CN102719355B - Cell vacuum absorption fixing device - Google Patents
Cell vacuum absorption fixing device Download PDFInfo
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- CN102719355B CN102719355B CN201210250586.8A CN201210250586A CN102719355B CN 102719355 B CN102719355 B CN 102719355B CN 201210250586 A CN201210250586 A CN 201210250586A CN 102719355 B CN102719355 B CN 102719355B
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/40—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pressure
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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
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/06—Bioreactors or fermenters specially adapted for specific uses for in vitro fertilization
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/12—Well or multiwell plates
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Abstract
The invention discloses a cell vacuum absorption fixing device comprising a comprehensive bottom plate, an ovum functional plate and a first sperm tank, wherein the ovum functional plate is provided with a concave ovum tank; the side of the comprehensive bottom plate is provided with a connector; and the ovum functional plate is also provided with a plurality of micropores formed in the bottom of the ovum tank. The cell vacuum absorption fixing device also comprises a pressure control loop comprising a diaphragm liquid pump, a pressure sensor, an electro-hydraulic servo valve and a liquid discharging pipe, wherein an inlet of the diaphragm liquid pump is connected with the electro-hydraulic servo valve, and an outlet of the diaphragm liquid pump is connected with the liquid discharging pipe; and the input end of the pressure sensor is connected between the connector and the electro-hydraulic servo valve, and the output end of the pressure sensor is connected to the input end of the electro-hydraulic servo valve. Cells are automatically fixed and released by means of the work of the diaphragm liquid pump, and the diaphragm liquid pump has the characteristics of high efficiency, high repeatability and the like.
Description
Technical field
The present invention relates to field of medical, relate in particular to a kind of cell vacuum adsorption fixing device.
Background technology
Micromanipulative technique (Micromanipulation Technique) refers under high power inverted microscope, utilize micromanipulator (Micromanipulator), control the mechanism that injection needle moves in field of microscope, be used for carrying out a kind of method of cell or body early embryo operation.The material that can be injected into cell includes: various organoids, kinases, histological chemistry's mark (such as horseradish peroxidase or fluorescent yellow), protein, metabolic substd, micro-magnetic head, ion, antibody, gene, molecular biological mRNA and DNA etc.Use this technology, also can realize the accurate conveying (Micro-perfusion in Graft After) of small amount for individual cells or one group of cell (skin rises to milliliter) medicament or medicine.
Modern biological, medical science, in the fields such as agricultural, cell injection has obtained application more and more widely, but experiment whole efficiency and success ratio are all not high, influence factor comprises that being injected cell catches with fixing, microinjection pin is accurately located, cytomorphosis is with broken, the operation links such as the control of microinjection amount, Chinese scholars is respectively from cell search and location, the fast feed of microinjection pin, the control of microinjection amount, image recognition processing, micro-operation locus is demarcated and is detected, improve in various degree the level of automation of cell injection, but integrated automation level is also very low.In addition the operation of cell injection at present need be asked and first use sticking pin by cell capture, then complete injection process with entry needle, conventionally such system needs two operators, and cell all needs with sticking pin absorption, fixing, efficiency ratio is lower, and in injection process, cell easily flows, and affects the success ratio of cell injection.Therefore study new cellular localization and stationary installation, microinjection pin fast feed method, microinjection amount is accurately controlled and automatic cytological injection operation technique and system architecture etc. are injected high success rate to realizing cell, high-throughput is significant.
Existing cell injecting systems is all the structure based on tradition " the simple eye formula of both hands ", operator is observed and is found object run cell by microscope ocular, then utilize the left and right operator that is installed on microscope both sides, wherein left hand is sticking pin, for catching of cell, the right hand is entry needle, completes the injection of cell.From cell injection feature, cell in micron dimension is suspended in nutrient solution conventionally, sticking pin moves or slight vibrations in nutrient solution, easily cause nutrient solution disturbance and drive cell position to change, cause the operations such as cell search and location very difficult, stability deficiency, in addition owing to can only fix a cell at every turn, causes cell injection conventional efficient low.Meanwhile, operator must have very strong professional technique technical ability, need through long-term strict training.The microinjection system that biotechnology circle generally uses is at present substantially by Japanese Narishige, and German Eppendorf and U.S. Sutter tri-companies capture, and existing cell injecting systems is still in the manual or semi-automatic stage.Therefore study the automatic identification of cell and the new device of fixed cell, change the injection system of traditional left and right operator, and then improve the core technology that cell injection efficiency is the application of popularization cell injecting systems.
