CN103196716B - Dialysis apparatus and dialysis process thereof in a kind of cell - Google Patents
Dialysis apparatus and dialysis process thereof in a kind of cell Download PDFInfo
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- CN103196716B CN103196716B CN201310071696.2A CN201310071696A CN103196716B CN 103196716 B CN103196716 B CN 103196716B CN 201310071696 A CN201310071696 A CN 201310071696A CN 103196716 B CN103196716 B CN 103196716B
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Abstract
The invention discloses dialysis apparatus and dialysis process thereof in a kind of cell, comprise electrode unit with dialysis and electrode jaw, electrode unit with dialysis is provided with first micro-intubate and second micro-intubate, central lumen is provided with in electrode jaw, electrode jaw is provided with the negative-pressure ward side pipe and micro-cannula sleeve that are communicated with central lumen, first micro-intubate and second micro-intubate to be inserted in micro-cannula sleeve simultaneously and are communicated with central lumen.The invention has the beneficial effects as follows, make that thin intracellular dialysis is easy and simple to handle, to change liquid efficiency high, long-term stable observation can be carried out.
Description
Technical field:
The invention belongs to medical instruments field, relate to a kind of electrode solution more changing device, dialysis apparatus and dialysis process thereof in especially a kind of cell.
Background technology:
Patch clamp technique to have reached its maturity and perfect since establishment, and application technique, people can analyse in depth from molecular level the character of memebrane protein (as ion channel, acceptor) and function etc.But in some experiments, technique receives certain restriction.As when studying the regulation and control to ion channel of cell membrane signal system, need in cell, apply suitable medicine; When the Current mechanism that a certain functional status of analysis to measure cell is corresponding, need to use liquid in corresponding electrode according to surveyed electric current.In these cases, only have the composition using custom-designed perfusion device to change liquid in electrode, and measure before this step and afterwards, strict contrast can be realized.This device is called as thin intracellular dialysis (perfusion) device.Thin intracellular dialysis principle is physical diffusion, and namely in electrode solution, material is spread in cell by eletrode tip.Early stage intracellular perfusion apparatus structure is complicated, noise is large, operation easier is large, experiment success rate is low, is difficult to universal in general electro physiology laboratory.Though the later stage improves to some extent, but still it is low to there is displacement efficiency, the shortcoming of poor stability.
Summary of the invention:
The object of the invention is to the shortcoming overcoming above-mentioned prior art, dialysis apparatus and dialysis process thereof in a kind of cell are provided, make that thin intracellular dialysis is easy and simple to handle, to change liquid efficiency high, long-term stable observation can be carried out.
The object of the invention is to solve by the following technical programs:
Dialysis apparatus in a kind of cell, comprise electrode unit with dialysis and electrode jaw, electrode unit with dialysis is provided with first micro-intubate and second micro-intubate, central lumen is provided with in electrode jaw, electrode jaw is provided with the negative-pressure ward side pipe and micro-cannula sleeve that are communicated with central lumen, first micro-intubate and second micro-intubate to be inserted in micro-cannula sleeve simultaneously and are communicated with central lumen.
Described electrode unit with dialysis comprises liquid storage tank, U-tube, waste collection pond, first micro-intubate, second micro-intubate, electrode jaw and three hole supports; One end of described U-tube is communicated with liquid storage tank by silicone tube, and the other end of U-tube is communicated with the first negative-pressure suction pipe, is provided with second micro-intubate in the bottom of U-tube; Top, described waste collection pond is provided with first micro-intubate; Described liquid storage tank, U-tube and waste collection pond are all fixed on three hole supports.
Described negative-pressure ward side pipe and the electrode jaw longitudinal axis are 90 ° of angles, and micro-cannula sleeve is arranged on the top of negative-pressure ward side pipe, and become 135 ° of angles with the electrode jaw longitudinal axis.
Described micro-cannula sleeve length is 1-1.5cm, and its external diameter is 1-2mm, and internal diameter is 1-1.5mm.
Top, described waste collection pond is provided with above top end opening, left side and is provided with left side opening, lower right-hand side is provided with right openings, and top end opening is communicated with first micro-intubate, and left side opening is connected with the second negative-pressure suction pipe, and right openings is connected with silicone tube.
Described waste collection pond is made up of transparent organic glass.
