CN101906379A - Device for precisely stretching visual cells under simulated in vivo environment - Google Patents
Device for precisely stretching visual cells under simulated in vivo environment Download PDFInfo
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- CN101906379A CN101906379A CN 201010222982 CN201010222982A CN101906379A CN 101906379 A CN101906379 A CN 101906379A CN 201010222982 CN201010222982 CN 201010222982 CN 201010222982 A CN201010222982 A CN 201010222982A CN 101906379 A CN101906379 A CN 101906379A
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
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Abstract
The invention discloses a device for precisely stretching visual cells under simulated in vivo environment, which relates to a device for mechanically loading cells in medical experimental instruments. The device of the invention comprises a basement membrane installing device for clamping a basement membrane, a supporting seat, an automatically stretching device for controlling the parallel movement time, frequency and amplitude of the basement membrane installing device, a visual observation device for observing the basement membrane in real time, a culture pond and a culture pond installing seat for placing the culture pond. Because the automatically stretching device of the invention can realize the purpose of continuously stretching cells through computer programming, the stretching precision is high, and the stretching range is large; the change of the cells and the simulated in vivo environment can be observed in real time in the loading process; and a basement membrane clamper can be used for simultaneously clamping some basement membranes. Thus, the device of the invention can be widely used as a device for mechanically loading cells in medical experimental instruments, and is an ideal loading device for carrying out mechanical stimulation response research on biological tissues and cells in biomechanics.
Description
Technical field
The present invention relates to a kind of accurate tensile loading device of cell that under simulated in vivo environment, can carry out Real Time Observation, belong to the technical field of cyto-mechanics.
Background technology
The biological effect of mechanical stimulation and their mechanism of action begin one's study in many cell types and tissue at present, these results of study have made people to they roles in the reparation of cytodifferentiation, genetic expression, cell and tissue growth and wound healing and function are integrated, have caused attention.Can influence cellular form as the periodicity strain, cause cytoskeleton to be reset.Different stress stimulations can cause the mesenchymal stem cells MSCs differentiation direction to change, and periodically loading to influence in the tissue engineering bone/cartilage cell proliferation etc.Yet, be how to be experienced and be passed in the cell for mechanical stress by cell, finally cause cell that a series of biological effects take place, its precise mechanism it be unclear that.
Therefore, need the specific external mechanics loading experimental apparatus of design to make the mechanics hormesis in institute's cultured cells, observation of cell is subjected to propagation, apoptosis, the differentiation behind the stress, and the variation of the expression of cyto-mechanics characteristic, gene, albumen and enzyme etc., this experimental technique is very a kind of rapidly research method of present biomechanics field development.But because body interior mechanics and biological environment are complicated unusually, the size of human body cell is between tens to tens microns, the thickness of cytolemma only has several nanometers to tens nanometer, therefore, can't directly use conventional macromechanics loading method and experimental technique; Setting up suitable loading culture model is the matter of utmost importance that cyto-mechanics faces.
For setting up the rational method of cell loading force, as far back as nineteen thirty-nine, scientist is just carrying out ground-breaking research aspect the mechanics loading of cultured cell in vitro.Through improving for many years and developing, developed the mechanical loading unit of multiple cultured cell in vitro.Be broadly divided into unicellular deceleration loading device, pressure conduction deceleration loading device, centrifugal force deceleration loading device, fluid shearing force loading device and substrate deformation deceleration loading device.
