CN111748474A - Elastic soft bottom equal stress strain cell dynamic culture device - Google Patents
Elastic soft bottom equal stress strain cell dynamic culture device Download PDFInfo
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Abstract
The invention discloses an elastic soft bottom equal stress strain cell dynamic culture device, which comprises a shell, wherein a cover plate is hinged to the opening end of the shell and is provided with a lock catch; the shell is used for accommodating at least one area, each area comprises a thrust assembly and a soft bottom equal stress strain six-hole plate, the thrust assembly comprises a jacking mechanism, a supporting plate and a partition plate, the jacking mechanism is connected with the shell through the supporting plate, and the partition plate is positioned above the jacking mechanism; the six-hole plate with the equal stress strain of the soft bottom comprises at least one culture chamber and is placed on a partition plate, and a variable-thickness equal stress strain film with the soft bottom is sealed at the bottom of the culture chamber; the jacking mechanism comprises a power mechanism, a transmission mechanism and an actuating mechanism, the transmission mechanism is driven by the power mechanism to drive the actuating mechanism to move up and down along the partition plate, and the jacking contact of the actuating mechanism enables the soft bottom equal stress strain film to generate equal stress strain. The cell culture quality of the invention has uniformity and stability, the accuracy of experimental results is high, each area can be independently controlled, the automation degree is high, and the supply requirement is met.
Description
Technical Field
The invention belongs to the field of biomedical engineering, and particularly relates to a dynamic culture device for elastic soft-bottom equal-stress strain cells.
Background
The cell culture technology is the most core and basic technology in biotechnology and has been widely applied to various fields such as biology, medicine, research and development of new drugs and the like. In the prior art, a stretching and tension culture system of a soft bottom membrane is usually adopted for in-vitro cell dynamic culture and amplification, but the thickness of the soft bottom membrane is consistent, the sizes of stress strains on cells at different parts are different during dynamic culture of the cells, so that the culture characteristics of the cells at different parts are different, the requirements on cell quality uniformity and stability are difficult to achieve, accurate experimental results are difficult to obtain for experiments such as biological materials and cell compatibility, the degree of automation is low, and the supply requirements of clinical medical cell treatment and medical experiments cannot be met.
Disclosure of Invention
The invention aims to solve the problems that the thickness of a soft bottom film of a cell dynamic culture unit is consistent, different cell culture characteristics of different parts are different due to uneven loading stress, the quality is difficult to achieve uniformity and stability, the accuracy of an experimental result is low, and the culture efficiency is low in the prior art, and provides an elastic soft bottom equal stress strain cell dynamic culture device.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
the invention provides a dynamic cell culture device with elastic soft bottom and equal stress strain, which comprises a shell,
the shell is provided with an upper opening, the side surface of the opening end is hinged with a cover plate, and a lock catch used for locking or unlocking the cover plate relative to the shell is arranged;
a housing containing at least one region, each region comprising:
the thrust assembly comprises a jacking mechanism, a supporting plate and a partition plate, the supporting plate is positioned on a bottom plate of the shell, the jacking mechanism is connected with the supporting plate, and the partition plate is horizontally arranged on the opening side of the shell and positioned above the jacking mechanism;
the six-hole plate with the soft bottom and the equal stress strain is placed on the partition plate and comprises at least one culture chamber for internally placing cells or biological materials, and the bottom of the culture chamber is sealed with a soft bottom and equal stress strain film with variable thickness;
the jacking mechanism comprises a power mechanism, a transmission mechanism and an actuating mechanism, the transmission mechanism can be driven by the power mechanism to drive the actuating mechanism to move up and down along the partition plate, the actuating mechanism is in jacking contact with the soft bottom equal stress strain film, and the soft bottom equal stress strain film generates equal stress strain under the action of jacking contact.
Preferably, the bottom plate of the shell is provided with a plurality of supporting legs, the outer side of the shell is also provided with air holes for gas exchange, and at least one joint fixing plate for connecting a cable is arranged; the cover plate is a transparent cover plate, and a limiting column for limiting all soft bottom equal-stress strain six-hole plates is arranged on the side close to the shell, and at least one limiting column is arranged.
Preferably, the six-hole plate with the soft bottom and equal stress strain further comprises an outer frame and a culture chamber cover plate, the culture chamber is arranged in the outer frame and is relatively fixed, the bottom surface of the outer frame is provided with a through groove corresponding to the center of each culture chamber, and the culture chamber cover plate is used for covering the culture chambers and can be buckled with the outer frame.
Preferably, the soft bottom equal stress strain film is a linear elastic circular film, the thickness is limited within the range of 0.5mm-2mm, and the calculation formula is as follows:
H=A*[(x2+y2)0.5-D/2]+B
in the formula, the plane coordinate of the circle center is (0,0), a plane coordinate system x-o-y is established by taking the circle center as an origin, and x and y are coordinates of positions away from the circle center; h is the film thickness in mm; d is the membrane diameter in mm; A. b is fitting coefficient, A is 6/D2,B=0.5。
Preferably, the transmission mechanism is a lead screw and nut mechanism or a gear and rack mechanism.
Preferably, the actuating mechanism comprises an ejector rod fixing plate which is driven by the transmission mechanism to move up and down, ejector rods with the same number as that of the culture chambers of the corresponding soft-bottom equal-stress strain six-hole plates are mounted on the ejector rod fixing plate, and at least one positioning pin shaft is further mounted on the ejector rod fixing plate.
Preferably, the positioning pin shaft and the positioning frame are mutually nested in a sliding mode to guide so that the ejector rod is aligned to the center of each soft bottom equal-stress strain film above the ejector rod, and the positioning frame is fixedly connected to the shell or the supporting plate.
