CN102636550A - Horseradish cell internal-external calcium ion exchange dynamic detection method - Google Patents
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- CN102636550A CN102636550A CN2012101140325A CN201210114032A CN102636550A CN 102636550 A CN102636550 A CN 102636550A CN 2012101140325 A CN2012101140325 A CN 2012101140325A CN 201210114032 A CN201210114032 A CN 201210114032A CN 102636550 A CN102636550 A CN 102636550A
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Abstract
The invention belongs to a method for researching the horseradish cell internal-external calcium ion (Ca<2+>) exchange dynamic by an electrochemical method. In the method, a nano capillary tube is prepared by taking a borosilicate capillary tube as a material, and the inner wall is subjected to silylation to obtain hydrophobicity; the 1,2-dichloroethane (DCE) solution containing Ca<2+> carrier is filled into the nano capillary tube with hydrophobic inner tube wall, as an organic phase; the horseradish cell solution is an aqueous phase; Ag/AgCl is used as a reference electrode in the aqueous phase, and Ag/AgTPBCl is used as a reference electrode in the organic phase; and the dynamic detection on the horseradish cell internal-external calcium ion (Ca<2+>) exchange is performed through a cyclic voltammetry by use of a nano-tube ion-selective microelectrode. The method provided by the invention is easy to operate and has high sensitivity, realizes perfect combination of the ion-selective electrode and the electrochemical detection technology, and provides a new method for researching the influence of metal ion on the property and function of an ion channel.
Description
Technical field
The present invention relates to the inside and outside dynamic assay method of calcium ion-exchanged of horseradish cell, belong to analytical chemistry and cell biology field.
Background technology
The existence of cell membrane makes that intracellular various material can stable existence and carry out the chemical reaction of life entity, can also optionally transport vital material of life entity such as Na simultaneously
+, K
+, Ca
2+, Cl
-Deng.Signal transmission in the life system and the ion between the cell are striden the film transportation has vital the contact, and the transportation of ion is accomplished by the various ion channels on the cell membrane usually.
Patch clamp technique is a kind of technology that reflects single on the cell membrane (or a plurality of) ion channel molecule activity with record through the gas current of ion channel.Yet vegetable cell diaphragm pincers detect and have difficulties, as go the ramp of wall living cells primary wall to cause to be difficult to form the high resistant sealing-in between plasma membrane and the microelectrode; More difficult sealing-in and detection after the cell membrane breakage; Patch-clamp detects needs rupture of membranes, causes experimental result to depart from actual value; Other ion interference is serious in the fluid of inside and outside cell.Rare earth damages vegetable cell after handling, and causes patch-clamp to realize continuous detecting to vegetable cell.
The non-damage micrometering technology is in real time, dynamically to measure live body or the ion of non-damage ground measurement material surface and the flow velocity and the direction of motion of molecule, is based on integrated sophisticated technologies of principle such as physics, chemistry, electricity, biology.This technology is not inquired into the inside and outside ion-exchange of cell, and simultaneously, this technological testing tool is expensive, uses not extensive in China.
ISE is a kind of can in the presence of different kinds of ions, mensuration the analysis to measure instrument of given ion activity in the solution with potential method, thereby be called electric potential type chemical sensor again.Electric charge (electronics and ion) is at liquid/liquid interface or to be referred to as two kinds of transfer reaction processes on the immiscible electrolyte solution interface are one of basic physical chemical processes.Little, nanoscale liquid/liquid interface is usually used in studying ion and the quick transfer reaction dynamics of speeding-up ion on liquid/liquid interface.
I in the patch-clamp data～V curve is the current value curve at each film potential place, inspired by this, according to the principle of ISE, and anode electrochemistry analysis means in addition, we have developed the inside and outside Ca of a kind of detection horseradish cell
2+The new method of exchange.Adopt this method research living cells surface C a
2+The variation of concentration, and then the variation of discussion ion channel character and function.This method has remedied the part shortcoming of traditional detection method patch-clamp mensuration vegetable cell.When carrying out cell tests, infinitely near cell, the pair cell not damaged.This method is the foreign matter especially detection of metallic ion cell membrane ion-exchange capacity with relevant cross discipline research method combination such as chemistry, cell biology, bioelectrochemistry, and new reference is provided.
