Device for improving gene mutation cloning inoculation efficiency of mouse lymphoma cells

Abstract

The invention relates to a device for improving gene mutation cloning inoculation efficiency of mouse lymphoma cells. The device comprises a shaking table, a liquid storage box and a multi-channel inoculation assembly, a liquid storage box is mounted on the shaking table and is used for driving the liquid storage box to shake so as to keep cell suspension in the liquid storage box uniform; the liquid storage box is used for storing a cell suspension and is provided with a plurality of liquid taking holes; the multi-channel inoculation assembly comprises a plurality of pipettor suction heads; the pipettor suction head can move and be inserted into the liquid taking hole so as to suck the cell suspension in the liquid storage box. According to the invention, the liquid storage box is shaken and uniformly mixed through the shaking table, so that the continuous uniform mixing of the cell suspension is realized, the consistency of the initial cell concentration of each parallel sample is fundamentally ensured, and the operation efficiency and the consistency are improved: the cell suspension in the liquid storage box is synchronously sucked by a plurality of pipettor suction heads of the multi-channel inoculation assembly and is inoculated into a porous plate; the original manual operation in several minutes is shortened to be completed in several seconds, so that the cell precipitation difference caused by the sequence of inoculation time is avoided.

Description

Technical Field

[0001] This invention relates to the field of experimental equipment technology, and in particular to a device for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells. Background Technology

[0002] In mutagenicity assays of the TK gene in mouse lymphoma L5178Y cells, determining plate colony efficiency is a crucial technical step. Current mouse lymphoma cell gene mutation assays rely entirely on manual methods for cell density measurement before and after the phenotypic expression phase. This involves precise counting using a hemocytometer, a series of complex gradient dilutions with cloning medium, thorough shaking, and the addition of the diluted cell suspension to each well of a 96-well plate using a multichannel pipette.

[0003] The mouse lymphoma cell gene mutation assay uses manual seeding and cell density adjustment. During manual seeding, cells gradually settle from the first well to the last due to gravity, leading to significant concentration differences between wells and inconsistent actual cell counts, affecting the accuracy of cloning efficiency and mutation frequency calculations. Because a large number of plates are seeded, multiple precise serial dilutions are required, which is cumbersome, time-consuming, inconsistent, and inefficient. These problems urgently need to be addressed. Summary of the Invention

[0004] This invention discloses a device for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells, aiming to solve the technical problems existing in the prior art.

[0005] The present invention adopts the following technical solution: This invention provides an apparatus for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells, comprising a shaker, a reservoir, and a multi-channel inoculation assembly; the reservoir is mounted on the shaker and used to shake the reservoir to keep the cell suspension inside the reservoir uniform; the reservoir is used to store the cell suspension and has multiple dispensing holes; the multi-channel inoculation assembly includes multiple pipette tips; the pipette tips are movable and can be inserted into the dispensing holes to aspirate the cell suspension in the reservoir.

[0006] In the device for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells according to the present invention, the shaker includes a mounting plate, a support, and a shaking assembly; the support and the shaking assembly jointly support the mounting plate, and the support is rotatably connected to the mounting plate; the shaking assembly is used to drive the mounting plate to swing back and forth around the rotation axis of the support; the mounting plate is used to mount the liquid storage box.

[0007] In the device for improving the efficiency of gene mutation clonal inoculation of mouse lymphoma cells according to the present invention, the shaking component includes a motor and a cam; the cam is fixed on the rotating shaft of the motor; the cam abuts against the bottom surface of the mounting plate.

[0008] In the device for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells according to the present invention, the motor is a forward and reverse stepper motor.

[0009] In the device for improving the efficiency of gene mutation cloning of mouse lymphoma cells according to the present invention, the cam includes a main body and a plurality of tooth blocks; the plurality of tooth blocks are circumferentially spaced around the main body, and the plurality of tooth blocks have different radial lengths; one end of each tooth block is fixed to the circumferential side of the main body, and the other end extends radially; the main body is disposed on the motor shaft.

