CN203546001U - Perfusion device for incubating living body brain slice or spinal marrow slice - Google Patents
Perfusion device for incubating living body brain slice or spinal marrow slice Download PDFInfo
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- CN203546001U CN203546001U CN201320647470.8U CN201320647470U CN203546001U CN 203546001 U CN203546001 U CN 203546001U CN 201320647470 U CN201320647470 U CN 201320647470U CN 203546001 U CN203546001 U CN 203546001U
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- hatching
- incubation
- perfusion device
- spinal cord
- dish
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- 230000012447 hatching Effects 0.000 claims description 53
- 210000000278 spinal cord Anatomy 0.000 claims description 37
- 230000035699 permeability Effects 0.000 claims description 16
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Images
Abstract
The utility model discloses a perfusion device for incubating a living body brain slice or a spinal marrow slice and belongs to the technical field of incubation of biological tissue slices. The perfusion device comprises an incubation disc and an incubation groove, and further comprises an input pipe, an output pipe and an incubation rack, wherein the input pipe and the output pipe are respectively communicated with the inner space of the incubation disc; the incubation rack is arranged in the incubation disc; the bottom and/or the side wall of the incubation rack are/is provided with hollow parts for communicating the inner space of the incubation disc; the incubation groove is formed in the incubation rack. With the adoption of the perfusion device, mixed gas or ACSF (Artificial Cerebral Spinal Fluid) liquid is continuously introduced into the living body brain slice or the spinal marrow slice to carry out substance exchange; the ACSF liquid is continuously replaced so that nutrient substances are continuously updated and metabolic wastes are continuously discharged; the living body brain slice or the spinal marrow slice is at the optimal environment all the time in a long-period incubation process and the activity is kept.
Description
Technical field
The utility model relates to a kind of biological tissue section and hatches equipment, particularly relates to a kind of perfusion device of hatching for live body brain sheet or spinal cord.
Background technology
1966, Yamamoto and McIlwain have recorded bioelectrical activity (Yamamoto C first on brain sheet, McIlwain H.Potentials evoked in vitro in preparations from the mammalian brain.Nature, 1966,210 (5040): 1055-1056), confirmed that cerebral tissue also can survive in vitro, and kept good active condition.The foundation of this method, has greatly promoted the process that Electrophysiology is studied.1989, Blanton combines (Yamamoto C by the patch clamp record of brain sheet electrophysiological recording and cell, McIlwain H.Electrical activities in thin sections from the mammalian brain maintained in chemically-defined media in vitro.J Neurochem.1966,13 (12): 1333-43), set up slice patch clamp recording technique, this becomes the milestone of neurobiology development.For connecting each other between neurone in researching neural network; Yuste R and Katz LC have been recorded to multidate information (Yuste R, the Katz LC.Control of postsynaptic Ca2+influx in developing neocortex by excitatory and inhibitory neurotransmitters.Neuron.1991Mar of calcium ion first in the brain sheet of rat and cat; 6 (3): 333-44), thereby created the interior method-functional type multi-neuron calcium imaging technique of neuron action potentials in a large number of a kind of energy real time record neural network.
Wherein, for the brain sheet of electrophysiological recording or functional calcium imaging or spinal cord, all need the activity that keeps very high, this is to test the essential condition of carrying out smoothly.Live body brain sheet, spinal cord need to be drawn materials by low temperature on live body, with being placed on, are filled with gas mixture (95%O
2, 5%CO
2) artificial cerebrospinal fluid (ACSF) in hatch, 30-35 ℃ of insulation, during to constantly be filled with gas mixture, this has effectively kept the activity of brain sheet.Currently for making the instrument of live body brain sheet, mostly be self-control, or replace with beaker.
In conventional technology, be generally that the ACSF liquid preparing is poured in large beaker at present, constantly pass into gas mixture, the incubation slot that fills brain sheet or spinal cord floats on ACSF liquid level top simultaneously.Someone improves the control of mixed gas flow by establish a plurality of holes of hatching at incubation slot center, although guaranteed the even of air-flow by improvement, but still can not make incubation slot fully enter ACSF liquid, keeps sufficient exchange of substance; Somebody makes the brain sheet can contact with liquid fully by filter structure is set, so that better keep brain sheet activity, but it installs each cell and be not communicated with mutually, ventilates and perfusion is all difficult to carry out.
And routine techniques is all traditional hatching, and does not relate to perfusion.Hatching in process of live body brain sheet or spinal cord, can constantly carry out exchange of substance with the external world, absorb extraneous nutritive substance, eliminating meta-bolites.Common ventilation is hatched and can be caused extraneous nutritive substance to reduce along with the prolongation of incubation time, and metabolic waste increases gradually, is unfavorable for keeping the activity of brain sheet.