Substrate deformation loading technique starts from the appearance of elastic cell's culture membrane.The Design of device thinking is as base material with elastica, by with cell seeding on elastica, produce controllable motion by device and make elastica generation deformation, and the cell of cultivating on elastomeric film surface is stressed because of the deformation of substrate, thereby realizes the possibility of cell growth rhythm research under the tensile stress.The afterburning mode of this deceleration loading device is comparatively suitable, and does not limit experimental period, is present a kind of comparatively ideal experimental facilities, can be used for various cells in vitro mechanics and loads, and is mainly used in the influence of research distraction force to cultured cell in vitro propagation and metabolism etc.Its loading technique mainly contains rectangular base stretching, circular-base stretching etc., and wherein the stretching of circular-base is more common.Existing circular-base stretching device generally adopts mimic channel to control, and stretching frequency, stimulates magnitude range limited, and after the device design finished, correlation parameter was difficult for change, and applicable surface is limited.From the measurement situation, under the 1Hz controlled frequency, owing to use restriction of silicone tube internal diameter and the parameter influence of peristaltic pump own, liquid flow rate is subjected to certain limitation, and film is difficult to reach the ratio of elongation more than 10%.But not the motion of planar rondure substrate stretch system by deformable circular-base applies strain, and the Flexercell loading system is the typical case of this class on-plane surface circular-base stretch system, can provide strain in a big way to stimulate; Yet in this device, the elasticity basement membrane is inhomogeneous in the strain distribution of different positions, and this has just brought systematic error to interpretation.And, developed internal milieu in view of the above and simulated device for precisely stretching visual cells at the loading of the required research of different cells or for frequency stretches or be continuously elongated.
Summary of the invention
The objective of the invention is weak point at existing cell loader, a kind of cell tensile loader of computer programming control is provided, have under simulated in vivo environment, can realize continuously elongated, and range of stretch is bigger, cell is stressed evenly, accurately control stretching amplitude and frequency can the cell growth situation be carried out Real Time Observation, also can stretch to many basilar membranes simultaneously.
For realizing above technical purpose, the present invention will take following technical scheme:
A kind of device for precisely stretching visual cells under simulated in vivo environment, comprise the basilar membrane erecting device that is used for the clamping basilar membrane, bearing, be used to spur the parallel traveling time of basilar membrane erecting device, the automatic stretching device of frequency and amplitude, the visual observation device that is used for the Real Time Observation basilar membrane, cultivation pool and the cultivation pool mount pad that is used to place cultivation pool, described basilar membrane erecting device comprises fixed link and stroke lever, the abutting end of described fixed link and stroke lever is installed the basilar membrane clamper all respectively, the basilar membrane clamping is installed between the two basilar membrane clampers, and fixed link is fixedlyed connected with bearing, stroke lever then can be supported on the bearing with moving horizontally, stroke lever is fixedlyed connected with the output terminal of automatic stretching device simultaneously, in addition, described cultivation pool mount pad is installed on the bearing, cultivation pool then places on the cultivation pool mount pad, and the line center-aligned of cultivation pool and fixed link and stroke lever, described visual observation device comprises CCD camera and the optical fiber that is connected with the CCD camera, and described optical fiber places the basilar membrane top.
Further, the controller that described automatic stretching device comprises tensilometer and is used for stretching time, stretching frequency and the stretch range of control stretching instrument, described tensilometer comprises piezoelectric ceramic motor, controller is then set up based on Work bench V5 interface.
Further, described bearing comprises the strongback of horizontal positioned and is installed at interval fixedly stand and slip stand on this strongback regularly, described fixed link is fixedlyed connected with fixing stand, stroke lever is supported in the slip stand movably, and the cultivation pool mount pad then is installed on the strongback.
Further, described stroke lever comprises the first stroke lever split and the second stroke lever split, the described first stroke lever split and the second stroke lever split link into an integrated entity by fixed connecting piece, and the first stroke lever split is erected in the slip stand, the basilar membrane clamper is installed in the end of this first stroke lever split simultaneously, the second stroke lever split then is connected with automatic stretching device, and on second stroke lever positioning adapter is installed.
Further, described basilar membrane clamper, comprise the first U-shaped plate and the second U-shaped plate, a described U andante is socketed in the second U-shaped plate, and the blind end of a U andante and the second U-shaped plate all is dull and stereotyped and is provided with, be equipped with on the first U-shaped plate simultaneously and be used for the union lever that is connected with stroke lever, the opening end of a described in addition U andante and the second U-shaped plate all is the outward flange setting, the first U-shaped plate and the second U-shaped plate tighten up and are connected by being connected fastening piece on both flange end, and basilar membrane is clamped between two blind ends of a U andante and the second U-shaped plate.
