CN102943041A - Oxygen continuously filling cell and tissue hatching device - Google Patents
Oxygen continuously filling cell and tissue hatching device Download PDFInfo
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- CN102943041A CN102943041A CN2012104769697A CN201210476969A CN102943041A CN 102943041 A CN102943041 A CN 102943041A CN 2012104769697 A CN2012104769697 A CN 2012104769697A CN 201210476969 A CN201210476969 A CN 201210476969A CN 102943041 A CN102943041 A CN 102943041A
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Abstract
The invention discloses an oxygen continuously filling cell and tissue hatching device which comprises a hatching cup, a support arranged in the hatching cup and an air filling device, wherein a plurality of placing supports in radial arrangement are arranged on the support, and tissue cups are arranged on the placing supports. The air filling device is provided with an air inlet pipe and a plurality of air outlet pipes which are connected with the air inlet pipe and are in radial arrangement surrounding the air inlet pipe, the plurality of air outlet pipes correspond to the plurality of placing supports one by one and are located below the placing supports, and a plurality of air outlet holes are arranged on the air outlet pipes at the positions corresponding to the placing supports. A liquid inlet circulation mechanism connected with the lower portion of a cavity arranged in the hatching cup is arranged on the outer lateral wall of the hatching cup and is provided with a liquid inlet connected with the outside. The oxygen continuously filling cell and tissue hatching device can be filled with oxygen continuously, improves oxygen concentration and avoids cell damage.
Description
Technical field
The present invention relates to the medical analysis technical field, particularly a kind of cell and tissue incubation's device that continues logical oxygen.
Background technology
It is to adopt the technology such as patch clamp, Laser Scanning Confocal Microscope to observe the variation of various ion(ic)currents, voltage to the cell near physiological status that the microcosmic bio-electro-physiologic learns a skill.Patch clamp namely adopts microelectrode to clamp down on cell voltage or electric current, carries out the researchs such as resting potential, action potential, ionic channel, neuroelectricity conduction, chemical mediator conduction.Laser Scanning Confocal Microscope then can be used for the special components such as the calcium in the viable cell, sodium, chlorine, hydrogen are carried out dynamic monitoring, makes the dynamic changing curve of these compositions.The object of research comprises all excitability cells, such as brain, spinal cord, peripheral nerve, cardiac muscle, sexual cell etc.Employed cell can be from the tissue block of animal, the primary cell of acute isolation or the cell strain of stable subculture.In order to simulate better the physiological status at body, improve experiment stability, avoid the factor affecting experimental results such as cytopathy, apoptosis, usually need in the damping fluid that continues logical oxygen, cultivate above-mentioned tissue block or cell, prolong tissue block and cytoactive.With respect to other somatocyte, neurocyte ischemic, anoxic namely enter irreversible apoptosis program more than 6 minutes, more need the culture condition and the environment that provide stable.Especially the neurone of brain sheet, spinal cord and the acute isolation of adult animals, the ability of tolerance anoxic is lower, and the experimenter slightly has carelessness namely to cause all neuronal deaths in the process of ex vivo treatment, can't study smoothly.And apparatus, current, bubble all may cause damage to cell to the collision of tissue block or primary cell, make it can't reach stable physiological level.Therefore, be necessary to select the incubation slot of sustainable logical oxygen to carry out the cultivation of tissue block and cell.
