CN113604358A

CN113604358A - Cell loading device capable of replacing plane force and pneumatic control method

Info

Publication number: CN113604358A
Application number: CN202110866791.6A
Authority: CN
Inventors: 徐振邦; 夏明一; 周成波
Original assignee: Changchun Institute of Optics Fine Mechanics and Physics of CAS
Current assignee: Changchun Institute of Optics Fine Mechanics and Physics of CAS
Priority date: 2021-07-29
Filing date: 2021-07-29
Publication date: 2021-11-05

Abstract

The invention provides a cell loading device capable of replacing plane force, which comprises a transparent cover, a culture plate, a loading platform assembly, a loading plate and a loading base, wherein the transparent cover, the culture plate, the loading platform assembly, the loading plate and the loading base are sequentially arranged from top to bottom; the culture plate is provided with a cell culture hole, the cell culture hole is used for bearing cells by arranging a loading membrane at the bottom of the hole, and the culture plate is isolated from the external environment by a transparent cover; the loading base is a cavity structure with an opening at the top; a culture plate packaging cavity; a loading plate is arranged in the cavity; the first loading platform and the second loading platform are selectively connected with the loading plate; when the first type of loading platform is arranged on the loading plate, the loading membrane is subjected to two-dimensional force, and when the second type of loading platform is arranged on the loading plate, the loading membrane is subjected to one-dimensional force. According to the invention, the loading membrane is arranged on the cell culture hole, so that the loading membrane can be conveniently and independently replaced; simple structure also is convenient for change loading platform. The loading platform assembly realizes the replacement of the stress condition of the loading membrane.

Description

Cell loading device capable of replacing plane force and pneumatic control method

Technical Field

The invention relates to the field of cell mechanics, in particular to a cell loading device capable of replacing plane force and a pneumatic control method.

Background

Simulating the stress condition of cells in vivo under in vitro experimental environment becomes the current trend of cell research. Because the action of mechanical factors and other factors on cells is difficult to distinguish in vivo cell mechanics experiments, at present, a cell loading device is mostly adopted in a laboratory to apply strain to the cells, different types of force loading in a common loading device need different loading plates, for example, in a plane force cell loading device, the loading of one-dimensional force and two-dimensional force needs to replace an integral loading plate, and if any part of the loading plate is damaged, the integral exchange is needed, so that the cost is higher.

It is therefore desirable to provide an alternative planar force cell loading plate to address the above problems.

Disclosure of Invention

The present invention provides a loading device for planar cells to solve the above problems.

In order to provide mechanical loading in a plane for cells to simulate the single stress condition in vivo, the invention adopts the following specific technical scheme:

a cell loading device capable of replacing plane force comprises a transparent cover, a culture plate, a loading platform assembly, a loading plate and a loading base which are sequentially arranged from top to bottom;

the culture plate comprises a cell culture hole and a loading membrane arranged at the bottom of the cell culture hole, wherein the loading membrane is used for bearing cells, and the cell culture hole is isolated from the external environment through the transparent cover;

the loading base is a cavity with an opening at the top;

the culture plate encapsulates the cavity;

the loading plate is arranged in the cavity;

the loading plate is provided with loading plate air holes penetrating through the thickness;

the loading platform assembly comprises a first loading platform and a second loading platform, and the first loading platform and the second loading platform are alternatively connected with the loading plate;

when the first loading platform is arranged on the loading plate and is positioned below the loading film, the loading film is subjected to two-dimensional force; when the second loading platform is arranged on the loading plate and is positioned below the loading film, the loading film is subjected to one-dimensional force.

The invention can obtain the following technical effects:

the culture plate of the loading device for the plane force cells provided by the invention is provided with a plurality of cell culture holes, and the loading membrane is integrated in the cell culture holes, so that compared with the prior art in which a whole loading membrane is used for separating the loading plate from the culture plate, the loading membrane can be conveniently and independently replaced according to the requirement; the loading plate is only required to be placed on the loading base, in other words, the loading plate and the loading base are movably disassembled, so that mechanical loading in one-dimensional and two-dimensional directions in a plane can be realized, and the mechanical loading can be realized only by replacing the loading platform in the loading plate; the device has small integral volume and compact and simple structure, and reduces the production cost. In addition, the device can be put into a cell constant temperature incubator for use, so that the cell culture with dynamic mechanical loading has feasibility. The detachable structure of the invention ensures that the processing procedure of the parts is simpler and the production cost is reduced. When the loading platform is damaged, only the loading platform needs to be replaced, so that the maintenance cost is lower, and the consumed time is less.

