CN112337372B

CN112337372B - Shaking mixing device for cell culture and use method

Info

Publication number: CN112337372B
Application number: CN202011135577.5A
Authority: CN
Inventors: 曾炳
Original assignee: Nanjing Bingpeng Biotechnology Co ltd
Current assignee: Nanjing Bingpeng Biotechnology Co ltd
Priority date: 2020-10-21
Filing date: 2020-10-21
Publication date: 2022-04-12
Anticipated expiration: 2040-10-21
Also published as: CN112337372A

Abstract

The invention discloses a shaking mixing device for cell culture and a using method thereof, relates to the technical field of cell culture, and aims to solve the problem that the existing shaking mixing device is single in shaking structure. The utility model discloses a culture dish, including base, sliding plate, shaking plate, eccentric wheel, box, fixing mechanism, the inside upper end of base is provided with the slip chamber, the internally mounted in slip chamber has the sliding plate, the inside of sliding plate is provided with the roll chamber, the inside in roll chamber is provided with first sector gear, the fixed shaking plate that is provided with in upper end of sliding plate, the inside both sides of shaking plate all are provided with the dead slot, the internally mounted in dead slot has the eccentric wheel, the top of shaking plate is provided with the backup pad, the inside upper end of backup pad is provided with the transmission chamber, the upper end of backup pad is installed and is placed the box, and places the box and rotate with the backup pad and be connected, the inner wall of placing the box is equipped with a plurality of fixing mechanism along annular evenly distributed, be provided with the culture dish between the fixing mechanism.

Description

Shaking mixing device for cell culture and use method

Technical Field

The invention relates to the technical field of cell culture, in particular to a shaking mixing device for cell culture and a using method thereof.

Background

Cell culture techniques refer to the growth of cells under in vitro conditions, during which the cells no longer form tissues (animals). A culture is a single cell or a population of cells. The cells are cultured in an artificial environment, the cells move or are influenced by other factors due to the change of the environment, the culture time is prolonged, the cells are singularized due to passage, the cell culture is also called as a cell cloning technology, the cell culture is an essential process for the whole bioengineering technology, and the cell culture is the large-scale cloning of the cells. The cell culture technology can lead one cell to be cultured into simple single cells or multi-cells with little differentiation in a large quantity.

The shaking mixing device is used in the cell culture process, and is culture performed by inoculating cells of microorganisms or animals and plants into a liquid culture medium and placing the cells on the shaking mixing device without stopping shaking, which is compared with static culture or surface culture of molds. In the case of bacterial cell culture, the contact with the medium components and the supply of oxygen are improved, and the growth is relatively uniform, and particularly, in the case of mold culture, no pellicle and no pellet formed by long hyphae are formed.

However, the existing shaking mixing device has a single structure, and in the shaking process, cells cannot be quickly and fully mixed with a liquid culture medium, so that the culture quality of the cells is influenced, and therefore the existing requirements are not met.

Disclosure of Invention

The present invention provides a shaking mixing device for cell culture and a use method thereof, so as to solve the problem of single shaking structure of the shaking mixing device in the background art.

In order to achieve the purpose, the invention provides the following technical scheme: a vibration mixing device for cell culture and a using method thereof comprise a base, wherein the upper end inside the base is provided with a sliding cavity, a sliding plate is arranged in the sliding cavity, a rolling cavity is arranged in the sliding plate, a first sector gear is arranged in the rolling cavity, the upper end of the sliding plate is fixedly provided with an oscillating plate, both sides of the inside of the oscillating plate are provided with empty grooves, an eccentric wheel is arranged in the hollow groove, a supporting plate is arranged above the vibrating plate, a plurality of second springs which are uniformly distributed along the ring shape are arranged between the supporting plate and the vibrating plate, the upper end of the inside of the supporting plate is provided with a transmission cavity, the upper end of the supporting plate is provided with a placing box, and the placing box is rotatably connected with the supporting plate, a plurality of fixing mechanisms which are uniformly distributed along the ring shape are arranged on the inner wall of the placing box, and culture dishes are arranged between the fixing mechanisms.