Incubation slot commonly used has three kinds during at present test.The first is the most frequently used, and the small beaker that namely directly uses 50ml fills with the buffered soln of imitation tissue juice as incubation slot, such as artificial cerebrospinal fluid, continues to pass into oxygen gas mixture.The second is Gibb brain sheet incubation slot, designed by Alasadair J doctor Gibb of London university, its structure is: 35mm Petri culture dish and lid thereof are removed the bottom, gauze is stuck between two remaining annulus, remove the end to end plastics tubing of two ends remainder with syringe shell, the annulus that sticks gauze is stuck in the middle part of 100ml beaker with plastics tubing.In plastics tubing, insert the tubule of 2-3mm internal diameter, fill with the Incubating Solution of buffering in the cup after, continue to pass into oxygen gas mixture through tubule.The advantage of this incubation slot is that the bubble of oxygen gas mixture from bottom to top promotes damping fluid and flows in plastics tubing, sluggish flow from top to bottom outside pipe, and the brain sheet is attached on the gauze by flowing fluid, is not easy to hike up.The third incubation slot is designed by James R doctor Moyer of Yale: the bottom of 24 hole culture dish is removed, and the polishing periphery is avoided sharp keen edge damage tissue; The wall of every culture hole all punches; Being beneficial to nutrient solution circulates; Whole 24 orifice plates half-and-half cut, and insert culture tank, and the nylon stocking support is used in the bottom, continue logical oxygen.
The incubation slot of prior art has the following disadvantages:
1, because the dissolution rate of oxygen is low, the bubble of the oxygen gas mixture that continues to pass into is many directly to leave the water surface after the rising, the solution that is difficult to keep whole system is the available oxygen state of saturation, less tissue or the cell of diffusing to downwards experimental results show that the Central Nervous System of The Adult Rat neurone of acute isolation is difficult to keep for a long time the normal physiological state in above incubation slot;
2, a plurality of tissue of inserting in the small beaker of first, second kind incubation slot or the single annulus card are easily assembled, and are easy to can't distinguish because of displacement in by the process of moving at liquid-flow, container;
3, the third incubation slot, liquid flow from top to bottom, make tissue bear downward gravity and hydrodynamicpressure and the buoyancy that makes progress and culture dish support, and pressure is larger, is easy to cause cell impaired.
This shows how a kind of incubating device is provided, can continue logical oxygen, improve oxygen concn, can avoid cell impaired simultaneously, this is the present technical issues that need to address in this area.
Summary of the invention
The object of the invention is to, a kind of cell and tissue incubation's device that continues logical oxygen is provided, can lead to constantly oxygen, improve oxygen concn, avoid cell impaired.
For solving above technical problem, technical scheme of the present invention is:
A kind of cell and tissue incubation's device that continues logical oxygen, comprise and hatch cup, be arranged on support and the breather of hatching in the cup, support is provided with a plurality of dispensers that arrange radially, lay the tissue cup on the described dispenser, described breather be provided with inlet pipe and with a plurality of escape pipes that arrange radially around inlet pipe of inlet pipe conducting, a plurality of escape pipes and a plurality of dispenser arrange correspondingly and escape pipe is positioned at the dispenser below, and the position corresponding with dispenser is provided with some production wells on the escape pipe; Described outer side wall of hatching cup is provided with the liquid feed liquor cycling mechanism with the bottom conducting of hatching glass internal cavities, and described liquid feed liquor cycling mechanism is provided with the fluid inlet with extraneous conducting.
Preferably, described support comprises central shank and a plurality of connection handles that arrange radially around central shank, and a plurality of dispensers are arranged at respectively the end of different connection handles.
Preferably, described central shank is provided with a through hole, and described inlet pipe is passed through hole.
Preferably, connection handle comprises long connection handle and short connection handle, and long connection handle and short connection handle are arranged alternately.
Preferably, be positioned on the long connection handle dispenser its be provided with connection bump near a side of hatching glass inwall, correspondingly be provided with the connection screens that cooperates with it on the described inwall of hatching cup.
Preferably, described inwall of hatching cup is provided with for the fixing mounting block of inlet pipe.
Preferably, described upper opening place of hatching cup is provided with a bowl cover, and described bowl cover is provided with the ventilating slit.
Preferably, described dispenser is toroidal frame.