Drawings

Fig. 1 is a schematic structural diagram of a loading device according to an embodiment of the present invention;

fig. 2 is an exploded view of a loading device according to an embodiment of the present invention;

FIG. 3 is a schematic diagram illustrating a pre-operation state of a first loading platform according to an embodiment of the present invention;

FIG. 4 is a schematic diagram illustrating a first loading platform according to an embodiment of the present invention in a working state;

FIG. 5 is a plan view of a first loading platform provided in an embodiment of the present invention;

FIG. 6 is a plan view of a second loading platform provided by the embodiment of the invention;

FIG. 7 is a schematic structural diagram of a transparent cover provided in an embodiment of the present invention;

FIG. 8 is a schematic front view of a culture plate according to an embodiment of the present invention;

FIG. 9 is a schematic view of the bottom structure of a culture plate according to an embodiment of the present invention;

FIG. 10 is a schematic structural diagram of a seal ring provided in accordance with an embodiment of the present invention;

FIG. 11 is a schematic structural diagram of a loading plate provided in an embodiment of the present invention;

FIG. 12 is a schematic structural diagram of a first loading platform provided in an embodiment of the present invention;

FIG. 13 is a schematic structural diagram of a second loading platform provided in the embodiment of the present invention;

FIG. 14 is a schematic front view of a loading base according to an embodiment of the present invention;

FIG. 15 is a schematic bottom view of a loading base according to an embodiment of the present invention;

FIG. 16 is a schematic diagram of a pneumatic system provided by an embodiment of the present invention;

fig. 17 is a flowchart illustrating a control method of a pneumatic system according to an embodiment of the present invention.

Wherein the reference numerals include: 11. a transparent cover; 12. culturing the plate; 13. a seal ring; 14. a load table assembly; 15. loading a base; 16. a loading plate;

111. chamfering the transparent cover;

121. chamfering the culture plate; 122. a cell culture well; 123. a rib plate on the front surface of the culture plate; 124. loading a film; 125. culturing a plate boss; 126. the rib plate on the back side of the culture plate;

131. an inner ring step; 132. the bottom of the sealing ring;

141. a first loading table; 142. a second loading table;

151. a boss of the air chamber; 152. a sealing ring mounting surface; 153. a positive pressure pump connection port; 154. a pressure sensor connector; 155. a negative pressure pump connector; 156. a mounting groove of a clamping tool;

161. a loading platform mounting hole; 162. loading plate air holes; 163. a grabbing port;

1411. a first loading table boss; 1412. a first loading surface; 1421. a second loading platform boss; 1422. a second loading surface;

61. a pneumatic device; 62. a pressure sensor; 63. a controller; 64. a cell loading device; 611. a negative pressure solenoid valve; 612. a negative pressure flow valve; 613. a negative pressure stabilizing bottle; 614. a negative pressure filter; 615. a negative pressure pump; 616. a positive pressure pump; 617. a positive pressure filter; 618. a positive pressure stabilizing bottle; 619. a positive pressure flow valve; 620. a positive pressure solenoid valve.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail below with reference to the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not to be construed as limiting the invention.

Referring to fig. 1-15, the loading device is used for providing in-plane mechanical loading for cells to simulate a single stress condition in vivo, and can realize multiple mechanical loading waveforms in two directions, as shown in fig. 1, assuming that the direction of an x-axis is a first direction, the direction of a y-axis is a second direction, and a z-axis is orthogonal to the x-axis and the y-axis.

As shown in FIGS. 1-2, the loading device mainly comprises a loading platform assembly 14 providing two kinds of loading platforms, a loading plate 16 for mounting the two kinds of loading platforms, and a transparent cover 11 for isolating from the external environment, a culture plate 12 for cell culture, and a loading base 15 for providing support and air chamber, which are sequentially arranged from top to bottom. The plate 12 may be provided with cell culture wells 122, six cell culture wells 122 being exemplified herein.

Wherein the cell culture wells 122 are used for carrying cells by arranging a loading membrane 124 at the bottom of the wells, and the cell culture wells 122 in the culture plate 12 are 6 separate cell culture chambers in which the cells are cultured. The cell culture hole 122 is a hollow cylinder with a top and a bottom removed, the inner wall surface of the cylinder is fixedly connected with an annular culture plate boss 125, the culture plate boss 125 extends to the radial inner part of the cylinder, the function of the culture plate boss 125 is to fix the loading membrane 124 on the cell culture hole 122, and the loading membrane 124 forms a seal with the periphery of the cell culture hole 122 positioned at the upper part of the culture plate boss 125. The cell culture hole 122 and the loading membrane 124 located under the projection 125 of the culture plate form a dome space in the shape of a bottle cap, which can be fitted over the top of the loading table to facilitate the positioning of the loading table. The culture plate front ribs 123 and the culture plate back ribs 126 in the culture plate 12 are connected between the culture plate main body and the cell culture holes 122 and between two adjacent cell culture holes, that is, the culture plate 12 is a flat plate on the plane of the loading membrane 124 as a whole, no gas flows through the thickness direction, so that the air chambers are formed below the culture plate 12 and the loading base 15.

