Membrane breaking device

Abstract

The utility model relates to the technical field of fine blasting, in particular to a membrane breaking device which comprises a bearing container, an exploder and a detonator, water is borne in the bearing container, cell tissues are placed in a waterproof bag filled with normal saline, the waterproof bag is soaked in the water, the exploder is located outside the bearing container, the detonator is located in the water in the bearing container, and the detonator is located in the water in the bearing container. And the exploder is connected with the detonator through a detonating cord. According to the membrane breaking device provided by the utility model, the detonator is detonated through the exploder, shock waves generated by explosion of the detonator are utilized, and energy is transmitted through medium water, so that the energy acts on cell tissues in the waterproof bag, and the aim of breaking cell membranes through the shock waves is fulfilled.

Description

TECHNICAL FIELD

[0001] The utility model relates to the fine blasting technical field, especially to a membrane breaking device. BACKGROUND

[0002] Cell membrane rupture is an important step in cell biology, biochemistry and molecular biology research.

[0003] The existing membrane breaking device for extracting cell target objects is a Thermofisher benchtop refrigerated high-speed centrifuge, which mainly comprises a driving system and a control system, a carbon brush-free maintenance-free frequency induction motor directly drives to complete rapid centrifugation or continuous centrifugation mode, a computer control system, and a large screen for digital display. UTILITY MODEL CONTENT

[0004] In order to solve the above technical problems or at least partially solve the above technical problems, the utility model provides a membrane breaking device, which can save a lot of centrifugation time and achieve the condition of extracting target objects.

[0005] The utility model provides a membrane breaking device, which comprises a bearing container, an initiator and a detonator, the bearing container bears water inside, cell tissues are placed in a waterproof bag containing physiological saline, the waterproof bag is soaked in water, the initiator is located outside the bearing container, the detonator is located in the water in the bearing container, and the initiator and the detonator are connected through an initiating line.

[0006] Optionally, a traction rod is horizontally arranged at the top opening of the bearing container, and the initiating line is wound on the traction rod.

[0007] Optionally, a limiting fixing device is connected to the top opening of the bearing container, and the limiting fixing device fixes the traction rod at the top opening of the bearing container.

[0008] Optionally, the limiting fixing device comprises two supports, the two supports are fixed to two symmetrical positions at the top opening of the bearing container, and the two ends of the traction rod are fixed to the supports.

[0009] Optionally, a traction rope is fixed to the waterproof bag, and the upper end of the traction rope is bound to the traction rod.

[0010] Optionally, the material of the bearing container is iron.

[0011] Optionally, the physiological saline is a 0.9% NaCl solution.

[0012] The technical scheme provided by the utility model embodiment has the following advantages compared with the prior art:

[0013] The utility model provides a kind of membrane breaking device, detonator is exploded by detonator, utilize the shock wave generated by detonator explosion, energy is transferred by medium water, make it act on the cell tissue in waterproof bag, complete the goal of shock wave breaking cell membrane.Shock wave breaks cell membrane mode as a novel cell rupture method, with broad application prospect, and is expected to play an important role in biological science research, shock wave as a new method of breaking cell membrane, with some unique characteristics, its principle is based on physical impact effect, shock wave is with high-energy short-time pulse waveform transmission energy shock wave, by the shock wave acting on cell, can produce highly compressed and membrane expansion force instantaneously, to cause cell membrane rupture. BRIEF DESCRIPTION OF DRAWINGS

[0014] Figure 1 The utility model provides a kind of membrane breaking device's structure schematic view.

[0015] Reference signs are explained as follows:

[0016] 1, bearing container;2, traction rod;3, detonation line;4, detonator;5, detonator;6, cell tissue;7, waterproof bag;8, fixing device. DETAILED DESCRIPTION

[0017] The utility model is described in detail in combination with the drawings, but it should be understood that the protection scope of the utility model is not limited by the specific embodiment.

[0018] In the description of the utility model, it should be understood that the orientation or positional relationship indicated by the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "axial", "radial", "circumferential" are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the technical scheme of the utility model and simplifying the description, and do not indicate or imply that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, so it cannot be understood as a limitation on the utility model.

