Pre-cooling plant tissue grinding and lysate mixing device
By using a liquid nitrogen pre-cooled plant tissue grinding and lysis buffer mixing device, the problem of RNA degradation caused by multiple sample transfers was solved, achieving efficient sample mixing and grinding at low temperatures, and improving the accuracy and safety of experimental results.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- NINGXIA UNIVERSITY
- Filing Date
- 2025-05-16
- Publication Date
- 2026-04-17
AI Technical Summary
Existing technologies require multiple sample transfers, increasing the risk of RNA exposure to room temperature, leading to degradation. Manual operation is not thorough and relies on low-temperature environments, posing a risk of aerosol contamination.
A pre-cooled plant tissue grinding and lysis buffer mixing device using liquid nitrogen pre-cooling is used. Liquid nitrogen is injected into the grinding equipment through a liquid nitrogen injection connection tube for pre-cooling, and the grinding rod is controlled by a drive motor to grind the plant sample. The mixing process is completed in a closed space, avoiding multiple transfers at room temperature.
This method enables rapid and thorough grinding and lysis of plant tissues under low-temperature conditions, reducing the risk of RNA degradation and improving the uniformity of grinding particle size and the accuracy of experimental results.
Smart Images
Figure CN224133058U_ABST
Abstract
Description
Technical Field
[0001] This utility model relates to the field of plant tissue grinding technology, specifically a pre-cooled plant tissue grinding and lysis solution mixing device. Background Technology
[0002] Plant cells have relatively tough structures such as cell walls. Grinding can initially disrupt plant tissue, exposing the cells. The surfactants and other components in the lysis buffer can further damage the cell membrane and cell wall, releasing the cell contents.
[0003] However, in actual operation, the inventors found that the following problems still exist: the existing technology requires multiple transfers of samples (such as transferring to centrifuge tubes after grinding and stratification after lysis), which increases the risk of RNA being exposed to room temperature environment and is prone to degradation. In addition, manual operation not only results in incomplete and uneven grinding of samples, but also requires a low temperature environment, which has strict requirements for experimental conditions and poses a risk of aerosol contamination.
[0004] Based on this, the present invention provides a pre-cooled plant tissue grinding and lysis solution mixing device. Utility Model Content
[0005] To address the shortcomings of existing technologies, this invention provides a pre-cooled plant tissue grinding and lysis solution mixing device, which has the advantages of using liquid nitrogen to pre-cool fresh plant tissue and rapidly grinding it into powder in liquid nitrogen, and then inverting the device to mix the powder with the lysis solution in a sealed space, thus solving the problems mentioned in the background technology.
[0006] This utility model provides the following technical solution: a pre-cooled plant tissue grinding and lysis solution mixing device, comprising a base plate and a connecting lower shell. A motor bracket is fixedly connected to the bottom of the connecting lower shell. A second drive motor is fixedly installed inside the motor bracket. A first grinding rod is fixedly connected to the output end of the second drive motor. A fixed shaft is fixedly connected to the top of the first grinding rod. A second grinding rod is fixedly connected to the top of the fixed shaft. A grinding equipment shell is fixedly installed inside the connecting lower shell. A receiving plate is fixedly connected to the surface of the grinding equipment shell. A filter screen is attached to the top of the receiving plate. A fixed funnel is fixedly connected to the top of the filter screen. A fixed centrifuge tube is fixedly connected to the top of the fixed funnel. An upper shell is fixedly connected to the top of the fixed centrifuge tube.
[0007] Preferably, a liquid nitrogen injection connection pipe is fixedly connected to the surface of the grinding equipment shell, and a fastening cap is threadedly connected to the surface of the liquid nitrogen injection connection pipe.
[0008] Preferably, a drive motor is fixedly installed at the center of the upper surface of the base plate, and the output shaft of the drive motor is fixedly connected to the connecting lower shell.
[0009] Preferably, the top of the base plate is fixedly connected to four sets of support legs, the top of the four sets of support legs is fixedly connected to a support bracket, and the lower shell is fitted inside the support bracket.
[0010] Preferably, the bottom end of the grinding equipment housing is attached to the output end of the second drive motor.
[0011] Preferably, the lower shell is threadedly connected to the upper shell.
[0012] Preferably, the surface of the connecting lower shell is provided with a mounting hole, and a liquid nitrogen injection connecting pipe is fixedly connected inside the mounting hole.
[0013] Preferably, a support plate is fixedly installed on the inner wall of the connecting lower shell.