Preferably, the plate 12 includes an outer frame, and the cell culture wells 122 are fixedly attached to the inside of the outer frame. The shape of the transparent cover 11 matches with the shape of the outer frame of the culture plate 12, and the transparent cover 11 is buckled on the culture plate 12 when in use to ensure that the cell culture hole 122 is isolated from the external environment. The present document takes a case whose outer frame is a rectangular parallelepiped as an example. More specifically, as shown in FIGS. 7 to 9, the outer frame of the culture plate 12 has a hollow rectangular shape in cross section which entirely surrounds the six cell culture wells 122, and the transparent cover 11 is a rectangular parallelepiped shell-shaped having an opening at the bottom. The transparent cover 11 is sleeved on the top of the culture plate 12, and when the culture plate is used, the transparent cover is pressed by the existing clamping tool in the field, so that the culture plate is also pressed immovably, the transparent cover is in contact connection with the culture plate, the transparent cover and the culture plate are not relatively displaced, and an additional sealing device is not needed between the transparent cover and the culture plate.

Preferably, a plate chamfer 121 is provided between two sides of the outer frame of the plate 12, i.e. a vertically placed side of the outer frame is chamfered. Similarly, a transparent cover chamfer 111 is arranged at the corresponding position of the transparent cover 11, so that one corner of the four corners of the culture plate 12 and the transparent cover 11 is cut off. The culture plate chamfer 121 is matched with the transparent cover chamfer 111, so that the transparent cover cannot move in the horizontal plane, and the transparent cover plays roles in blocking environmental pollution and protecting cells.

Preferably, the outer frame of the culture plate 12 has an L-shaped cross section in the vertical plane, and a step surface is formed on the outer frame to facilitate the overlapping of the bottom surface of the transparent cover 11 on the step surface, thereby further protecting the culture plate.

Wherein the loading base 15 as shown in figures 13-14 is a cavity with an opening at the top for providing support and air space. The loading base 15 is a hollow cuboid, and the culture plate 12 encapsulates a cavity.

Preferably, the culture plate 12 is connected to the loading base 15 in a sealing manner by a sealing ring 13. As shown in fig. 10, the cross-sectional shape of the seal ring 13 is L-shaped, and an inner ring step 131 is formed inside the seal ring. The bottom surface of the culture plate 12 is matched with the inner ring step 131 of the sealing ring 13 to play a role of sealing; the sealing ring mounting surface 152 of the loading base 15 cooperates with the sealing ring bottom 132 of the sealing ring 13 to provide sealing. More specifically, a circle of sealing boss is arranged on the top of the loading base 15, i.e. the sealing ring mounting surface 152, so that a step surface is formed on the inner wall surfaces of the sealing ring mounting surface 152 and the sealing boss for mounting a sealing ring with a matched shape. The inner wall surface of the sealing ring surrounds the outer surface and the bottom surface of the culture plate, the inner wall surface of the sealing boss surrounds the outer wall surface of the sealing ring, and the bottom surface of the sealing ring is lapped on the loading base. So that the packing 13 is sandwiched between the bottom surface of the culture plate 12 and the top surface of the loading base 15, and between the outer surface of the culture plate 12 and the inner wall surface of the seal projection. Because the cell loading device is in a negative pressure state in the device when in work, the sealing ring can play a role in sealing without being compressed; in addition, as will be readily appreciated by those skilled in the art, the loading device generally includes a clamping tool disposed on the transparent cover, the transparent cover is pressed down to be close to the culture plate, and the seal ring can be pressed by the culture plate when the culture plate is pressed.

Load plate 16 is a generally flat plate that is disposed within the interior of the load base. The grabbing opening 163 of the loading plate 16 is arc-shaped, so that the loading plate 16 can be conveniently taken out of the loading base 15, a smooth air passage is provided, and the grabbing opening of the loading plate is provided with a round angle so as to prevent a hand from being scratched by a sharp corner in operation; the load table mounting holes 161 of the load plate 16 are distributed in a2 × 3 array. A first load table protrusion 1411 and a second load table protrusion 1421, which will be described later, may be provided on both load tables to be inserted into the load table mounting hole 161. The detachable connection of the two loading platforms and the loading plate is realized in an inserting mode.

More preferably, since the loading platform mounting hole 161 is an oblong, i.e., a kidney-shaped hole, the two types of loading platforms do not rotate around the third direction, i.e., the axial direction of the loading platform, during the mechanical loading; load plate air holes 162 of load plate 16 are symmetrically distributed on the front face of load plate 16 to provide air passages for the device and to reduce the mass of load plate 16.

