Device for promoting protoplast fusion
By designing a cell fusion device containing a protective shell and a multi-layer filter, the problems of complex operations and lack of protection in the prior art are solved, and efficient and safe protoplast cell fusion and filtration are achieved, simplifying the operation process and improving the cells Fusion frequency.
Patent Information
- Application Number
- CN202420720427.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-04-09
- Publication Date
- 2024-12-13
- Estimated Expiration
- 2034-04-09
AI Technical Summary
Existing cell fusion devices are complex in operation and lack external protection, which makes protoplast cells susceptible to contamination and collision damage.
A device including a culture tank, a protective shell, an electrode system, an injection channel, a filter device and a fusion chamber were designed to protect cells through a multi-layer filter net and a closed space, and the cell fusion was performed by electrofusion method, and through the electrode system and filter devices to achieve efficient impurity filtration and fusion of cells.
The operating process is simplified, the frequency and safety of cell fusion is improved, the risks of contamination and collision are reduced, and the screening efficiency is accelerated through multi-layer filtration, and subsequent cell culture is facilitated.
Smart Images
Figure CN222160189U_ABST
Abstract
Description
Technical Field
[0001] The utility model relates to the technical field of protoplasts, in particular to a device for promoting protoplast fusion. Background Art
[0002] Protoplasts have cell totipotency. Given appropriate culture conditions, they can regenerate cell walls, divide and proliferate until they develop into complete plants. The fusion of protoplast cells can combine specific genes or expression products in cells of different types or sources to generate new cells or organisms, providing new technologies for the field of cell engineering. The main methods currently used for protoplast cell fusion are Sendai virus induction, PEG induction, and electrofusion induction. Electrofusion is an efficient, repeatable, and non-toxic technology. The operation requires the uniform addition of protoplast solution at a constant temperature. During the electrofusion process, cells are usually placed in a transparent fusion chamber or microslide to clearly observe the cell fusion process under a microscope. Protoplast electrofusion requires the use of a cell fusion instrument, but the cell fusion instruments currently on the market have operational complexity problems and are not very closed. In addition, most cell fusion instruments are placed directly on the workbench for use without protecting the outside, and protoplast cells are susceptible to collision damage and contamination.
[0003] Therefore, how to propose a device for promoting protoplast fusion that can reduce the probability of contamination and collision during cell fusion has become an urgent problem to be solved by those skilled in the art. Utility Model Content
[0004] The utility model aims to provide a device for promoting protoplast fusion, so as to solve the problems that the existing device lacks external protection and has complicated operation.
[0005] In order to solve the above technical problems, the utility model adopts the following technical solutions:
[0006] The utility model discloses a device for promoting protoplast fusion, comprising a culture tank, a culture dish, a protective shell, an electrode system, an injection channel, a filter device and a fusion chamber; the protective shell is provided with the electrode system, the internal top end of the protective shell is provided with the injection channel, the bottom end of the injection channel is communicated with the filter device, the middle part of the filter device is communicated with the fusion chamber, one side of the bottom end of the protective shell is open, the culture tank is movably arranged in the opening, the culture tank is provided with a culture dish, the culture dish is filled with culture fluid, the bottom end of the filter device is located above the culture tank, the bottom end of the fusion chamber is located inside the culture dish, and the electrode system is electrically connected to a power supply device.
[0007] Preferably, the electrode system includes a switch, a first electrode and a second electrode; the first electrode and the second electrode are arranged in parallel on the inner wall of the protective shell, the switch is set on the outer wall of the protective shell, the first electrode and the second electrode are electrically connected to an external power supply device after passing through the switch, and the power supply device supplies sinusoidal alternating current.
[0008] Preferably, the injection channel is V-shaped, a first injection hole and a second injection hole are provided on the top plate of the protective shell, and the bottom end of the injection channel is connected to the top end of the filtration channel in the filtration device.
[0009] Preferably, the filtering device includes a large-pore filter, a small-pore filter, a collection box and a filter channel; the filter channel is an L-shaped structure, the large-pore filter is arranged on the upper part of the filter channel, the small-pore filter is arranged in the middle part of the filter channel, the bottom end opening of the filter channel is arranged on the side wall of the protective shell, and the collection box is slidably connected to the bottom end of the filter channel.