Therefore,, for above-mentioned technical problem, be necessary to provide a kind of cell vacuum adsorption fixing device in batches, to overcome above-mentioned defect.
Summary of the invention
In view of this, the object of the present invention is to provide a kind of cell vacuum adsorption fixing device, this cell vacuum adsorption fixing device has high precision, the features such as high-level efficiency and high repeatability.
For achieving the above object, the invention provides following technical scheme:
A kind of cell vacuum adsorption fixing device, it comprises comprehensive egative film, be installed on the ovum functional sheet on comprehensive egative film, be formed at the first sperm groove on comprehensive egative film, described ovum functional sheet is provided with the ovum groove of spill, the side of described comprehensive egative film is provided with a joint, on described ovum functional sheet, be also provided with the some micropores that are formed at ovum trench bottom, described cell vacuum adsorption fixing device also comprises pressure control circuit, described pressure control circuit comprises diaphragm type liquid pump, pressure transmitter, the electrohydraulic servo valve being connected with joint and liquid discharge pipe, the import of described diaphragm type liquid pump is connected with electrohydraulic servo valve, described diaphragm type liquid pump outlet is connected with liquid discharge pipe, the input terminus of described pressure transmitter is connected between joint and electrohydraulic servo valve, the output terminal of described pressure transmitter is connected to the input terminus of electrohydraulic servo valve.
Preferably, in above-mentioned cell vacuum adsorption fixing device, described comprehensive egative film is provided with second rectangular tank in order to the first rectangular tank of ovum functional sheet to be installed and to be positioned at the first rectangular tank and further to cave in of depression, described the second rectangular tank is vacuum chamber, is provided with the passage that makes liquid communication between described vacuum chamber and joint.
Preferably, in above-mentioned cell vacuum adsorption fixing device, described cell vacuum adsorption fixing device also comprises the second sperm groove being formed on comprehensive egative film and the raceway groove that connects the first sperm groove and the second sperm groove.
Preferably, in above-mentioned cell vacuum adsorption fixing device, the width range of described raceway groove is 80 ~ 120 μ m.
Preferably, in above-mentioned cell vacuum adsorption fixing device, the diameter range of described micropore is 10 ~ 500 μ m.
Preferably, in above-mentioned cell vacuum adsorption fixing device, described joint is connected to electrohydraulic servo valve by silicone tube, and the other end of described electrohydraulic servo valve is connected to the import of diaphragm type liquid pump by silicone tube.
Preferably, in above-mentioned cell vacuum adsorption fixing device, described ovum groove is square.
Can find out from technique scheme, the cell vacuum adsorption fixing device of the embodiment of the present invention relies on the work of diaphragm type liquid pump to fix or release cells, repeatable high; And device can carry out fixing of cell in batches, and working efficiency is high; The output terminal of described pressure transmitter is received the input terminus of electrohydraulic servo valve, forms closed loop, accurately controls the pressure of liquid in pipeline.
Accompanying drawing explanation
In order to be illustrated more clearly in the embodiment of the present invention or technical scheme of the prior art, to the accompanying drawing of required use in embodiment or description of the Prior Art be briefly described below, apparently, accompanying drawing relevant of the present invention in the following describes is only some embodiments of the present invention, for those of ordinary skills, do not paying under the prerequisite of creative work, can also obtain according to these accompanying drawings other accompanying drawing.
Fig. 1 is the schematic perspective view of cell vacuum adsorption fixing device of the present invention;
Fig. 2 is the schematic diagram of pressure control circuit in cell vacuum adsorption fixing device of the present invention;
Fig. 3 is the enlarged view of micropore on ovum functional sheet in cell vacuum adsorption fixing device of the present invention;
Fig. 4 is the enlarged view of raceway groove between two sperm grooves in cell vacuum adsorption fixing device of the present invention.
Wherein: 1, comprehensive egative film; 2, ovum functional sheet; 3, ovum groove; 4, micropore; 5, the first sperm groove; 6, raceway groove; 7, the second sperm groove; 8, vacuum chamber; 9, joint; 10, diaphragm type liquid pump; 11, electrohydraulic servo valve; 12, pressure transmitter; 13, liquid discharge pipe.