[0019] Cell membrane rupture is an important step in cell biology, biochemistry and molecular biology research. Traditional cell membrane rupture methods include physical crushing, chemical crushing, thermal lysis, enzyme treatment, etc. These methods have certain limitations in some cases, such as high requirement for cell integrity, difficulty in applying to specific tissue types or inability to efficiently release target molecules. Therefore, researchers have been looking for new cell rupture methods to meet different experimental needs.

[0020] Currently, the device for extracting target substances from cells by breaking down membranes is the Thermofisher benchtop refrigerated high-speed centrifuge. Its structure mainly consists of a drive system and a control system. The brushless, maintenance-free frequency induction motor directly drives the centrifugation to complete rapid or continuous centrifugation modes. The specific technical problem with this device is that it is difficult to extract target substances from certain tissue cells, such as mouse tail cells. Repeated and long-term centrifugation operations are required, which is inefficient and the results are not ideal.

[0021] Therefore, this utility model provides a membrane breaking device that can save a lot of centrifugation time and meet the conditions for extracting the target substance.

[0022] At least one embodiment of this utility model provides a membrane rupture device, comprising: a support container, a detonator, and a detonator. The support container contains water, the cell tissue is placed in a waterproof bag containing physiological saline, the waterproof bag is immersed in water, the detonator is located outside the support container, the detonator is located in the water inside the support container, and the detonator and the detonator are connected by a detonating wire.

[0023] In the membrane-breaking device provided in the above-described embodiment of the present invention, a detonator is detonated by an initiator, and the shock wave generated by the detonator explosion is used to transfer energy through the medium water, so that it acts on the cell tissue inside the waterproof bag, thereby achieving the goal of breaking the cell membrane by shock wave.

[0024] The present invention will be described below through several specific embodiments. To keep the following description of the embodiments of the present invention clear and concise, detailed descriptions of known functions and components may be omitted. When any component of an embodiment of the present invention appears in more than one drawing, the component may be represented by the same reference numerals in each drawing.

[0025] refer to Figure 1 , Figure 1 This is a schematic diagram of the structure of a membrane-breaking device provided in an embodiment of the present invention, as shown below. Figure 1 As shown, this utility model embodiment provides a membrane breaking device, including: a carrying container 1, an initiator 4 and a detonator 5. The carrying container 1 contains water, the cell tissue 6 is placed in a waterproof bag 7 containing physiological saline, the waterproof bag 7 is immersed in water, the initiator 4 is located outside the carrying container 1, the detonator 5 is located in the water inside the carrying container 1, and the initiator 4 and the detonator 5 are connected by a detonating wire 3.

[0026] It should be understood that the carrying container 1 for containing water needs to be able to withstand the work done by the shock wave in water without structural damage, and there is no specific strength value requirement, the function of the carrying container 1 is only to charge the function of the shock wave medium of the detonator explosion - water, which can be disposable or reusable, and the minimum selection requirement of the carrying container 1 is that the carrying container 1 cannot be blown open after the detonator explosion, and the cell is still in the water, which is basically the same as before the experiment, and the deformation of the carrying container 1 is not important. The waterproof bag 7 in the embodiment of the utility model is a common waterproof packaging belt, and the material is not limited. The distance between the waterproof bag 7 and the detonator 5 needs to be adjusted according to the difficulty of breaking the membrane of different cell tissues. There is no strict requirement for whether the detonator 5 and the cell tissue 6 are in a horizontal straight line relationship.

[0027] The energy of the shock wave can be adjusted according to the needs of different cells, and the energy of the shock wave is adjusted by adjusting the detonation speed of the detonator 5, the distance under water, the distance between the detonator 5 and the cell tissue 6 and the like. By adjusting the energy size and frequency of the shock wave, the accurate rupture of the cell membrane can be realized. The energy size and frequency are closely related to the cell membrane. For example, for cell tissues prone to membrane rupture, lower energy and frequency size should be used. Excessive energy will damage the extraction and capture of target molecules, so different parameters need to be adjusted according to different cell tissues, so as to realize the accurate rupture of the cell membrane and the efficient release of the target molecules. This method is suitable for various cell types, including protists, eukaryotic cells and bacteria.