[0010] Preferably, the fusion chamber includes a fusion channel, a transfer tube and a hose; the fusion channel is arranged obliquely, the upper end of the fusion channel is connected to the filtration channel, the upper end opening of the fusion channel is located at the small-pore filter, the lower end of the fusion channel is rotatably connected to the transfer tube, the other end of the transfer tube is fixedly connected to the top end of the hose, and the bottom end of the hose is located inside the culture dish.
[0011] Preferably, a handle is provided on the outer side of the culture tank.
[0012] Preferably, a handle is provided on the outer side of the collection box.
[0013] Compared with the prior art, the beneficial technical effects of the utility model are:
[0014] The utility model discloses a device for promoting protoplast fusion, comprising a culture tank, a protective shell, an electrode system, an injection channel, a filtering device and a fusion chamber; a culture dish is arranged in the culture tank, the electrode system comprises a first electrode, a second electrode and a switch; the injection channel is provided with a first injection hole and a second injection hole; the filtering device comprises a filtering channel, a large-pore filter screen, a small-pore filter screen and a collection box; the fusion chamber comprises a fusion channel, a transfer tube and a hose;
[0015] 1) Simple structure, easy operation, high fusion frequency using electrofusion method and no chemical toxicity;
[0016] 2) A protective shell and a culture tank are provided to form a closed space, which is beneficial to protect the internal structure from damage and prevent the protoplasts from being contaminated by the outside world during the fusion process. The culture tank can be withdrawn at any time to replace the internal culture dish;
[0017] 3) A filtering device is provided to filter the extracted protoplasts for impurities through a multi-layer filter screen, which can speed up the screening efficiency and reduce the operation steps;
[0018] 4) A fusion channel is provided in the fusion chamber, and the protoplasts are electrically fused in the fusion channel. The fused protoplasts are then placed in a culture dish for culture.
[0019] The utility model discloses a device for promoting protoplast fusion, which has a simple structure and is easy to operate. It adopts an electric fusion method with high fusion frequency and no chemical toxicity. A protective shell and a culture tank are provided to form a closed space, so as to protect the internal structure from being damaged and prevent the protoplasts from being polluted by the outside world. A filtering device is provided to filter the protoplasts through a multi-layer filtering net to screen the protoplast cells of suitable size, thereby accelerating the screening efficiency and reducing the operation steps. The protoplasts are electrically fused in the fusion channel, and the culture dish at the bottom can directly collect the fused protoplasts for subsequent culture. BRIEF DESCRIPTION OF THE DRAWINGS
[0020] The utility model will be further described below in conjunction with the accompanying drawings.
[0021] Figure 1 This is a schematic diagram of the device for promoting protoplast fusion of the utility model (perspective);
[0022] Figure 2 This is a top view of the device for promoting protoplast fusion of the utility model;
[0023] Figure 3 This is a schematic diagram of the connection between the filtration device and the fusion chamber of the utility model;
[0024] Figure 4 It is a schematic diagram of the culture tank of the utility model.
[0025] Description of reference numerals: 1. culture tank; 2. culture dish; 3. protective housing; 4. electrode system; 5. injection channel; 6. filtering device; 7. fusion chamber;
[0026] 4-1, switch; 4-2, including a first electrode; 4-3, a second electrode;
[0027] 5-1, first injection hole; 5-2, second injection hole;
[0028] 6-1, large-pore filter; 6-2, small-pore filter; 6-3, collection box; 6-4, filter channel;
[0029] 7-1, fusion channel; 7-2, transfer tube; 7-3, hose. DETAILED DESCRIPTION
[0030] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.
[0031] like Figure 1-4 As shown, a device for promoting protoplast fusion comprises a culture tank 1, a culture dish 2, a protective shell 3, an electrode system 4, an injection channel 5, a filter device 6 and a fusion chamber 7; the protective shell 3 is provided with the electrode system 4, the inner top of the protective shell 3 is provided with the injection channel 5, the bottom end of the injection channel 5 is connected with the filter device 6, the middle part of the filter device 6 is connected with the fusion chamber 7, one side of the bottom end of the protective shell 3 is open, the culture tank 1 is movably arranged in the opening, the culture tank 1 is provided with a culture dish 2, the culture dish 2 is filled with culture fluid, the bottom end of the filter device 6 is located above the culture tank 1, the bottom end of the fusion chamber 7 is located inside the culture dish 2, and the electrode system is electrically connected to a power supply device.