More preferably, the first loading table 141 or the second loading table 142 is interference-mounted on the loading plate 16. It is easy to think of the interference that two kinds of loading platforms can be inserted into the loading plate and can also be pulled out of the loading platform mounting hole under the action of human force.

Preferably, the loading base 15 is provided with an air chamber boss 151 extending toward the inside of the cavity, and the loading plate 16 is movably overlapped on the air chamber boss 151. The air chamber bosses 151 may be provided on a pair of inner wall surfaces of the loading base which are arranged oppositely. The top surface of the air chamber boss 151 is engaged with the bottom surface of the loading plate 16, and the periphery of the loading plate is engaged with the periphery of the loading base to play a role in positioning, so that the loading plate cannot move in the first direction and the second direction. In addition, the air chamber bosses 151 raise the load plate 16 out of contact with the lowermost surface of the load base, thereby forming air chambers, i.e., air passages.

Gas entering the gas chamber from the positive pressure pump connection port 153 can exit from the negative pressure pump connection port 155; the positive pressure pump connecting port 153 is connected with a pipeline port of the positive pressure pump and used as an air inlet of the device, the negative pressure pump connecting port 155 is connected with a pipeline port of the negative pressure pump and used as an air outlet of the device, and the pressure sensor connecting port 154 is connected with a pressure sensor which is used for monitoring the pressure in the device in real time; the clamping tool mounting groove 156 in the loading base 15 is used for mounting a clamping tool. The positive pressure pump connection port 153, the negative pressure pump connection port 155, the positive pressure pump, the negative pressure pump, the pressure sensor connection port 154 and the pressure sensor form a pneumatic system, and the details can be described later. Compared with the prior art that the gas in the gas chamber is singly controlled by only the negative pressure pump, the gas pressure of various waveforms, such as static state, positive rotation, E heart type, P heart type, triangle, rectangle and various custom waveforms, can be simulated by controlling the relationship between the total output gas pressure value of the positive pressure pump and the negative pressure pump and the time, so that the stress condition of cells can be simulated more abundantly. The device for loading cells with alternative planar forces, see fig. 1-14, comprises a mechanical structure part, a pneumatic system and a control system, wherein the mechanical structure part is connected with the pneumatic part, and the control system controls the pneumatic system to provide the mechanical structure part with the expected positive and negative pressure, thereby simulating the air pressure of various waveforms. The mechanical structure part comprises a transparent cover 11, a culture plate 12, a sealing ring 13, a loading platform assembly 14 for assisting in forming the loading, a loading base 15 and a loading plate 16. The mechanical and pneumatic parts are referred to the above description and will not be described in detail. The control system is prior art in the field of pneumatic pumps and will not be described in detail here.

Wherein the loading platform assembly 14 comprises a first loading platform 141 and a second loading platform 142, the first loading platform 141 and the second loading platform 142 are respectively detachable from the loading plate 16 to enable them to be alternatively connected to the loading plate 16, and the detachment can be achieved by detachable connection means. The first loading stage 141 or the second loading stage 142 is positioned below the loading film 124, the loading film 124 is subjected to a two-dimensional force when the first loading stage 141 is disposed on the loading plate 16, and the loading film 124 is subjected to a one-dimensional force when the second loading stage 142 is disposed on the loading plate 16. The first and second loading surfaces 1412, 1422 are located in close proximity to the loading membrane 124 of the growth plate 12, and are considered to be in contact with each other before loading, but do not generate force, and when the device starts loading, the loading membrane 124 is pressed against the first loading surface 1412 of the first loading stage 141 and the second loading surface 1422 of the second loading stage, and the loading membrane 124 is subjected to tensile deformation, so that the cells on the loading membrane are subjected to strain force. The structure of the first loading table 141 or the second loading table 142 may be referred to in the art, and the following structure may be preferred.

Preferably, the loading platform of the first loading platform 141 is a cylinder, the diameter of the cylinder is smaller than the inner diameter of the projection 125 of the culture plate, so that the loading membrane 124 is subjected to a two-dimensional force, and the cylinder is placed in the aforementioned dome space. The first loading platform 141 and the loading plate are detachably connected, which can be referred to in the prior art of detaching connection in the mechanical field. The cylinder may also be provided as a first load table body having a top surface that serves as the first load surface 1412, a bottom surface that contacts the top surface of load plate 16, and a first load table boss 1411 of a type that mates with load table mounting hole 161, provided below the first load table body.