Embodiment
The invention discloses a kind of cell vacuum adsorption fixing device, this cell vacuum adsorption fixing device has high precision, the features such as high-level efficiency and high repeatability.
This cell vacuum adsorption fixing device comprises comprehensive egative film, be installed on ovum functional sheet on comprehensive egative film, be formed at the first sperm groove on comprehensive egative film.Described ovum functional sheet is provided with the ovum groove of spill.The side of described comprehensive egative film is provided with a joint.On described ovum functional sheet, be also provided with the some micropores that are formed at ovum trench bottom.Described cell vacuum adsorption fixing device also comprises pressure control circuit, and described pressure control circuit comprises diaphragm type liquid pump, pressure transmitter, the electrohydraulic servo valve and the liquid discharge pipe that are connected with joint.The import of described diaphragm type liquid pump is connected with electrohydraulic servo valve, described diaphragm type liquid pump outlet is connected with liquid discharge pipe, the input terminus of described pressure transmitter is connected between joint and electrohydraulic servo valve, and the output terminal of described pressure transmitter is connected to the input terminus of electrohydraulic servo valve.
Further, described comprehensive egative film is provided with second rectangular tank in order to the first rectangular tank of ovum functional sheet to be installed and to be positioned at the first rectangular tank and further to cave in of depression, described the second rectangular tank is vacuum chamber, is provided with the passage that makes liquid communication between described vacuum chamber and joint.
Further, described cell vacuum adsorption fixing device also comprises the second sperm groove being formed on comprehensive egative film and the raceway groove that connects the first sperm groove and the second sperm groove.
Further, the width range of described raceway groove is 80 ~ 120 μ m.
Further, the diameter range of described micropore is 10 ~ 500 μ m.
Further, described joint is connected to electrohydraulic servo valve by silicone tube, and the other end of described electrohydraulic servo valve is connected to the import of diaphragm type liquid pump by silicone tube.
Further, described ovum groove is square.
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is described in detail, obviously, described embodiment is only the present invention's part embodiment, rather than whole embodiment.Based on the embodiment in the present invention, the every other embodiment that those of ordinary skills obtain under the prerequisite of not making creative work, belongs to the scope of protection of the invention.
As shown in Figure 1 to Figure 3, cell vacuum adsorption fixing device comprises comprehensive egative film 1, is installed on ovum functional sheet 2 on comprehensive egative film 1, is formed at the first sperm groove 5 on comprehensive egative film 1.The size of comprehensive egative film 1 is according to general slide glass, and only thickness increases a little, is applicable to most microscopes.Ovum functional sheet 2 is provided with ovum groove 3 spill and that be square.Ovum functional sheet 2 and comprehensive egative film 1 are all injection molded, nontoxic as raw material take medical polycarbonate (PC), and light transmission is good, is applicable to micrurgy.The side of comprehensive egative film 1 is provided with a joint 9, is also provided with the some micropores 4 that are formed at ovum groove 3 bottoms on ovum functional sheet 2, and the diameter range of micropore 4 is 10 ~ 500 μ m.Micropore 4 can break into laser-beam drilling machine, so arranges, and is convenient to process the through hole that diameter is different, and then can operates for different ovum.In the cell vacuum adsorption fixing device of the embodiment of the present invention, can be as required, do the quantity of portalling and be: 1,2,3 × 3 arrays, 4 × 4 arrays, 5 × 5 arrays even more (size of pitch of holes, ovum groove 3 all can change because of needs).As shown in Figure 3, take 5 × 5 as example, so just can once realize multiple, in batches cell is fixing, has significantly improved the efficiency of injection., between array hole, can demarcate by auxiliary positioning mark meanwhile, make the fixing cell position of this device clear and definite, the system of being convenient to catches and does not need to search for randomly.