Preferably, a first motor is installed to first sector gear's below, and first motor is located the inside intermediate position department of base, and the output shaft of first motor passes through the shaft coupling transmission with first sector gear and is connected, roll the intracavity portion front and back end and all be provided with the tooth's socket, and the tooth's socket is connected with first sector gear meshing, the ring circle central angle of first sector gear is one hundred eighty degrees.

Preferably, the front end and the rear end of the sliding plate are both fixedly provided with sliding blocks, the sliding blocks are connected with the base in a sliding mode, the lower end of the sliding plate is provided with two groups of rollers in a rotating mode, the rollers are connected with the base in a sliding mode, the two sides of the sliding plate are both provided with first springs, and the two ends of each first spring are fixedly connected with the sliding plate and the base respectively.

Preferably, a second motor is installed on one side of the empty groove, the second motor is located inside the vibration plate, the output end of the second motor is fixedly connected with the eccentric wheel, the length from one end of the eccentric wheel to the circle center is greater than the length from the other end of the eccentric wheel to the circle center, and one end of the eccentric wheel is attached to the supporting plate.

Preferably, the rotation axis is installed to the intermediate position department of transmission intracavity portion, and the one end of rotation axis passes through bearing frame fixed connection with the backup pad, the other end of rotation axis with place box fixed connection, the outside welded fastening of rotation axis has drive gear, and drive gear is located the inside in transmission chamber, one side fixed mounting of transmission intracavity portion has the third motor, the output shaft fixed mounting of third motor has second sector gear, and the ring circle central angle of the tooth of second sector gear is eighty-five degrees, second sector gear is connected with the drive gear meshing.

Preferably, fixed establishment comprises sleeve, movable rod, clamp splice and third spring, the one end of movable rod is located telescopic inside, and the other end of movable rod extends to telescopic outside, the other end and the clamp splice fixed connection of movable rod, the both ends of third spring respectively with sleeve and clamp splice fixed connection, the clamp splice laminates with the culture dish mutually.

Preferably, the upper end of placing the box is installed the closing cap, and the closing cap with place box screw-thread fit, the lower extreme bonding of closing cap has the sponge, and the sponge is filled in the inside of placing the box, the fixed handle that is provided with in upper end of closing cap.

The use method of the shaking mixing device for cell culture comprises the following steps:

firstly, fixing a culture dish, placing the culture dish at the middle position in a placing box, enabling a clamping block at one end of a movable rod to be tightly attached to the outside of the culture dish under the elastic reset of a third spring, covering a sealing cover after the fixing is finished, and enabling sponge in the culture dish to clamp the culture dish;

step two, starting the first motor, forming a reciprocating cycle of the transverse movement of the sliding plate in the sliding cavity along with the continuous rotation of the first sector gear, wherein the speed of the reciprocating cycle is adjusted according to the rotating speed of the first motor, so that the culture dish fixed above also synchronously moves along with the transverse reciprocating motion of the sliding plate;

thirdly, driving the eccentric wheels to rotate at a high speed by starting the second motor, wherein the end with the larger radius is in contact with the bottom of the supporting plate in the rotating process, and the end with the smaller radius is not in contact with the bottom of the supporting plate to form intermittent collision, and the supporting plate is collided at a high frequency to generate vibration under the mutual interval collision of the two eccentric wheels;

step four, starting a third motor, and forming rapid mixing between cells and a liquid culture medium in the culture dish under rapid change of acceleration and deceleration when a gear ring on a second sector gear drives a transmission gear to rotate;

and step five, opening the sealing cover, taking out the culture dish, and finishing integral vibration mixing.