Preferably, the upper surface of the partition board is provided with a boss for transverse limiting of the soft bottom equal stress strain six-hole plate, the upper surface of the partition board is also provided with concave liquid storage tanks which are aligned with the culture chambers of the corresponding soft bottom equal stress strain six-hole plate or the corresponding ejector rods on the ejector rod fixing plate and have the same number, and the centers of the liquid storage tanks are provided with through holes.
Preferably, the ejector rod is in sealing sliding fit with the through hole.
Preferably, the shell or the supporting plate is also provided with a limiting mechanism for limiting each actuating mechanism.
Compared with the prior art, the invention has the beneficial effects that: the multichannel cell dynamic culture system is combined with a novel soft-bottom equal-stress-strain elastic membrane to perform mechanical application, frequency, displacement and stress-strain regulation and control of the equal-stress-strain dynamic culture unit, each unit can independently operate, an even stress-strain culture environment is provided for dynamic culture and cell amplification, and a test and detection system of an equal-stress-strain field is provided for experiments such as cell compatibility of biological materials. The uniformity and stability of the cell culture quality are high, the experimental result is more accurate, the automation degree is high, and the supply requirements of clinical medical cell treatment and medical experiments are met.
Drawings
FIG. 1 is a schematic view of a first perspective structure of the present invention;
FIG. 2 is a schematic view of a second perspective structure of the present invention;
FIG. 3 is a cross-sectional view A-A of the present invention;
FIG. 4 is a cross-sectional view B-B of the present invention;
FIG. 5 is a rear view of the present invention;
FIG. 6 is a cross-sectional view C-C of the present invention;
FIG. 7 is a D-D cross-sectional view of the present invention;
FIG. 8 is a schematic structural view of a six-hole plate with equal stress strain and soft bottom according to the present invention;
FIG. 9 is a structural diagram of a dynamic culture control system for equal stress strain of soft foundation in example 2 of the present invention;
fig. 10 is a system hardware configuration diagram in embodiment 2 of the present invention;
FIG. 11 is a comparison graph of the radial distribution of displacement at different thicknesses in example 3 of the present invention (center force loading 10 kg);
FIG. 12 is a comparison graph of stress distribution in the radial direction at different thicknesses in example 3 of the present invention (center force loading 10 kg);
FIG. 13 is a comparison graph of the radial distribution of strain at different thicknesses (center force loading) for example 3 of the present invention;
FIG. 14 is a comparison graph of radial distribution of displacement at different thicknesses in example 3 of the present invention (center ejection height 10 mm);
FIG. 15 is a comparison graph of stress distribution in the radial direction at different thicknesses in example 3 of the present invention (center lift height 10 mm);
FIG. 16 is a comparison graph of the radial distribution of strain at different thicknesses (center lift height 10mm) for example 3 of the present invention;
FIG. 17 is a comparison graph of radial distribution of displacement at different thicknesses in example 3 of the present invention (center ejection height 5 mm);
FIG. 18 is a comparison graph of stress distribution in the radial direction at different thicknesses in example 3 of the present invention (center lift-out height 5 mm);
FIG. 19 is a comparison graph of the radial distribution of strain at different thicknesses (center lift-off height 5mm) for example 3 of the present invention;
FIG. 20 is a graph of the thickness variation profile across the membrane in example 3 of the present invention;
FIG. 21 is a radial stress profile (center force loading) of the film in accordance with example 3 of the present invention;
FIG. 22 is a radial distribution plot of strain on the membrane (center force loading) in example 3 of the present invention;
FIG. 23 is a radial stress distribution (center protrusion height 10mm) of the film in example 3 of the present invention;
FIG. 24 is a radial strain distribution plot (center lift height 10mm) on a film in example 3 of the present invention;
FIG. 25 is a radial stress distribution (center lift-off height 5mm) of the film in example 3 of the present invention;
FIG. 26 is a radial strain distribution plot (center lift-off 5mm) on a film in example 3 of the present invention;
FIG. 27 is a top view of a soft underlayer isostress strained film in example 3 of the present invention;
FIG. 28 is a front view of a soft underlayer isostress strained film in example 3 of the present invention.
Description of reference numerals: 1. a housing; 2. locking; 3. a six-hole plate with soft bottom and equal stress strain; 4. a transparent cover plate; 5. a limiting column; 6. air holes are formed; 7. a lower motor sealing cover; 8. a support plate; 9. a thrust bearing; 10. a first fixing hole; 11. a movable block; 12. a screw rod; 13. a ball bearing support frame; 14. a second fixing hole; 15. a positioning frame; 16. a top rod; 17. a mandril fixing plate; 18. a support bar; 19. an upper positioning hole; 20. mounting a sensor fixing frame; 21. a limiting column; 22. a lower sensor mount; 23. a lower positioning hole; 24. a thrust bearing support frame; 25. a coupling; 26. electrifying a sealing cover of the machine; 27. positioning blocks; 28. a partition plate; 29. a partition plate fixing hole; 30. a base plate; 31. a feed screw nut; 32. a ball bearing; 33. a left positioning pin shaft; 34. a right positioning pin shaft; 35. a support plate fixing hole; 36. a motor; 37. supporting legs; 38. a joint fixing plate; 39. a motor drive cable; 40. a sensor signal line; 41. a motor signal line; 42. a joint fixing plate mounting hole; 43. a first region; 44. a horizontal positioning wall; 45. a second region; 46. a vertical positioning wall; 47. a third region; 48. a liquid storage tank; 49. a fourth region; 50. a left locating rack fixing hole; 51. a mounting hole of the ejector rod fixing plate; 52. a right positioning frame fixing hole; 53. a housing bottom plate fixing hole; 54. a culture chamber; 55. a culture chamber cover plate; 56. a soft bottom iso-stress strain film; 57. an outer frame; 58. a motor bracket; 59. an upper limit sensor and a lower limit sensor; 60. positioning the shaft sleeve; 61. and positioning shaft sleeve fixing holes.
