CN201372287Y - Living tissue specimen and cell perfusion device adaptable to functional study - Google Patents
Living tissue specimen and cell perfusion device adaptable to functional study Download PDFInfo
- Publication number
- CN201372287Y CN201372287Y CN200920126738U CN200920126738U CN201372287Y CN 201372287 Y CN201372287 Y CN 201372287Y CN 200920126738 U CN200920126738 U CN 200920126738U CN 200920126738 U CN200920126738 U CN 200920126738U CN 201372287 Y CN201372287 Y CN 201372287Y
- Authority
- CN
- China
- Prior art keywords
- perfusion
- bath
- cell
- sample
- bath tank
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Disclosed are a living tissue specimen and a cell perfusion device which are adaptable to functional study. The device comprises a perfusion fluid inputting system, a specimen perfusion bath tank and a perfusion fluid collecting system, wherein the perfusion fluid inputting system and the perfusion fluid collecting system are respectively connected to the specimen perfusion bath tank via perfusion fluid flowing pipelines, and the specimen perfusion bath tank is a structure composed of three independent but communicated small chambers formed on completely transparent polytetrafluoroethylene materials. A record bath tank is the middle one of the specimen perfusion bath tank, an input trough and an output trough of perfusion fluid are respectively disposed on two sides of the record bath tank, and an entire cover glass is fixed onto the bottom of the specimen perfusion bath tank. By adopting the three independent but communicated small chambers of the specimen perfusion bath tank, the device can precisely control extra-cellular fluid volume, ensure stable flow rate of the perfusion fluid, and facilitate detection of cell secretion activity. The specimen perfusion bath tank is designed into an open structure and has succinct design, low cost, easy disassembly and replacement and capability of being compatible with all microscope stages so as to meet the requirements on visualized electrophysiology functional record to tissue cells.
Description
Technical field
The utility model relates to medical biotechnology and Neuroscience Research field, be mainly used in the section of vitro culture biological tissue, culturing cell are carried out continous perfusion, keep tissue and cytoactive, realize the experimental installation of emiocytosis activity and electrophysiology functional study.
Background technology
In modern medicine biology especially Neuroscience Research, biological tissue or cells in vitro culture technique are one of experimental techniques of very important and widespread use, and it is that significant contribution has been made in cytology, genetics, virusology, immunologic research and application.The tissue culture technique purpose that development in recent years is got up is to make tissue break away from body and survive, and has higher requirement to culture technique.A lot of scholars improve and attempt at aspects such as the vessel of cultivating method of operating, cultured tissue and substratum, and the technology that vitro tissue is cultivated more is tending towards perfect, and cells growth activity is improved.
But existing culture technique mainly based under aseptic condition to the observation of morphological features such as tissue or cell survival, growth, growth, differentiation.Therefore in this closed system, the environment that imitation live body environment etc. can be controlled arbitrarily can be set, cell or tissue is remained under the situation of said cultivation position and supply with substratum, not only can prevent to pollute, and can realize the vitro culture of cell or tissue expeditiously.The major defect that the culture system of this present widespread use exists just is the closure of system.Carry out the research of histiocytic function, especially electrophysiological function if desired and when detecting, can not effectively write down observation, also be difficult to the external environment of accurate control tissue or cell.According to the report of document, and the exploration of investigator in real work, set up and thisly can be used for biopsy cell's Function detection, especially carry out the continous perfusion system that the patch clamp electrophysiology detects.In present perfusion device, compare as Chinese patent application " isolated organ constant temperature perfusion instrument " (application number 02280295.9), " a kind of new tissue perfusion device " (application number 00216922.3), " a kind of intracellular perfusion liquid changing device " (application number 96236386.3) etc., many improvement have been arranged.Deficiency such as overcome in the past that the device systems design is complicated, systems bulky is difficult to carry out electromagnetic shielding and the perfusion tankage is bigger.Especially present perfusion system is difficult in real time, dynamically detects the micro-cytokine or the neurotransmitter of emiocytosis and release because capacity is bigger, can not be equipped with compatible all microscopic examinations.And can bring bigger electrical signal to disturb based on the constant temperature perfusion of heating at the cell platform, be difficult to write down the bioelectrical activity of cell itself.