Preferably, the loading stage of the second loading stage 142 is an elongated shape placed in the radial direction of the loading film 124, so that the loading film 124 is subjected to a one-dimensional force. Preferably, the width of the loading platform is smaller than the diameter of the loading membrane, so that two sides of the loading membrane are not contacted with the loading platform; the loading table is placed at a diameter of the loading film passing through the center of the circle such that the middle portion of the loading table is in contact with the loading film. The loading membrane 124 and the wall of the culture well 122 form a dome space with a downward opening, and the two ends of the loading platform in the length direction are arc surfaces matching the shape of the dome space. The loading table and the dome are in interference fit in space, so that gas is prevented from entering the loading film 124 from two arc surfaces of the loading table. The second loading platform 142 and the loading plate are detachably connected, as can be seen in the prior art of detaching connection in the mechanical field. The elongated load applying table may be provided as a second load applying table body, a top surface of which is a second load applying surface 1422, a bottom surface of which contacts the top surface of the load applying plate 16, and a second type of load applying table protrusion 1421 which is engaged with the load applying table mounting hole 161 may be provided under the second load applying table body.

The working principle of the device can be clearly understood with reference to fig. 3-6. FIG. 3 is a schematic view showing the pre-operation state of the first loading platform, in which the loading membrane 124 of the culture plate 12 is located at a close distance from the loading surface 1412 of the first loading platform 141, and the two are considered to be in contact with each other, but no stress is generated. Fig. 4 is a schematic view of the loading platform in operation, when the device cavity is under negative pressure, the loading membrane around the loading surface 1412 of the first loading platform 141 is subjected to downward suction, and the loading membrane on the loading surface 1412 is subjected to tension along the radius of the loading surface, as shown in the force diagram of the first loading platform in operation in a top view in fig. 5. The second loading platform works on a similar principle to the first loading platform, the top view stress of the second loading platform during working is shown in fig. 6, and unlike the first loading platform, the loading film on the loading surface 1422 of the second loading platform 142 is tensioned along a direction perpendicular to the linear side of the loading platform, that is, the loading film on the loading surface 1412 of the first loading platform 141 is subjected to a two-dimensional force, and the loading film on the loading surface 1422 of the second loading platform 142 is subjected to a one-dimensional force. When the pneumatic system provides air pressure with different waveforms to the mechanical part of the device under the control of the control system, the loading membrane is subjected to forces with different waveforms, and finally the cells are subjected to forces with different waveforms, and the related control part can refer to the prior art.

In a preferred embodiment of the invention, the loading means further comprises a pneumatic system for loading the chamber with pressure.

Referring to fig. 16-17, as shown, a pneumatic system for cell loading includes: a pneumatic device 61 for adjusting the pressure in the cell loading device, a pressure sensor 62 for monitoring the pressure in the cell loading device in real time, and a controller 63 for receiving, processing and sending signals.

The pneumatic device 61 is connected to the loading base 15, and includes a negative pressure pump 615, a negative pressure filter 614, a negative pressure stabilizing bottle 613, a negative pressure flow valve 612, and a negative pressure solenoid valve 611, which are connected in sequence by a pipe joint and a gas pipe, and the negative pressure solenoid valve 611 is connected to the negative pressure pump connection port 155.

The pneumatic device 61 further comprises a positive pressure pump 616, a positive pressure filter 617, a positive pressure stabilizer 618, a positive pressure flow valve 619 and a positive pressure solenoid valve 620 which are sequentially connected by a pipe joint and a gas pipe, wherein the positive pressure solenoid valve 620 is connected with the positive pressure pump connecting port 153.

The positive pressure pump connection port 153 and the negative pressure pump connection port 155 of the cell loading device are respectively connected with the positive pressure electromagnetic valve and the negative pressure electromagnetic valve through pipe joints and air pipes.

The positive pressure pump 616 and the negative pressure pump 615 are air sources, the pneumatic device 61 provides a required power source by the air sources, pure, dry and stable air flow is obtained from the air flow in the air sources through a filter and a pressure stabilizing bottle, then the air flow is adjusted through a flow valve and a solenoid valve to obtain air flow with required flow, and finally the air flow enters the loading base 15. The air from the positive pressure pump or the negative pressure pump firstly passes through the corresponding filter and then enters the corresponding pressure stabilizing bottle, so that moisture and impurities in the air are prevented from being accumulated in the pressure stabilizing bottle; the solenoid valves connected in sequence are closer to the loading base 15 than the flow valves because: the electromagnetic valve is used for opening or cutting off the air path, the flow valve is used for adjusting the size of the opening of the air path, and when the electromagnetic valve is closed, any action of the flow valve has no influence on the pressure in the loading base 15.

The pressure sensor 62 is connected to a pressure sensor connection port 154 of the loading base 15, and can monitor the pressure inside the loading base 15 in real time and convert the pressure into an electrical signal to be transmitted to the controller 63.

The controller 63 is connected to two flow valves, two solenoid valves and also to the pressure sensor 62. The controller 63 receives the electric signal from the pressure sensor 62, processes the waveform of the target pressure and converts it into a corresponding electric signal, and transmits a control signal to the flow valve and the solenoid valve in the pneumatic device 61.