Summary of the invention
The purpose of this invention is to provide the inside and outside Ca of a kind of new horseradish cell
2+The detection method of exchange, have highly sensitive, accuracy good, selectivity is strong, simple to operate, advantage such as analysis speed is fast.Cyclic voltammetry of the present invention detects little current method, has realized that rare earth ion is to vegetable cell surfaces externally and internally Ca
2+Continuous detecting.
The inside and outside Ca of electricity consumption chemical cycle voltammetric measuring horseradish cell of the present invention
2+The method of exchange is to be the material preparation nanotube capillaries with the borosilicate kapillary, and carries out the inwall silanization and make it become hydrophobicity; In the nanotube capillaries of inner tubal wall hydrophobization, charge into and contain Ca
2+The DCE solution of carrier is organic phase; The horseradish cell solution is a water, uses Ag/AgCl to be the aqueous phase contrast electrode, and Ag/AgTPBCl is a contrast electrode in the organic phase, utilizes cyclic voltammetry to Ca inside and outside the horseradish cell
2+Dynamic continuous detecting is carried out in exchange.
The cyclic voltammetry experiment of nanotube capillaries adopts two electrode systems in the such scheme, carries out in the electrolytic cell in shielded box.
The invention also discloses said method and measuring rare earth ion pair cell surface C a
2+Application in the migration influence.Specifically: with the borosilicate kapillary is the material preparation nanotube capillaries, and carries out the inwall silanization and make it become hydrophobicity; In the nanotube capillaries of inner tubal wall hydrophobization, charge into and contain Ca
2+The DCE solution of carrier is organic phase; The horseradish cell solution is a water, uses Ag/AgCl to be the aqueous phase contrast electrode, and Ag/AgTPBCl is a contrast electrode in the organic phase, utilizes cyclic voltammetry to Ca inside and outside the horseradish cell
2+Exchange detects; In cell solution, drip the finite concentration rare earth ion subsequently, hatch 5 min, carry out cyclic voltammetric and measure.After obtaining the processing of variable concentrations rare earth ion, cell surface Ca
2+Cyclic voltammetry curve in the liquid/liquid interface migration.
The innovation part of this patent is:
That the full cell record figure of patch-clamp record representes is the inside and outside K of cell
+The channel current (comprising steady-state current and tail current) of exchange, along with the adding of specimen, the K that cell is inside and outside
+Variation has taken place in exchange, is reflected in the channel current change, and presents dose-effect relationship, and the variation of channel current has reflected rare earth ion pair cell K
+The influence of channel function.(Fig. 2)
We are through the surface C a of cyclic voltammetry cell
2+The exchange situation obtains closed cyclic voltammetric current curve.Along with the adding of rare earth ion, variation has taken place in the cyclic voltammetric current curve electric current difference of cell, also demonstrates dose-effect relationship simultaneously.Ca in the variation of electric current difference and the test cell solution
2+The proportional relation of concentration, and Ca
2+The variation of concentration is because the inside and outside Ca of cell
2+Exchange capacity changes and causes.Through repeatedly with patch-clamp data comparative experiments checking, we find: the variation of cyclic voltammetric electric current difference can correspondingly reflect rare earth ion pair cell Ca
2+The influence of channel function.
Bibliographical information is difficult to realize Ca inside and outside the vegetable cell through patch clamp technique
2+Exchange and the specific detection of function of passage.Therefore, we select for use cyclic voltammetry that the horseradish cell is carried out specific continuous detection of dynamic, and the reflection rare earth ion is to Ca inside and outside the horseradish cell
2+Exchange and the influence of channel function, and realized the horseradish cell is carried out dynamic continuous detecting.