Summary of the invention
Key problem in technology to be solved in the utility model is to be not easy to realize that at existing organ, tissue or intracellular perfusion system thermostatic control or temperature heating bring bigger power frequency electrical signal to disturb easily, be difficult to realize part defectives such as emiocytosis detection and electrophysiological function record, open, constant temperature, constant current, the low capacity perfusion system of proposition outside conventional cell or tissue is cultivated, realize the real-time collecting extracellular fluid, detection of dynamic biopsy cell secretory activity can be provided, be particularly useful for carrying out the electrophysiological function record of tissue or culturing cell.
The technical scheme of being taked is for achieving the above object:
Design a kind of biological tissue and cell continous perfusion device that is applicable to functional study, this perfusion device comprises perfusate input system, water bath with thermostatic control system, sample perfusion bath, microscope adaptation unit and perfusate gathering system.Perfusate input system and perfusate gathering system are connected the input slot and the output magazine of sample perfusion bath respectively by the perfusion liquid flow conduits.The design of sample perfusion bath is the core of this device, make by the polytetrafluoroethylmaterial material that full impregnated is bright, sample perfusion bath is made of the cell that three independences still communicate, and middle bigger cell is the record bath, and the cell of both sides is respectively applied for the input slot and the output magazine of perfusion liquid.The volume of cell has determined this perfusion device to collect the capacity of liquid in the middle of the sample perfusion bath, therefore is designed to 225 μ L, really to be convenient to improve the concentration that needs to detect composition in the collection liquid.Three bottom lands of sample perfusion bath are the whole piece cover glass fixedly.
Further, this device is provided with the microscope adaptation unit, it is the key that this perfusion device can have versatility, and the microscope adaptation unit is the pre-opened hole form fit on profile and the microscope stage, and the centre is useful on the groove of embedded placement sample perfusion bath.
In addition, for cell or brain sheet are remained in the relative constant temperature system, the pipeline that described perfusate input system connects sample perfusion bath is provided with the water bath with thermostatic control system, general needs according to electric Physiological Experiment, temperature is set in 25-32 ℃, set by the accessory water-bath, guarantee that the temperature of system does not have drift.
Also have height to design, not only can be used for inverted microscope in conjunction with the design that embeds the microscope adaptation unit and observe, also can be used for just putting the microscopical observation of water immersion objective at 3mm with sample perfusion bath.
The perfusion liquid flow conduits is all used plastics or Glass tubing, wraps the shield effectiveness that is formed similar BNC cable by the aluminium foil of one deck ground connection, prevents to continue the mobile perfusate and forms the power frequency interference that " antenna " brought.
Liquid is flowed out by input system according to the perfusion rate of setting, and after the heating of water bath with thermostatic control system, enters sample perfusion bath, reclaims perfusate by the perfusate gathering system at last and detects for the analysis of cells secretory activity.This device adds through the saturated extracellular fluid of oxygen by the perfusate input system, and the flow rate of liquid that needs of basis of design experiment, so the perfusion rate of this device depends on the adjusting of perfusate input system.
This device by sample perfusion bath three independences but the cell that communicates is accurately controlled extracellular fluid volume, guarantee that the perfusate rate of flow of fluid is stable, be convenient to detect the emiocytosis activity; Sample perfusion bath is designed to open structure; Design succinct, with low cost, easy dismounting and change, can compatible all microscope stages, reach the visual requirement of carrying out the electrophysiological function record of histocyte; To the perfusion liquid flow conduits reasonably shielding can be used for electrophysiological recording with ground connection, in conjunction with the volume control of sample perfusion bath, make this device can be from the form to the function, Real-time and Dynamic is observed the relation between biological function activity and the secretory activity.
Description of drawings
Fig. 1 tissue and cell perfusion device structural representation
Fig. 2 A is the front view of sample perfusion bath
Fig. 2 B is the A-A sectional view of Fig. 2 A
The master of Fig. 3 A and Fig. 3 B microscope adaptation unit looks and sectional view
Fig. 4 A is the front view that sample perfusion bath cooperates in conjunction with the microscope adaptation unit
Fig. 4 B is the B-B sectional view of Fig. 4 A
Embodiment
Below in conjunction with accompanying drawing and concrete working process this device construction is elaborated.