The pneumatic device can adjust the pressure in the loading base 15 to pressurize, decompress and pressure-maintaining the device. The control method monitors the pressure in the cell loading device at any time and feeds the pressure back to the controller, error compensation is added compared with open-loop control, so that the control precision is higher, pressure relief operation is carried out on the cell loading device before the system finishes working, the device cannot be operated under pressure, and the safety and the reliability of the system during experiment are ensured.

A method of controlling a pneumatic system for cell loading, as shown in fig. 17, comprising the steps of:

A1、

s10, starting a pneumatic system;

s20, inputting a waveform of the required pressure within the time from the initial time to the termination time, uniformly dividing the time from the initial time to the termination time into a plurality of time periods according to a preset width value, setting a target pressure for each time period by the controller 13, wherein the target pressure is used for fitting the pressure corresponding to the time period; for example, the desired pressure waveform is a sine curve, the waveform is sequentially divided equally in time, and the average of the pressures for each time period may be selected as the target pressure. Converting the corresponding target pressure into an electric signal, wherein the preset width value is preferably 10 ms;

a2, S30, the controller 63 judges whether the target pressure of the current time period is increased or decreased compared with the target pressure of the previous time period; if the current time period is the first time period, the target pressure in the current time period is increased or decreased compared with the pressure of 0 value, and it is easy to think that the target pressure is not related to the pressure value in the loading base 15;

A3、

s311, if the pressure is increased, closing the negative pressure electromagnetic valve 611 and the negative pressure flow valve 612, and opening the positive pressure electromagnetic valve 620 and the positive pressure flow valve 619;

s312, if the pressure is reduced, closing the positive pressure electromagnetic valve 620 and the positive pressure flow valve 619, and opening the negative pressure electromagnetic valve 611 and the negative pressure flow valve 612;

a4, according to the judgment result of A3 or A7, adjusting the opening of the positive pressure flow valve 619 or the negative pressure flow valve 612 as required to adjust the pressure in the loading base 15;

a5, the controller 13 finishes executing the current time period and judges whether the current time period reaches the termination time, that is, whether the loading process is finished, if yes, the step A8 is executed; if not, executing the step A6;

a6, the controller 63 executes the next time slot and judges whether the target pressure of the time slot of the step A5 needs to be changed, if yes, the step A2 is executed; if not, step a7 is executed, if the pressure does not change, the valve is controlled according to the actual and expected pressure difference;

a7, the controller 63 receives the electric signal from the pressure sensor 62, compares it with the electric signal converted from the target pressure in the time period in step a6 to obtain the difference between the electric signal of the pressure sensor 62 and the electric signal of the target pressure in the time period in step a6, and if the absolute value of the difference is less than or equal to a predetermined error, then step a5 is executed;

if the difference is smaller than zero and the absolute value of the difference is larger than the preset error, the pressure is increased, the negative pressure electromagnetic valve 611 and the negative pressure flow valve 612 are closed, the positive pressure electromagnetic valve 620 and the positive pressure flow valve 619 are opened, and the step A4 is executed;

if the difference is greater than zero and the absolute value of the difference is greater than the predetermined error, the pressure is reduced, the positive pressure electromagnetic valve 620 and the positive pressure flow valve 619 are closed, the negative pressure electromagnetic valve 611 and the negative pressure flow valve 612 are opened, and step a4 is executed;

a8, unloading the pressure in the loading base 15, opening the device to observe the cells and stopping the pneumatic system.

The example is described as a linear waveform diagram, in which the time varies from the initial time 0 to the end time 20ms, the pressure is kept at 2KPa, the preset width value is 10ms, and the preset error epsilon is 0.1 KPa.

B1、

S10, starting a pneumatic system;

s20, inputting a waveform of required pressure, and sequentially dividing time into a first time period and a second time period in sequence, wherein the width of each time period is 10 ms; assigning a target pressure to each time segment to fit a design value of the pressure for the time segment, where the pressure for each time segment is 2 KPa; the controller 63 converts the target pressure corresponding to each time period into an electric signal;

b2, S30 and the controller 63 judge that the current time period is the first time period, and the target pressure of the current time period is increased compared with the 0-value pressure;

b3 and S311, if pressurization is needed, closing the negative pressure electromagnetic valve 611 and the negative pressure flow valve 612, and opening the positive pressure electromagnetic valve 620 and the positive pressure flow valve 619;

b4, adjusting the opening of the positive pressure flow valve 619 to adjust the pressure in the loading base 15;

b5, the controller 63 finishes executing the current time period, determines that the ending time is not reached at present, that is, the second time period needs to be executed, and replaces the step B6;