Further, described fixed link and stroke lever are installed in fixedly stand and the slip stand by stop screw respectively.
According to above technical scheme, can realize following beneficial effect:
1, the present invention adopts parallel the moving of automatic stretching device pulling basilar membrane erecting device, and parallel traveling time, frequency and the amplitude of this basilar membrane erecting device controlled, promptly the basilar membrane that clamping is installed on the basilar membrane erecting device carries out the automatic control of stretching time, stretching frequency and stretch range, can realize the continuously elongated of pair cell simultaneously.Hence one can see that, and the present invention is dynamic axial basilar membrane stretching system, and in the basilar membrane elastic range, its strain is unlimited adjustable, and the scope of application is wider.
2, the present invention has also added the visual observation device, the optical fiber of this visual observation device by linking to each other with the CCD camera, the variation of the observation basilar membrane cell of can in loading procedure, carry out regularly, fixing a point, cultivate the unit and need not migratory cell, avoided the contaminated possibility of culture membrane and cell.
3, the present invention has considered internal milieu mimic problem in design, and cell culture unit is placed CO
2Experimentize in the incubator, to obtain experimental result more accurately.Under this experimental program, also can together put into incubator to control group, make stretching loading group be in identical culture environment, satisfy the different experiments demand.
4, the present invention has also designed the basilar membrane clamper especially, and its biggest advantage is to press from both sides several basilar membranes simultaneously, and the stressed even and loading convenience of film has been simplified schedule of operation.Simple, the low price of this apparatus structure can be widely used as the cyto-mechanics deceleration loading device of medical experiment instrument, is the desirable deceleration loading device that biological tissue and cell is carried out the mechanical stimulation response investigations in biomechanics.
Description of drawings
Fig. 1 is a structural representation of the present invention;
Fig. 2 is the cell culture unit synoptic diagram;
Wherein, controller 1, lead 2, tensilometer 3, stroke lever 4, the first stroke lever split 41, the second stroke lever split 42, positioning adapter 5, set screw 6, slip stand 7, stop screw 8, cultivation pool 9, basilar membrane clamper 10, cultivation pool mount pad 11, fixed link 12, fixedly stand 13, strongback 14, optical fiber 15, CCD camera 16, cell culture incubator 17, cultivation pool top cover 18, basilar membrane 19, nutrient solution 20, clamper loose slot 21.
Embodiment
Accompanying drawing discloses the structural representation of a preferred embodiment of the invention without limitation, explains technical scheme of the present invention below with reference to accompanying drawing.
As depicted in figs. 1 and 2, device for precisely stretching visual cells under simulated in vivo environment of the present invention, comprise the basilar membrane erecting device that is used for clamping basilar membrane 19, bearing, be used to spur the parallel traveling time of basilar membrane erecting device, the automatic stretching device of frequency and amplitude, the visual observation device that is used for Real Time Observation basilar membrane 19, cultivation pool 9 and the cultivation pool mount pad 11 that is used to place cultivation pool 9, described basilar membrane erecting device comprises fixed link 12 and stroke lever 4, the abutting end of described fixed link 12 and stroke lever 4 is installed basilar membrane clamper 10 all respectively, the basilar membrane clamping is installed between the two basilar membrane clampers 10, and fixed link 12 is fixedlyed connected with bearing, stroke lever 4 then can be supported on the bearing with moving horizontally, therefore, basilar membrane erecting device and bearing form firm rigid foundation, can guarantee the steady of drawing process, stroke lever 4 is fixedlyed connected with the output terminal of automatic stretching device simultaneously, described automatic stretching device comprises tensilometer 3 and the stretching time that is used for control stretching instrument 3, the controller 1 of stretching frequency and stretch range, described tensilometer 3 comprises piezoelectric ceramic motor, controller 1 is then set up based on Work bench V5 interface, therefore, the present invention can realize the continuously elongated of pair cell by computer programming, stroke lever 4 can move around under the driving of tensilometer 3 in the horizontal direction, be that stroke lever 4 makes horizontal rectilinear motion for the cell culture unit of clamping stretched film the transmission of power of tensilometer 3, so that the cell that adheres to is loaded stretching, in addition, described cultivation pool mount pad 11 is installed on the bearing, cultivation pool 9 then places on the cultivation pool mount pad 11, and the line center-aligned of cultivation pool 9 and fixed link 12 and stroke lever 4, described visual observation device comprises CCD camera 16 and the optical fiber 15 that is connected with CCD camera 16, and described optical fiber 15 places basilar membrane 19 tops.