The present invention adopts following technical scheme:
Electrode preparation: with borosilicate kapillary (external diameter 1.0 mm, internal diameter 0.58 mm, L=10 cm) is the material preparation nanotube capillaries; Through regulating 5 parameter (Heat of P-2000 Puller; Filament, Velocity, Delay; Pull), the inside and outside caliber of preparation is than the kapillary (patch-type) that is 2.Before use, whether kapillary need smooth through BX-41 optical microscope (Olympus) trial inspection its profile, the particularly mouth of pipe.The mouth of pipe irregularity and the mouth of pipe are not that circular kapillary all can not use.
In the experiment, we need inject DCE organic solution in kapillary, and the borosilicate kapillary is hydrophilic, and outer water gets in the pipe and the accuracy of influence experiment in order to prevent to manage, must elder generation it to be become hydrophobic.The method of kapillary inside surface hydrophobization is: be immersed in trimethyl chlorosilane solution to the nanotube capillaries tip; Continue a moment (capillary effect makes trimethyl chlorosilane get into kapillary); To get into trimethyl chlorosilane capillaceous from the pipe rear end with syringe then slowly promotes toward pipe end; Make it arrive tip, can not retain bubble in the kapillary and make it in pipe, keep about 30 min, then from managing the rear end unnecessary trimethyl chlorosilane sucking-off.Nanotube capillaries behind the silanization leaves standstill a night, dries subsequent use.
The inside and outside Ca of horseradish cell
2+Exchange detects:
The cyclic voltammetry experiment of nanotube capillaries adopts two electrode systems, carries out in the electrolytic cell in shielded box.The aqueous phase contrast electrode is Ag/AgCl electrode (diameter is that the filamentary silver of 0.20 mm is made anode, and diameter is that the platinum filament of 1 mm is made negative electrode, and the WS of electrolysis KCl makes the Ag/AgCl electrode), and supporting electrolyte is used LiCl; Contrast electrode in the organic phase (is dissolved in a small amount of TBATPBCl in the DCE solution with Ag/AgTPBCl; Diameter is that the filamentary silver of 0.2 mm is an anode, and diameter is that the platinum filament of 1 mm is a negative electrode, the DCE solution of electrolysis TBATPBCl; Can prepare the Ag/AgTPBCl electrode), supporting electrolyte is used TBATPBCl.BAS 100B electrochemical workstation (U.S. Bioanalytical Systems company) is used to write down the cyclic voltammogram of experiment.(25 ± 2 oC) all at room temperature carried out in all experiments.
In the nanotube capillaries of inner tubal wall hydrophobization, charge into and contain the Calcium ionophore II DCE solution of (having another name called: N, N, N ', N '-Fourth Ring hexyl-3-oxa-glutaramide), utilize cyclic voltammetry to detect Ca in the solution
2+Change in concentration, the expression of electrochemical cell design is following:
Ag|AgCl|CaCl
2||10mM?TBATPBCl+?0.5～1?mM?Ca
2+?ionophore?II?|Ag|AgTPBCl
In order to confirm Ca
2+Typical curve (Fig. 3), we adopt titrimetry, to variable concentrations Ca
2+The cyclic voltammetric detection has been carried out in transfer at liquid/liquid interface.With Ca
2+Concentration (x) and corresponding dissufion current (y) mapping obtain electric current and Ca
2+The relational expression of concentration is Ca inside and outside the follow-up cell
2+The exchange quantitative Analysis provides the basis.
In experimentation, select to do a lot of work in the concentration of Calcium ionophore, confirm that finally 0.5～1 mM is than suitable concn.In the testing process, voltage range is set also has difficulties, through groping repeatedly, finally confirm, the horseradish mesophyll cell be provided with voltage for-300 mV to 400 mV, the horseradish root-tip cells be provided with voltage for-200 mV to 700 mV.
Infinitely near the cell in the electrolytic cell, utilize cyclic voltammetry to detect cell surface Ca capillary electrode
2+Concentration.