Detailed Description
The technical solutions of the present invention are further described in detail below with reference to the drawings and examples, which should not be construed as limiting the present invention.
Example 1:
as shown in figures 1-8, the dynamic cell culture device with elastic soft bottom and equal stress strain comprises a shell,
the shell is provided with an upper opening, the side surface of the opening end is hinged with a cover plate, and a lock catch used for locking or unlocking the cover plate relative to the shell is arranged;
a housing containing at least one region, each region comprising:
the thrust assembly comprises a jacking mechanism, a supporting plate and a partition plate, the supporting plate is positioned on a bottom plate of the shell, the jacking mechanism is connected with the supporting plate, and the partition plate is horizontally arranged on the opening side of the shell and positioned above the jacking mechanism;
the six-hole plate with the soft bottom and the equal stress strain is placed on the partition plate and comprises at least one culture chamber for internally placing cells or biological materials, and the bottom of the culture chamber is sealed with a soft bottom and equal stress strain film with variable thickness;
the jacking mechanism comprises a power mechanism, a transmission mechanism and an actuating mechanism, the transmission mechanism can be driven by the power mechanism to drive the actuating mechanism to move up and down along the partition plate, the actuating mechanism is in jacking contact with the soft bottom equal stress strain film, and the soft bottom equal stress strain film generates equal stress strain under the action of jacking contact.
Wherein, the casing includes shell 1 and bottom plate 30, and shell bottom plate fixed orifices 53 is furnished with to the bottom of shell 1, and the screw passes through shell bottom plate fixed orifices 53 and is connected the fixed casing that forms to have an upper shed with bottom plate 30, and the weight reduction when considering when guaranteeing casing intensity can select to set up the strengthening rib on the casing, if set up on the inner wall of shell 1. The side face of the opening end of the shell 1 is connected with the cover plate 4 through a hinge, meanwhile, the side face of the opening end of the shell 1 and the outer side of the cover plate 4 are also provided with the lock catches 2, and the cover plate 4 can be closed or opened relative to the shell 1 through locking or unlocking the lock catches 2. When the cover plate 4 is locked relative to the shell 1, the shell 1 and the cover plate 4 form a sealed chamber, so that the internal structure can be protected from being polluted or damaged; when the cover plate 4 is opened, the stress strain six-hole plate 3 with a soft bottom and the like can be conveniently taken out or placed to carry out experimental observation or dynamic culture on cells or biological materials in the six-hole plate.
Wherein, the casing is inside including four independently controlled regions, be first region 43 respectively, the second region 45, third region 47 and fourth region 49, every region includes thrust subassembly and soft end six orifice plates 3 of equal stress strain again, the thrust subassembly includes climbing mechanism, backup pad 8 and baffle 28, backup pad 8 is fixed respectively on bottom plate 30, climbing mechanism all fixes the side at the backup pad 8 of longitudinal arrangement, place the casing opening side in the baffle 28 level, and be connected with locating piece 27 on the shells inner wall, baffle 28 is located the climbing mechanism top. All the supporting plates 8 are rectangular plates with different sizes, and can also be arranged by adopting an L-shaped plate or other plane structures for installing the jacking mechanism. As shown in fig. 7, the present embodiment includes five rectangular support plates 8 arranged in a vertical and horizontal direction, and further includes a fixing bar 8f, four vertical support plates 8 are respectively denoted by 8a, 8c, 8d, 8e, and one horizontal support plate 8 is denoted by 8 b. The fixing strip 8f is used for connecting two plates 8b and 8c, supporting plate fixing holes 35 connected with the corresponding supporting plates 8 are distributed on each supporting plate 8, the inner cavity of the shell 1 is equally divided into four mutually isolated spaces by the transverse supporting plate 8b and the two longitudinal supporting plates 8c and 8d, and the first area 43, the second area 45, the third area 47 and the fourth area 49 are respectively positioned in the spaces and are mutually isolated. During installation, the longitudinal support plate 8d is connected with the transverse support plate 8b through a screw, the other longitudinal support plate 8c is connected with the transverse support plate 8b through the fixing strip 8f, the installation accuracy of the longitudinal support plate 8c and the longitudinal support plate 8d is guaranteed, a slotted hole for walking or providing a tool installation space can be formed in the transverse support plate 8b, and the other two longitudinal support plates 8a and 8e are distributed in a reverse symmetrical mode and are fixedly connected with the transverse support plate 8b through the support plate fixing holes 35 in a threaded mode. The whole supporting plate 8 can make the structure more stable, and the shell can surround four areas to be protected from the outside.
It should be noted that the first area 43, the second area 45, the third area 47 and the fourth area 49 may be arranged in a symmetrical or array manner or in any other longitudinal and transverse arrangement manner, the partitions 28 may be independent individuals or connected with each other to form one or more modules, the support plates 8 may be combined in any number or shape according to the distribution of the areas, for example, one support plate 8 may be used for each area alone or a plurality of areas share the same support plate 8, etc.