Compared with the prior art, the invention has the beneficial effects that:

1. the invention is characterized in that a sliding cavity provided with a sliding plate is arranged in a base, when the sliding plate provided with a culture dish above moves transversely, a first motor is started, a first sector gear is fixedly arranged on an output shaft of the first motor, a gear ring on the first sector gear is distributed in a sector shape, the first sector gear is arranged in a rolling cavity in the sliding plate, the gear ring of the first sector gear is matched with tooth grooves at two sides of the rolling cavity, the first sector gear is driven to rotate by the first motor, the tooth groove at one side of the rolling cavity moves towards one side of the sliding cavity under the meshing action of the first sector gear, the external gear ring contacts the tooth groove at the other side of the rolling cavity along with the continuous rotation of the first sector gear, at the moment, the tooth groove at one side of the rolling cavity is not meshed with the first sector gear, and when the tooth groove at the other side of the rolling cavity is meshed, because the directions are opposite, the sliding plate moves towards one side of the sliding cavity under the rotation of the first sector gear, a reciprocating cycle of the transverse movement of the sliding plate is formed in the sliding cavity along with the continuous rotation of the first sector gear, the speed of the reciprocating cycle is adjusted according to the rotating speed of the first motor, and then the culture dish fixed above the sliding plate synchronously moves along with the transverse reciprocating movement of the sliding plate, so that the mixing efficiency between the internal cells and the liquid culture medium is accelerated.

2. In the invention, the two sides in the oscillating plate are both provided with the empty grooves, the eccentric wheels are arranged in the empty grooves, by starting the second motor, the output shaft of the second motor drives the eccentric wheel to rotate at high speed, and because the lengths from the two ends of the eccentric wheel to the circle center are different, the end with larger radius is contacted with the bottom of the supporting plate in the rotating process, and the end with smaller radius is not contacted with the bottom of the supporting plate, thereby forming intermittent collision, under the mutual interval collision of the two eccentric wheels, the supporting plate is collided by high frequency to generate vibration, when the bulge of one of them eccentric wheel contacted the backup pad in addition, the bulge of another eccentric wheel then was located and is close to the one end that shakes the board, so under mutually supporting of two eccentric wheels, under the prerequisite of making backup pad high-frequency vibration, kept holistic equilibrium, avoided causing and shake inhomogeneous problem between inside cell of culture dish and the liquid medium.

3. The invention is characterized in that a transmission cavity is arranged in a support plate, an output shaft of a third motor drives a second sector gear to rotate by starting a third motor, a part of a toothed ring of the second sector gear is meshed with a transmission gear with a rotating shaft arranged on one side, the toothed ring on the transmission gear is meshed with the toothed ring on the second sector gear, the transmission gear can be driven to rotate along with the rotation of the second sector gear, the part of the second sector gear without the toothed ring idles with the transmission gear, when the toothed ring on the second sector gear drives the transmission gear to rotate, a placing box with a culture dish arranged above is driven to rotate through the rotating shaft, when the transmission gear idles with the second sector gear, the rotating shaft loses driving force, and simultaneously gradually decelerates under the pressure of the placing box, and the transmission gear drives the rotating shaft to rotate again along with the rotation of the second sector gear, and then make and place the box and accelerate once more, under the fast switch of acceleration and deceleration, form the flash mixed between inside cell of culture dish and the liquid medium, improved holistic shock mixing rate.

4. According to the invention, under the mutual cooperation of the sliding plate, the oscillating plate and the supporting plate, the culture dish fixed in the supporting plate can rapidly mix the cells and the liquid culture solution in the transverse reciprocating direction, the vertical reciprocating direction and the variable-speed rotation, so that on one hand, the oscillating mixing efficiency is improved, on the other hand, the oscillating mixing effect between the cells is improved, and the optimal culture of the cells is ensured.