Summary of the invention
Based on this, the purpose of this utility model is to overcome the defect of prior art, a kind of perfusion device of hatching for live body brain sheet or spinal cord is provided, can realize Perifusion, continuous replacing along with liquid, nutritive substance is constantly updated, and metabolic waste is constantly discharged, and is beneficial to the activity that keeps brain sheet or spinal cord.
For achieving the above object, the utility model is taked following technical scheme:
A perfusion device of hatching for live body brain sheet or spinal cord, comprises and hatches dish and incubation slot; Also comprise input tube, output tube and hatch frame; Described input tube and output tube are communicated with respectively the internal space of hatching dish, described in hatch to be placed on and hatch in dish, and this hatches the bottom of frame and/or sidewall and is provided with and is communicated with the hollow-out parts of hatching dish internal space, described incubation slot is placed in hatches on frame.
Adopt above-mentioned perfusion device, in hatching the process of live body brain sheet or spinal cord, live body brain sheet or spinal cord are put into incubation slot, again incubation slot is put into and hatched on frame, owing to hatching bottom and/or the sidewall of frame, be provided with the hollow-out parts that dish internal space is hatched in connection, therefore, when constantly inputting gas mixture or ACSF liquid by input tube, live body brain sheet or spinal cord that gas mixture or ACSF liquid can see through in this hollow-out parts and incubation slot carry out exchange of substance, and by the meta-bolites of generation along with ACSF liquid is discharged by output tube, be conducive to keep the activity of brain sheet or spinal cord.
Therein in an embodiment, described in hatch frame and comprise connection section and at least two side direction supports; Described side direction support is strip, described connecting portion props up top of the trellis in side direction, side direction support connects as one by connection section, described side direction mid-stent is also provided with the supporting bracket of horizontal expansion, the common barrel-like structure with container cavity that forms of this side direction support, connection section and supporting bracket, described incubation slot is placed in this container cavity.Adopt said structure, the ACSF liquid that can make to hatch in dish carries out sufficient exchange of substance by bottom and the surrounding of incubation slot with it.
In an embodiment, described side direction support is at least two therein, and described connection section is circular; Described incubation slot is barrel-shaped, and the top of this incubation slot is provided with the fixed part radially extending along incubation slot, and this fixed part is provided with the buckling groove with connection section clamping.By buckling groove is connected in to fixed part, incubation slot can be fixed, avoid producing mobile.
In an embodiment, the external diameter of described incubation slot and the internal diameter of described container cavity match therein.Avoid incubation slot with flowing of ACSF liquid, to produce in the horizontal direction in container cavity mobile.
Therein in an embodiment, described in hatch frame and be at least two, the connection section of respectively hatching frame is connected as a single entity.Can as required, insert one or more incubation slots and hatch.
In an embodiment, this perfusion device also comprises permeability cell, on this permeability cell, is evenly equipped with breather hole therein, and described permeability cell is located at and is hatched in dish, and described input tube is located at this permeability cell of open communication of hatching in dish.So that make by input tube enter hatch the gas mixture of dish or ACSF liquid can be in hatching dish by discharge, unlikely generation is large impact power too.
In an embodiment, described permeability cell is along hatching the setting of tray bottom edge therein.Gas mixture or ACSF liquid are evenly discharged by the surrounding of hatching dish, can better carry out exchange of substance.
Therein in an embodiment, this perfusion device also comprises hatches lid and gas output tube, and described gas output tube connects hatches lid, described in hatch the size of lid and hatch dish size and match.By hatching the use of lid, can reduce the whole pollution that is subject to external substance in process of hatching.
Therein in an embodiment, described in hatch dish and hatch lid that to be lucifuge hatch dish and hatch lid (as manufactured by brown glass or brown polystyrene material), be convenient to live body brain sheet or the spinal cord lucifuge in immunostaining and operate.
In an embodiment, described incubation slot consists of framework and the nylon membrane or the semi-permeable membranes that are covered on framework therein.Live body brain sheet in permission incubation slot or spinal cord and mixed gas and ACSF liquid carry out exchange of substance, and routine can be used 100 object nylon membranes, when having dyeing demand, can use 0.4 μ m semi-permeable membranes.
Compared with prior art, the utlity model has following beneficial effect:
Perfusion device of hatching for live body brain sheet or spinal cord of the present utility model, be provided with perfusion system, can make live body brain sheet or spinal cord and the gas mixture constantly passing into or ACSF liquid carry out exchange of substance, along with ACSF liquid is constantly changed, nutritive substance is constantly updated, metabolic waste is constantly discharged, effectively guaranteed brain sheet or spinal cord in hatching process for a long time always in best environment, keep its activity.And this device sets ventilation hatch and two functions of Perifusion, met the different demand of experimenter.