The six-hole plate 3 with the equal stress strain of the soft bottom is placed on the partition plate 28 and comprises six annular culture chambers 54, the culture chambers 54 can be square or in other shapes, and the bottoms of the culture chambers 54 are sealed with the variable-thickness equal stress strain films 56 with the soft bottom. The culture chamber 54 is used for placing cells and biological materials. Compared with the equal-thickness membrane, the variable-thickness soft-bottom equal-stress strain membrane 56 can generate uniform stress strain when the center is subjected to jacking load, so that the culture environment of cells or biological materials is more stable, and the accuracy of experimental results is improved.
The jacking mechanism comprises a power mechanism, a transmission mechanism and an execution mechanism, the power mechanism comprises a motor 36, the motor 36 is fixed on a motor support 58, and the motor support 58 is fixed with the support plate 8 through screws. In consideration of the service life and safety of the motor 36, the motor 36 can be covered by the lower motor sealing cover 7 and the upper motor sealing cover 26, so that the safety problem caused by the moisture of the motor 36 can be avoided. The transmission mechanism can be driven by the motor 36 to drive the actuating mechanism to move up and down relative to the partition plate 28, and the actuating mechanism is contacted with the soft bottom equal stress strain film 56 in a jacking manner when ascending, so that the soft bottom equal stress strain film 56 generates equal stress strain, cells and biological materials in the culture chamber 54 are in a uniform stress strain culture environment, the obtained cell culture quality has good uniformity and stability, and the experimental result is more accurate.
In the embodiment, a plurality of supporting legs are arranged below a bottom plate of a shell, air holes for gas exchange are formed in the outer side of the shell, and at least one connector fixing plate for connecting a cable is arranged on the outer side of the shell; the cover plate is a transparent cover plate, and a limiting column for limiting all soft bottom equal-stress strain six-hole plates is arranged on the side close to the shell, and at least one limiting column is arranged.
Wherein, four supporting legs 37 are installed to bottom plate 30 lower surface four corners and are convenient for the leveling make the device more stable, and the outer side is close to the six orifice plates 3 of soft end isostress strain about the shell 1 open end position and has still opened bleeder vent 6, plays gas exchange's effect, avoids the cell in the culture room 54 to be in oxygen deficiency or acidic environment growth influence culture quality or cultivation efficiency. Meanwhile, the outer side of the shell 1 is further fixed with two joint fixing plates 38 through joint fixing plate mounting holes 42, the joint fixing plates 38 are all provided with motor driving cables 39, sensor signal lines 40 and motor signal lines 41 for being externally connected with control equipment or power supply equipment, and the two joint fixing plates 38 can be arranged at the outer side of the shell 1 close to the lower part to reduce wiring distance. The cover plate 4 is of a transparent structure, so that the cultivation dynamics in the internal soft-bottom equal-stress-strain six-hole plate can be conveniently and visually observed, the limiting columns 5 are arranged on the plane of the cover plate 4 close to the shell 1, after the cover plate 4 is locked, the effect of limiting the upper part and the lower part of each soft-bottom equal-stress-strain six-hole plate 3 is achieved, and the accuracy of a cultivation result is prevented from being influenced by uneven stress caused by movement.
The number and the installation position of the support legs 37 and the joint fixing plate 38 may be selected or not provided according to actual requirements. The air holes 6 can also be arranged on the whole outer ring side of the opening end of the shell 1, and one or more air holes can be adopted. The cover plate 4 can also be limited by adopting a mode of directly contacting with the soft bottom equal stress strain six-hole plate 3.
In this embodiment, the six-hole plate with soft bottom and equal stress strain further comprises an outer frame and a culture chamber cover plate, the outer frame is arranged in the culture chamber and is relatively fixed, the bottom surface of the outer frame is provided with through grooves corresponding to the centers of the culture chambers, and the culture chamber cover plate is used for covering the culture chambers and can be buckled with the outer frame.
In this embodiment, the soft-bottom equal-stress strain film is a linear elastic circular film, the thickness limit range is 0.5mm-2mm, and the calculation formula is as follows:
H=A*[(x2+y2)0.5-D/2]+B
in the formula, the plane coordinate of the circle center is (0,0), a plane coordinate system x-o-y is established by taking the circle center as an origin, and x and y are coordinates of positions away from the circle center; h is the film thickness in mm; d is the membrane diameter in mm; A. b is fitting coefficient, A is 6/D2,B=0.5。
The six-hole plate 3 with equal stress and strain at the soft bottom comprises six annular culture chambers 54 and the equal stress and strain membranes 56 at the soft bottom corresponding to the culture chambers 54, and further comprises a square outer frame 57 and a culture chamber cover plate 55 which can be buckled with the outer frame 57, wherein the culture chamber cover plate 55 can adopt a transparent structure, so that the observation is convenient. Six culture chambers 54 are arranged in an outer frame 57 at equal intervals and fixed with the outer frame 57 into a whole, through grooves with the same inner diameter and corresponding to the centers of the six culture chambers are formed in the bottom surface of the outer frame 57, and a culture chamber cover plate 55 plays roles of ventilating, shielding bacteria and protecting the culture chambers 54, so that the culture chambers 54 are prevented from being polluted, leaked or damaged by cells or biological materials in the culture chambers 54.
The isostress strain elastic membrane 56 may be changed in size and shape according to the actual application, so that the curved surface structure of the isostress strain elastic membrane 56 and the structure of the incubation chamber 54 may be changed accordingly. The six-well plate 3 with equal stress strain of soft bottom can also be arranged by connecting and fixing a plurality of culture chambers 54 or by directly fastening the cover 55 of the culture chamber with each culture chamber 54.
In this embodiment, the transmission mechanism is a screw nut mechanism or a rack and pinion mechanism.