Preferably, described liquid feed liquor cycling mechanism is to be arranged on the hollow cup ear of hatching on glass outer side wall, described hollow cup ear comprises the upright arm of hollow, the upper oblique arm of hollow, the lower oblique arm of hollow, the upper end of described upright arm is provided with opening, as liquid feeding end, described upper oblique arm one end be connected with the top in the outside of hatching cup and with hatch a glass internal cavities conducting, the other end is downward-sloping be connected with the outer wall of upright arm and with the conducting of upright arm internal cavities, described lower oblique arm one end be connected with the bottom in the outside of hatching cup and with hatch a glass internal cavities conducting, the other end be inclined upwardly be connected with the outer wall of upright arm and with the conducting of upright arm internal cavities.
Preferably, the cup end of described tissue cup is screen cloth.
Compared with prior art, cell and the tissue incubation's device that continues logical oxygen of the present invention, hatch cup owing to comprising, be arranged on support and the breather of hatching in the cup, support is provided with a plurality of dispensers that arrange radially, lay the tissue cup on the described dispenser, thisly separate the mode that different tissue cups is hatched tissue or cell, can avoid a plurality of tissue to assemble or in the process that liquid-flow, container are moved, be shifted; Since breather be provided with inlet pipe and with a plurality of escape pipes that arrange radially around inlet pipe of inlet pipe conducting, a plurality of escape pipes and a plurality of dispenser arrange correspondingly and escape pipe is positioned at the dispenser below, the position corresponding with dispenser is provided with some production wells on the escape pipe, the breather of this structure can provide the environment of oxygen enrichment effectively, make the solution of whole system be the available oxygen state of saturation, the oxygen that is shed by escape pipe fully upwards diffuses to tissue or the cell in the tissue cup on the corresponding dispenser, to keep for a long time its normal physiological state; Be provided with liquid feed liquor cycling mechanism with the bottom conducting of hatching glass internal cavities owing to hatching the outer side wall of cup, described liquid feed liquor cycling mechanism is provided with the fluid inlet with extraneous conducting, this is so that hatch glass liquid stream from lower to upper, tissue and cell only bear self downward gravity, hydrodynamicpressure under undirected, buoyancy and culture dish support upwards arranged in addition, reduce tissue and cell impaired, can keep the circulation of slow liquid stream simultaneously.
Description of drawings
Fig. 1 is that the present invention continues the cell of logical oxygen and the structural representation of tissue incubation's device (not comprising the tissue cup);
Fig. 2 is the vertical view of Fig. 1 medium-height trestle;
Fig. 3 is the schematic diagram of breather among Fig. 1;
Fig. 4 is that the A of breather among Fig. 3 is to sectional view;
Fig. 5 is the structural representation of tissue cup.
Embodiment
Below in conjunction with drawings and the specific embodiments, cell and the tissue incubation's device that continues logical oxygen of the present invention is elaborated.
Referring to Fig. 1 ~ Fig. 5, the cell that continues logical oxygen in the present embodiment and tissue incubation's device comprise hatch cup 1, be arranged on support 2 and the breather 3 of hatching in the cup 1, be arranged on hatch on the cup sidewall, as the hollow cup ear 4 of liquid feed liquor cycling mechanism.
Hatch cup 1 and be cylindrical, hatch glass position of inwall middle and upper part along 6 the screens (not shown)s that are circumferentially with of inwall, be used for laying the support 2 of tissue cup.Hatch and also be provided with two mounting blocks on glass inwall, this mounting block is half arc convex (not shown), is used for the fixedly inlet pipe of breather, and two and half arc convex are positioned at hatches At The Heights different on glass inwall.The position of hatching close upper opening on glass inwall also is provided with the groove (not shown), is used for holding bowl cover 1a.Hatch a glass outer wall and be labeled as and establish one every 200ml and walk crosswise scale, wherein the scale of 1000ml is longer, and outer wall corresponding to screens be mark the 1st, 2,3,4,5, No. 6 respectively, so that the coding of dividing tissue piece.