[0010] In the device for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells according to the present invention, the liquid storage box includes a box body and a box lid; the top of the box body is an open square structure with graduations; the box lid is placed on the top opening of the box body and has the liquid dispensing hole.

[0011] In the device for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells according to the present invention, when the liquid storage box is in a horizontal position, the liquid dispensing hole is set at an acute angle A with the vertical direction.

[0012] In the device for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells according to the present invention, when the liquid storage box is in a horizontal position, the orifices on both sides of the liquid dispensing hole do not overlap vertically.

[0013] In the device for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells according to the present invention, the multi-channel inoculation assembly further includes a first guide rail, a second guide rail, and a slider; the second guide rail is slidably connected to the first guide rail and is arranged perpendicular to the first guide rail to be close to or away from the liquid storage box; the slider is slidably connected to the second guide rail; a plurality of pipette tips are mounted on the slider to be slidably inserted into the liquid dispensing hole via the slider.

[0014] The device for improving the inoculation efficiency of mouse lymphoma cell gene mutation clones according to the present invention further includes a splash shield; the splash shield can cover the shaker, the liquid storage box and the multi-channel inoculation assembly.

[0015] In the device for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells according to the present invention, an ultraviolet lamp is provided inside the splash shield.

[0016] The technical solution adopted in this invention can achieve the following beneficial effects: This invention primarily provides a device for improving the inoculation efficiency of mouse lymphoma cell gene mutation clones. By shaking and mixing the reservoir via a shaker, continuous mixing of the cell suspension is achieved, fundamentally ensuring consistent initial cell concentration for each parallel sample. This significantly improves the accuracy and reproducibility of experimental data, enhancing operational efficiency and consistency. Furthermore, multiple pipette tips from the multi-channel inoculation assembly simultaneously aspirate the cell suspension from the reservoir and inoculate it into a multi-well plate, reducing the original manual operation of several minutes to within seconds. This avoids differences in cell precipitation caused by the timing of inoculation, making it particularly suitable for large-scale drug screening experiments. Attached Figure Description

[0017] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below, forming part of the present invention. The illustrative embodiments of the present invention and their descriptions explain the present invention and do not constitute an improper limitation of the present invention. In the accompanying drawings: Figure 1 This is a schematic diagram of the device for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells according to the present invention; Figure 2 This is an exploded structural diagram of a device for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells according to the present invention. Figure 3 This is a schematic diagram of the shaker and liquid storage box of the present invention; Figure 4 This is a schematic diagram of the cam structure of the present invention.

[0018] Explanation of reference numerals in the attached figures: 1. Shaker; 11. Mounting plate; 12. Support; 13. Shaking assembly; 131. Motor; 132. Cam; 1321. Main body; 1322. Tooth block; 2. Liquid storage box; 21. Liquid dispensing port; 22. Box body; 23. Box cover; 3. Multi-channel inoculation assembly; 31. Pipette tip; 32. First guide rail; 33. Second guide rail; 34. Slider; 4. Splash shield; 5. Ultraviolet lamp; 6. Tray. Detailed Implementation

[0019] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below in conjunction with specific embodiments and corresponding drawings. In the description of this invention, it should be noted that the term "or" is generally used to include the meaning of "and / or," unless otherwise expressly indicated.

[0020] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "installation," "connection," and "linking" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or a magnetic connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal communication between two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances. Furthermore, in the description of this application, the terms "first," "second," etc., are used only for distinguishing descriptions and should not be construed as indicating or implying relative importance. In the description of this invention, "a plurality of" means at least two, such as two, three, or more, unless otherwise explicitly specified.

[0021] Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0022] To address the problems existing in the prior art, this application provides a device for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells.

[0023] like Figure 1 and Figure 2 As shown, an apparatus for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells includes a shaker 1, a reservoir 2, and a multi-channel inoculation assembly 3. The reservoir 2 is mounted on the shaker 1 to shake the reservoir 2 and keep the cell suspension inside the reservoir 2 uniform. The reservoir 2 is used to store the cell suspension and has multiple dispensing holes 21. The multi-channel inoculation assembly 3 includes multiple pipette tips 31. The pipette tips 31 can move and be inserted into the dispensing holes 21 to aspirate the cell suspension in the reservoir 2.