In this embodiment, the actuator includes a top rod fixing plate driven by the transmission mechanism to move up and down, the top rod fixing plate is provided with top rods with the same number as that of the culture chambers of the corresponding soft bottom equal stress strain six-hole plate, and the top rod fixing plate is further provided with at least one positioning pin.
The transmission mechanism is a screw and nut mechanism and comprises a screw 12 and a screw nut 31, and the screw nut 31 can move up and down under the driving of the screw 12. Two ends of the screw rod 12 are provided with a thrust bearing 9 and a ball bearing 32, the thrust bearing 9 is fixed in the thrust bearing support frame 24, the ball bearing 32 is fixed in the ball bearing support frame 13, and the thrust bearing support frame 24 and the ball bearing support frame 13 are fixedly connected with the support plate 8 through the first fixing hole 10 and the second fixing hole 14 by screws. The motor 36 is connected with the screw rod 12 through the coupler 25, and the screw rod nut 31 on the screw rod 12 is fixedly connected with the actuating mechanism. The actuating mechanism comprises a movable block 11, a support rod 18 is arranged on the movable block 11, the support rod 18 is fixedly connected with a mandril fixing plate 17 through a mandril fixing plate mounting hole 51 by screws, and six mandrils 16 which are arranged at equal intervals and respectively aligned with six culture chambers 54 of the soft bottom equal stress strain six-hole plate 3 are arranged on the mandril fixing plate 17. The feed screw nut 31 is fixedly connected with the movable block 11 through a screw, the feed screw nut mechanism drives the ejector rod fixing plate 17 and the ejector rod 16 on the ejector rod fixing plate to move up and down under the driving of the motor 36 and simultaneously lift up and contact or keep away from the corresponding soft bottom equal stress strain film 56, and the culture environment can be changed by adjusting the stroke or the frequency.
It should be noted that the transmission mechanism may also adopt a rack and pinion mechanism or other mechanisms for performing linear motion transmission, such as a hydraulic transmission mechanism, a cam mechanism, and a link mechanism.
In this embodiment, the positioning pin and the positioning frame are slidably nested to guide the ejector rod to align with the center of each soft bottom equal-stress strain film above the ejector rod, and the positioning frame is fixedly connected to the housing or the supporting plate.
Wherein, all install locating rack 15 on locating block 27's the lateral wall and the longitudinal support board 8c or locating block 27's the lateral wall and the longitudinal support board 8d in each region, adorn respectively in the left and right sides, left locating rack fixed orifices 50 has been seted up on left side locating rack 15, right locating rack fixed orifices 52 has been seted up on right side locating rack 15, shell 1 and longitudinal support board 8c or 8d go up to open the mounting hole that corresponds with left locating rack fixed orifices 50 and right locating rack fixed orifices 52, two location axle sleeve fixed orifices 61 have still been seted up on each locating rack 15, location axle sleeve 60 is fixed through location axle sleeve fixed orifices 61 and locating rack 15 threaded connection. Two positioning pin shafts, namely a left positioning pin shaft 33 and a right positioning pin shaft 34, are arranged at two ends of the ejector rod fixing plate 17, and the left positioning pin shaft 33 and the right positioning pin shaft 34 are respectively embedded into the positioning shaft sleeves 60 in the positioning frame 15, so that the up-and-down movement guiding positioning and stabilizing effects are achieved, and the running precision and stability of the ejector rod 16 are ensured.
The top rod may be guided to be aligned with the center of each of the soft bottom equal stress strain films above the top rod by using a guide rail slider mechanism.
In this embodiment, the partition plate has a boss on the upper surface thereof for laterally limiting the soft-bottom iso-stress-strain six-hole plate, and also has concave liquid storage tanks aligned with the culture chambers of the corresponding soft-bottom iso-stress-strain six-hole plate or the corresponding ejector rods on the ejector rod fixing plates and having the same number, and through holes are formed in the centers of the liquid storage tanks.
As shown in fig. 6, the partition plate 28 is located above the ejector rod fixing plate 17, the partition plates 28 in each region are integrated and fixed to the housing 1, the transverse support plate 8b, the longitudinal support plate 8c, and the longitudinal support plate 8d through the partition plate fixing holes 29, the partition plate 28 is provided with bosses including horizontal positioning walls 44 and vertical positioning walls 46 which are distributed in a cross shape, and the horizontal positioning walls 44 or the vertical positioning walls 46 may also adopt other distribution forms for transversely limiting the soft bottom equal stress strain six-hole plate 3. Six rectangular concave liquid storage tanks 48 which are aligned with the culture chambers of the soft-bottom equal-stress-strain six-hole plate and have the same number are also arranged on the surface of the partition plate 28, a boss is arranged in the center of each liquid storage tank 48 and penetrates through the liquid storage tanks 48 through the through holes, and the liquid storage tanks 48 can also be cylindrical or other polygonal grooves. When the membrane of the soft bottom equal stress strain six-hole plate 3 is broken, the cell or other biological material liquid can directly flow into the liquid storage groove 48 without polluting and corroding other structures. And the boss at the center of the liquid storage groove 48 can block the flowing-out cells or other biological material liquid in the concave part of the liquid storage groove 48, so that the clapboard 28 can be conveniently cleaned directly or disassembled for cleaning.
In this embodiment, the ejector pin is in sealing sliding fit with the through hole.
Wherein, the ejector rod 16 is matched with the through hole at the center of the liquid storage tank 48 in a sliding way to form a sliding sealing structure. When the membrane of the six-hole plate 3 with the soft bottom and equal stress strain is broken while the ejector rod 16 moves up and down, the cell or other biological material liquid cannot flow out of the liquid storage tank 48 along the ejector rod 16 or the through hole on the liquid storage tank 48 to pollute or corrode other structures.