In the present embodiment, the volume of hatching cup is 2000ml, height 15cm, and internal diameter is respectively 10cm, wall thickness 2mm.6 screens are positioned at hatches 10cm At The Height on glass inwall.Two and half arc-shaped protrusions lay respectively at hatches glass inwall 5cm and 10cm At The Height, and the groove that holds bowl cover is positioned at hatches glass cup inwall from upper limb 3mm place.
Support 2 comprises central shank 21 and 12 link spans that arrange radially around central shank 21, the annular dispenser 24 that link span comprises connection handle and is positioned at the connection handle end, wherein 6 long connection handles 22,6 short connection handles 23, long connection handle and short connection handle are arranged alternately; Be positioned at the dispenser on the long connection handle, it is provided with projection 25 near a side of hatching glass inwall, is used for hatching the clamping matching on glass inwall support being fixed; The centre of central shank 21 also is provided with a through hole 21a, is used for the inlet pipe of breather is passed through; In the present embodiment, the long 0.5cm of central shank, middle through hole internal diameter is 0.25cm.Dispenser in the present embodiment is used for laying the tissue cup; In the present embodiment, each connection handle and dispenser all are in same plane, in other embodiments, also can make each connection handle and stagger up and down and be not in conplane structure.In the present embodiment, the dispenser of annular, its external diameter 2.0cm, internal diameter 1.8cm.
The profile of tissue cup 5 is trapezoidal, and in the present embodiment, it highly is 1.5cm, and internal diameter suitable for reading is 2.0cm, and end opening internal diameter 1.6cm allows the tissue cup to insert in the annular tissue glass stand of hatching in the frame, and the tissue cup cup end 51 is 6 purpose standard nylon screen clothes.Tissue cup upper limb the projection 52 at the vertical cup end is arranged and be parallel to glass at the bottom of projection 53, tweezers easy to use or mosquito forceps take out or insert tissue glass.The tissue cup is used for holding tissue block or cell paster.
Hatch the cup ear 4 that arranges on the outer side wall of cup and be the hollow cup ear of similar " K " shape, this hollow cup ear is as liquid feed liquor cycling mechanism, it comprises upright arm 41, upper oblique arm 42, lower oblique arm 43, upper oblique arm 42 1 ends with hatch glass outer side wall top and be connected, the other end is downward-sloping to be connected with upright arm 41 middle parts, lower oblique arm 43 1 ends with hatch a glass outer side wall bottom and be connected, the other end is connected with the upright arm middle part, upper oblique arm 42, lower oblique arm 43, upright arm 41 is hollow structure, the internal cavities conducting of upper oblique arm 42 and upright arm 41, the internal cavities conducting of lower oblique arm 43 and upright arm 41, upright arm with hatch the cup contour, its upper end is provided with opening, stretches into for liquid-inlet pipe, and this opening part is provided with ear lid (not shown); Cup ear internal diameter 2cm, the length of upper oblique arm and lower oblique arm is 8cm, and upper oblique arm and lower oblique arm all intersect with upright arm, and upright arm is with to hatch cup contour, and opening suitable for reading allows to insert liquid-inlet pipe, and the upright arm inwall is provided with groove from upper limb 3mm place, covers for holding ear.The hollow cup ear allows the liquid in the whole incubating device slowly to circulate under the drive of air-flow or outer liquid feeding stream, enters oblique arm on glass ear, upright arm, lower oblique arm by hatching glass top, returns and hatches in the cup.
Also the liquid in-out pipe can be set, progressively replace liquid in the incubation slot when liquid in-out pipe was hatched for the long period, for selecting device, be similar to the intravenous infusion pipeline, flexible pipe can enter the ear lid.Bowl cover diameter 10cm, there is the circular hole of an internal diameter 0.6cm center, can be by the inlet pipe of breather, bowl cover 1a blow-by, it on be a plurality of height alternately, the mutual incomplete secter pat of connection, be conducive to the gas discharge, avoid simultaneously extraneous particulate waft into.