Described bearing comprises the strongback 14 of horizontal positioned and is installed at interval fixedly stand 13 and slip stand 7 on this strongback 14 regularly, described fixed link 12 is fixedlyed connected with fixing stand 13, stroke lever 4 is supported in the slip stand 7 movably, and cultivation pool mount pad 11 then is installed on the strongback 14.
Described cultivation pool mount pad 11 is supported on the bearing by an experimental machine tool lifting table.Be that cultivation pool mount pad of the present invention 11 is Height Adjustable, guaranteed universality of the present invention effectively.
Described stroke lever 4 comprises the first stroke lever split and the second stroke lever split, the described first stroke lever split and the second stroke lever split link into an integrated entity by fixed connecting piece, and the first stroke lever split is erected in the slip stand 7, basilar membrane clamper 10 is installed in the end of this first stroke lever split simultaneously, the second stroke lever split then is connected with automatic stretching device, and on second stroke lever 4 positioning adapter 5 is installed.
Described basilar membrane clamper 10, comprise the first U-shaped plate and the second U-shaped plate, a described U andante is socketed in the second U-shaped plate, and the blind end of a U andante and the second U-shaped plate all is dull and stereotyped and is provided with, be equipped with on the first U-shaped plate simultaneously and be used for the union lever that is connected with stroke lever 4, union lever passes cultivation pool top cover 18 backs and is connected with stroke lever 4, cultivation pool top cover 18 is formed by two splits, be that union lever can be along with the translation between two splits of cultivation pool top cover 18 of moving of stroke lever 4, be equivalent to that the gap forms clamper loose slot 21 between two splits of cultivation pool top cover 18, the opening end of a described in addition U andante and the second U-shaped plate all is the outward flange setting, make win U-shaped plate and the second U-shaped plate tighten up and be connected by being connected fastening piece on both flange end, basilar membrane 19 is clamped between two blind ends of a U andante and the second U-shaped plate.Therefore, in prior art, directly basilar membrane 19 is placed on the platform and compares, basilar membrane clamper 10 of the present invention is several basilar membranes of clamping 19 simultaneously, and each to open base film stressed evenly and load conveniently, simplified schedule of operation.
Described fixed link 12 is installed in the fixedly stand 13 by stop screw 8, simultaneously stroke lever 4 also be by stop screw 8 location and installation in slip stand 7, therefore, in the vertical direction, can be by the adjusting of stop screw 8, make stroke lever 4 running fix in slip stand 7, fixed link 12 is running fix in fixing stand 13 then.