In this embodiment, the housing or the support plate is further provided with a limiting mechanism for limiting each actuating mechanism.
In order to ensure the safety and precision of the up-and-down movement of the actuating mechanism of the jacking mechanism, a limiting mechanism is further arranged beside each region, if an upper limiting sensor 59 and a lower limiting sensor 59 are arranged on the shell 1, the upper limiting sensor 59 and the lower limiting sensor 59 comprise an upper limiting sensor and a lower limiting sensor, the upper limiting sensor is connected with the upper sensor fixing frame 20, the lower limiting sensor is connected with the lower sensor fixing frame 22, the upper sensor fixing frame 20 and the lower sensor fixing frame 22 can be fixed through screw connection through an upper positioning hole 19 and a lower positioning hole 23 on the transverse supporting plate 8b, kidney-shaped grooves for position adjustment are formed in the two fixing frames, and the upper limiting sensor or the lower limiting sensor can be respectively adjusted to be locked and fixed after being in a. The limiting column 21 is fixedly connected with the supporting rod 18, the limiting column 21 is an induction structure of an upper limiting sensor 59 and a lower limiting sensor 59, the upper limiting sensor 59 and the lower limiting sensor 59 can adopt position sensors such as photoelectric sensors or travel switches, the upper sensor fixing frame 20 and the lower sensor fixing frame 22 can also be arranged on other supporting plates 8, and the limiting column 21 is arranged at a corresponding position along with the change of the positions of the upper limiting sensor 59 and the lower limiting sensor 59.
When the cell dynamic culture is carried out, the motor 36 works to drive the screw rod 12 to rotate, the screw rod nut 31 drives the support rod 18 to move up and down while moving up and down under the driving of the screw rod 12, the limit column 21 is fixedly connected with the support rod 18, the limit column 21 moves up and down simultaneously, and when the limit column 21 senses an upper limit sensor or a lower limit sensor, the forward and reverse rotation of the motor 36 is switched or the motor 36 is controlled to stop rotating. The limiting mechanism can control the stroke of the jacking mechanism to adapt to different loading requirements of a cell culture environment and carry out limiting protection.
Example 2:
as shown in FIGS. 1 to 10, this example includes a dynamic cell culture apparatus with elastic soft bottom and other stress strain similar to that of example 1, and a control box electrically connected to the culture apparatus. As shown in fig. 9, the control box comprises a power circuit, a display screen, an electric meter, a driving circuit and a single chip circuit, wherein the power circuit is used for supplying power to the display screen, the electric meter, the driving circuit and the single chip circuit, the single chip circuit controls the motor to drive the screw nut transmission mechanism in the culture box to act through the driving circuit, so that the six-hole plate with equal stress strain at the soft bottom is subjected to equal stress strain to perform dynamic culture of cells or biological materials, and meanwhile, the single chip circuit can perform real-time information interaction with the display screen and the electric meter.
Further, as shown in fig. 10, the single chip microcomputer circuit adopts an STM32 single chip microcomputer, the driving circuit is driven by a 4-way motor, the motor is a 4-way stepping motor, for example, a 4-way servo stepping motor is adopted, and a photoelectric sensor is adopted for limiting, and the display screen is a touch screen. The control box is supplied with power by 220V mains supply and carries out electric leakage or short circuit protection through an air switch and a circuit breaker. The AC-DC conversion is carried out through the power management circuit, and required voltages are provided for 4-path motor drive, 4-path stepping motors, an STM32 single chip microcomputer, the photoelectric sensor and the touch screen, for example, 24V direct current voltage is provided for the motors, 5V voltage is provided for the touch screen and the like. The STM32 single chip microcomputer drives and controls 4 paths of stepping motors through 4 paths of motors to drive a lead screw nut transmission mechanism in the culture device to move so that the six stress-strain pore plates with equal stress strain at the soft bottom and the like generate equal stress strain to carry out dynamic culture on cells or biological materials, and simultaneously receives photoelectric sensor signals to carry out position detection. The touch screen can display dynamic information and the like of the power consumption and the transmission mechanism in real time and carry out real-time information interaction with the STM32 single chip microcomputer, and meanwhile, the touch screen can also control the STM32 single chip microcomputer circuit through touch control or key operation to control the start and stop of the motor 36 of the culture device and the like.
The working principle of the embodiment is as follows:
1) the transparent cover plate 4 is opened, the soft-bottom equal stress strain six-hole plate 3 filled with the cell or other biological material culture sample is placed on the partition plate 28, and the soft-bottom equal stress strain six-hole plate 3 is transversely limited by the horizontal positioning wall 44 and the vertical positioning wall 46. After the transparent cover plate 4 is closed, the limiting column 5 on the cover plate 4 can limit the soft bottom equal stress strain six-hole plate 3 up and down, and the shell 1 and the transparent cover plate 4 are locked firmly by the lock catch 2;
2) parameters are set on the display screen, and the motor 36 in the button control culture device is started to drive the ejector rod 16 to act to apply load to the soft bottom equal stress strain film 56 to generate a preset equal stress strain environment for dynamic culture. The operation mode has two modes of continuous culture and timing culture and can be set before operation;
3) the transparent cover plate 4 is opened as required, the six-hole plate 3 with equal stress and strain at the soft bottom is taken out and placed in an aseptic and ultra-clean environment, the culture chamber cover plate 55 is opened to take out an internal sample for detection, then the culture chamber cover plate 55 is covered, the six-hole plate 3 with equal stress and strain at the soft bottom is placed back into the dynamic culture device for the stress and strain cells at the elastic soft bottom and the like to be dynamically cultured together with other samples, if the samples in the six-hole plate 3 with equal stress and strain at the soft bottom in a certain area all complete parameter detection, the six-hole plate 3 with equal stress and strain at the soft bottom is moved out, the six-hole plate 3 with equal stress and strain at the soft bottom in other areas can continue to be.