[0014] Compared with the prior art, the present invention has the following beneficial effects:
[0015] This pre-cooled plant tissue grinding and lysis buffer mixing device injects liquid nitrogen into the grinding equipment shell via an external liquid nitrogen injection connection tube to pre-cool the plant samples inside. Simultaneously, a second drive motor controls the first grinding rod, a fixed shaft, and the second grinding rod to grind and pulverize the pre-cooled plant samples. A filter screen is installed at the bottom of the upper shell to prevent insufficiently ground sample tissue. A fixed centrifuge tube containing lysis buffer is installed at the top of the upper shell cavity. After smoothly connecting the lower and upper shells, the device is inverted, allowing the sample ground in the grinding equipment shell to enter the fixed centrifuge tube containing lysis buffer in the upper shell for mixing. The fresh plant tissue is rapidly ground into powder using liquid nitrogen pre-cooling. The device is then inverted to mix the powder with the lysis buffer in a sealed space. This entire process avoids multiple sample transfers exposed to room temperature and operates entirely in a low-temperature liquid nitrogen environment, resulting in better particle size from mechanical grinding. Attached Figure Description
[0016] Figure 1 This is a schematic diagram of the overall structure of the device of this utility model;
[0017] Figure 2 This utility model Figure 1 Internal structure diagram;
[0018] Figure 3 This utility model Figure 2 A schematic diagram of a partial structure;
[0019] Figure 4 This utility model Figure 3 A partial structural diagram.
[0020] In the picture:
[0021] 1. Base plate; 101. Support leg; 102. Support bracket; 103. Drive motor one;
[0022] 2. Connecting lower shell; 201. Liquid nitrogen injection connecting pipe; 202. Fastening cover; 203. Motor bracket; 204. Drive motor two; 205. Grinding equipment shell; 206. Support plate; 207. Grinding rod one; 208. Fixed shaft; 209. Grinding rod two;
[0023] 3. Connect the upper shell; 301. Secure the centrifuge tubes; 302. Secure the funnel;
[0024] 4. Filter screen. Detailed Implementation
[0025] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.
[0026] To make the above-mentioned objectives, features and advantages of this utility model more apparent and understandable, the utility model will be further described in detail below with reference to the accompanying drawings and specific embodiments.
[0027] The main purposes of mixing plant tissue grinding and lysis buffer are as follows:
[0028] Cell disruption: Plant cells have relatively tough cell walls, which are often difficult to break down directly with simple lysis solutions. Grinding, through mechanical force such as the crushing, beating, or impact of steel balls in a grinder, breaks down the cells in the plant tissue into a powder, increasing the contact area between the cells and the lysis solution. This allows the lysis solution to penetrate the cell more fully, creating conditions for subsequent cell lysis and the release of cell contents.
[0029] Release of cellular contents: After cell lysis, the cells are mixed with a lysis buffer. The various components in the lysis buffer then release intracellular substances. For example, surfactants in the lysis buffer can disrupt the lipid bilayer of the cell membrane, releasing intracellular nucleic acids, proteins, organelles, and other substances into the solution for subsequent extraction, separation, and analysis.
[0030] Protecting biomolecules: The formulation of lysis buffers varies depending on the experimental purpose, but they usually contain components that can protect biomolecules. For example, when extracting DNA or RNA, EDTA in the lysis buffer can chelate metal ions, inhibit the activity of nucleases, and prevent DNA or RNA from being degraded; when extracting proteins, protease inhibitors in the lysis buffer can prevent proteases from hydrolyzing the target protein, maintaining the integrity and activity of the protein.
[0031] Maintaining a suitable reaction environment: The composition of the lysis buffer can also adjust the pH, ionic strength, and other properties of the mixed system, providing a suitable reaction environment for subsequent experimental operations. For example, certain enzymatic reactions or analytical assays require specific pH values and ion concentrations to ensure enzyme activity and smooth reaction. Thorough mixing with the lysis buffer through grinding allows the cell contents to remain stable in a suitable environment, which helps improve the accuracy and reproducibility of experimental results.
[0032] Specific operational steps for mixing plant tissue grinding and lysis buffer:
[0033] Preparation: Select suitable plant tissues, such as leaves, roots, and stems, and pre-treat them according to the experimental purpose, such as washing and chopping. At the same time, prepare experimental equipment and reagents such as mortars, pestles, centrifuge tubes, and lysis buffer.
[0034] Grinding plant tissues:
[0035] Manual grinding: Place the plant tissue in a pre-cooled mortar and pestle, add an appropriate amount of liquid nitrogen to rapidly freeze and brittle the tissue. Then, gently tap the tissue in the liquid nitrogen with a pestle to break it into small pieces. Next, when the liquid nitrogen is about to evaporate, use the pestle to grind the tissue sample, replenishing the liquid nitrogen as needed, until the tissue is ground into powder.