As shown in Figures 1 and 2, cell vacuum adsorption fixing device also comprises pressure control circuit.Pressure control circuit comprises diaphragm type liquid pump 10, pressure transmitter 12, the electrohydraulic servo valve 11 and the liquid discharge pipe 13 that are connected with joint 9.The import of diaphragm type liquid pump 10 is connected with electrohydraulic servo valve 11, the outlet of diaphragm type liquid pump 10 is connected with liquid discharge pipe 13, the input terminus of pressure transmitter 12 is connected between joint 9 and electrohydraulic servo valve 11, and the output terminal of pressure transmitter 12 is connected to the input terminus of electrohydraulic servo valve 11.Joint 9 is connected to electrohydraulic servo valve 11 by silicone tube, and the other end of electrohydraulic servo valve 11 is connected to the import of diaphragm type liquid pump 10 by silicone tube.Fix or release cells by 10 work of diaphragm type liquid pump, repeatable high; And device can carry out fixing of cell in batches, and working efficiency is high; And the output terminal of pressure transmitter is received the input terminus of electrohydraulic servo valve, form closed loop, accurately control the pressure of liquid in pipeline.
Pressure transmitter 12 has encapsulated signal amplifier, the input terminus of pressure transmitter 12 is connected between the joint 9 and electrohydraulic servo valve 11 of comprehensive egative film 1, measure liquid pressure, electrical signal after amplifying is passed to electrohydraulic servo valve 11, and with the force value comparison setting in advance, specify the action of electrohydraulic servo valve 11 to carry out pressure compensation, thereby the pressure of control loop accurately continuously carry out cell and fix under the prerequisite that guarantees damaging cells not.
Comprehensive egative film 1 is provided with second rectangular tank in order to the first rectangular tank of ovum functional sheet 2 to be installed and to be positioned at the first rectangular tank and further to cave in of depression, described the second rectangular tank is vacuum chamber 8, between described vacuum chamber 8 and joint 9, is provided with the passage that makes liquid communication.
As shown in Figure 4, cell vacuum adsorption fixing device also comprises the second sperm groove 7 being formed on comprehensive egative film 1 and the raceway groove 6 that connects the first sperm groove 5 and the second sperm groove 7.The width range of raceway groove 6 is 80 ~ 120 μ m., two sperm grooves are interconnected, and splash into the first sperm groove 5 solution that contains sperm, because sperm has mobility, swimming across the sperm that raceway groove 6 enters the second sperm groove 7 must be that vitality is stronger, and the sperm catching is wherein injected, so arrange, can improve survival rate of embryo.
The principle of work of the cell vacuum adsorption fixing device of the embodiment of the present invention is: the quality of cell (for example ovum) is minimum, when there being a very little power near it, just can make its motion, therefore in the time that working, extracts by diaphragm type liquid pump 10 the interior liquid of vacuum chamber 8 out, at micropore, 4 places produce negative pressure, and near cell is subject to suction to draw close finally and be fixed to micropore 4.
The cell vacuum adsorption fixing device of the embodiment of the present invention relies on the work of diaphragm type liquid pump to fix or release cells, repeatable high; And device can carry out fixing of cell in batches, and working efficiency is high; The output terminal of described pressure transmitter is received the input terminus of electrohydraulic servo valve, forms closed loop, accurately controls the pressure of liquid in pipeline.
To those skilled in the art, obviously the invention is not restricted to the details of above-mentioned one exemplary embodiment, and in the situation that not deviating from spirit of the present invention or essential characteristic, can realize the present invention with other specific form.Therefore, no matter from which point, all should regard embodiment as exemplary, and be nonrestrictive, scope of the present invention is limited by claims rather than above-mentioned explanation, is therefore intended to all changes that drop in the implication and the scope that are equal to important document of claim to include in the present invention.Any Reference numeral in claim should be considered as limiting related claim.
In addition, be to be understood that, although this specification sheets is described according to embodiment, but be not that each embodiment only comprises an independently technical scheme, this narrating mode of specification sheets is only for clarity sake, those skilled in the art should make specification sheets as a whole, and the technical scheme in each embodiment also can, through appropriately combined, form other embodiments that it will be appreciated by those skilled in the art that.