[0024] This invention provides a device for improving the inoculation efficiency of gene mutation clones of mouse lymphoma cells. A shaker 1 mixes the cell suspension in the reservoir 2, ensuring continuous mixing and fundamentally guaranteeing consistent initial cell concentrations for each parallel sample. This significantly improves the accuracy and reproducibility of experimental data, enhancing operational efficiency and consistency. Multiple pipette tips 31 of the multi-channel inoculation assembly 3 simultaneously aspirate the cell suspension from the reservoir 2 and inoculate it into a multi-well plate, reducing the original manual operation of several minutes to within seconds. This avoids differences in cell precipitation caused by the timing of inoculation, making it particularly suitable for large-scale drug screening experiments.

[0025] In some preferred embodiments, such as Figures 1-3As shown, the shaker 1 includes a mounting plate 11, a support 12, and a shaking assembly 13. The support 12 and the shaking assembly 13 jointly support the mounting plate 11, and the support 12 is rotatably connected to the mounting plate 11. The shaking assembly 13 is used to drive the mounting plate 11 to swing back and forth around the axis of rotation of the support 12. The mounting plate 11 is used to mount the liquid storage box 2. In this way, the existence of mixing dead zones can be avoided, especially when a square liquid storage box 2 is selected, effectively avoiding mixing dead zones, ensuring that the cell suspension in each part of the liquid storage box 2 is uniform and consistent, and the mixing action is relatively gentle, reducing the impact on the cells in the cell suspension.

[0026] Specifically, taking the mounting plate 11 as horizontal as an example, the rotation axis between the support 12 and the mounting plate 11 is horizontally set, and the shaking component 13 is set on one side of the rotation axis. The shaking component 13 drives the mounting plate 11 to swing up and down around the rotation axis, thereby shaking the cell suspension in the storage box 2 evenly.

[0027] Preferably, the support 12 and the swaying assembly 13 are arranged opposite each other on both sides of the mounting plate 11 along its length, and the upper limit of the sway of the mounting plate 11 is higher than the axis of rotation and the lower limit is against the axis of rotation, so that the cell suspension in the liquid storage box 2 flows back and forth from one side to the other.

[0028] In some preferred embodiments, the mounting plate 11 is magnetically connected to the bottom surface of the liquid storage box 2; thereby facilitating assembly and disassembly.

[0029] In some preferred embodiments, the shaking component 13 is a magnetic stirring device.

[0030] In some preferred embodiments, such as Figure 3 As shown, the rocking assembly 13 includes a motor 131 and a cam 132; the cam 132 is fixed on the shaft of the motor 131; the cam 132 abuts against the bottom surface of the mounting plate 11.

[0031] Preferably, the motor 131 is a forward and reverse stepper motor. Based on this, combined with the different curvatures of the wheel surface of the cam 132 and the forward and reverse stepper rotation, different amplitudes of oscillation can be achieved to adapt to different mixed needs.

[0032] More preferably, such as Figure 4As shown, the cam 132 includes a main body 1321 and multiple toothed blocks 1322; the multiple toothed blocks 1322 are circumferentially spaced around the main body 1321, and the lengths of the multiple toothed blocks 1322 in the radial direction are different; one end of each toothed block 1322 is fixed to the circumferential side of the main body 1321, and the other end extends radially; the main body 1321 is mounted on the rotating shaft of the motor 131; when shaking the liquid storage box 2, a single toothed block 1322 keeps in contact with the liquid storage box 2 and rotates in both directions. Based on the different radial lengths of the multiple toothed blocks 1322, different swing amplitudes are achieved when driving the liquid storage box 2 to swing, thereby achieving different mixing effects and improving the applicability range.