[0036] Using a grinder: For some tough plant tissues or those requiring large-scale processing, a plant tissue grinder can be used. Cut the plant tissue into small segments and place them in centrifuge tubes containing grinding balls and an appropriate amount of lysis buffer. Insert the centrifuge tubes into the grinder's adapter, secure them, set the grinder's parameters, and start the grinder to grind.
[0037] Add lysis buffer and mix: Once the tissue has been ground into powder or reached the desired level of grinding, immediately add an appropriate amount of lysis buffer. If grinding by hand, add the lysis buffer directly to the mortar and pestle and continue grinding to ensure the tissue powder and lysis buffer are fully mixed until a uniform paste is formed. If using a grinder, after grinding, remove the centrifuge tube, open the cap, and then agitate or vortex the centrifuge tube appropriately to ensure the lysis buffer and ground tissue are fully in contact and mixed.
[0038] Subsequent processing: The mixed sample is centrifuged to remove insoluble impurities, and the supernatant is collected for subsequent experimental analysis, such as DNA or RNA extraction or protein separation.
[0039] Existing techniques require multiple sample transfers (such as transferring to centrifuge tubes after grinding and stratifying after lysis), which increases the risk of RNA exposure to room temperature and degradation. In addition, manual operation not only results in incomplete and uneven grinding of samples, but also requires a low-temperature environment, which places stringent requirements on experimental conditions and poses a risk of aerosol contamination.
[0040] Please see Figure 1-4 A pre-cooled plant tissue grinding and lysis solution mixing device includes a base plate 1 and a connecting lower shell 2. A motor bracket 203 is fixedly connected to the bottom of the connecting lower shell 2. A second drive motor 204 is fixedly installed inside the motor bracket 203. A grinding rod 207 is fixedly connected to the output end of the second drive motor 204. A fixed shaft 208 is fixedly connected to the top of the first grinding rod 207. A second grinding rod 209 is fixedly connected to the top of the fixed shaft 208. A grinding equipment shell 205 is fixedly installed inside the connecting lower shell 2. A receiving plate 206 is fixedly connected to the surface of the grinding equipment shell 205. A filter screen 4 is attached to the top of the receiving plate 206. A fixed funnel 302 is fixedly connected to the top of the filter screen 4. A fixed centrifuge tube 301 is fixedly connected to the top of the fixed funnel 302. An upper shell 3 is fixedly connected to the top of the fixed centrifuge tube 301. Liquid nitrogen is injected into the grinding equipment shell 205 through an external liquid nitrogen injection pipe via a liquid nitrogen injection connection pipe 201. The plant samples inside are pre-cooled, and the grinding rod 207, fixed shaft 208, and grinding rod 209 are controlled by the drive motor 204 to grind and pulverize the pre-cooled plant samples. A filter screen 4 is set at the bottom of the upper shell 3 to block insufficiently ground sample tissue. A fixed centrifuge tube 301 is set at the top of the upper shell 3 cavity. The fixed centrifuge tube 301 contains lysis solution. After the lower shell 2 and the upper shell 3 are stably connected, the device is inverted so that the sample ground by the grinding device shell 205 enters the fixed centrifuge tube 301 with lysis solution in the upper shell 3 to form a mixture. Liquid nitrogen is used for pre-cooling to quickly grind the fresh plant tissue into powder in liquid nitrogen. The device is then inverted so that the powder and lysis solution are mixed in a sealed space to complete the mixing. The whole operation can avoid multiple sample transfers exposed to room temperature environment, and the operation is carried out in the low temperature environment of liquid nitrogen, resulting in better particle size of mechanical grinding.
[0041] The surface of the grinding equipment housing 205 is fixedly connected to a liquid nitrogen injection connection pipe 201, and the surface of the liquid nitrogen injection connection pipe 201 is threadedly connected to a fastening cap 202.
[0042] Among them, a drive motor 103 is fixedly installed at the center of the upper surface of the base plate 1, and the output shaft of the drive motor 103 is fixedly connected to the connecting lower shell 2.
[0043] The base plate 1 has four sets of support legs 101 fixedly connected to its top, and the support brackets 102 are fixedly connected to the top of the four sets of support legs 101. The lower shell 2 is attached to the inside of the support brackets 102.
[0044] The bottom end of the grinding equipment housing 205 is attached to the output end of the drive motor 204.