Fig. 1 is tissue and cell perfusion device structural representation, and device comprises perfusate input system 1, water bath with thermostatic control system 2, sample perfusion bath 3, microscope adaptation unit 4 and perfusate gathering system 5.Perfusate input system 1 and perfusate gathering system 5 are connected the input slot 31 and the output magazine 33 of sample perfusion bath 3 respectively by the perfusion liquid flow conduits.The pipeline that perfusate input system 1 connects sample perfusion bath 3 is provided with water bath with thermostatic control system 2.Sample perfusion bath 3 is placed in the microscope adaptation unit 4 intermediary grooves with embedded mode.
Fig. 2 A and Fig. 2 B are the main sectional view of sample perfusion bath 3.Sample perfusion bath 3 is to form three independences but structure that the cell that communicates constitutes on the bright polytetrafluoroethylmaterial material of full impregnated, middle bigger cell is record bath 32, the cell of both sides is respectively applied for the input slot 31 and the output magazine 33 of perfusion liquid, and the bottom land of sample perfusion bath 3 is fixed with the cover glass of 24 * 50mm.The flow-guiding channel of the design by the bottom between the input slot 31 of the record bath 32 of sample perfusion bath 3 and both sides and output magazine 33 is communicated with, and makes medicine can act on sample rapidly, and liquid stabilising in the cell in the middle of not influencing.
When using native system, between the bottom of sample perfusion bath 3 and microscope adaptation unit 4, fix the cover glass 6 of 24 * 50mm size, three cells of sample perfusion bath 3 are communicated in the bottom, and tissue slice or culturing cell place on the interior cover glass of middle cell.Liquid is entered by the left side input slot 31 of sample perfusion bath 3, and two design undercuts through the bottom are full of intermediate record bath 32, and then flow into right side output magazine 33.Place glass capillary (external diameter 2mm) the drainage output perfusate of internal diameter 1.5mm at right side output magazine 33 edges.Treat liquid three cells continue to charge cut flow stable after, liquid capacities can be controlled in 225 μ L in the intermediate record bath 32.Because three cells are with the bottom circulating liquid, can guarantee still can change liquid fast, up hill and dale under the liquid capacity is little outside, the histocyte less relatively at flow velocity the situation, do not stay dead space.Sample perfusion bath 3 is open systems, especially can be suitable for patch clamp and other electric Physiological Experiment unicellular, the brain sheet.In the experiment relevant with electric physiology, sample perfusion bath 3 applied three cells change liquid system, can reduce the interior ion interference of cell groove and the interference of perfusion liquid levels instability generation.Reference electrode in the experimental record is placed in the output magazine 33 on right side, near collecting liquid, also can reduce the drift that the reference electrode polarizing voltage is introduced.
Fig. 3 A and Fig. 3 B are the main sectional view of microscope adaptation unit 4, microscope adaptation unit 4 is the pre-opened hole form fit on profile and the microscope stage, there is the sample of placement perfusion bath 3 centre, makes native system be fit to the fixedly Stage microscope of widening of various microscope stages and electric Physiological Experiment needs.
Fig. 4 A and Fig. 4 B are that sample perfusion bath 3 is in conjunction with the main sectional view of microscope adaptation unit 4 in application.The height of sample perfusion bath 3 is 3mm, can embed fully in the microscope adaptation unit 4 intermediary grooves.This device is that open active somatic cell is observed perfusion system, can be contained in fully to be inverted or just to put on the microscope, also can be used for the infrared differential-interference contrast microscope of water immersion objective, carries out the form observation of active somatic cell under various different conditions.Microscope adaptation unit 4 can conveniently provide with various drug delivery systems, pump and link to each other, and connects sample perfusion bath 3, because the design of bottom diversion trench makes medicine can act on sample rapidly, and the interior liquid stabilising of cell in the middle of not influencing.