The electrochemical cell expression that uses is following:
Ag|AgCl|extracellular?fluid||10mM?TBATPBCl+?0.5～1?mM?Ca
2+?ionophore?II|Ag|AgTPBCl
In cell solution, drip finite concentration La (III) subsequently, hatch 5 min, carry out cyclic voltammetric and measure.After obtaining variable concentrations La (III) processing, cell surface Ca
2+At the cyclic voltammetry curve of liquid/liquid interface migration, the result sees Fig. 4 and Fig. 5.Can know by Fig. 4 and Fig. 5, after variable concentrations La (III) handles, cell surface Ca
2+Migration at liquid/liquid interface has all obtained good cyclic voltammetric response.
The present invention has following advantage: according to the present invention, the rare earth La (III) of realizing variable concentrations is to vegetable cell surface C a
2+The METHOD FOR CONTINUOUS DETERMINATION of influence; Change Ca
2+Carrier concn can be realized different plants, different blades, the inside and outside Ca of the cell of different growth periods
2+The detection of exchange.The present invention is by means of cyclic voltammetry, and easy instrument is measured.Realize Ca inside and outside the cell according to the present invention
2+The detection of exchange is significant to the research of various membrane ion-exchange capacities.
Description of drawings
Fig. 1 is the device synoptic diagram.
The horseradish mesophyll cell K that Fig. 2 variable concentrations rare earth La (III) is handled
+The full cell record figure of channel current cyclic voltammogram; (a: horseradish mesophyll cell; B: through the horseradish mesophyll cell of 30 μ M La (III) processing; C: through the horseradish mesophyll cell of 80 μ M La (III) processing).
Fig. 3 variable concentrations Ca
2+Cyclic voltammogram (calcium ion concentration is 0.05 mM)
The inside and outside Ca of the horseradish mesophyll cell that Fig. 4 variable concentrations rare earth La (III) is handled
2+The exchange detection curve; (a: horseradish mesophyll cell; B: through the horseradish mesophyll cell of 30 μ M La (III) processing; C: through the horseradish mesophyll cell of 80 μ M La (III) processing).
The inside and outside Ca of the horseradish root-tip cells that Fig. 5 variable concentrations rare earth La (III) is handled
2+The exchange detection curve; (a: horseradish root-tip cells; B: through the horseradish root-tip cells of 30 μ M La (III) processing; C: through the horseradish root-tip cells of 80 μ M La (III) processing).
Embodiment
(1) nanotube capillaries draws
With borosilicate kapillary (external diameter 1.0 mm, internal diameter 0.58 mm, L=10 cm) is the material preparation nanotube capillaries; Be provided with and draw appearance (P-2000 Puller) parameter (Heat, Filament, Velocity; Delay; Pull), draw kapillary, obtain most advanced and sophisticated inside and outside caliber than the kapillary that is 2.0.(capillary pipe length is 10 cm, breaks from the centre, obtains the two one cuspidated nanotube capillaries of end)
(2) nanotube capillaries inwall hydrophobization is handled
Be immersed in trimethyl chlorosilane solution to the nanotube capillaries tip; Continue a moment (capillary effect makes trimethyl chlorosilane get into kapillary); To get into trimethyl chlorosilane capillaceous from the pipe rear end with syringe then slowly promotes toward pipe end; Make it arrive tip, can not retain bubble in the kapillary and make it in pipe, keep about 30 min, then from managing the rear end unnecessary trimethyl chlorosilane sucking-off.Nanotube capillaries behind the silanization leaves standstill a night, dries subsequent use.
(3) electrode preparation
The preparation of Ag/AgCl: diameter is that the filamentary silver of 0.25 mm is made anode, and diameter is that the platinum filament of 1 mm is made negative electrode, the WS of electrolysis KCl.
The preparation of Ag/AgTPBCl: be dissolved in a spot of tetraphenylarsonium chloride boron TBuA (TBATPBCl) in the DCE solution, diameter be the filamentary silver of 0.25 mm as anode, diameter be the platinum filament of 1 mm as negative electrode, the DCE solution of electrolysis TBATPBCl.