Ear lid diameter is 2cm, does not need to change the constantly tight capping cup ear of liquid.
The working process of the present embodiment is as follows:
Liquid enters lower oblique arm from upright arm, enter the bottom of hatching cup from lower oblique arm again, liquid slowly enters into from the bottom to top and hatches cup, liquid in the whole incubating device slowly circulates under the drive of air-flow or outer liquid feeding stream, enter oblique arm on the hollow cup ear, upright arm, lower oblique arm by hatching glass top, return and hatch in the cup; The inlet pipe external gas source, oxygen enters from inlet pipe, then enter and be positioned at the escape pipe of hatching the cup bottom, outwards shed by the production well on the escape pipe, because production well just in time is positioned at the dispenser below corresponding with escape pipe, then the oxygen that sheds of the production well of escape pipe is supplied with tissue cup on the dispenser thus because each dispenser arranges an escape pipe accordingly, so oxygen can be evenly, cell paster and the tissue block in providing organization's cup constantly.
Compare with incubating device of the prior art, the cell and the tissue incubation's device that continue logical oxygen of the present embodiment have following advantage:
1, the environment of effective supply oxygen enrichment, the solution of keeping whole system is the available oxygen state of saturation, and oxygen fully upwards diffuses to tissue or cell, to keep for a long time its normal physiological state;
2, keep the circulation of slow liquid stream, hatch glass liquid stream from lower to upper, tissue and cell only bear self downward gravity, and undirected lower hydrodynamicpressure has buoyancy and culture dish support upwards in addition, and minimizing tissue and cell are impaired;
3, separate different tissue cups and hatch tissue or cell, avoid a plurality of tissue to assemble or be shifted in by the process of moving at liquid-flow, container;
4, equipment is detachable, and is succinct convenient;
Only be preferred implementation of the present invention below, should be pointed out that above-mentioned preferred implementation should not be considered as limitation of the present invention, protection scope of the present invention should be as the criterion with the claim limited range.For those skilled in the art, without departing from the spirit and scope of the present invention, can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1. cell and tissue incubation's device that continues logical oxygen, it is characterized in that, comprise and hatch cup, be arranged on support and the breather of hatching in the cup, support is provided with a plurality of dispensers that arrange radially, lay the tissue cup on the described dispenser, described breather be provided with inlet pipe and with a plurality of escape pipes that arrange radially around inlet pipe of inlet pipe conducting, a plurality of escape pipes and a plurality of dispenser arrange correspondingly and escape pipe is positioned at the dispenser below, and the position corresponding with dispenser is provided with some production wells on the escape pipe; Described outer side wall of hatching cup is provided with the liquid feed liquor cycling mechanism with the bottom conducting of hatching glass internal cavities, and described liquid feed liquor cycling mechanism is provided with the fluid inlet with extraneous conducting.
2. cell and the tissue incubation's device that continues logical oxygen as claimed in claim 1 is characterized in that, described support comprises central shank and a plurality of connection handles that arrange radially around central shank, and a plurality of dispensers are arranged at respectively the end of different connection handles.
3. cell and the tissue incubation's device that continues logical oxygen as claimed in claim 2 is characterized in that, described central shank is provided with a through hole, and described inlet pipe is passed through hole.
4. cell and the tissue incubation's device that continues logical oxygen as claimed in claim 2 is characterized in that, connection handle comprises long connection handle and short connection handle, and long connection handle and short connection handle are arranged alternately.
5. cell and the tissue incubation's device that continues logical oxygen as claimed in claim 4, it is characterized in that, be positioned on the long connection handle dispenser its be provided with connection bump near a side of hatching glass inwall, correspondingly be provided with the connection screens that cooperates with it on the described inwall of hatching cup.