(24 * 50mm) between sample perfusion bath 3 and microscope adaptation unit 4, and the two unites use at first to use a small amount of vaseline paste fixed cap slide during use.Tissue slice or culturing cell place in the sample perfusion bath 3.(general experiment need be 2～4mL/min) to the flow velocity that tissue or cell perfusate are set as required through input system 1, process water bath with thermostatic control system 2 heats and is the temperature (generally being set at 25～28 ℃) of needs, enter sample perfusion bath 3, reclaim perfusate by perfusate gathering system 4 at last and detect for the analysis of cells secretory activity.Auxiliary unit when water bath with thermostatic control system 2 imports for perfusate is isolated fully with perfusate, needs also can remove according to experimental study.The perfusate input system continues to feed the gas mixture of 95% oxygen and 5% carbonic acid gas, uses thermostatic control system 2 cell or brain sheet are remained in the relative constant temperature system, unites and uses constant temperature and oxygenation can fully keep the sample activity.Perfusate gathering system 5 is used the drainage of constant current peristaltic pump, can realize fast, in real time, regularly collect perfusate, for biochemistry detection emiocytosis or metabolism situation.
Claims (7)
1, be applicable to the biological tissue sample and the cell perfusion device of functional study, described device comprises perfusate input system (1), sample perfusion bath (3) and perfusate gathering system (5); Perfusate input system (1) and perfusate gathering system (5) are connected the input slot and the output magazine of sample perfusion bath (3) respectively by the perfusion liquid flow conduits; It is characterized in that: described sample perfusion bath (3) is to form three independences but structure that the cell that communicates constitutes on the bright polytetrafluoroethylmaterial material of full impregnated, middle bigger cell is record bath (32), the cell of both sides is respectively the input slot (31) and the output magazine (32) of perfusion liquid, and three bottom lands of sample perfusion bath (3) are the whole piece cover glass fixedly.
2, biological tissue sample and the cell perfusion device that is applicable to functional study according to claim 1, it is characterized in that, also be provided with microscope adaptation unit (4), microscope adaptation unit (4) is the pre-opened hole form fit on profile and the microscope stage, and the centre is useful on the groove that embeds sample perfusion bath (3).
3, biological tissue sample and the cell perfusion device that is applicable to functional study according to claim 1 and 2, it is characterized in that the record bath (32) of described sample perfusion bath (3) and the input slot (31) of both sides and output magazine (33) are communicated with by the flow-guiding channel of the design of bottom.
4, biological tissue sample and the cell perfusion device that is applicable to functional study according to claim 1 and 2, it is characterized in that described perfusate input system (1) is provided with water bath with thermostatic control system (2) with the perfusion liquid flow conduits that is connected sample perfusion bath (3).
5, biological tissue sample and the cell perfusion device that is applicable to functional study according to claim 1 and 2 is characterized in that the height of sample perfusion bath (3) is 3mm.
6, biological tissue sample and the cell perfusion device that is applicable to functional study according to claim 1 and 2 is characterized in that the volume of described sample perfusion bath (3) intermediary record bath (32) is designed to 225 μ L.
7, biological tissue sample and the cell perfusion device that is applicable to functional study according to claim 1 and 2 is characterized in that described perfusion liquid flow conduits all uses plastics or Glass tubing, and bag is by the aluminium foil of one deck ground connection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200920126738U CN201372287Y (en) | 2009-03-23 | 2009-03-23 | Living tissue specimen and cell perfusion device adaptable to functional study |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200920126738U CN201372287Y (en) | 2009-03-23 | 2009-03-23 | Living tissue specimen and cell perfusion device adaptable to functional study |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN201372287Y true CN201372287Y (en) | 2009-12-30 |
Family
ID=41498374