[0032] Specifically, the electrode system 4 includes a switch 4-1, a first electrode 4-2 and a second electrode 4-3; the first electrode 4-2 and the second electrode 4-3 are arranged in parallel on the inner wall of the protective shell 3, and the switch 4-1 is set on the outer wall of the protective shell 3. The first electrode 4-2 and the second electrode 4-3 are electrically connected to an external power supply device after passing through the switch 4-1, and the power supply device supplies sinusoidal alternating current; after turning on the switch 4-1, an electric field is formed between the first electrode 4-2 and the second electrode 4-3, and the sinusoidal alternating current wave causes the protoplasts in the fusion channel 7-1 to be polarized through a unique bidirectional electrophoresis effect, and arranges the cells in strings, completing the formation of cell strings before electrical fusion, and the DC pulse voltage induces the cell membrane to produce tiny pores, so that the adjacent and closely attached cell membranes fuse with each other.
[0033] Specifically, the injection channel 5 is V-shaped, and a first injection hole 5-1 and a second injection hole 5-2 are provided on the top plate of the protective shell 3. The bottom end of the injection channel 5 is connected to the top end of the filtration channel 6-4 in the filtration device 6; the protoplasts to be fused are added through different channels, so that the proportion of addition is easily mastered.
[0034] Specifically, Figure 3As shown, the filtering device 6 includes a large-pore filter 6-1, a small-pore filter 6-2, a collection box 6-3 and a filtering channel 6-4; the filtering channel 6-4 is an L-shaped structure, the large-pore filter 6-1 is arranged on the upper part of the filtering channel 6-4, the small-pore filter 6-2 is arranged in the middle part of the filtering channel 6-4, the bottom end opening of the filtering channel 6-4 is arranged on the side wall of the protective shell 3, and the bottom end of the filtering channel 6-4 is slidably connected with the collecting box 6-3; the large-pore filter 6-1 intercepts larger impurities, the small-pore filter 6-2 intercepts protoplasts, and small impurities pass through the small-pore filter 6-2 and fall into the collecting box 6-3, thereby realizing impurity filtration, accelerating screening efficiency and reducing operation steps.
[0035] Specifically, the fusion chamber 7 includes a fusion channel 7-1, a transfer tube 7-2 and a hose 7-3; the fusion channel 7-1 is arranged obliquely, the upper end of the fusion channel 7-1 is connected to the filtration channel 6-4, the upper end opening of the fusion channel 7-1 is located at the small hole filter 6-2, the lower end of the fusion channel 7-1 is rotatably connected to the transfer tube 7-2, the other end of the transfer tube 7-2 is fixedly connected to the top of the hose 7-3, and the bottom end of the hose 7-3 is located inside the culture dish 2; the filtered protoplasts enter the inclined fusion channel 7-1 under the action of gravity, and the fusion of the cell membrane is achieved under the action of the electrode system 3; the fused protoplasts fall into the culture dish through the hose 7-3 for cultivation.
[0036] Specifically, a handle is provided on the outer side of the culture tank 1; after the culture tank 1 is pulled out by the handle, the culture dish 2 can be replaced.
[0037] Specifically, a handle is provided on the outer side of the collection box 6-3; after the collection box 6-3 is pulled out by the handle, the cleaning operation can be carried out.