[0045] The lower shell 2 is threadedly connected to the upper shell 3. The lower shell 2 and the upper shell 3 are connected by a screw thread and are divided into upper and lower layers and are detachable. The lower shell 2 can be rotated by the drive motor 103, which in turn drives the plant sample inside to rotate. The external rotation of the lower shell 2 can drive the internal grinding media and the material being ground to make a circular motion, which promotes the uniform distribution of the material in the cavity of the grinding equipment shell 205 and avoids local accumulation. The grinding component composed of the internal rotating grinding rod 207, the fixed shaft 208 and the grinding rod 209 directly grinds the material, which can more accurately control the degree of grinding. The combination of the two can greatly improve the grinding efficiency.
[0046] The surface of the lower shell 2 is provided with a mounting hole, and a liquid nitrogen injection connection pipe 201 is fixedly connected inside the mounting hole.
[0047] A support plate 206 is fixedly installed on the inner wall of the lower shell 2.
[0048] Working principle: The lower shell 2 and the upper shell 3 are connected by a screw thread and are divided into two layers that are detachable. The lower shell 2 can be rotated by the drive motor 103, which in turn drives the internal plant sample to rotate. Liquid nitrogen is injected into the grinding device shell 205 through the external liquid nitrogen pipe using the liquid nitrogen injection connecting pipe 201 to pre-cool the internal plant sample. At the same time, the grinding rod 207, the fixed shaft 208 and the grinding rod 209 are controlled by the drive motor 204 to grind and pulverize the pre-cooled plant sample. The bottom layer of the upper shell 3 is equipped with a filter screen 4 to block insufficiently ground sample tissue. The top layer of the upper shell 3 cavity is equipped with a fixed centrifuge tube 301, which contains lysis solution. After the lower shell 2 and the upper shell 3 are stably connected, the device is inverted so that the sample ground by the grinding device shell 205 enters the fixed centrifuge tube 301 with lysis solution in the upper shell 3 and forms a mixture.
[0049] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.
[0050] Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the present invention, the scope of which is defined by the appended claims and their equivalents.
Claims
1. A pre-cooled plant tissue grinding and lysis solution mixing device, characterized in that, The device includes a base plate (1) and a connecting lower shell (2). A motor bracket (203) is fixedly connected to the bottom of the connecting lower shell (2). A second drive motor (204) is fixedly installed inside the motor bracket (203). A first grinding rod (207) is fixedly connected to the output end of the second drive motor (204). A fixed shaft (208) is fixedly connected to the top of the first grinding rod (207). A second grinding rod (209) is fixedly connected to the top of the fixed shaft (208). A grinding equipment shell (205) is fixedly installed inside the lower shell (2). A receiving plate (206) is fixedly connected to the surface of the grinding equipment shell (205). A filter screen (4) is attached to the top of the receiving plate (206). A fixed funnel (302) is fixedly connected to the top of the filter screen (4). A fixed centrifuge tube (301) is fixedly connected to the top of the fixed funnel (302). A connecting upper shell (3) is fixedly connected to the top of the fixed centrifuge tube (301).
2. A pre-chilled plant tissue grinding and lysing buffer mixing device according to claim 1, wherein: The surface of the grinding equipment housing (205) is fixedly connected to a liquid nitrogen injection connection pipe (201), and a fastening cap (202) is threadedly connected to the surface of the liquid nitrogen injection connection pipe (201).
3. A pre-chilled plant tissue grinding and lysing buffer mixing device as defined in claim 1, wherein: A drive motor (103) is fixedly installed at the center of the upper surface of the base plate (1), and the output shaft of the drive motor (103) is fixedly connected to the connecting lower shell (2).
4. A pre-chilled plant tissue grinding and lysing buffer mixing device as defined in claim 1, wherein: The top of the base plate (1) is fixedly connected to four sets of support legs (101), and the top of the four sets of support legs (101) is fixedly connected to support brackets (102). The lower shell (2) is attached to the inside of the support brackets (102).
5. A pre-chilled plant tissue grinding and lysing buffer mixing device as defined in claim 1, wherein: The bottom end of the grinding equipment housing (205) is attached to the output end of the second drive motor (204).
6. A pre-chilled plant tissue grinding and lysing buffer mixing device as defined in claim 1, wherein: The lower shell (2) is threadedly connected to the upper shell (3).
7. A pre-chilled plant tissue grinding and lysing buffer mixing device as defined in claim 1, wherein: The surface of the lower shell (2) is provided with an installation hole, and a liquid nitrogen injection connection pipe (201) is fixedly connected inside the installation hole.
8. A pre-chilled plant tissue grinding and lysing buffer mixing device as defined in claim 1, wherein: A support plate (206) is fixedly installed on the inner wall of the connecting lower shell (2).