Novel method for manufacturing fish otolith specimen

Abstract

The invention provides a novel method for preparing a fish otolith specimen, and belongs to the field of biological specimen preparation. Comprising the steps of specimen fixation, head skin dissection and segmentation, head muscle dissection and segmentation, head skeleton exposure and dissection, muscle dissection, dehydration, degreasing, polymer infiltration, fixation, solidification and the like. More importantly, the spatial position relation and the connection mode of the otolith, the skull, the brain, the nerve and the muscle system are displayed in situ. The otolith can be processed in a non-destructive mode, the otolith can be taken out or put back, the flexible display effect that the same specimen can display the details of the otolith and can also display overall dissection association is achieved, and the teaching and scientific research value of the specimen is greatly improved.

Description

Technical Field

[0001] This invention belongs to the field of biological specimen preparation, and in particular relates to a novel method for preparing fish otolith specimens. Background Technology

[0002] Existing methods for preparing otoliths in fish cannot adequately showcase their location and physiological structure within the specimen, nor can they establish a physiological connection with the specimen itself. This approach completely destroys the specimen's original structure and fails to reflect its intrinsic structure. Further research is needed to develop a method that can simultaneously display the otolith specimen and demonstrate the relationship between the otoliths and the fish's body. Summary of the Invention

[0003] In view of this, the purpose of this invention is to provide a novel method for preparing fish otolith specimens.

[0004] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a novel method for preparing fish otolith specimens, comprising the following steps: 1) After immersing the fish specimen in formalin solution for 20-60 days, retain the skin at the edges of each fin, the skin on the back and abdomen, and peel off the remaining skin to obtain a specimen with the skin removed; 2) Dissect the head muscles, skull, and muscle tissue on one side of the specimen after the skin has been removed to obtain the dissected specimen; 3) The dissected specimens were dehydrated, degreased, impregnated with polymers, fixed and solidified to obtain novel fish otolith specimens; The polymer includes silica gel; The fixation includes embedding stainless steel bars within the specimen; The curing process involves using a curing agent to cure the material in an environment with a temperature of 40~50℃. Step 2) After the specimen's head muscles and skull are dissected and the skin is peeled off, the otoliths can be non-destructively removed from their anatomical location and / or returned.

[0005] Preferably, the fish specimen includes large yellow croaker.

[0006] Preferably, step 3) involves dehydration and degreasing, which includes soaking the dissected specimen in acetone of varying concentrations. The volume concentration of acetone is 80%~100%; The temperature for dehydrating acetone is -30 to -20°C, and the dehydration time is 30 to 90 days. The acetone temperature during the defatting process is 16~26℃, and the defatting time is 30~90 days.

[0007] Preferably, the curing agent is in a gaseous state.

[0008] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a novel method for preparing fish otolith specimens, including specimen fixation, dissection and segmentation of the head skin, dissection and segmentation of the head muscles, exposure and dissection of the head bones, muscle dissection, dehydration, degreasing, polymer infiltration, fixation, and curing. This invention, for the first time, integrates the display of otoliths into the overall anatomical structure of the fish. Through layered dissection, it not only displays the otoliths themselves, but more importantly, it demonstrates in situ the spatial relationships and connections between the otoliths and the skull, brain, nervous system, and muscular system, enabling viewers to systematically understand how otoliths participate in physiological processes such as hearing and balance. The otolith processing in this invention is non-destructive; the otoliths can be removed or reinserted, achieving a flexible display effect where the same specimen can showcase both otolith details and overall anatomical connections, greatly enhancing the teaching and research value of the specimen. This invention also employs bioplasticization technology to process the dissected specimen, enabling it to be dried, odorless, and tactilely preserved for a long time without losing its original shape, color, and texture details, resulting in a visually intuitive and durable display effect. In addition, by preserving part of the skin and dissecting the muscles and bones layer by layer, the specimen presents a clear anatomical layer from the epidermis to the deep bones and from the whole to the parts, combining artistry and scientific value. Attached Figure Description

[0009] Figure 1 Image of a specimen of large yellow croaker; Figure 2 Image of a fish specimen showing subcutaneous structures after removing the head skin; Figure 3 An image of a fish specimen showing its muscles after the skin has been removed. Figure 4 An image of a fish specimen showing the skull structure in order to dissect the head muscles; Figure 5 To dissect the skull, locate the otoliths, and display the surrounding structures, an image of the fish specimen was obtained; Figure 6 An image showing the effect of otoliths; Figure 7 An image of a fish specimen showing deep structures such as vertebrae and blood vessels through dissection of one side of the body's muscles. Detailed Implementation

[0010] This invention provides a novel method for preparing fish otolith specimens, comprising the following steps: 1) After immersing the fish specimen in formalin solution for 20-60 days, retain the skin at the edges of each fin, the skin on the back and abdomen, and peel off the remaining skin to obtain a specimen with the skin removed; 2) Dissect the head muscles, skull, and muscle tissue on one side of the specimen after the skin has been removed to obtain the dissected specimen; 3) The dissected specimens were dehydrated, degreased, impregnated with polymers, fixed and solidified to obtain novel fish otolith specimens; In this invention, the fish specimen preferably includes a bony fish, more preferably a large yellow croaker, and the fish specimen is a whole fish, including gills and internal organs. The method for processing the fish specimen is described in the specimen processing method of plastination technology.

[0011] In this invention, after the head muscles and skull of the specimen are dissected and peeled off, the otoliths can be nondestructively removed from their anatomical location and / or returned.