5. In the invention, six fixing mechanisms are arranged in the placing box in an annular array and each fixing mechanism consists of a sleeve, a movable rod, a clamping block and a third spring, when the culture dish is fixed, the culture dish is placed at the middle position in the placing box, the clamping block at one end of the movable rod is tightly attached to the outside of the culture dish under the elastic reset of the third spring, the fixing mode is simple and quick to operate, can quickly fix culture dishes with different specifications, meanwhile, the stability is higher, the sponge is adhered and fixed at the lower end of the sealing cover and is filled in the residual space inside the culture dish and the placing box, when wholly shaking and mixing, the elastic construction of sponge can be inseparable laminate in the culture dish, avoid the upper cover of culture dish to drop because of vibrations and lead to spilling of inside cell and liquid culture solution, and the protectiveness obtains further improvement.

Drawings

FIG. 1 is a schematic view of the overall structure of the present invention;

FIG. 2 is an overall corrosion view of the present invention;

FIG. 3 is a top view of the interior of the storage box of the present invention;

FIG. 4 is a top view of the interior of the slide plate of the present invention;

FIG. 5 is a schematic view of the eccentric wheel structure of the present invention;

fig. 6 is a schematic view of the internal structure of the transmission chamber of the present invention.

In the figure: 1. a base; 2. a sliding plate; 3. a vibrating plate; 4. a support plate; 5. placing the box; 6. a culture dish; 7. sealing the cover; 8. a first motor; 9. a sliding cavity; 10. a roller; 11. a first spring; 12. a second motor; 13. an empty groove; 14. an eccentric wheel; 15. a second spring; 16. a transmission cavity; 17. a third motor; 18. a rotating shaft; 19. a bearing seat; 20. a fixing mechanism; 21. a sponge; 22. a sleeve; 23. a movable rod; 24. a clamping block; 25. a third spring; 26. a slider; 27. a rolling chamber; 28. a tooth socket; 29. a first sector gear; 30. a handle; 31. a transmission gear; 32. a second sector gear.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.

Referring to fig. 1-6, an embodiment of the present invention is shown: a vibration mixing device for cell culture and a using method thereof comprise a base 1, wherein the upper end inside the base 1 is provided with a sliding cavity 9, the sliding cavity 9 is internally provided with a sliding plate 2, the sliding plate 2 is internally provided with a rolling cavity 27, the rolling cavity 27 is internally provided with a first sector gear 29, the upper end of the sliding plate 2 is fixedly provided with a vibration plate 3, two sides inside the vibration plate 3 are respectively provided with an empty groove 13, the inside of the empty groove 13 is provided with an eccentric wheel 14, a supporting plate 4 is arranged above the vibration plate 3, a plurality of second springs 15 which are uniformly distributed along an annular shape are arranged between the supporting plate 4 and the vibration plate 3, the upper end inside the supporting plate 4 is provided with a transmission cavity 16, the upper end of the supporting plate 4 is provided with a placing box 5, the placing box 5 is rotatably connected with the supporting plate 4, the inner wall of the placing box 5 is provided with a plurality of fixing mechanisms 20 which are uniformly distributed along the annular shape, a culture dish 6 is arranged between the fixing mechanisms 20, under the mutual cooperation of sliding plate 2, shake board 3 and backup pad 4, make and be fixed in the inside culture dish 6 of backup pad 4 under horizontal reciprocating direction, perpendicular reciprocating direction and variable speed rotation, make fast mixing between cell and the liquid culture liquid fast, the shock mixing efficiency that improves on the one hand, on the other hand has improved the shock mixing effect between the two, ensures the optimization of cell and cultivates.

Further, a first motor 8 is installed below the first sector gear 29, the first motor 8 is located at a middle position inside the base 1, an output shaft of the first motor 8 is in transmission connection with the first sector gear 29 through a coupler, tooth grooves 28 are formed in front and rear ends inside the rolling cavity 27, the tooth grooves 28 are in meshing connection with the first sector gear 29, a central angle of a toothed ring of the first sector gear 29 is one hundred eighty degrees, the first sector gear 29 is fixedly installed on the output shaft of the first motor 8, toothed rings on the first sector gear 29 are distributed in a sector shape, the first sector gear 29 is installed in the rolling cavity 27 inside the sliding plate 2, the first sector gear 29 is driven to rotate by the first motor 8, the tooth grooves 28 on one side of the rolling cavity 27 are in meshing action with the first sector gear 29, the sliding plate 2 is moved towards one side of the sliding cavity 9, and along with the continuous rotation of the first sector gear 29, the external toothed ring contacts the toothed groove 28 on the other side of the rolling chamber 27, at this time, the toothed groove 28 on one side of the rolling chamber 27 is not meshed with the first sector gear 29, when the toothed groove is meshed with the toothed groove 28 on the other side of the rolling chamber 27, the sliding plate 2 is moved towards the first side of the sliding chamber 9 under the rotation of the first sector gear 29 due to the opposite direction, and a reciprocating cycle of the sliding plate 2 moving transversely is formed in the sliding chamber 9 along with the continuous rotation of the first sector gear 29.