Claims (4)
1. a cell vacuum adsorption fixing device, it is characterized in that: it comprises comprehensive egative film, be installed on the ovum functional sheet on comprehensive egative film, be formed at the first sperm groove on comprehensive egative film, described ovum functional sheet is provided with the ovum groove of spill, the side of described comprehensive egative film is provided with a joint, on described ovum functional sheet, be also provided with the some micropores that are formed at ovum trench bottom, described cell vacuum adsorption fixing device also comprises pressure control circuit, described pressure control circuit comprises diaphragm type liquid pump, pressure transmitter, the electrohydraulic servo valve being connected with joint and liquid discharge pipe, the import of described diaphragm type liquid pump is connected with electrohydraulic servo valve, described diaphragm type liquid pump outlet is connected with liquid discharge pipe, the input terminus of described pressure transmitter is connected between joint and electrohydraulic servo valve, the output terminal of described pressure transmitter is connected to the input terminus of electrohydraulic servo valve, described comprehensive egative film is provided with second rectangular tank in order to the first rectangular tank of ovum functional sheet to be installed and to be positioned at the first rectangular tank and further to cave in of depression, described the second rectangular tank is vacuum chamber, between described vacuum chamber and joint, be provided with the passage that makes liquid communication, described cell vacuum adsorption fixing device also comprises the second sperm groove being formed on comprehensive egative film and the raceway groove that connects the first sperm groove and the second sperm groove, the width range of described raceway groove is 80～120 μ m.
2. cell vacuum adsorption fixing device according to claim 1, is characterized in that: the diameter range of described micropore is 10～500 μ m.
3. cell vacuum adsorption fixing device according to claim 1, is characterized in that: described joint is connected to electrohydraulic servo valve by silicone tube, the other end of described electrohydraulic servo valve is connected to the import of diaphragm type liquid pump by silicone tube.
4. cell vacuum adsorption fixing device according to claim 1, is characterized in that: described ovum groove is square.
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| CN201210250586.8A CN102719355B (en) | 2012-07-19 | 2012-07-19 | Cell vacuum absorption fixing device |
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| CN201210250586.8A CN102719355B (en) | 2012-07-19 | 2012-07-19 | Cell vacuum absorption fixing device |
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| CN102719355B true CN102719355B (en) | 2014-07-09 |
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| CN104099237A (en) * | 2014-06-30 | 2014-10-15 | 浙江星博生物科技有限公司 | Sperm optimal selection device based on microtubule passages and optimal selection method |
| CN108660072B (en) * | 2018-05-18 | 2023-07-18 | 苏州大学 | Oocyte micro-operation system and method based on auxiliary support of targeting region |
| CN115431076B (en) * | 2022-10-08 | 2023-08-18 | 广东长盈精密技术有限公司 | Thin-wall workpiece clamp |
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| WO2001019978A1 (en) * | 1999-09-14 | 2001-03-22 | Cornell Research Foundation, Inc. | Microfabrication of a nuclear transfer array for high-throughput animal cloning |
| CN1572877A (en) * | 2003-05-21 | 2005-02-02 | 富士通株式会社 | System and apparatus for injecting substance into cell |
| EP1762608A1 (en) * | 2005-09-09 | 2007-03-14 | Fujitsu Limited | Petri dish for trapping cell |
| CN102242055A (en) * | 2011-06-03 | 2011-11-16 | 博奥生物有限公司 | Method for evaluating sperm activity and screening sperms and special microfluidic chip device for same |
| CN202688315U (en) * | 2012-07-19 | 2013-01-23 | 苏州大学 | Cell vacuum adsorption fixing device |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101705185A (en) * | 2009-11-24 | 2010-05-12 | 广州海太光电生物科技有限公司 | Anaerobic and microaerophilic gas treatment device |
| CN202279825U (en) * | 2011-10-20 | 2012-06-20 | 天津开发区合普工贸有限公司 | Simulated experiment device for cell low-pressure hypoxia condition |
| CN102517209A (en) * | 2011-11-24 | 2012-06-27 | 中国人民解放军第四军医大学 | Multichannel cell stretch stress loading control meter |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001019978A1 (en) * | 1999-09-14 | 2001-03-22 | Cornell Research Foundation, Inc. | Microfabrication of a nuclear transfer array for high-throughput animal cloning |
| CN1572877A (en) * | 2003-05-21 | 2005-02-02 | 富士通株式会社 | System and apparatus for injecting substance into cell |
| EP1762608A1 (en) * | 2005-09-09 | 2007-03-14 | Fujitsu Limited | Petri dish for trapping cell |
| CN102242055A (en) * | 2011-06-03 | 2011-11-16 | 博奥生物有限公司 | Method for evaluating sperm activity and screening sperms and special microfluidic chip device for same |
| CN202688315U (en) * | 2012-07-19 | 2013-01-23 | 苏州大学 | Cell vacuum adsorption fixing device |
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