b6, the controller 63 replaces the second time period, and judges whether the value of the target pressure in the second time period compared with the target pressure in the previous time period needs to be changed, if not, the opening sizes of the positive pressure electromagnetic valve 620 and the positive pressure flow valve 619 are not changed, and step B7 is replaced;

b7, the controller 63 receives the electric signal from the pressure sensor 62, compares the electric signal with the target pressure in the second time period, and replaces the step B8 if the difference is larger than the preset error;

b8, judging whether pressure is required to be increased or reduced according to the difference controller, and adjusting the opening of the positive pressure flow valve 619 or the negative pressure flow valve 612 as required to adjust the pressure in the loading base 15 so that the difference is smaller than or equal to a preset error;

b9, the controller 63 judges whether the ending time is reached at present, the system has already executed the second time period to reach the ending time, and replaces the step B10;

b10, unloading the pressure in the loading base 15, opening the device to observe cells conveniently, and stopping the pneumatic system.

The pneumatic system provides various waveform pressures to simulate the single stress condition of cells under different environments; the control method monitors the pressure in the cell loading device at any time and feeds the pressure back to the controller, and compared with open-loop control, the control method has the advantages that error compensation is increased, and the control precision is higher; before the system finishes working, the pressure relief operation can be carried out on the cell loading device, so that the device can not be operated under pressure, and the safety and the reliability when the system is used for carrying out experiments are ensured

In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.

While embodiments of the present invention have been shown and described above, it should be understood that the above embodiments are exemplary and should not be taken as limiting the invention. Variations, modifications, substitutions and alterations of the above-described embodiments may be made by those of ordinary skill in the art without departing from the scope of the present invention.

The above embodiments of the present invention should not be construed as limiting the scope of the present invention. Any other corresponding changes and modifications made according to the technical idea of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. A cell loading device capable of replacing plane force is characterized by comprising a transparent cover (11), a culture plate (12), a loading platform assembly (14), a loading plate (16) and a loading base (15) which are arranged from top to bottom in sequence;

the culture plate (12) comprises a cell culture hole (122) and a loading membrane (124) arranged at the bottom of the cell culture hole (122), the loading membrane (124) is used for loading cells, and the cell culture hole (122) is isolated from the external environment through the transparent cover (11);

the loading base (15) is a cavity with an opening at the top;

the culture plate (12) encloses the cavity;

the loading plate (14) is arranged in the cavity;

the loading plate (16) is provided with loading plate air holes (162) penetrating through the thickness;

the loading platform assembly (14) comprises a first loading platform (141) and a second loading platform (142), and the first loading platform (141) and the second loading platform (142) are alternatively connected with the loading plate (16);

when the first loading platform (141) is arranged on the loading plate (16) and is positioned below the loading film (124), the loading film (124) is subjected to two-dimensional force; when the second loading platform (142) is arranged on the loading plate (16) and is positioned below the loading film (124), the loading film (124) is subjected to one-dimensional force.

2. A device for loading cells with alternative planar forces according to claim 1, characterized in that the loading base (15) is provided with a gas chamber boss (151) extending towards the inside of the cavity, and the loading plate (16) is lapped on the gas chamber boss (151).

3. An alternative planar force cell loading device according to claim 1, wherein the walls of the cell culture wells (122) extend below the loading membrane (124) to form a dome space below the loading membrane (124);

the first loading platform (141) is a cylinder and is arranged in the dome space, and the diameter of the cylinder is smaller than that of the loading membrane (124);

the second type loading platform (142) is in a strip shape and is arranged in the round cover space, the strip shape is arranged along the radial direction of the loading film (124), and two ends of the second type loading platform (142) are sealed with the round cover space.

4. The device for loading cells with an alternative planar force according to claim 1, wherein the loading plate (16) is provided with a loading plate mounting hole (161), the first loading plate (141) comprises a first loading plate body and a first loading plate boss (1411) having a shape matching the loading plate mounting hole (161), a bottom surface of the first loading plate body contacts a top surface of the loading plate (16), and the first loading plate boss (1411) is inserted into the loading plate mounting hole (161);

the second type of load table (142) includes a second load table body whose bottom surface contacts the top surface of the load plate (16), and a second type of load table boss (1421) that is shape-matched to the stage mounting hole (161), the second load table boss (1421) being inserted into the stage mounting hole (161).

5. The device for cellular loading with alternating planar forces according to claim 4, wherein the loading platform mounting hole (161) is a kidney-shaped hole.

6. An alternative planar force cell loading device according to claim 4, wherein the walls of the cell culture wells (122) extend below the loading membrane (124) to form a dome space below the loading membrane (124);

the first loading platform main body is a cylinder and is arranged in the dome space, and the diameter of the cylinder is smaller than that of the loading membrane (124);

7. A device for loading cells with alternative planar forces according to claim 1, characterized in that the loading plate (16) is provided with gripping openings (163) at its edges.