[0033] In some preferred embodiments, the liquid storage box 2 includes a box body 22 and a box lid 23; the box body 22 has an open, graduated square structure at the top, which facilitates control of the added amount; the box lid 23 covers the top opening of the box body 22 and has a dispensing hole 21. The design of the box lid 23 reduces the risk of contamination during aseptic operation.

[0034] Preferably, the box body 22 and the box lid 23 are magnetically connected.

[0035] Preferably, the multiple liquid collection holes 21 are arranged in a straight line.

[0036] In some preferred embodiments, such as Figure 3 As shown, when the reservoir 2 is in a horizontal position, the dispensing hole 21 is set at an acute angle A to the vertical direction. Based on this, the probability of cell suspension splashing out of the reservoir 2 during shaking can be reduced, and the residue of cell suspension can be reduced when the cell suspension is aspirated. That is, when the multi-channel inoculation component 3 is extracting, the reservoir 2 is higher on one side and lower on the other, and the cell suspension gathers on the lower side. At this time, it is more conducive to completely extracting the cell suspension. That is, the top opening of the dispensing hole 21 is closer to the shaking component 13, and the dispensing hole 21 is set near the edge of one side of the reservoir 2 so that the bottom of the multi-channel inoculation component 3 is located at the lowest point in the reservoir 2 after insertion.

[0037] Preferably, when the liquid storage box 2 is in a horizontal position, the openings on both sides of the liquid dispensing hole 21 do not overlap vertically; thereby further reducing the probability of cell suspension splashing.

[0038] In some preferred embodiments, such as Figure 1 and Figure 2As shown, the multi-channel inoculation assembly 3 also includes a first guide rail 32, a second guide rail 33, and a slider 34. The second guide rail 33 is slidably connected to the first guide rail 32 and is set perpendicular to the first guide rail 32 to be close to or away from the reservoir 2. The slider 34 is slidably connected to the second guide rail 33. Multiple pipette tips 31 are mounted on the slider 34 to slide into the dispensing hole 21. Based on this, precise positioning of the pipette tips 31 and the dispensing hole 21 is achieved. At the same time, based on the simultaneous aspiration of multiple pipette tips 31, "one-click" synchronous and equal-volume inoculation of cell suspension into all wells of the multi-well plate is achieved, completely solving the time difference and cell sedimentation problems caused by manual inoculation sequence. Preferably, the first guide rail 32, the second guide rail 33, and the slider 34 can all be selected from existing movable devices, such as linear slider rail mechanisms, to achieve automated operation.

[0039] In some preferred embodiments, the pipette tip 31 can be selected from existing devices, such as a multichannel pipette, for example, the Research Plus pipette.

[0040] In some preferred embodiments, such as Figure 1 and Figure 2 As shown, it also includes a splash guard 4; the splash guard 4 can cover the shaker 1, the liquid storage box 2, and the multi-channel inoculation assembly 3. The splash guard 4 can ensure ventilation and prevent external contamination during operation.

[0041] In some preferred embodiments, such as Figure 1 and Figure 2 As shown, an ultraviolet lamp 5 is installed inside the splash shield 4; the ultraviolet lamp 5 can be used for local sterilization before use.

[0042] In some preferred embodiments, such as Figure 1 and Figure 2 As shown, the shaker 1, the liquid storage box 2, and the multi-channel inoculation assembly 3 are mounted on the tray 6; the tray 6 and the splash guard 4 are fitted together to form a receiving space.

[0043] In some preferred embodiments, the reservoir 2 is a container with a heating function, such as a heating rod or heating wire provided on the bottom surface of the container 22 to heat the cell suspension inside and maintain cell activity.

[0044] The working method of the device of the present invention is as follows: Preparation: Slide open the splash guard 4 and place the cell suspension reservoir 2 on the shaker 1.

[0045] Mixing and Temperature Control: Turn off the splash guard. Turn on the power and set the stirring speed (ensuring the cells are evenly suspended but not foamy) and temperature (37°C). Begin continuous and gentle mixing and temperature control of the cell suspension.

[0046] Simultaneous inoculation: Place a sterile multiwell plate (such as a 96-well plate) in the predetermined position in front of the reservoir 2.