6. cell and the tissue incubation's device that continues logical oxygen as claimed in claim 1 is characterized in that, described inwall of hatching cup is provided with for the fixing mounting block of inlet pipe.
7. cell and the tissue incubation's device that continues logical oxygen as claimed in claim 1 is characterized in that, described upper opening place of hatching cup is provided with a bowl cover, and described bowl cover is provided with the ventilating slit.
8. cell and the tissue incubation's device that continues logical oxygen as claimed in claim 1 is characterized in that, described dispenser is toroidal frame.
9. cell and the tissue incubation's device that continues logical oxygen as claimed in claim 1, it is characterized in that, described liquid feed liquor cycling mechanism is to be arranged on the hollow cup ear of hatching on glass outer side wall, described hollow cup ear comprises the upright arm of hollow, the upper oblique arm of hollow, the lower oblique arm of hollow, the upper end of described upright arm is provided with opening, as liquid feeding end, described upper oblique arm one end be connected with the top in the outside of hatching cup and with hatch a glass internal cavities conducting, the other end is downward-sloping be connected with the outer wall of upright arm and with the conducting of upright arm internal cavities, described lower oblique arm one end be connected with the bottom in the outside of hatching cup and with hatch a glass internal cavities conducting, the other end be inclined upwardly be connected with the outer wall of upright arm and with the conducting of upright arm internal cavities.
10. cell and the tissue incubation's device that continues logical oxygen as claimed in claim 1 is characterized in that, the cup end of described tissue cup is screen cloth.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210476969.7A CN102943041B (en) | 2012-11-21 | 2012-11-21 | Oxygen continuously filling cell and tissue hatching device |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210476969.7A CN102943041B (en) | 2012-11-21 | 2012-11-21 | Oxygen continuously filling cell and tissue hatching device |
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| Publication Number | Publication Date |
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| CN102943041A true CN102943041A (en) | 2013-02-27 |
| CN102943041B CN102943041B (en) | 2014-11-12 |
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| CN201210476969.7A Active CN102943041B (en) | 2012-11-21 | 2012-11-21 | Oxygen continuously filling cell and tissue hatching device |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101745440A (en) * | 2008-12-09 | 2010-06-23 | 北京量质科技有限公司 | Incubation chamber and wrapped moving incubation device |
| CN202189051U (en) * | 2011-07-14 | 2012-04-11 | 董方 | Chip cup for preventing fluid countercurrent |
| CN102445553A (en) * | 2010-09-30 | 2012-05-09 | 广州阳普医疗科技股份有限公司 | Blending incubation device |
| CN202458366U (en) * | 2011-12-16 | 2012-10-03 | 马劼 | Novel brain slice incubation instrument |
| CN202945246U (en) * | 2012-11-21 | 2013-05-22 | 陈子怡 | Cell and tissue incubation device for replenishing oxygen continuously |
-
2012
- 2012-11-21 CN CN201210476969.7A patent/CN102943041B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101745440A (en) * | 2008-12-09 | 2010-06-23 | 北京量质科技有限公司 | Incubation chamber and wrapped moving incubation device |
| CN102445553A (en) * | 2010-09-30 | 2012-05-09 | 广州阳普医疗科技股份有限公司 | Blending incubation device |
| CN202189051U (en) * | 2011-07-14 | 2012-04-11 | 董方 | Chip cup for preventing fluid countercurrent |
| CN202458366U (en) * | 2011-12-16 | 2012-10-03 | 马劼 | Novel brain slice incubation instrument |
| CN202945246U (en) * | 2012-11-21 | 2013-05-22 | 陈子怡 | Cell and tissue incubation device for replenishing oxygen continuously |
Non-Patent Citations (1)
| Title |
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| 葛求富等: "评价小鼠脑片缺血性损伤及药物保护作用定量方法的改进", 《浙江大学学报（医学版）》 * |
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| CN102943041B (en) | 2014-11-12 |
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