This perfusion device also, by adopting the frame of hatching of structure uniqueness, can guarantee that brain sheet or spinal cord fully contact with Incubating Solution, keep the activity of brain sheet or spinal cord to greatest extent; And can carry out hatching of many increments bases simultaneously.
This perfusion device is also by the use of permeability cell, and the air-flow, the flow that make to hatch in dish are even, for hatching, creates envrionment conditions stably.
This perfusion device also can adopt light tight material to make, thereby can carry out the experiment that immunostaining etc. needs lucifuge.
Accompanying drawing explanation
Fig. 1 is perfusion device overall appearance schematic diagram described in embodiment;
Fig. 2 is perfusion device internal structure schematic diagram described in embodiment;
Fig. 3 is for hatching dish and hatching frame vertical view;
Fig. 4 has put into the vertical view of incubation slot in Fig. 3;
Fig. 5 is single incubation slot and hatches shelf structure schematic diagram;
Fig. 6 covers schematic diagram for hatching.
Wherein: 1. hatch dish; 2. incubation slot; 21. fixed parts; 3. input tube; 4. output tube; 5. hatch frame; 51. side direction supports; 52. connection sections; 53. supporting brackets; 6. hatch lid; 7. gas output tube; 8. permeability cell.
Embodiment
Below in conjunction with the drawings and specific embodiments, describe the present invention in detail.
A perfusion device of hatching for live body brain sheet or spinal cord, comprise hatch dish 1, incubation slot 2, input tube 3, output tube 4, hatch frame 5, hatch cover 6, gas output tube 7 and permeability cell 8, described input tube 3 and output tube 4 are communicated with respectively the internal space of hatching dish 1, described hatch frame 5 be placed in hatch dish 1 in, the described frame 5 of hatching comprises connection section 52 and four side direction supports 51, described side direction support 51 is strip, described connection section 52 is positioned at side direction and props up top of the trellis, side direction support connects as one by connection section 52, described side direction support 51 middle parts are also provided with the supporting bracket 53 of horizontal expansion, this side direction support 51, the common barrel-like structure with container cavity that forms of connection section 52 and supporting bracket 53, described incubation slot is placed in this container cavity, the external diameter of this incubation slot 2 and the internal diameter of described container cavity match.
Described connection section 52 is circular; Described incubation slot 2 is barrel-shaped, and the top of this incubation slot 2 is provided with the fixed part 21 radially extending along incubation slot, and this fixed part 21 is provided with the buckling groove with connection section 52 clampings.
Described to hatch frame 5 be five, be evenly distributed in and hatch in dish 1, and the connection section 52 of respectively hatching frame 5 is linked as an individuality, makes a plurality of framves 5 of hatching form an integral body.
On described permeability cell 8, be evenly equipped with breather hole, this permeability cell 8 is located at and is hatched in dish 1, and along hatching dish 1 bottom margin setting, and described input tube 3 is located at this permeability cell 8 of open communication of hatching in dish.
Described gas output tube 7 connects to hatch and cover 6, described in hatch and cover 6 size and coil 1 size and match with hatching.Can be fixed on and hatch on dish 1, will hatch dish 1 and cover.
Described hatch dish 1 and hatch cover 6 to be lucifuge hatches dish and hatch lid (as manufactured by brown glass or brown polystyrene material), be convenient to lucifuge in immunostaining of live body brain sheet or spinal cord and operate.
Described incubation slot 2 consists of framework and the nylon membrane or the semi-permeable membranes that are covered on framework.Live body brain sheet in permission incubation slot or spinal cord and mixed gas and ACSF liquid carry out exchange of substance, and routine can be used 100 object nylon membranes, when having dyeing demand, can use 0.4 μ m semi-permeable membranes.
While adopting above-mentioned perfusion device to carry out hatching of live body brain sheet or spinal cord, live body brain sheet or spinal cord are put into incubation slot 2, again incubation slot 2 is put into and hatched frame 5, owing to hatching bottom and the sidewall of frame 5, be all directly communicated with the internal space of hatching dish, therefore, when constantly inputting gas mixture or ACSF liquid by input tube 3, gas mixture or ACSF liquid can with incubation slot 2 in live body brain sheet or spinal cord carry out exchange of substance, and by the meta-bolites of generation along with ACSF liquid is discharged by output tube 4, be conducive to keep the activity of brain sheet or spinal cord.