Example 3:
as further illustrated in FIGS. 11-28 for the isostrained elastic film 56 of example 1, the isostrained elastic film 56 was modeled and analyzed using COMSOL finite element analysis software. Under the determined parameters and constraint conditions, the upper surface of the iso-stress-strain elastic membrane 56 is ensured to have the same anisotropic stress and strain, and the thickness distribution of the iso-stress-strain elastic membrane 56 is calculated. And 2D-membrane units are adopted to establish a membrane structure, so that the parameterized definition of the thickness is realized conveniently. The membrane diameter D was 35mm, the membrane thickness H varied: 0.5mm-2mm, material parameters of the film: the elastic modulus is 1.2GPa, the Poisson ratio is 0.48, and the material is linear elastic material, such as transparent silica gel material, and polymer material such as gel and the like can also be adopted. The initial displacement of membrane structure is 0, and the membrane circumference is fixed, and there are two kinds of load application methods at circular membrane center: the first is loading of central force, the acting force is upward vertically, and the magnitude of the force is 10 kg; the second and third are center displacement loading with center ejection heights of 10mm and 5mm, respectively.
1) Calculation of equal thickness
The diameter D of the film is 35mm, four thicknesses of 0.5mm, 1mm, 1.5mm and 2mm in the range of 0.5mm-2mm of the film thickness H are taken for simulation analysis, the result of numerical simulation and the actual result are in an order of magnitude range, the model is accurate and reliable, and the analysis result is as follows:
a. the central force is loaded, and the acting force is 10 kg. As shown in fig. 11-13, which are comparative graphs of the radial distribution of displacement, stress and strain along the film at different thicknesses, the graphs correspond to the thickness of 1mm, 1.5mm and 2mm from top to bottom at 5mm on the abscissa in each graph. When the thickness is 0.5mm, the maximum displacement of the membrane is reduced from 37.4mm to 3.4mm, namely the ejection height of the membrane ranges from 3.4mm to 37.4mm under the working condition, and the maximum displacement of the membrane structure is too large, so that the analysis of the thickness is excluded;
b. the center displacement is loaded, and the center ejection height is 10 mm. As shown in fig. 14-16, which are graphs comparing the radial distribution of displacement, stress and strain along the film at different thicknesses, the graphs corresponding to the thicknesses of 0.5mm, 1mm, 1.5mm and 2mm are sequentially from top to bottom at 5mm on the abscissa in fig. 14, and the graphs corresponding to the thicknesses of 2mm, 1.5mm, 1mm and 0.5mm are sequentially from top to bottom at 5mm on the abscissa in fig. 15 and 16;
c. the center displacement is loaded, and the center ejection height is 5 mm. As shown in fig. 17-19, which are graphs comparing the radial distribution of displacement, stress and strain along the film at different thicknesses, fig. 17 shows the thicknesses of 0.5mm, 1mm, 1.5mm and 2mm from top to bottom at 5mm on the abscissa, and fig. 18 and 19 show the corresponding curves of the thicknesses of 2mm, 1.5mm, 1mm and 0.5mm from top to bottom at 5mm on the abscissa.
In the equal thickness simulation analysis, the numerical simulation results of different thicknesses in three cases are shown as follows: when the center of the membrane structure is subjected to an applied load, the displacement, the stress and the strain at the center are all large and gradually decrease along the direction from the center to the periphery, and the displacement, the stress and the strain also show a decreasing trend along with the increase of the thickness. From the results, it is known that when the film thickness is uniform, the stress and strain on the film cannot be made uniform in the range of 0.5mm to 2mm, and particularly, the difference between the stress and strain on the film is significant in the range of 5mm from the center of the film. It follows that in order to have the same stress and strain on the film, it is not feasible to have a uniform thickness.
2) Variable thickness optimization calculation
According to the simulation result of the equal thickness, the stress and the strain of the central area are larger, and the stress and the strain are smaller as the central area is farther away, so that the thickness change of the variable thickness structure ensures that the thickness of the central area is larger and the continuous gradient change of the central area is smaller, and the thickness limit range of 0.5mm-2mm is required to be met. Thus, again with a film diameter D of 35mm, assuming a thickness H, with plane coordinates at the center of the circle being (0,0), and the origin of the plane coordinate system x-o-y at the center of the circle, a thickness-variation expression is established as:
H=A*[(x2+y2)0.5-35/2]+B
in the formula, the plane coordinate of the circle center is (0,0), a plane coordinate system x-o-y is established by taking the circle center as an origin, and x and y are coordinates of positions away from the circle center; h is the film thickness in mm, and A, B is the fitting coefficient; a is 6/(35 × 35), and B is 0.5.
According to the formula, the change curved surface of the thickness of the iso-stress strain elastic membrane 56 in the X and Y directions is obtained, as shown in fig. 20. The three-dimensional model is shown in fig. 27 and 28.
The optimized variable-thickness membrane structure is adopted for simulation analysis, and the analysis result is as follows:
a. the central force is loaded, and the acting force is 10 kg. FIGS. 21-22 show the radial distribution of strain and stress on the film, respectively;
b. the center displacement is loaded, and the center ejection height is 10 mm. 23-24, which are radial profiles of strain, stress on the membrane, respectively;
c. the center displacement is loaded, and the center ejection height is 5 mm. As shown in fig. 25-26, the strain and stress on the film are plotted along the radial direction, respectively.