[0028] The utility model provides a kind of membrane breaking device, detonator is detonated by initiator, utilize the shock wave generated by detonator explosion, energy is transferred by medium water, make it act on the cell tissue in waterproof bag, complete the goal of shock wave breaking cell membrane. Shock wave breaks cell membrane method as a novel cell rupture method, with broad application prospect, and is expected to play an important role in biological science research, shock wave as a new method of breaking cell membrane, with some unique features, its principle is based on physical impact effect, shock wave is with high-energy short-time pulse waveform transmission energy shock wave, by the shock wave acting on cell, can produce highly compressed and membrane expansion force instantaneously, so as to cause cell membrane rupture. Shock wave breaks cell membrane method compared with traditional method has the following characteristics: high efficiency, no additive, suitable for a variety of cells, low damage and controllability.

[0029] Specifically, the carrying container 1 is provided with a traction rod 2 across the top opening, and the detonating cord 3 is wound around the traction rod 2. The traction rod 2 can avoid the contact between the detonator 5 and the side wall of the carrying container 1, so that the blasting force of the detonator 5 can be better transmitted to the cell tissue 6. In addition, the traction rod 2 can better adjust the distance between the detonator 5 and the cell tissue 6 and the depth of the detonator 5.

[0030] The distance between the detonator 5 and the cell tissue 6 is adjusted according to the difficulty of breaking the cell membrane of different cell tissues 6, and there is no definite corresponding relationship, which is not distinguished according to the cell diameter, but distinguished according to the difficulty of breaking the membrane of the cell tissue 6. For example, the liver tissue cells of a mouse are easy to break the membrane and extract target molecules, and compared with this, the tail tissue cells of a mouse are difficult to extract target molecules.

[0031] Optionally, the top opening of the bearing container 1 is connected with a limiting fixing device 8, the limiting fixing device 8 fixes the traction rod 2 at the top opening of the bearing container 1, so that the traction rod 2 can be fixed at the corresponding position, and the position of the detonator 5 is further ensured, and the blasting force of the detonator 5 can be better transmitted to the cell tissue 6.

[0032] Referring again to Figure 1 , the limiting fixing device 8 comprises two supports, the two supports are fixed at two symmetrical positions of the top opening of the bearing container 1, and the two ends of the traction rod 2 are fixed with the supports. Specifically, perforations can be formed on the supports, and the traction rod 2 can be fixed by being sequentially inserted through the perforations on the two supports. The supports can be fixed to the bearing container 1 by bolts, and the specific structure of the supports is not limited as long as the traction rod 2 can be fixed.

[0033] Specifically, the waterproof bag 7 is fixed with a traction rope, and the upper end of the traction rope is bound to the traction rod 2. The waterproof bag 7 can be stably hung through the traction rope, so that the position of the cell tissue 6 is fixed, and the shock wave generated by the explosion of the detonator 5 can be transmitted through the medium water to act on the cell tissue 6 in the waterproof bag 7.

[0034] Specifically, in the embodiment of the utility model, the bearing container 1 can be a square container or a barrel-shaped container.

[0035] Optionally, the material of the bearing container 1 is iron, and it can also be other materials.

[0036] Optionally, the physiological saline is a 0.9% NaCl solution.

[0037] Example 1

[0038] Device assembly

[0039] Place the rectangular iron barrel flat, install a set of limiting fixing devices on the barrel wall, pass the traction rod 2 through the round holes on the limiting fixing devices to complete the position fixing, then pour enough water into the rectangular iron barrel, wind the detonating cord 3 on the traction rod 2, one end of which is connected with the detonator 5 and is placed below the water surface at a certain distance through the detonating cord 3, and the other end is connected with the initiator 4.

[0040] Cell tissue filling

[0041] Take the appropriate amount of cell tissue 6 placed in a waterproof sealed sleeve containing a sufficient amount of normal saline (0.9% NaCl solution), through the fine line connection into the water below.

[0042] Adjust the parameters of each detonator 5 initiation, finally through the initiator 4 detonation, complete the shock wave broken cell membrane experiment.

[0043] The experiment selected mouse myocardial tissue cells, study the cell lysis under different conditions:

[0044] I, the experimental steps of the blast shock wave:

[0045] 1, the detonating tube 5 tube is placed in the center of the barrel;

[0046] 2, at a distance of 510 cm, 15 cm, 20 cm from the detonator 5 to place the treated sealed myocardial cells;

[0047] 3, the detonator 5 connection initiator 4 detonation, complete the experiment.