Therefore, device for precisely stretching visual cells under simulated in vivo environment of the present invention mainly comprises Controlling System, mechanical system, Real Time Observation system.Controlling System adopts computer control, by stretching frequency, stretch range, the stretching time of programming Control tensilometer 3.Mechanical system mainly is made of tensilometer 3, stroke lever 4, slip stand 7, fixed link 12, fixedly stand 13, strongback 14 and cell culture unit.The Real Time Observation system is a CCD camera 16, connects optical fiber 15 on it.Controlling System is arranged on outside the slip stand 7, and tensilometer 3 is fixed in an end of slip stand 7, connects by lead 2 between Controlling System and the tensilometer 3.Tensilometer 3 is that power set links to each other with stroke lever 4, and stroke lever 4 makes horizontal rectilinear motion for the cell culture unit of clamping stretched film the transmission of power of tensilometer 3, so that the cell that adheres to is loaded stretching.One end of stroke lever 4 is fixedlyed connected with tensilometer 3 output shafts, and the other end and positioning adapter 5 are affixed.The other end of stroke lever 4 passes 5 of positioning adapters and rides in the slip stand 7, connects basilar membrane clamper 10.Stroke lever 4 can move around under the driving of tensilometer 3 in the horizontal direction, and in the vertical direction can pass through stop screw 8 running fix in slip stand 7.Fixed link 12 links to each other with fixing stand 13, and in the vertical direction can be by stop screw 8 running fix in fixing stand 13.Slip stand 7 is fixedlyed connected with strongback 14 by set screw 6 respectively with the two ends of fixedly stand 13.Therefore, fixed link 12, stroke lever 4, fixedly stand 13, slip stand 7 constitute firm rigid foundation with strongback 14, can guarantee the steady of drawing process.An end that is attached with the basilar membrane 19 of cell is connected with fixed link 12 by clamper, the other end is connected with stroke lever 4 by basilar membrane clamper 10, on the adjustable for height cultivation pool mount pad 11 below the basilar membrane 19, be placed with semi closed cultivation pool 9, on this semi closed cultivation pool 9 cultivation pool top cover 18 be installed.The culture membrane that is attached with cell carries out unilateral stretching by stroke lever 4 by slip stand 7 under the effect of power system, and can pull down stretching motion from frame easily and partly carry out microscopic examination by backing out set screw 6.Also can carry out microscopic examination, to understand the growing state of cell by being suspended from the optical fiber 15 that links to each other with CCD camera 16 above the culture dish.Mechanical system in whole device except that tensilometer 3 and Real Time Observation system all place cell culture incubator 17.
During cultivation, cell culture unit is placed CO
2In the incubator, clamping has the basilar membrane clamper 10 of basilar membrane 19 to be dipped in the nutrient solution 20 to experimentize, and with simulated in vivo environment, obtains experimental result more accurately.Under this experimental program, also can together put into described CO to control group
2Incubator makes stretching loading group be in identical culture environment, satisfies the different experiments demand.
Claims (6)
1. device for precisely stretching visual cells under simulated in vivo environment, it is characterized in that, comprise the basilar membrane erecting device that is used for the clamping basilar membrane, bearing, be used to spur the parallel traveling time of basilar membrane erecting device, the automatic stretching device of frequency and amplitude, the visual observation device that is used for the Real Time Observation basilar membrane, cultivation pool and the cultivation pool mount pad that is used to place cultivation pool, described basilar membrane erecting device comprises fixed link, stroke lever and basilar membrane clamper, the abutting end of described fixed link and stroke lever is installed the basilar membrane clamper all respectively, the basilar membrane clamping is installed between the two basilar membrane clampers, and fixed link is fixedlyed connected with bearing, stroke lever then can be supported on the bearing with moving horizontally, stroke lever is fixedlyed connected with the output terminal of automatic stretching device simultaneously, in addition, described cultivation pool mount pad is installed on the bearing, cultivation pool then places on the cultivation pool mount pad, and the line center-aligned of cultivation pool and fixed link and stroke lever, described visual observation device comprises CCD camera and the optical fiber that is connected with the CCD camera, and described optical fiber places the basilar membrane top.
2. according to the described device for precisely stretching visual cells under simulated in vivo environment of claim 1, it is characterized in that, the controller that described automatic stretching device comprises tensilometer and is used for stretching time, stretching frequency and the stretch range of control stretching instrument, described tensilometer comprises piezoelectric ceramic motor, and controller is then set up based on Work bench V5 interface.
3. according to the described device for precisely stretching visual cells under simulated in vivo environment of claim 1, it is characterized in that, described bearing comprises the strongback of horizontal positioned and is installed at interval fixedly stand and slip stand on this strongback regularly, described fixed link is fixedlyed connected with fixing stand, stroke lever is supported in the slip stand movably, and the cultivation pool mount pad then is installed on the strongback.
4. according to the described device for precisely stretching visual cells under simulated in vivo environment of claim 3, it is characterized in that, described stroke lever comprises the first stroke lever split and the second stroke lever split, the described first stroke lever split and the second stroke lever split link into an integrated entity by fixed connecting piece, and the first stroke lever split is erected in the slip stand, the basilar membrane clamper is installed in the end of this first stroke lever split simultaneously, the second stroke lever split then is connected with automatic stretching device, and on second stroke lever positioning adapter is installed.