[0012] In this invention, the dissected specimen is dehydrated, degreased, impregnated with polymers, fixed, and solidified to obtain a novel fish otolith specimen. The dehydration and degreasing include soaking the dissected specimen in acetone of varying concentrations. The acetone volume concentration is preferably 80%~100%, more preferably 85%~100%, and even more preferably 90%~99.9%. The gradient is preferably increased by 1%~2% each time, more preferably 1.2%~1.8%, and even more preferably 1.5%. The acetone temperature during dehydration is preferably -30~-20℃, more preferably -28~-22℃, and even more preferably -25℃. The dehydration time is preferably 10~50 days, more preferably 20~40 days, and even more preferably 30 days. The acetone temperature during degreasing is preferably 16~26℃, more preferably 20~24℃, and even more preferably 22℃. The degreasing time is preferably 10~50 days, more preferably 20~40 days, and even more preferably 30 days.

[0013] In this invention, the polymer preferably comprises silica gel.

[0014] In this invention, the fixation includes embedding stainless steel ribs inside the specimen and adjusting the specimen's shape to conform to physiological morphology while also taking aesthetics into account.

[0015] In this invention, the curing is preferably performed using a curing agent, which is preferably in a gaseous state. The curing temperature is 40-50°C, preferably 42-48°C, more preferably 45°C, and the curing time is preferably 20 days. The curing agent is preferably Hoffen S6. The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0016] Example 1

[0017] I. Selection of Subject: In this example, the large yellow croaker is used. The large yellow croaker belongs to the Sciaenidae family, and its otolith structure is representative among fish. Specifically, a mature large yellow croaker is selected, such as... Figure 1 As shown.

[0018] 2. Specimen fixation: Immerse the specimen in a 10% formalin solution for fixation. The fixation time depends on the size and type of the specimen, but is 30 days, in order to allow for later dissection and display.

[0019] III. Specimen Skin Dissection and Separation: The skin along the edges of each fin, as well as parts of the dorsal and ventral skin, is preserved. The remaining skin is completely removed to expose the specimen's muscle structure. First, the head skin is removed to reveal the subcutaneous structures, such as... Figure 2 As shown. Then remove the body skin to reveal the muscles, as... Figure 3 As shown.

[0020] IV. Anatomical dissection of the head muscles of the specimen, revealing the composition of the skull, such as... Figure 4 As shown, locate the skull.

[0021] 5. Dissect the skull to locate the otoliths and display the surrounding structures, such as... Figure 5 As shown. Figure 6 The otolith specimens are removed without any destructive opening. They can be displayed individually or returned to the whole specimen for display.

[0022] VI. Anatomical dissection of one side of the body muscles reveals deep structures such as vertebrae and blood vessels. See details. Figure 7 .

[0023] VII. The dissected specimens undergo dehydration, degreasing, and polymer impregnation treatment. The steps are as follows: ① Place the specimen in acetone at a low temperature (-25℃) to remove its own water for one month. ② After dehydration, place it in acetone at 22℃ to remove fat for one month, gradually increasing the acetone concentration from 90% to 99.9%, increasing the concentration by 1.5% every 2-3 days until reaching 99.9%. ③ After dehydration and degreasing, immerse the specimen in silica gel, using a displacement method to replace the acetone remaining in the specimen (see "Bioplasticization Technology" for details). This allows for long-term preservation while maximizing the display of biological specimen details.

[0024] 8. Fixing and Restoring the Specimen. The steps are as follows: ① Embed suitable stainless steel reinforcing bars inside the specimen and adjust its shape to conform to physiological morphology while also considering aesthetics. ② Use fine steel needles or pins to fix the fins and other free tissues, keeping them in their normal positions. ③ Use the specimen's own tissue to repair any damage caused by previous procedures, ensuring it achieves the correct physiological morphology. (For detailed steps, please refer to "Bioplasticization Technology")

[0025] 9. The specimen is then cured and cleaned. The steps are as follows: ① Place the restored specimen in a sealed room and add a curing agent (Hoffen S6). Use an air pump or similar tool to condense the curing agent into a gaseous state and heat it appropriately to 45°C. Through the combined action of the agent and temperature, the specimen curing process is completed, achieving the goal of long-term preservation. ② After curing, remove the steel needles and other items used to fix the specimen, and use a scalpel or similar tool to carefully clean the surface of the specimen, making it more presentable. The specimen is now complete. (For detailed steps, please refer to "Bioplasticization Technology").

[0026] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A novel method for preparing fish otolith specimens, characterized in that, Includes the following steps: 1) After immersing the fish specimen in formalin solution for 20-60 days, retain the skin at the edges of each fin, the skin on the back and abdomen, and peel off the remaining skin to obtain a specimen with the skin removed; 2) Dissect the head muscles, skull, and muscle tissue on one side of the specimen after the skin has been removed to obtain the dissected specimen; 3) The dissected specimens were dehydrated, degreased, impregnated with polymers, fixed and solidified to obtain novel fish otolith specimens; The polymer includes silica gel; The fixation includes embedding stainless steel bars within the specimen; The curing process involves using a curing agent to cure the material in an environment with a temperature of 40~50℃. Step 2) After the specimen's head muscles and skull are dissected and the skin is peeled off, the otoliths can be non-destructively removed from their anatomical location and / or returned.

2. The method according to claim 1, characterized in that, The fish specimens include large yellow croaker.

3. The method according to claim 1, characterized in that, Step 3) The dehydration and degreasing process includes soaking the dissected specimen in acetone of varying concentrations. The volume concentration of acetone is 80%~100%; The temperature for dehydrating acetone is -30 to -20°C, and the dehydration time is 30 to 90 days. The acetone temperature during defatting is 16~26℃, and the defatting time is 30~90 days.

4. The method according to claim 1, characterized in that, The curing agent is in a gaseous state.

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