[0048] In particular, the diameter of the iron barrel is 60 cm, pour the water depth of about 30 cm, detonator 5 and myocardial cells are placed in the middle of the water, about 15 cm or so.

[0049] II, protein concentration detection steps:

[0050] 1, cell crushing: using high pressure homogenizer by electric operation, the cells in the appropriate buffer homogenate quickly broken, so that the release of cell contents.

[0051] 2, centrifugal separation of protein: centrifugal separation by centrifuge, get the supernatant containing protein components.

[0052] 3, concentration determination: using enzyme label instrument by colorimetric method to determine the protein concentration. Some common protein quantification method such as Bradford method, BCA method can be detected by enzyme label instrument.

[0053] In the use of enzyme label instrument to determine the protein concentration, containing protein samples and specific color reagent mixed, after the reaction produces colored material. Enzyme label instrument by measuring the absorbance of colored material at a specific wavelength, according to the standard curve to calculate the concentration of protein, as shown in table 1 below.

[0054] Table 1: cell lysis results

[0055] 10 cm 15 cm 20 cm Homogenate 0.812 0.932 0.978 Explosion + homogenate 0.858 0.857 1.056 Explosion 0.902 0.799 0.750

[0056] Remarks:

[0057] 1, homogenate refers to only enzyme-linked immunoassay instrument (enzyme label instrument) treatment;Explosion + homogenate refers to the combined effect of explosion shock wave and enzyme label instrument;Explosion refers to only the treatment of detonation shock wave.

[0058] 2, the experiment exists 10cm, 15cm, 20cm three kinds of distance, each distance has three groups of experiments, and the experimental data are the average of three groups.

[0059] 3, the data results are the protein BCA results after dilution ten times. The numerical value is the absorbance value, and then the protein concentration is calculated according to the calculation. The higher the absorbance, the higher the protein concentration, and the more complete the lysis.

[0060] 4, the homogenate and explosion + homogenate group are treated in the enzyme label instrument for 5-10min (generally, the membrane breaking centrifugation effect is basically finished after 5min).

[0061] Experimental conclusion:

[0062] 1, from the three different distances, it can be found that the detonation shock wave has the best membrane breaking effect at the distance of 10cm compared with the other two types.

[0063] 2, the pretreatment of detonation shock wave can make the homogenate show more complete lysis effect in the subsequent enzyme label instrument detection.

[0064] From the above, it can be found that the membrane breaking device provided by the utility model is an effective and feasible method for detonation shock wave membrane breaking, which is helpful for more complete lysis of cells and extraction of protein concentration.

[0065] The above utility model is only several specific embodiments of the utility model, but the utility model embodiments are not limited to this, any changes that can be thought of by those skilled in the art should fall within the protection scope of the utility model.

Claims

1. A membrane breaking device, characterized by, The application relates to a device for detonating a cell tissue (6) in water, comprising: a bearing container (1) containing water, the cell tissue (6) being placed in a waterproof bag (7) containing physiological saline, the waterproof bag (7) being soaked in water; a detonator (4) located outside the bearing container (1); a detonating fuse (3) connecting the detonator (4) and a detonator (5) located in the water in the bearing container (1).

2. A membrane breaking device as claimed in claim 1, characterized in that A traction rod (2) is horizontally arranged at the top opening of the bearing container (1), and the detonating fuse (3) is wound around the traction rod (2).

3. A membrane breaking device as claimed in claim 2, characterized in that A limiting fixing device (8) is connected to the top opening of the bearing container (1), and the limiting fixing device (8) fixes the traction rod (2) to the top opening of the bearing container (1).

4. A membrane breaking device as claimed in claim 3, characterized in that The limiting fixing device (8) comprises two supports, the two supports are respectively fixed to two symmetrical positions of the top opening of the bearing container (1), and the two ends of the traction rod (2) are respectively fixed to the supports.

5. A membrane breaking device as claimed in claim 2, wherein, A traction rope is fixed to the waterproof bag (7), and the upper end of the traction rope is bound to the traction rod (2).

6. A membrane breaking device as claimed in claim 1, wherein, The material of the bearing container (1) is iron.

7. The membrane breaking device of claim 1, wherein, The physiological saline is a 0.9% NaCl solution.

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