(4) experimental provision builds
Before the detection, in the nanotube capillaries that step (2) hydrophobization is handled, contain Ca through syringe injection a certain amount of (nanotube capillaries volume 2/3)
2+1 of selectivity carrier, 2-ethylene dichloride (DCE) organic solution.
Ag/AgTPBCl electrode one end is inserted into above-mentioned kapillary (organic phase) to most advanced and sophisticated; Eletrode tip after the assembling is inserted in the cell liquid (water); The Ag/AgTPBCl electrode that is exposed at the nanotube capillaries outside is clamped with the connection of electrochemical workstation, and fixing.Ag/AgCl electrode one end is inserted in the cell solution (water) (2 mL), and the other end is clamped with the connection of electrochemical workstation, and fixing.(device synoptic diagram such as Fig. 1)
(5) cyclic voltammetry detects little electric current
Said cyclic voltammetry detects horseradish cell surface Ca
2+Concrete experimentation go up to accomplish at BAS 100B electrochemical workstation (U.S. Bioanalytical Systems company).The cyclic voltammetry experiment of nanotube capillaries adopts two electrode systems, carries out in the electrolytic cell in shielded box.Ca in the nanotube capillaries
2+The selectivity carrier is to Ca
2+Little electric current of forming of translocation, regulate test parameter (voltage range is swept speed, sensitivity), detect this little electric current through cyclic voltammetry.Through little change in current, confirm to migrate to the Ca of near interface
2+The variation of concentration, and then the inside and outside Ca of definite cell
2+Exchange capacity.
The separation and purification of horseradish mesophyll cell is according to document (Sarhan F, Cesar D, High yield isolation of mesophyll protoplasts from wheat; Barley and rye [J]; Physiologia Plantarum, 1988,72 (2): 337-342) carry out.Get the horseradish spire and be cut into small pieces (1 mm * 2 mm), be placed in the little double dish that fills 5 mL enzyme liquid, make a catalase liquid removing lower epidermis and be paved with one deck, enzymolysis 3-4 h under 25-28 ° of C.Enzyme liquid is dissolved in the mannitol solution of 0.8 M by 1.0% cellulase (R-10) and 0.1 pectase (Y-23), and sweet mellow wine is the homeo-osmosis agent.With suction pipe enzymolysis liquid is drawn in the 5 mL centrifuge tubes, makes cell settlement stay deposition with centrifugal 5 min of 600 rpm.Observation can be seen a large amount of sphaerocysts under common inversion optical microscope.
Infinitely near the horseradish mesophyll cell in the electrolytic cell, utilize cyclic voltammetry to detect cell surface Ca capillary electrode
2+Concentration.The expression of electrochemical cell design is following:
Ag|AgCl|extracellular?fluid||10mM?TBATPBCl+1?mM?Ca
2+?ionophore?II?|Ag|AgTPBCl
During mensuration, be provided with voltage for-300 mV to 400 mV, sweeping speed is 20 mV/s, measurement sensitivity is 10 nA, pair cell solution carries out cyclic voltammetric and detects that (Fig. 3 a) obtains good cyclic voltammetric response.
In cell solution, drip finite concentration Rare Earth Lanthanum La (III) subsequently, hatch 5 min, carry out cyclic voltammetric and measure.After obtaining variable concentrations La (III) processing, cell surface Ca
2+At the cyclic voltammetry curve of liquid/liquid interface migration, the result sees Fig. 3 (b-c).Can know by Fig. 3 (b-c), after variable concentrations La (III) handles, cell surface Ca
2+Migration at liquid/liquid interface has all obtained good cyclic voltammetric response.Compare the horseradish mesophyll cell solution C a that 30 μ M La (III) handle with control cells
2+Density loss, show that 30 μ M La (III) handle after, Ca
2+Increase by the intracellular amount of the outer entering of horseradish mesophyll cell.When cell is handled cell surface Ca through 80 μ M La (III)
2+The concentration ratio control cells is low, but is higher than the cell that 80 μ M La (III) handle, show that high concentration La (III) handles after, horseradish mesophyll cell Ca
2+Interior flow increases, and recruitment is handled cell less than 30 μ M La (III).Can judge rare earth La (III) pair cell Ca thus
2+The influence of exchange capacity.