8. An alternative planar force cell loading device according to claim 1, further comprising a sealing ring (13) interposed between the culture plate (12) and the loading base (15);

a sealing boss surrounding the sealing ring (13) is arranged at the top of the loading base (15);

the cross section of the sealing ring (13) is L-shaped, so that the sealing ring (13) is clamped between the bottom surface of the culture plate (12) and the top surface of the loading base (15), and is clamped between the outer surface of the culture plate (12) and the inner wall surface of the sealing boss.

9. The device for loading cells capable of replacing planar force as claimed in claim 1, wherein said culture plate (12) comprises an outer frame and said cell culture holes (122) fixed in said outer frame, said outer frame is a rectangular frame structure, said transparent cover (11) is sleeved on the top of said culture plate (12);

and a culture plate chamfer (121) is arranged between two side faces of the outer frame, and a transparent cover chamfer (111) matched with the culture plate chamfer (121) is arranged on the transparent cover (11).

10. A device for loading cells with an alternative planar force according to claim 1, characterised in that it further comprises a pneumatic system comprising a pneumatic device (61), a pressure sensor (62) and a controller (63);

the air chamber is connected with a positive pressure pump connecting port (153) and a negative pressure pump connecting port (155) in parallel;

the pneumatic device (61) comprises a negative pressure pump (615), a negative pressure filter (614), a negative pressure stabilizing bottle (613), a negative pressure flow valve (612) and a negative pressure electromagnetic valve (611) which are connected in sequence, and the negative pressure electromagnetic valve (611) is connected with the negative pressure pump connecting port (155);

the pneumatic device (61) further comprises a positive pressure pump (616), a positive pressure filter (617), a positive pressure stabilizing bottle (618), a positive pressure flow valve (619) and a positive pressure electromagnetic valve (620) which are sequentially connected, and the positive pressure electromagnetic valve (620) is connected with the positive pressure pump connecting port (153);

the pressure sensor (62) is used for monitoring the pressure value in the air chamber and converting the pressure value into an electric signal to be transmitted to the controller (63);

the controller (63) is respectively electrically connected with the negative pressure flow valve (612), the negative pressure electromagnetic valve (611), the positive pressure flow valve (619) and the positive pressure electromagnetic valve (620).

11. A method of pneumatic system control by the pneumatic system of claim 10, comprising the steps of:

a1, starting the pneumatic system, inputting a required pressure waveform from initial time to termination time, averagely dividing the initial time to the termination time into a plurality of time periods according to a preset width value, and setting a target pressure for each time period, wherein the target pressure is used for fitting the pressure of the corresponding waveform in the time period; the controller (63) converting the value of the target pressure into an electrical signal;

a2, the controller (63) judges whether the target pressure in the current time period is increased or reduced compared with the target pressure in the last time period; if the current time period is the first time period, judging whether the target pressure in the current time period is increased or reduced compared with the pressure of 0 value;

a3, if the pressure is increased, closing the negative pressure electromagnetic valve (611) and the negative pressure flow valve (612), and opening the positive pressure electromagnetic valve (620) and the positive pressure flow valve (619); if the pressure is reduced, closing the positive pressure electromagnetic valve (120) and the positive pressure flow valve (619), and opening the negative pressure electromagnetic valve (611) and the negative pressure flow valve (612);

a4, according to the judgment result of the step A3 or the step A7, adjusting the opening of the corresponding positive pressure flow valve (619) or negative pressure flow valve (612) as required to adjust the pressure in the air chamber;

a5, the controller (63) judges whether the current time is the termination time, if yes, the step A8 is executed; if not, executing the step A6;

a6, the controller (63) executes the next time period, judges whether the target pressure of the current time period and the target pressure of the time period of the step A5 are changed, if yes, executes the step A2; if not, executing the step A7;

a7, the controller (63) receiving the electric signal from the pressure sensor (62), comparing with the target pressure converted electric signal for the time period in the step A6, if the absolute value of the difference is less than or equal to a predetermined error, executing the step A5;

if the difference is smaller than zero and the absolute value of the difference is larger than a preset error, judging that the pressure is increased, closing the negative pressure electromagnetic valve (611) and the negative pressure flow valve (612), opening the positive pressure electromagnetic valve (620) and the positive pressure flow valve (619), and executing the step A4;

if the difference is greater than zero and the absolute value of the difference is greater than a preset error, determining that the pressure is reduced, closing the positive pressure electromagnetic valve (620) and the positive pressure flow valve (619), opening the negative pressure electromagnetic valve (611) and the negative pressure flow valve (612), and executing the step A4;

a8, unloading the pressure in the air chamber.