Further, the front end and the rear end of sliding plate 2 are all fixed and are provided with slider 26, and slider 26 and base 1 sliding connection, the lower extreme of sliding plate 2 rotates and is provided with two sets of gyro wheels 10, and gyro wheel 10 and base 1 sliding connection, the both sides of sliding plate 2 all are provided with first spring 11, and the both ends of first spring 11 respectively with sliding plate 2 and base 1 fixed connection, can assist sliding plate 2 to slide, reduce the noise that the removal produced, first spring 11 can avoid moving inertia too big.

Furthermore, a second motor 12 is installed at one side of the empty slot 13, the second motor 12 is located inside the oscillation plate 3, the output end of the second motor 12 is fixedly connected with an eccentric wheel 14, the length from one end of the eccentric wheel 14 to the center of a circle is greater than the length from the other end of the eccentric wheel 14 to the center of a circle, one end of the eccentric wheel 14 is attached to the support plate 4, because the lengths from the two ends of the eccentric wheel 14 to the center of a circle are different, the end with the larger radius is contacted with the bottom of the support plate 4 in the rotating process, and the end with the smaller radius is not contacted with the bottom of the support plate 4, so that intermittent collision is formed, under the mutual interval collision of the two eccentric wheels 14, the support plate 4 is vibrated by high-frequency collision, and when the protruding part of one of the eccentric wheel 14 is contacted with the support plate 4, the protruding part of the other eccentric wheel 14 is located at the end close to the oscillation plate 3, so under the mutual matching of the two eccentric wheels 14, the supporting plate 4 keeps the overall balance under the premise of high-frequency vibration.

Furthermore, a rotating shaft 18 is installed at the middle position inside the transmission cavity 16, one end of the rotating shaft 18 is fixedly connected with the support plate 4 through a bearing seat 19, the other end of the rotating shaft 18 is fixedly connected with the placing box 5, a transmission gear 31 is fixedly welded outside the rotating shaft 18, the transmission gear 31 is located inside the transmission cavity 16, a third motor 17 is fixedly installed at one side inside the transmission cavity 16, a second sector gear 32 is fixedly installed at an output shaft of the third motor 17, a central angle of a toothed ring of the second sector gear 32 is eighty-five degrees, the second sector gear 32 is meshed with the transmission gear 31 and connected with the transmission gear 31, the transmission gear 31 can be driven to rotate along with the rotation of the second sector gear 32, and the part of the second sector gear 32 without the toothed ring idles with the transmission gear 31, when the transmission gear 31 is driven to rotate by the toothed ring on the second sector gear 32, through the rotation axis 18, drive the top and be equipped with placing box 5 of culture dish 6 and rotate, when drive gear 31 and second sector gear 32 take place the idle running, rotation axis 18 loses drive power, and the while is slowing down gradually under the pressure of placing box 5, along with the rotation of second sector gear 32, drive gear 31 drives the rotation axis once more and advances to rotate, and then the order is placed box 5 and is accelerated once more, under the rapid change with the speed reduction with higher speed, form the flash mixed between 6 inside cells of culture dish and the liquid medium.