Described U-tube and silicone tube junction are provided with the second threeway, and U-tube and the first negative-pressure suction pipe junction are provided with the first threeway.
The dialysis process of dialysis apparatus in described cell, in accordance with the following steps: before (1) starts experiment, U-tube is started, with the use of the first threeway and the second threeway, make liquid in liquid storage tank be full of second micro-cannula tip, exist with bubble-free in second micro-intubate and be as the criterion and do not flow;
(2) to select after suitable cell with conventional electrodes liquid, conventional practices clamp cells under the microscope, carry out record;
(3) the second three-way cock connecting liquid storage tank is opened;
(4) the second negative-pressure suction pipe on the left of waste collection pond being opened, by increasing negative pressure, making liquid in liquid storage tank flow in glass microelectrode; Judge that electrode solution changes effect by the fluid drips number in observation waste collection pond and speed, 3 ~ 4 can make electrode solution update all;
(5), after above-mentioned steps completes, close waste collection pond second negative-pressure suction pipe and the second threeway, namely whole dialysis system is in stand-by state.
The invention has the beneficial effects as follows, make that thin intracellular dialysis is easy and simple to handle, to change liquid efficiency high, long-term stable observation can be carried out.
Accompanying drawing illustrates:
Fig. 1 is electrode jaw structural representation of the present invention;
Fig. 2 is electrode unit with dialysis structural representation of the present invention;
Wherein: 1 is electrode jaw; 1-1 is central lumen; 1-2 is negative-pressure ward side pipe; 2 is micro-cannula sleeve; 3 is waste collection pond; 4 is U-tube; 5 is liquid storage tank; 6 is three hole supports; 7 is silicone tube; 8 is first micro-intubate; 9 is second micro-intubate; 10 is the first threeway; 11 is the second threeway; 12 is the first negative-pressure suction pipe; 13 is the second negative-pressure suction pipe.
Embodiment:
Below in conjunction with accompanying drawing, the present invention is described in further detail:
See Fig. 1 and Fig. 2, dialysis apparatus in a kind of cell, comprise electrode unit with dialysis and electrode jaw, electrode unit with dialysis is provided with first micro-intubate and second micro-intubate, central lumen is provided with in electrode jaw, electrode jaw is provided with the negative-pressure ward side pipe and micro-cannula sleeve that are communicated with central lumen, first micro-intubate and second micro-intubate to be inserted in micro-cannula sleeve simultaneously and are communicated with central lumen.
Described electrode unit with dialysis comprises liquid storage tank, U-tube, waste collection pond, first micro-intubate, second micro-intubate, electrode jaw and three hole supports; One end of described U-tube is communicated with liquid storage tank by silicone tube, and the other end of U-tube is communicated with the first negative-pressure suction pipe, is provided with second micro-intubate in the bottom of U-tube; Top, described waste collection pond is provided with first micro-intubate; Described liquid storage tank, U-tube and waste collection pond are all fixed on three hole supports.
Described negative-pressure ward side pipe and the electrode jaw longitudinal axis are 90 ° of angles, and micro-cannula sleeve is arranged on the top of negative-pressure ward side pipe, and become 135 ° of angles with the electrode jaw longitudinal axis.
Described micro-cannula sleeve length is 1cm, and its external diameter is 1.5mm, and internal diameter is 1mm.
Top, described waste collection pond is provided with above top end opening, left side and is provided with left side opening, lower right-hand side is provided with right openings, and top end opening is communicated with first micro-intubate, and left side opening is connected with the second negative-pressure suction pipe, and right openings is connected with silicone tube.
Described waste collection pond is made up of transparent organic glass.
Described U-tube and silicone tube junction are provided with the second threeway, and U-tube and the first negative-pressure suction pipe junction are provided with the first threeway.