Embodiment 2
The cyclic voltammetry experiment of nanotube capillaries adopts two electrode systems, carries out in the electrolytic cell in shielded box.The aqueous phase contrast electrode is Ag/AgCl electrode (diameter is that the filamentary silver of 0.20 mm is made anode, and diameter is that the platinum filament of 1 mm is made negative electrode, and the WS of electrolysis KCl makes the Ag/AgCl electrode), and supporting electrolyte is used LiCl; Contrast electrode in the organic phase (is dissolved in a small amount of TBATPBCl in the DCE solution with Ag/AgTPBCl; Diameter is that the filamentary silver of 0.2 mm is an anode, and diameter is that the platinum filament of 1 mm is a negative electrode, the DCE solution of electrolysis TBATPBCl; Can prepare the Ag/AgTPBCl electrode), supporting electrolyte is used TBATPBCl.BAS 100B electrochemical workstation is used to write down the cyclic voltammogram of experiment.(25 ± 2 oC) at room temperature carried out in experiment.
The separation and purification of horseradish root-tip cells is according to document (Sarhan F, Cesar D, High yield isolation of mesophyll protoplasts from wheat; Barley and rye [J]; Physiologia Plantarum, 1988,72 (2): 337-342) carry out.Get the horseradish tip of a root and be cut into small pieces (1 mm * 2 mm), be placed in the little double dish that fills 5 mL enzyme liquid, make a catalase liquid removing lower epidermis and be paved with one deck, enzymolysis 3-4 h under 25-28 ° of C.Enzyme liquid is dissolved in the mannitol solution of 0.8 M by 1.0% cellulase (R-10) and 0.1 pectase (Y-23), and sweet mellow wine is the homeo-osmosis agent.With suction pipe enzymolysis liquid is drawn in the 5 mL centrifuge tubes, makes cell settlement, abandon supernatant, stay deposition with centrifugal 5 min of 600 rpm.Observation can be seen a large amount of sphaerocysts under common inversion optical microscope.
In the nanotube capillaries of inner tubal wall hydrophobization, charge into and contain Ca
2+The DCE solution of carrier infinitely near the horseradish root-tip cells in the electrolytic cell, utilizes cyclic voltammetry to detect cell surface Ca capillary electrode
2+Concentration.The expression of electrochemical cell design is following:
Ag|AgCl|extracellular?fluid||10mM?TBATPBCl+0.5?mM?Ca
2+?ionophore?II?|Ag|AgTPBCl
During mensuration, be provided with voltage for-200 mV to 700 mV, sweeping speed is 20 mV/s, measurement sensitivity is 10 nA, pair cell solution carries out cyclic voltammetric and detects that (Fig. 4 a) obtains good cyclic voltammetric response.
In cell solution, drip finite concentration rare earth La (III) subsequently, hatch 5 min, carry out cyclic voltammetric and measure.After obtaining variable concentrations La (III) processing, cell surface Ca
2+At the cyclic voltammetry curve of liquid/liquid interface migration, the result sees Fig. 4 (b-c).Can know by Fig. 4 (b-c), after variable concentrations La (III) handles, cell surface Ca
2+Migration at liquid/liquid interface has all obtained good cyclic voltammetric response.After 30 μ M La (III) handle, horseradish root-tip cells Ca
2+Exchange capacity strengthens, and the Ca of the horseradish root-tip cells that 80 μ M La (III) handle
2+Exchange capacity also strengthens to some extent, but the amplitude that increases is less than the processing of 30 μ M La (III).