Further, fixed establishment 20 is by sleeve 22, the movable rod 23, clamp splice 24 and third spring 25 constitute, the one end of movable rod 23 is located sleeve 22's inside, and the other end of movable rod 23 extends to sleeve 22's outside, the other end and the clamp splice 24 fixed connection of movable rod 23, the both ends of third spring 25 respectively with sleeve 22 and clamp splice 24 fixed connection, clamp splice 24 laminates with culture dish 6 mutually, this kind of fixed mode easy operation is swift, can carry out quick fixed to the culture dish 6 of different specifications, stability is higher simultaneously.

Further, place the upper end of box 5 and install closing cap 7, and closing cap 7 with place 5 screw-thread fit of box, the lower extreme bonding of closing cap 7 has sponge 21, and sponge 21 fills in the inside of placing box 5, the fixed handle 30 that is provided with in upper end of closing cap 7, when wholly shaking the mixture, the elastic construction of sponge 21 can be inseparable laminating in culture dish 6, avoid the upper cover of culture dish 6 to drop because of vibrations and lead to scattering of inside cell and liquid culture solution, the protectiveness obtains further improvement.

firstly, fixing a culture dish 6, placing the culture dish at the middle position in a placing box 5, enabling a clamping block 24 at one end of a movable rod 23 to be tightly attached to the outside of the culture dish 6 under the elastic reset of a third spring 25, covering a sealing cover 7 after the fixing is finished, and enabling a sponge 21 in the culture dish to clamp the culture dish 6;

step two, starting the first motor 8, forming a reciprocating cycle of the transverse movement of the sliding plate 2 in the sliding cavity 9 along with the continuous rotation of the first sector gear 29, wherein the speed of the reciprocating cycle is adjusted according to the rotating speed of the first motor 8, and then the culture dish 6 fixed above also synchronously moves along with the transverse reciprocating motion of the sliding plate 2;

thirdly, the second motor 12 is started to drive the eccentric wheels 14 to rotate at a high speed, one end with the larger radius is in contact with the bottom of the supporting plate 4 in the rotating process, the other end with the smaller radius is not in contact with the bottom of the supporting plate 4, intermittent collision is formed, and the supporting plate 4 is collided at a high frequency to generate vibration under the mutual interval collision of the two eccentric wheels 14;

step four, starting the third motor 17, and forming rapid mixing between cells and the liquid culture medium in the culture dish 6 under rapid change of acceleration and deceleration when the gear ring on the second sector gear 32 drives the transmission gear 31 to rotate;

and step five, opening the sealing cover 7, taking out the culture dish 6, and finishing integral vibration mixing.

It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.