[0038] The use process of the utility model is as follows:
[0039] First, the device is powered and the switch 4-1 is turned on. At this time, the first electrode 4-2 and the second electrode 4-3 form an electric field, and protoplasts are added to the injection channel 5 through the first injection hole 5-1 and the second injection hole 5-2, and the protoplasts downwardly enter the filtration channel 6-4; the large-pore filter 6-1 intercepts impurities with larger shapes, and the small-pore filter 6-2 intercepts protoplasts, and impurities with small shapes pass through the small-pore filter 6-2 and fall into the collection box 6-3, thereby filtering impurities; under the action of gravity, the protoplasts staying on the small-pore filter 6-2 will flow into the fusion channel 7-1 chamber, and during the flow process, the protoplasts are in a high-voltage direct current pulse electric field of a certain value, causing reversible breakdown of the cell membrane, thereby inducing the protoplasts in contact with each other to fuse; the fused protoplasts flow through the tube wall of the hose 7-3 to the culture dish 2 inside the culture tank 1 for cultivation;
[0040] After the protoplasts are cultured in the culture dish 2 to meet the requirements, the culture tank 1 is pulled out using the handle to remove and replace the culture dish 2. When the side wall of the culture tank 1 contacts the hose 7-3, the transfer tube 7-2 rotates and does not hinder the movement of the culture tank 1. When the culture tank 1 is moved away, the transfer tube 7-2 automatically resets under the action of gravity and does not affect the next use.
[0041] When there are a lot of debris in the collection box 6-3, the collection box 6-3 can be easily pulled out using the handle to clean or replace the collection box 6-3.
[0042] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.
[0043] The embodiments described above are only descriptions of the preferred methods of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should fall within the protection scope determined by the claims of the present invention.
Claims
1. A device for promoting protoplast fusion, characterized in that: The invention comprises a culture tank (1), a culture dish (2), a protective shell (3), an electrode system (4), an injection channel (5), a filter device (6) and a fusion chamber (7); the protective shell (3) is provided with the electrode system (4); the injection channel (5) is provided at the top of the inner part of the protective shell (3); the bottom end of the injection channel (5) is connected to the filter device (6); the middle part of the filter device (6) is connected to the fusion chamber (7); one side of the bottom end of the protective shell (3) is open; the culture tank (1) is movably arranged in the opening; a culture dish (2) is provided inside the culture tank (1); the culture dish (2) is filled with culture fluid; the bottom end of the filter device (6) is located above the culture tank (1); the bottom end of the fusion chamber (7) is located inside the culture dish (2); and the electrode system is electrically connected to a power supply device.
2. The device for promoting protoplast fusion according to claim 1, characterized in that: The electrode system (4) comprises a switch (4-1), a first electrode (4-2) and a second electrode (4-3); the first electrode (4-2) and the second electrode (4-3) are arranged in parallel on the inner wall of the protective shell (3), the switch (4-1) is arranged on the outer wall of the protective shell (3), and the first electrode (4-2) and the second electrode (4-3) are electrically connected to an external power supply device after passing through the switch (4-1), and the power supply device supplies sinusoidal alternating current.
3. The device for promoting protoplast fusion according to claim 1, characterized in that: The injection channel (5) is V-shaped, and a first injection hole (5-1) and a second injection hole (5-2) are provided on the top plate of the protective shell (3). The bottom end of the injection channel (5) is connected to the top end of the filter channel (6-4) in the filter device (6).
4. The device for promoting protoplast fusion according to claim 1, characterized in that: The filtering device (6) comprises a large-pore filter (6-1), a small-pore filter (6-2), a collection box (6-3) and a filtering channel (6-4); the filtering channel (6-4) is an L-shaped structure, the large-pore filter (6-1) is arranged at the upper part of the filtering channel (6-4), the small-pore filter (6-2) is arranged in the middle part of the filtering channel (6-4), the bottom end opening of the filtering channel (6-4) is arranged on the side wall of the protective shell (3), and the bottom end of the filtering channel (6-4) is slidably connected to the collecting box (6-3).
5. The device for promoting protoplast fusion according to claim 4, characterized in that: The fusion chamber (7) comprises a fusion channel (7-1), a transfer tube (7-2) and a hose (7-3); the fusion channel (7-1) is arranged obliquely, the upper end of the fusion channel (7-1) is connected to the filtering channel (6-4), the upper end opening of the fusion channel (7-1) is located at the small hole filter (6-2), the lower end of the fusion channel (7-1) is rotatably connected to the transfer tube (7-2), the other end of the transfer tube (7-2) is fixedly connected to the top end of the hose (7-3), and the bottom end of the hose (7-3) is located inside the culture dish (2).
6. The device for promoting protoplast fusion according to claim 1, characterized in that: A handle is provided on the outer side of the culture tank (1).
7. The device for promoting protoplast fusion according to claim 4, characterized in that: A handle is provided on the outer side of the collecting box (6-3).