5. according to the described device for precisely stretching visual cells under simulated in vivo environment of claim 1, it is characterized in that, described basilar membrane clamper, comprise the first U-shaped plate and the second U-shaped plate, a described U andante is socketed in the second U-shaped plate, and the blind end of a U andante and the second U-shaped plate all is dull and stereotyped and is provided with, be equipped with on the first U-shaped plate simultaneously and be used for the union lever that is connected with stroke lever, the opening end of a described in addition U andante and the second U-shaped plate all is the outward flange setting, the first U-shaped plate and the second U-shaped plate tighten up and are connected by being connected fastening piece on both flange end, and basilar membrane is clamped between two blind ends of a U andante and the second U-shaped plate.
6. according to the described device for precisely stretching visual cells under simulated in vivo environment of claim 1, it is characterized in that described fixed link and stroke lever are installed in fixedly stand and the slip stand by stop screw respectively.
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| CN 201010222982 CN101906379A (en) | 2010-07-09 | 2010-07-09 | Device for precisely stretching visual cells under simulated in vivo environment |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103966094A (en) * | 2014-05-16 | 2014-08-06 | 中国人民解放军第三军医大学野战外科研究所 | Culture device applied to cell tensile stress and method |
| CN105628601A (en) * | 2015-12-21 | 2016-06-01 | 天津理工大学 | Rolling load application control device for relevant cartilage experiment based on digital image |
| CN105950466A (en) * | 2016-05-13 | 2016-09-21 | 上海健康医学院 | Endangium experimental apparatus capable of correcting stress relaxation of elastic base |
| CN106867888A (en) * | 2017-03-20 | 2017-06-20 | 北京理工大学 | The at the uniform velocity single axis of symmetry tensile cell mechanics device that can be observed in place in real time |
| CN106995780A (en) * | 2011-05-05 | 2017-08-01 | 安派科生物医学科技有限公司 | Tumour cell detector |
| CN112128335A (en) * | 2020-11-25 | 2020-12-25 | 常州市凯迪电器股份有限公司 | Linear actuator |
| CN115980131A (en) * | 2022-10-25 | 2023-04-18 | 南开大学 | Planar mechanical controllable junction cracking technology based on flexible material |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106995780A (en) * | 2011-05-05 | 2017-08-01 | 安派科生物医学科技有限公司 | Tumour cell detector |
| CN106995780B (en) * | 2011-05-05 | 2019-11-22 | 安派科生物医学科技有限公司 | Tumour cell detector |
| CN103966094A (en) * | 2014-05-16 | 2014-08-06 | 中国人民解放军第三军医大学野战外科研究所 | Culture device applied to cell tensile stress and method |
| CN105628601A (en) * | 2015-12-21 | 2016-06-01 | 天津理工大学 | Rolling load application control device for relevant cartilage experiment based on digital image |
| CN105628601B (en) * | 2015-12-21 | 2018-06-08 | 天津理工大学 | A kind of rolling load for cartilage digital picture related experiment applies control device |
| CN105950466A (en) * | 2016-05-13 | 2016-09-21 | 上海健康医学院 | Endangium experimental apparatus capable of correcting stress relaxation of elastic base |
| CN106867888A (en) * | 2017-03-20 | 2017-06-20 | 北京理工大学 | The at the uniform velocity single axis of symmetry tensile cell mechanics device that can be observed in place in real time |
| CN112128335A (en) * | 2020-11-25 | 2020-12-25 | 常州市凯迪电器股份有限公司 | Linear actuator |
| CN112128335B (en) * | 2020-11-25 | 2021-03-16 | 常州市凯迪电器股份有限公司 | Linear actuator |
| CN115980131A (en) * | 2022-10-25 | 2023-04-18 | 南开大学 | Planar mechanical controllable junction cracking technology based on flexible material |
| CN115980131B (en) * | 2022-10-25 | 2023-08-01 | 南开大学 | Flexible material-based planar mechanical controllable cracking method |
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Application publication date: 20101208 |