a direction perpendicular to the sagittal suture 21, the tail of the curved hemostatic forceps 5 is provided with a finger ring, which can be pulled apart according to the principle of lever, so that the parietal bone 24 above the brain tissue 3 of the rat is evenly separated into two parts along the sagittal suture 21; the connection between the brain tissue 3 and the nerves of the skull base is clamped off, and the brain tissue 3 is completely taken out, after the parietal bone 24 is separated into two parts, the brain tissue 3 is exposed, at this time, the brain tissue 3 is located on one side of the skull base, a relatively sharp tool at the front end can be used to clamp off the nerve connection between the brain tissue 3 and the skull base, after the brain tissue 3 and the skull base are completely disconnected, the brain tissue 3 can be directly taken out without damaging the integrity of the brain tissue 3.

[0039] In some alternative embodiments, the clamping off of the connection between the brain tissue 3 and the nerves of the skull base and the complete taking out of the brain tissue 3 can include using the sharp head 51 detachably connected to the curved hemostatic forceps 5 to clamp off the connection between the brain tissue 3 and the nerves of the skull base, that is, installing the sharp head 51 at the front end of the curved hemostatic forceps 5 and using the sharp head 51 to clamp off the nerve connection between the brain tissue 3 and the skull base; the sharp head 51 is detached from the curved hemostatic forceps 5, the brain tissue 3 is taken out using the curved hemostatic forceps 5, and the complete brain tissue 3 is separated from the skull base, detaching the sharp head 51 from the curved hemostatic forceps 5 can protect the brain tissue 3 from being injured by the sharp head 51 during the process of being taken out and separated from the skull base, so as to further ensure the integrity of the brain tissue 3 itself, and using the curved hemostatic forceps 5 multiple times can enable the operator to prepare less equipment during the operation of obtaining the brain tissue 3 to complete the experiment.

[0040] In some alternative embodiments, the curved hemostatic forceps 5 can include a main body and an arc-shaped portion 52 provided at the front end of the main body, and the use of the curved hemostatic forceps 5 to take out the brain tissue 3 and separate the complete brain tissue 3 from the skull base can include touching the skull base with the front end of the arc-shaped portion 52 at this time, the front end of the curved hemostatic forceps 5 and the sharp head 51 have been detached, the nerve connection between the brain tissue 3 and the skull base has been completely clamped off, and the inside of the arc-shaped portion 52 is used to contact the brain tissue 3, when the arc-shaped portion 52 contacts the brain tissue 3, the inside of the arc-shaped portion 52 contacts the brain tissue 3, the brain tissue 3 is relatively soft and can be in contact with more parts of the inside of the arc-shaped portion 52, so that the operation is more convenient and the integrity of the brain tissue 3 itself can be better ensured, and then the complete brain tissue 3 is pried out of the skull base.

[0041] Preferably, after the curved hemostat 5 is fitted with a spiked head 51 to sever the nerve connection between the brain tissue 3 and the skull base, since part of the nerve connection between the brain tissue 3 and the skull base is located at the contact point between the brain tissue 3 and the skull base, the brain tissue 3 presses down on this part of the nerve connection, making it inconvenient to sever it directly. At this time, the curved part 52 of the curved hemostat 5 is used to contact the brain tissue 3. Through the contact between the curved part 52 and the brain tissue 3, the position of the brain tissue 3 is moved, exposing the nerves at the connection point between the brain tissue 3 and the skull base. Then, the spiked head 51 is used to sever these nerve connections, and then the intact brain tissue 3 is pulled out of the skull base.

[0042] See Figure 2 and Figure 3 As shown, the step of using tissue scissors to cut the occipital bone 26 from the foramen magnum of the rat along the sagittal suture 21 to the V-shaped point 22 can include cutting the occipital bone 26 from the foramen magnum of the rat along the sagittal suture 21 to a distance of 0.5 to 0.8 cm from the V-shaped point 22, or cutting the occipital bone 26 from the foramen magnum exactly to the V-shaped point 22, but not exceeding the V-shaped point 22. Exceeding the V-shaped point 22 will cut the parietal bone 24. The brain tissue 3 is located below the parietal bone 24. When cutting the bone with tissue scissors, bone fragments will be generated. After falling, the bone fragments may fall onto the surface of the brain tissue 3, affecting the integrity of the brain tissue 3.

[0043] Preferably, the step of using tissue scissors to cut the occipital bone 26 from the foramen magnum of the rat along the sagittal suture 21 to the front of the zygomatic point 22 may further include: using tissue scissors to cut open the skin of the cranial vault along the sagittal suture 21. In this embodiment, tissue scissors can be used to cut only the skin of the cranial vault along the sagittal suture 21. In some other embodiments, the skin on the occipital bone 26, the cranial vault, and the frontal bone 23 may also be cut off, or even all the skin covering the skull may be removed. The fascia on the cranial vault is cleaned. This can be done by cleaning the fascia at all the areas where the skin was cut off, or only cleaning the fascia on the cranial vault. Cleaning the fascia on the bone surface ensures that when the parietal bone 24 is subsequently opened, there is no resistance from the fascia preventing the parietal bone 24 from being opened. After the fascia is cleaned, alcohol or other suitable disinfectants can be used to clean the areas that need to be treated, in order to achieve asepticity in the brain retrieval process as possible.