Claims (7)
1. the inside and outside Ca of an easy horseradish cell
2+The exchange dynamic testing method is characterized in that may further comprise the steps:
(1) is the material preparation nanotube capillaries with the borosilicate kapillary, and carries out the inwall silanization and make it become hydrophobicity;
(2) in the nanotube capillaries of inner tubal wall hydrophobization, charge into and contain Ca
2+
The DCE solution of carrier is organic phase; The horseradish cell solution is a water, uses Ag/AgCl to be the aqueous phase contrast electrode, and Ag/AgTPBCl is a contrast electrode in the organic phase, utilizes cyclic voltammetry to Ca inside and outside the horseradish cell
2+
Detection of dynamic is carried out in exchange.
2. method according to claim 1 is characterized in that said borosilicate kapillary external diameter 1.0 mm internal diameters 0.58 mm, L=10 cm.
3. method according to claim 2 is characterized in that the cyclic voltammetry experiment of nanotube capillaries adopts two electrode systems, carries out in the electrolytic cell in shielded box.
The described method of claim 1 measure rare earth metal to the horseradish cell inside and outside Ca
2+Application in the migration influence.
5. application according to claim 4 is characterized in that: with the borosilicate kapillary is the material preparation nanotube capillaries, and carries out the inwall silanization and make it become hydrophobicity; In the nanotube capillaries of inner tubal wall hydrophobization, charge into and contain Ca
2+The DCE solution of carrier is organic phase; The horseradish cell solution is a water, uses Ag/AgCl to be the aqueous phase contrast electrode, and Ag/AgTPBCl is a contrast electrode in the organic phase, utilizes cyclic voltammetry to Ca inside and outside the horseradish cell
2+Exchange detects; In cell solution, drip the finite concentration rare earth ion subsequently, hatch 5 min, carry out cyclic voltammetric and measure.
6. after obtaining the processing of variable concentrations rare earth ion, the inside and outside Ca of cell
2+Cyclic voltammetry curve in the liquid/liquid interface migration.
7. according to claim 4 and 5 described application, it is characterized in that said rare earth metal is a Rare Earth Lanthanum.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114180851A (en) * | 2021-11-25 | 2022-03-15 | 中山大学 | Method for forming stable micro-nano liquid/liquid interface through silanization of inner wall of tip of glass tube |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030132109A1 (en) * | 2001-11-30 | 2003-07-17 | Andrew Bullen | Pipette configurations and arrays thereof for measuring cellular electrical properties |
| CN101076600A (en) * | 2004-05-03 | 2007-11-21 | 惠氏公司 | Rapid pouring system and memberane tongs technology using interface room system with high-flux and low-volume requirement |
-
2012
- 2012-04-18 CN CN2012101140325A patent/CN102636550A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030132109A1 (en) * | 2001-11-30 | 2003-07-17 | Andrew Bullen | Pipette configurations and arrays thereof for measuring cellular electrical properties |
| CN101076600A (en) * | 2004-05-03 | 2007-11-21 | 惠氏公司 | Rapid pouring system and memberane tongs technology using interface room system with high-flux and low-volume requirement |
Non-Patent Citations (3)
| Title |
|---|
| DONGPING ZHAN等: "Scanning Electrochemical Microscopy. 58. Application of a Micropipet-Supported ITIES Tip to Detect Ag+ and Study Its Effect on Fibroblast cells", 《ANALYTICAL CHEMISTRY》, vol. 79, no. 14, 15 July 2007 (2007-07-15) * |
| 薛亚东: "肿瘤细胞表面聚糖的电化学检测与细胞成像分析", 《中国博士学位论文全文数据库医药卫生科技辑》, 15 July 2011 (2011-07-15) * |
| 贾莉君: "离子选择微电极测定植物细胞跨膜电位和液泡中硝酸根离子活度的方法研究", 《中国优秀硕士学位论文全文数据库工程科技Ⅰ辑》, 15 February 2007 (2007-02-15) * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114180851A (en) * | 2021-11-25 | 2022-03-15 | 中山大学 | Method for forming stable micro-nano liquid/liquid interface through silanization of inner wall of tip of glass tube |
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Application publication date: 20120815 |