Claims

1. The utility model provides a cell culture is with shaking mixing arrangement, includes base (1), its characterized in that: the upper end of the interior of the base (1) is provided with a sliding cavity (9), a sliding plate (2) is installed in the sliding cavity (9), sliding blocks (26) are fixedly arranged at the front end and the rear end of the sliding plate (2), the sliding blocks (26) are in sliding connection with the base (1), two groups of rollers (10) are rotatably arranged at the lower end of the sliding plate (2), the rollers (10) are in sliding connection with the base (1), first springs (11) are arranged on two sides of the sliding plate (2), two ends of each first spring (11) are respectively and fixedly connected with the sliding plate (2) and the base (1), a rolling cavity (27) is arranged in the sliding plate (2), a first sector gear (29) is arranged in the rolling cavity (27), a first motor (8) is installed below the first sector gear (29), and the first motor (8) is located in the middle position of the interior of the base (1), an output shaft of the first motor (8) is in transmission connection with the first sector gear (29) through a coupler, tooth grooves (28) are formed in the front end and the rear end of the inner portion of the rolling cavity (27), the tooth grooves (28) are in meshed connection with the first sector gear (29), the central angle of a gear ring of the first sector gear (29) is one hundred eighty degrees, an oscillating plate (3) is fixedly arranged at the upper end of the sliding plate (2), hollow grooves (13) are formed in two sides of the inner portion of the oscillating plate (3), an eccentric wheel (14) is installed inside the hollow grooves (13), a supporting plate (4) is arranged above the oscillating plate (3), a second motor (12) is installed on one side of each hollow groove (13), the second motor (12) is located inside the oscillating plate (3), the output end of the second motor (12) is fixedly connected with the eccentric wheel (14), and the length from one end of the eccentric wheel (14) to the circle center is greater than that from the other end of the eccentric wheel (14) to the circle center, one end of the eccentric wheel (14) is attached to the supporting plate (4), a plurality of second springs (15) which are uniformly distributed along an annular shape are arranged between the supporting plate (4) and the vibration plate (3), a transmission cavity (16) is arranged at the upper end inside the supporting plate (4), a placing box (5) is installed at the upper end of the supporting plate (4), the placing box (5) is rotatably connected with the supporting plate (4), a rotating shaft (18) is installed at the middle position inside the transmission cavity (16), one end of the rotating shaft (18) is fixedly connected with the supporting plate (4) through a bearing seat (19), the other end of the rotating shaft (18) is fixedly connected with the placing box (5), a transmission gear (31) is welded and fixed to the outer portion of the rotating shaft (18), the transmission gear (31) is located inside the transmission cavity (16), a third motor (17) is fixedly installed at one side inside the transmission cavity (16), the output shaft fixed mounting of third motor (17) has second sector gear (32), and the ring gear central angle of second sector gear (32) is eighty-five degrees, second sector gear (32) are connected with drive gear (31) meshing, the inner wall of placing box (5) is equipped with a plurality of fixed establishment (20) along annular evenly distributed, be provided with culture dish (6) between fixed establishment (20), fixed establishment (20) comprise sleeve (22), movable rod (23), clamp splice (24) and third spring (25), the one end of movable rod (23) is located the inside of sleeve (22), and the other end of movable rod (23) extends to the outside of sleeve (22), the other end and the clamp splice (24) fixed connection of movable rod (23), the both ends of third spring (25) respectively with sleeve (22) and clamp splice (24) fixed connection, the clamping block (24) is attached to the culture dish (6).

2. The shaking mixing device for cell culture according to claim 1, wherein: place the upper end of box (5) and install closing cap (7), and closing cap (7) with place box (5) screw-thread fit, the lower extreme bonding of closing cap (7) has sponge (21), and sponge (21) fill in the inside of placing box (5), the fixed handle (30) that is provided with in upper end of closing cap (7).

3. The method for using the shaking mixing device for cell culture according to any one of claims 1-2, wherein: the method comprises the following steps:

firstly, fixing a culture dish (6), placing the culture dish at the middle position inside a placing box (5), enabling a clamping block (24) at one end of a movable rod (23) to be tightly attached to the outside of the culture dish (6) under the elastic reset of a third spring (25), covering a sealing cover (7) after the fixing is finished, and enabling a sponge (21) inside to clamp the culture dish (6);

step two, starting the first motor (8), forming a reciprocating cycle of the transverse movement of the sliding plate (2) in the sliding cavity (9) along with the continuous rotation of the first sector gear (29), wherein the speed of the reciprocating cycle is adjusted according to the rotating speed of the first motor (8), and then the culture dish (6) fixed above also synchronously moves along with the transverse reciprocating motion of the sliding plate (2);

thirdly, driving the eccentric wheel (14) to rotate at a high speed by starting the second motor (12), wherein one end with a larger radius is in contact with the bottom of the supporting plate (4) in the rotating process, and the other end with a smaller radius is not in contact with the bottom of the supporting plate (4) to form intermittent collision, and under the mutual interval collision of the two eccentric wheels (14), the supporting plate (4) is collided at a high frequency to generate vibration;

step four, starting a third motor (17), and forming rapid mixing between cells and a liquid culture medium in the culture dish (6) under rapid change of acceleration and deceleration when a gear (31) is driven to rotate by a gear ring on a second sector gear (32);

and step five, opening the sealing cover (7), taking out the culture dish (6) and finishing integral shaking and mixing.