[0044] Preferably, before cutting the skin of the top of the skull with tissue scissors along the direction of the sagittal suture 21 and cleaning the fascia on the top of the skull, the procedure may further include anesthetizing and euthanizing the rat, and severing the cervical spinal cord of the rat. That is, after anesthetizing and euthanizing the rat using an appropriate method, and after confirming that the rat has completely lost its responsiveness, clamping the rat's neck with forceps and quickly severing the cervical spinal cord of the rat, so as to minimize the animal's pain as much as possible.

[0045] In the description of the present application, it should be noted that the terms "upper", "lower", and the like are used for indicating 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 do not indicate or imply 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 on the present application. Unless otherwise explicitly specified and limited, the terms "mounting", "connection", "connecting" should be understood broadly, for example, can be fixed connection, can also be detachable connection, or integral connection; can be mechanical connection, can also be electrical connection; can be directly connected, can also be indirectly connected through an intermediate medium, can be internal communication of two elements. For those skilled in the art, the specific meaning of the above terms in the present application can be understood according to the specific circumstances.

[0046] It should be noted that in the present application, relational terms such as "first" and "second" and the like are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply that there is any such actual relationship or order between these entities or operations. Moreover, the terms "comprise", "include" or any other variant thereof are intended to cover non-exclusive inclusion, so that a process, method, article or apparatus including a series of elements includes not only those elements, but also other elements not explicitly listed, or other elements inherent in such a process, method, article or apparatus. Without more limitations, the element defined by the statement "comprising a" does not exclude the presence of other identical elements in the process, method, article or apparatus including the element.

[0047] The above is only a specific embodiment of the present application, which enables those skilled in the art to understand or implement the present application. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present application. Therefore, the present application will not be limited to these embodiments shown herein, but will conform to the widest scope consistent with the principles and novel features applied herein.

Claims

1. A method for taking out a brain of an experimental rat, characterized by, It comprises the following steps: After the rat is anesthetized, it is executed, and the cervical spinal cord of the rat is cut off; The skull skin is cut open along the sagittal suture (21) direction using tissue scissors, and the fascia on the skull is cleaned; The occipital bone (26) is cut to the inion (22) from the foramen magnum of the rat along the sagittal suture (21) direction using tissue scissors; The frontal bone (23) is vertically punctured to the skull base at 0.5-0.8 cm from the anterior fontanel (25) to the frontal bone (23) direction of the rat along the extension direction of the sagittal suture (21) using the curved hemostatic forceps (5). The parietal bone (24) is pried open using the curved hemostatic forceps (5), and the complete brain tissue (3) is taken out.

2. The experimental rat brain removal method according to claim 1, wherein The frontal bone (23) is vertically punctured to the skull base at 0.5-0.8 cm from the anterior fontanel (25) to the frontal bone (23) direction of the rat along the extension direction of the sagittal suture (21) using the curved hemostatic forceps (5), and further comprising: Lock the curved hemostatic forceps (5).

3. The experimental rat brain taking method of claim 2, characterized in that: The jaw of the curved hemostatic forceps (5) is provided with a sharp head (51).

4. The experimental rat brain removal method according to claim 1, wherein The parietal bone (24) is pried open using the curved hemostatic forceps (5), and the complete brain tissue (3) is taken out, comprising: Unlock the curved hemostatic forceps (5), pry open the curved hemostatic forceps (5) along the direction perpendicular to the sagittal suture (21), and evenly divide the parietal bone (24) above the brain tissue (3) of the rat into two parts along the sagittal suture (21); Clamp off the connection between the brain tissue (3) and the nerves of the skull base, and completely take out the brain tissue (3).

5. The experimental rat brain removal method according to claim 4, wherein Clamp off the connection between the brain tissue (3) and the nerves of the skull base, and completely take out the brain tissue (3), comprising: Use the sharp head (51) detachably connected to the curved hemostatic forceps (5) to clamp off the connection between the brain tissue (3) and the nerves of the skull base; Remove the sharp head (51) from the curved hemostatic forceps (5), use the curved hemostatic forceps (5) to take out the brain tissue (3), and make the complete brain tissue (3) separate from the skull base.

6. The experimental rat brain removal method according to claim 5, wherein The curved hemostatic forceps (5) comprise a main body and an arc-shaped portion (52) arranged at the front end of the main body; Use the curved hemostatic forceps (5) to take out the brain tissue (3), and make the complete brain tissue (3) separate from the skull base, comprising: Touch the skull base with the front end of the arc-shaped portion (52), and use the inner side of the arc-shaped portion (52) to contact the brain tissue (3), and scrape out the complete brain tissue (3) from the skull base.

7. The experimental rat brain removal method of claim 1, wherein, Use tissue scissors to cut the occipital bone (26) to the inion (22) from the foramen magnum of the rat along the sagittal suture (21) direction, comprising: Use tissue scissors to cut the occipital bone (26) to the inion (22) from the foramen magnum of the rat along the sagittal suture (21) direction, comprising: Cut the occipital bone (26) to a distance of 0.5-0.8 cm from the inion (22) along the sagittal suture (21) direction using tissue scissors.

Images