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200920126738U Expired - Fee Related CN201372287Y (en) | 2009-03-23 | 2009-03-23 | Living tissue specimen and cell perfusion device adaptable to functional study |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN201372287Y (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102174365A (en) * | 2010-12-30 | 2011-09-07 | 唐明 | Constant temperature perfusion experimental table of cells or tissues |
| CN102879557A (en) * | 2012-10-11 | 2013-01-16 | 南京大学 | Microscopic sample fixing device used for plant electrophysiological experiments |
| CN106940333A (en) * | 2017-04-17 | 2017-07-11 | 山西医科大学 | The outer Electrophysiology research perfusion system of isolated cells |
| CN108490165A (en) * | 2018-03-21 | 2018-09-04 | 厦门大学附属第医院 | Constant temperature patch-clamp cell record system |
| CN110514855A (en) * | 2018-09-27 | 2019-11-29 | 重庆微浪生物科技有限公司 | It is a kind of to change liquid method, changing liquid plate and its purposes in cell dyeing and particle washing |
| CN110583626A (en) * | 2019-09-26 | 2019-12-20 | 四川大学华西医院 | Multichannel constant-temperature perfusion system and application method thereof |
-
2009
- 2009-03-23 CN CN200920126738U patent/CN201372287Y/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102174365A (en) * | 2010-12-30 | 2011-09-07 | 唐明 | Constant temperature perfusion experimental table of cells or tissues |
| CN102174365B (en) * | 2010-12-30 | 2013-01-09 | 唐明 | Constant temperature perfusion experimental table of cells or tissues |
| CN102879557A (en) * | 2012-10-11 | 2013-01-16 | 南京大学 | Microscopic sample fixing device used for plant electrophysiological experiments |
| CN106940333A (en) * | 2017-04-17 | 2017-07-11 | 山西医科大学 | The outer Electrophysiology research perfusion system of isolated cells |
| CN108490165A (en) * | 2018-03-21 | 2018-09-04 | 厦门大学附属第医院 | Constant temperature patch-clamp cell record system |
| CN110514855A (en) * | 2018-09-27 | 2019-11-29 | 重庆微浪生物科技有限公司 | It is a kind of to change liquid method, changing liquid plate and its purposes in cell dyeing and particle washing |
| CN110583626A (en) * | 2019-09-26 | 2019-12-20 | 四川大学华西医院 | Multichannel constant-temperature perfusion system and application method thereof |
| CN110583626B (en) * | 2019-09-26 | 2021-07-30 | 四川大学华西医院 | Multichannel constant-temperature perfusion system and application method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN201372287Y (en) | Living tissue specimen and cell perfusion device adaptable to functional study | |
| CN103719075B (en) | Isolated organ preserving device and preserving method thereof | |
| CN106047691A (en) | 3D cell culture system | |
| WO2011063218A3 (en) | Cell collecting devices and methods for collecting cells | |
| WO2011105818A2 (en) | Cell culture unit and cell culture device including the same | |
| CN201999944U (en) | Continuous cell perfusion device based on living cell experiment | |
| CN104946528A (en) | Primary hepatic tissue cell separation system and method | |
| CN205313526U (en) | Organize perfusion groove suitable for morphology is observed and functional study | |
| CN108685946A (en) | A method of improving colon cancer PDX model success rates | |
| CN102174365B (en) | Constant temperature perfusion experimental table of cells or tissues | |
| CN103305419A (en) | Extraction and perfusion culture system for MSCs (Mesenchymal Stem Cells) | |
| US6602701B2 (en) | Three-dimensional cell growth assay | |
| Dailey et al. | Maintaining live cells and tissue slices in the imaging setup | |
| CN210090269U (en) | In vitro microvascular endothelial barrier function detection equipment | |
| CN2799572Y (en) | Automatic separation and culturing device for biological sample | |
| US20040253716A1 (en) | Bioreactor for cultivating cells on a matrix | |
| CN110108622B (en) | Isolated microvascular endothelial barrier function detection method and device | |
| CN204608020U (en) | The micro-culture apparatus of lymphocyte micronuclei experiment and lymphocyte micronuclei experiment test kit | |
| CN109913372A (en) | A kind of culture systems and method of living tissue | |
| CN210085471U (en) | Culture system of living tissue | |
| CN105385598B (en) | Human cerebral arteriovenous malformations biomechanical model and its vitro construction method | |
| KR20210075591A (en) | Cell culture apparatus using mixed gas injection | |
| CN212247072U (en) | Micro-fluidic device for in-vitro 3D cell and tissue culture | |
| CN212476743U (en) | T cell preparation facilities of high-efficient expression P53 oncostatin | |
| CN204731065U (en) | A kind of specimen sample device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20091230 Termination date: 20150323 |
|
| EXPY | Termination of patent right or utility model |