When the optimized variable-thickness membrane structure is adopted, the stress and strain distribution on the membrane becomes uniform under three conditions, the maximum stress and the maximum strain of the variable-thickness membrane structure adopted under the same load or displacement are increased compared with the maximum stress and the maximum strain of the structure with the uniform thickness of 2mm, and the integral stress and strain become more uniform.
In conclusion, the equal-thickness membrane structure is difficult to realize equal-stress strain, and the calculation and analysis of the optimized variable-thickness expression result show that the stress and the strain on the optimized variable-thickness membrane structure are uniform, and under the condition of the same stress strain, the jacking stroke of the variable-thickness membrane structure is smaller, the reaction is more sensitive, the efficiency is higher, and the application requirement of the equal-stress strain culture environment is met.
The above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same, and those skilled in the art can make various corresponding changes and modifications according to the present invention without departing from the spirit and the essence of the present invention, but these corresponding changes and modifications should fall within the protection scope of the appended claims.
Claims (10)
1. The utility model provides a soft end isostress strain cell dynamic culture device of elasticity, includes the casing, its characterized in that:
the shell is provided with an upper opening, the side surface of the opening end is hinged with a cover plate, and a lock catch used for locking or unlocking the cover plate relative to the shell is arranged;
the housing, accommodates at least one area, each area including:
the thrust assembly comprises a jacking mechanism, a supporting plate and a partition plate, the supporting plate is positioned on a bottom plate of the shell, the jacking mechanism is connected with the supporting plate, and the partition plate is horizontally arranged on the opening side of the shell and positioned above the jacking mechanism;
the six-hole plate with the soft bottom and the equal stress strain is placed on the partition plate and comprises at least one culture chamber for internally placing cells or biological materials, and the bottom of the culture chamber is sealed with a soft bottom and equal stress strain film with variable thickness;
the jacking mechanism comprises a power mechanism, a transmission mechanism and an execution mechanism, the transmission mechanism can be driven by the power mechanism to drive the execution mechanism to move up and down along the partition plate, the execution mechanism is in jacking contact with the soft bottom equal stress strain film, and the soft bottom equal stress strain film generates equal stress strain under the action of the jacking contact.
2. The dynamic cell culture device with elastic soft bottom and equal stress strain as claimed in claim 1, wherein: the bottom plate of the shell is provided with a plurality of supporting legs, the outer side of the shell is also provided with air holes for gas exchange, and at least one joint fixing plate for connecting a cable is arranged; the cover plate is a transparent cover plate, a limiting column for limiting all the soft bottom equal-stress-strain six-hole plates is arranged on the side, close to the shell, and at least one limiting column is arranged.
3. The dynamic cell culture device with elastic soft bottom and equal stress strain as claimed in claim 1, wherein: the six-hole plate with the soft bottom and equal stress strain further comprises an outer frame and a culture chamber cover plate, the culture chamber is arranged in the outer frame and is relatively fixed, the bottom surface of the outer frame is provided with a through groove corresponding to the center of each culture chamber, and the culture chamber cover plate is used for covering the culture chambers and can be buckled with the outer frame.
4. The dynamic cell culture device with elastic soft bottom and equal stress strain as claimed in claim 1, wherein: the soft bottom equal stress strain film is a linear elastic circular film, the thickness limit range is 0.5mm-2mm, and the calculation formula is as follows:
H=A*[(x2+y2)0.5-D/2]+B
in the formula, the plane coordinate of the circle center is (0,0), a plane coordinate system x-o-y is established by taking the circle center as an origin, and x and y are coordinates of positions away from the circle center; h is the film thickness in mm; d is the membrane diameter in mm; A. b is fitting coefficient, A is 6/D2,B=0.5。
5. The dynamic cell culture device with elastic soft bottom and equal stress strain as claimed in claim 1, wherein: the transmission mechanism is a screw nut mechanism or a gear rack mechanism.
6. The dynamic cell culture device with elastic soft bottom and equal stress strain as claimed in claim 1, wherein: the actuating mechanism comprises an ejector rod fixing plate driven by the transmission mechanism to move up and down, ejector rods with the same number as that of culture chambers of the corresponding soft-bottom equal-stress strain six-hole plate are mounted on the ejector rod fixing plate, and at least one positioning pin shaft is further mounted on the ejector rod fixing plate.
7. The dynamic cell culture device with elastic soft bottom and equal stress strain as claimed in claim 6, wherein: the positioning pin shaft and the positioning frame are mutually nested in a sliding mode to guide, so that the ejector rod is aligned to the center of each soft bottom equal-stress strain film above the ejector rod, and the positioning frame is fixedly connected to the shell or the supporting plate.
8. The dynamic cell culture device with elastic soft bottom and equal stress strain as claimed in claim 6, wherein: the upper surface of the partition board is provided with a boss for transversely limiting the soft bottom equal stress strain six-hole plate, the upper surface of the partition board is also provided with concave liquid storage tanks which are aligned with the culture chambers of the corresponding soft bottom equal stress strain six-hole plate or the corresponding ejector rods on the ejector rod fixing plate and have the same number, and the centers of the liquid storage tanks are provided with through holes.
9. The dynamic cell culture device with elastic soft bottom and equal stress strain as claimed in claim 8, wherein: the ejector rod is in sealing sliding fit with the through hole.
10. The dynamic cell culture device with elastic soft bottom and equal stress strain as claimed in claim 1, wherein: and the shell or the supporting plate is also provided with a limiting mechanism for limiting each actuating mechanism.
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