Intracellular dialysis device is formed primarily of two parts: one is electrode jaw part, and two is electrode solution unit with dialysis (Fig. 2 shows).Electrode jaw part is that transformation forms on patch-clamp pipette holder (patch clamp pipette holder) basis, get electrode jaw, in the top of negative-pressure ward side pipe (be 90 ° angles with the clamper longitudinal axis), to become 135 ° of angles with electrode bare terminal end, with 2mm bit drills one side opening, straight-through clamper central lumen.Inserting one and be about 1cm, the fine steel tube (No. 18 injection needle external diameter 1.5mm, available) that external diameter is suitable with institute's hole diameter, around seal with " 302 " tackifier, fix, using during in order to inserting micro-intubate.Electrode unit with dialysis is made up of four parts: micro-intubate, waste collection pond, liquid storage tank, U-tube.The micro-intubate (inserting in micro-cannula sleeve) inserted in glass microelectrode utilizes micropipet rifle head to form manual drawing of spirit lamp flame heating, generally choose 100 μ l heads of pipette, the micro-cannula outer diameter drawn about 100 μm, internal diameter about 50 μm, in each device, standby micro-intubate two, is called first micro-intubate, second micro-intubate.The organic glass that the making in waste collection pond selects transparency high, is beneficial to monitoring and changes liquid effect.About this collecting pit and above respectively open flatly: left side opening connects negative pressure source, and lower right opening connects the silicone tube for removal waste fluid, and upper opening is connected with first micro-intubate.Disposable polyethylene syringe needle tube (5ml) selected by liquid storage tank, after pulling out inner core, closes for subsequent use by lower end scorification.Liquid storage tank is fixed on support, and top is drawn silicone tube and passed therethrough, and is connected with " U " pipe upper end one side opening.Being prepared as follows of U-tube: the plastic tube of cut-off footpath 1.5mm curves " U " type, an osculum is opened on the top of lower end arch section, after micro-intubate (second micro-intubate) the thicker side be drawn into another one is connected closely, with the sealing of " 302 " tackifier (this micro-intubate need be inserted in glass microelectrode through micro-cannula sleeve equally); " U " pipe upper end open, a side joint liquid storage tank, a side joint negative pressure suction device, all arranges a three links: link of trade, travel and post switch in junction.Supporting and the placement of intracellular dialysis device: after each part mentioned above is ready to complete, two micro-intubate inserted along micro-cannula sleeve, determine the degree of depth inserted according to the length of glass microelectrode, second micro-intubate outline is longer than first micro-intubate.Three hole supports are prepared in addition with organic glass, can by placed on it to waste collection pond, " U " pipe and liquid storage tank, stentplacement, near experimental record electrode jaw, is mated use.Change the basic step of electrode solution: before (1) starts experiment, start U-tube, with the use of three-way device, make liquid in liquid storage tank be full of second micro-cannula tip (confirming that wherein bubble-free exists), and do not flow.(2) select under the microscope with conventional electrodes liquid after suitable cell, conventional practices clamp cells, carries out record.(3) three-way cock connecting liquid storage tank is opened.(4) by negative pressure source switch opens on the left of waste collection pond, by increasing negative pressure, liquid in liquid storage tank is flowed in glass microelectrode.Judge that electrode solution changes effect by the fluid drips number in observation waste collection pond and speed, usual 3 ~ 4 (about 1/s) can make electrode solution update all.After above-mentioned steps completes, close waste collection pond negative pressure source and liquid storage tank threeway, namely whole dialysis system is in stand-by state.Later replacing glass electrode and routine experimentation almost just the same, if desired, also can as required at experimental session, by opening and closing threeway, coordinating negative-pressure ward, changing electrode solution at any time.
The above, it is only preferred embodiment of the present invention, not any pro forma restriction is done to the present invention, although the present invention discloses as above with preferred embodiment, but and be not used to limit the present invention, any those skilled in the art, do not departing within the scope of technical solution of the present invention, when the method and technology contents that can utilize above-mentioned announcement are made a little change or be modified to the Equivalent embodiments of equivalent variations, in every case be the content not departing from technical solution of the present invention, according to any simple modification that technical spirit of the present invention is done above embodiment, equivalent variations and modification, still belong in the scope of technical solution of the present invention.