[0047] The operator manually or by pressing a button controls the slider 34 to descend as a whole. During the descent, the pipette tip 31 passes through the synchronous and vertical insertion hole 21 of the reservoir 2 to draw cell suspension; then the slider 34 moves the pipette tip 31 upward and, through the second guide rail 33, moves the pipette tip 31 above the multi-well plate. The slider 34 then moves the pipette tip 31 downward into the well of the multi-well plate, releasing the liquid in the pipette tip 31, completing a one-time synchronous addition of samples to all wells, thus completing the inoculation.

[0048] Sterilization: After use, close the splash guard 4 and turn on the ultraviolet lamp 5 to disinfect the inside of the device by irradiation at regular intervals.

[0049] The embodiments of the present invention have been described above with reference to the accompanying drawings. However, the present invention is not limited to the specific embodiments described above. The specific embodiments described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of the present invention without departing from the spirit and scope of the claims, and all of these forms are within the protection scope of the present invention.

Claims

1. A device for improving the inoculation efficiency of gene-mutant clones of mouse lymphoma cells, characterized in that, Includes shaker, reservoir, and multi-channel inoculation assembly; The liquid storage box is installed on the shaker to drive the liquid storage box to shake, so as to keep the cell suspension inside the liquid storage box uniform. The storage box is used to store cell suspensions and has multiple dispensing holes; The multichannel inoculation assembly includes multiple pipette tips; the pipette tips are movable and can be inserted into the dispensing holes to aspirate the cell suspension in the reservoir.

2. The device for improving the inoculation efficiency of mouse lymphoma cell gene mutation clones according to claim 1, characterized in that, The shaking table includes a mounting plate, a support, and a shaking assembly; The support and the rocking assembly together support the mounting plate, and the support is rotatably connected to the mounting plate; The swaying component is used to drive the mounting plate to oscillate back and forth around the rotation axis of the support. The mounting plate is used to mount the liquid storage box.

3. The device for improving the inoculation efficiency of gene-mutant clones of mouse lymphoma cells according to claim 2, characterized in that, The rocking assembly includes a motor and a cam; The cam is fixed on the shaft of the motor; The cam abuts against the bottom surface of the mounting plate.

4. The device for improving the inoculation efficiency of gene-mutant clones of mouse lymphoma cells according to claim 3, characterized in that, The motor is a forward and reverse rotating stepper motor.

5. The device for improving the inoculation efficiency of gene-mutant clones of mouse lymphoma cells according to claim 1, characterized in that, The liquid storage box includes a box body and a box lid; The top of the box is an open, square structure with markings; The lid is placed over the top opening of the box body and has the liquid dispensing hole.

6. The device for improving the inoculation efficiency of gene-mutant clones of mouse lymphoma cells according to claim 1, characterized in that, When the liquid storage box is in a horizontal position, the liquid dispensing hole is set at an acute angle A with the vertical.

7. The device for improving the inoculation efficiency of mouse lymphoma cell gene mutation clones according to claim 6, characterized in that, When the liquid storage box is in a horizontal position, the openings on both sides of the liquid dispensing hole do not overlap vertically.

8. The device for improving the inoculation efficiency of mouse lymphoma cell gene mutation clones according to claim 1, characterized in that, The multi-channel inoculation assembly also includes a first guide rail, a second guide rail, and a slider; The second guide rail is slidably connected to the first guide rail and is set perpendicular to the first guide rail to be close to or away from the liquid storage box; The slider is slidably connected to the second guide rail; Multiple pipette tips are mounted on the slider to slide into the dispensing port via the slider.

9. The device for improving the inoculation efficiency of mouse lymphoma cell gene mutation clones according to claim 1, characterized in that, It also includes a splash guard; The splash guard can cover the shaker, the liquid storage box, and the multi-channel inoculation assembly.

10. The device for improving the inoculation efficiency of gene-mutant clones of mouse lymphoma cells according to claim 9, characterized in that, An ultraviolet lamp is installed inside the splash guard.

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