The above embodiment has only expressed several embodiment of the present utility model, and it describes comparatively concrete and detailed, but can not therefore be interpreted as the restriction to the utility model the scope of the claims.It should be pointed out that for the person of ordinary skill of the art, without departing from the concept of the premise utility, can also make some distortion and improvement, these all belong to protection domain of the present utility model.Therefore, the protection domain of the utility model patent should be as the criterion with claims.
Claims (10)
1. a perfusion device of hatching for live body brain sheet or spinal cord, comprises and hatches dish and incubation slot; It is characterized in that, also comprise input tube, output tube and hatch frame; Described input tube and output tube are communicated with respectively the internal space of hatching dish, described in hatch to be placed on and hatch in dish, and this hatches the bottom of frame and/or sidewall and is provided with and is communicated with the hollow-out parts of hatching dish internal space, described incubation slot is placed in hatches on frame.
2. perfusion device of hatching for live body brain sheet or spinal cord according to claim 1, is characterized in that, described in hatch frame and comprise connection section and at least two side direction supports; Described side direction support is strip, described connecting portion props up top of the trellis in side direction, side direction support connects as one by connection section, described side direction mid-stent is also provided with the supporting bracket of horizontal expansion, the common barrel-like structure with container cavity that forms of this side direction support, connection section and supporting bracket, described incubation slot is placed in this container cavity.
3. perfusion device of hatching for live body brain sheet or spinal cord according to claim 2, is characterized in that, described side direction support is at least two, and described connection section is circular; Described incubation slot is barrel-shaped, and the top of this incubation slot is provided with the fixed part radially extending along incubation slot, and this fixed part is provided with the buckling groove with connection section clamping.
4. perfusion device of hatching for live body brain sheet or spinal cord according to claim 2, is characterized in that, the external diameter of described incubation slot and the internal diameter of described container cavity match.
5. perfusion device of hatching for live body brain sheet or spinal cord according to claim 2, is characterized in that, described in hatch frame and be at least two, the connection section of respectively hatching frame is connected as a single entity.
6. perfusion device of hatching for live body brain sheet or spinal cord according to claim 1, it is characterized in that, also comprise permeability cell, on this permeability cell, be evenly equipped with breather hole, described permeability cell is located at and is hatched in dish, and described input tube is located at this permeability cell of open communication of hatching in dish.
7. perfusion device of hatching for live body brain sheet or spinal cord according to claim 6, is characterized in that, described permeability cell is along hatching the setting of tray bottom edge.
8. perfusion device of hatching for live body brain sheet or spinal cord according to claim 1, is characterized in that, also comprises and hatches lid and gas output tube, and described gas output tube connects hatches lid, described in hatch the size of lid and hatch dish size and match.
9. perfusion device of hatching for live body brain sheet or spinal cord according to claim 8, is characterized in that, described in hatch dish and hatch lid that to be lucifuge hatching dish and hatching lid.
10. perfusion device of hatching for live body brain sheet or spinal cord according to claim 1, is characterized in that, described incubation slot consists of framework and the nylon membrane or the semi-permeable membranes that are covered on framework.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201320647470.8U CN203546001U (en) | 2013-10-18 | 2013-10-18 | Perfusion device for incubating living body brain slice or spinal marrow slice |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201320647470.8U CN203546001U (en) | 2013-10-18 | 2013-10-18 | Perfusion device for incubating living body brain slice or spinal marrow slice |
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| CN203546001U true CN203546001U (en) | 2014-04-16 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104713907A (en) * | 2015-01-29 | 2015-06-17 | 新乡医学院 | In vivo brain slice nerve network oscillation recording system and recording method thereof |
| CN107418891A (en) * | 2017-09-20 | 2017-12-01 | 四川省人民医院 | A kind of perfusion type bladder bacterial biof iotalm dynamic is incubated simulation system |
| CN107858283A (en) * | 2017-10-12 | 2018-03-30 | 中国人民解放军第四军医大学 | A kind of neuromodulation method and device |
-
2013
- 2013-10-18 CN CN201320647470.8U patent/CN203546001U/en not_active Expired - Lifetime
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104713907A (en) * | 2015-01-29 | 2015-06-17 | 新乡医学院 | In vivo brain slice nerve network oscillation recording system and recording method thereof |
| CN104713907B (en) * | 2015-01-29 | 2017-09-22 | 新乡医学院 | Brain slice in vitro neutral net vibrates record system and the method for record |
| CN107418891A (en) * | 2017-09-20 | 2017-12-01 | 四川省人民医院 | A kind of perfusion type bladder bacterial biof iotalm dynamic is incubated simulation system |
| CN107858283A (en) * | 2017-10-12 | 2018-03-30 | 中国人民解放军第四军医大学 | A kind of neuromodulation method and device |
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Granted publication date: 20140416 |