Claims (7)
1. dialysis apparatus in a cell, it is characterized in that: comprise electrode unit with dialysis and electrode jaw, electrode unit with dialysis is provided with first micro-intubate and second micro-intubate, central lumen is provided with in electrode jaw, electrode jaw is provided with the negative-pressure ward side pipe and micro-cannula sleeve that are communicated with central lumen, first micro-intubate and second micro-intubate to be inserted in micro-cannula sleeve simultaneously and are communicated with central lumen; Described electrode unit with dialysis comprises liquid storage tank, U-tube, waste collection pond, first micro-intubate, second micro-intubate and three hole supports; One end of described U-tube is communicated with liquid storage tank by silicone tube, and the other end of U-tube is communicated with the first negative-pressure suction pipe, is provided with second micro-intubate in the bottom of U-tube; Top, described waste collection pond is provided with first micro-intubate; Described liquid storage tank, U-tube and waste collection pond are all fixed on three hole supports; Top, described waste collection pond is provided with above top end opening, left side and is provided with left side opening, lower right-hand side is provided with right openings, and top end opening is communicated with first micro-intubate, and left side opening is connected with the second negative-pressure suction pipe, and right openings is connected with silicone tube.
2. dialysis apparatus in cell as claimed in claim 1, is characterized in that: described negative-pressure ward side pipe and the electrode jaw longitudinal axis are 90 ° of angles.
3. dialysis apparatus in cell as claimed in claim 1, is characterized in that: described micro-cannula sleeve is arranged on the top of negative-pressure ward side pipe, and becomes 135 ° of angles with the electrode jaw longitudinal axis.
4. dialysis apparatus in cell as claimed in claim 1, is characterized in that: described micro-cannula sleeve length is 1-1.5cm, and its external diameter is 1-2mm, and internal diameter is 1-1.5mm.
5. dialysis apparatus in cell as claimed in claim 2, is characterized in that: described waste collection pond is made up of transparent organic glass.
6. dialysis apparatus in cell as claimed in claim 2, it is characterized in that: described U-tube and silicone tube junction are provided with the second threeway, U-tube and the first negative-pressure suction pipe junction are provided with the first threeway.
7. the dialysis process of dialysis apparatus in cell as claimed in claim 6, is characterized in that, in accordance with the following steps:
(1) before starting experiment, start U-tube, with the use of the first threeway and the second threeway, make liquid in liquid storage tank be full of second micro-cannula tip, exist with bubble-free in second micro-intubate and be as the criterion and do not flow;
(2) to select after suitable cell with conventional electrodes liquid, conventional practices clamp cells under the microscope, carry out record;
(3) the second three-way cock connecting liquid storage tank is opened;
(4) the second negative-pressure suction pipe on the left of waste collection pond being opened, by increasing negative pressure, making liquid in liquid storage tank flow in glass microelectrode; Judge that electrode solution changes effect by the fluid drips number in observation waste collection pond and speed, 3 ~ 4 can make electrode solution update all;
(5), after above-mentioned steps completes, close waste collection pond second negative-pressure suction pipe and the second threeway, namely whole dialysis system is in stand-by state.
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| CN201310071696.2A CN103196716B (en) | 2013-03-07 | 2013-03-07 | Dialysis apparatus and dialysis process thereof in a kind of cell |
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| CN201310071696.2A CN103196716B (en) | 2013-03-07 | 2013-03-07 | Dialysis apparatus and dialysis process thereof in a kind of cell |
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| CN103196716B true CN103196716B (en) | 2015-08-12 |
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| CN108956506A (en) * | 2018-09-19 | 2018-12-07 | 北京大学 | A kind of glyoxal or methyl-glyoxal standard gas generating apparatus and method |
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| CN2289239Y (en) * | 1996-10-31 | 1998-08-26 | 中国人民解放军第四军医大学 | Intracellular perfusion and fluid-changing device |
| CN2465866Y (en) * | 2001-02-21 | 2001-12-19 | 华中科技大学 | Glass electrode holder of membrane tongs |
| EP1474188A4 (en) * | 2002-01-25 | 2008-01-16 | Biocure Inc | Coaxial delivery device |
| US20050137471A1 (en) * | 2003-12-18 | 2005-06-23 | Hans-Peter Haar | Continuous glucose monitoring device |
| CN2798089Y (en) * | 2005-04-14 | 2006-07-19 | 李国华 | Apparatus for diaphragm forceps intracellular dialysis |
| WO2007078463A1 (en) * | 2005-12-22 | 2007-07-12 | The Trustees Of Columbia University In The City Of New York | Systems and methods for intravascular cooling |
| CN101703817B (en) * | 2009-11-23 | 2012-07-18 | 浙江大学 | Jugular vein microdialysis probe and probe guiding device |
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