CN202330238U - Multifunctional probe for single cell detection - Google Patents
Multifunctional probe for single cell detection Download PDFInfo
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- CN202330238U CN202330238U CN2011202686436U CN201120268643U CN202330238U CN 202330238 U CN202330238 U CN 202330238U CN 2011202686436 U CN2011202686436 U CN 2011202686436U CN 201120268643 U CN201120268643 U CN 201120268643U CN 202330238 U CN202330238 U CN 202330238U
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- microelectrodes
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Abstract
The utility model relates to a multifunctional probe for single cell detection, which comprises temperature measurement, film potential measurement, ion channel detection, pH detection and ion concentration detection. The probe comprises a temperature measuring film thermocouple (11) in a cell and three microelectrodes. The three microelectrodes include a film potential measurement and ion channel detection microelectrode (12), a PH value measurement microelectrode (13) and an ion concentration detection microelectrode (14). The three microelectrodes are all conical and cling to each other, and the cone tips of the three microelectrodes are located in the same direction. The temperature measuring film thermocouple (11) is composed of two metal films and an insulation layer, wherein the two metal films form temperature measuring knots at the tip ends on the outside of the three microelectrodes, and other portions are insulated. The multifunctional probe for single cell detection is capable of achieving common measurement of multiple parameters and is favorable for better studying structure and performance in the subcellular fraction level.
Description
Technical field
The utility model belongs to the multiprobe of unicellular detection range, the particularly making of the film thermocouple on glass tube surface, and responsive microelectrode of PH and ion concentration microelectrode are made.
Background technology
People have been deep into unicellular, monomolecular level to the observation and the research of biological phenomena, how in such range scale, to obtain useful biological physics and chemistry information each research field has all been proposed new requirement.The application of unicellular detection technique becomes one of main flow field of modern analysis research gradually.Can carry out the focus and emphasis that molecular probe that real-time, online, original position, live body detect and superminiature biology sensor become people's research.
General biophysics Neher of research institute of wherein German horse and Sakmann have created patch clamp technique in 1976, this is a kind ofly to reflect cell membrane single (or technology of a plurality of ion channel molecule activities) with the gas current of record through ion channel.Later on because the establishment of gigohm nurse impedance method for sealing and the establishment of several method.This technology has been lighted the fire of revolution of the Physiologic Studies of cell and molecular level, and huge onward impulse has been brought to life science in it and gene clone technology horse and horse.The contribution that this is great makes Neher and Sakmann obtain Nobel's physiology and the medical science prize in 1991 years.Development so far; Become the conventional method of modern cellular electrophysiologicalsensor; It not only can be used as the instrument of basic biomedical research; And directly or indirectly be the clinic study service, patch clamp technique is widely used in multidisciplinary area researches such as nerve (brain) science, cardiovascular science, pharmacology, cell biology, Pathological Physiology, traditional Chinese medicine and pharmacy, plant cell physiology, physiology of exercise at present.
Based on patch clamp technique; In detection of cells film gas current, measure other bio signal; Like intracellular temperature, pH value, ion concentration etc.; Then can be the single celled life process of research more physical and chemical parameter is provided,, also can be used as drug screening instrument efficiently is provided for unicellular fundamental research provides research tool.
Summary of the invention
Technical matters: the purpose of the utility model is to provide a kind of multiprobe that is used for unicellular detection, can detect the unicellular probe of multiple physiological signal simultaneously, with research cell life process new research tool is provided for measuring unicellular physical and chemical parameter.
Technical scheme: the multiprobe that is used for unicellular detection of the utility model comprises temperature-measuring thin film thermopair and three microelectrodes in the cell; Described three microelectrodes are: film potential is measured and ion channel detects microelectrode; PH value is measured microelectrode, and ion concentration detects microelectrode; Said three microelectrodes all are conical, and three microelectrodes abut against together, and the epiconus of three microelectrodes is positioned at same direction; Said temperature-measuring thin film thermopair is made up of two kinds of metallic films and insulation course, and wherein two kinds of metallic films are at the most advanced and sophisticated formation in the outside of three microelectrodes thermometric knot, remainder mutually insulated.
Three microelectrodes draw through the glass tube thermal treatment that contains tube core in three and form.
Said pH value is measured microelectrode, adopts the responsive liquid film of PH, PH sensing glass or solid-state PH sensor to realize.Said ion concentration measurement microelectrode adopts ion-sensitive glass or ion sensitive polymer film.
Described temperature-measuring thin film thermopair, but the target of employing sputter or vapor deposition; Wherein select platinum-tungsten, platinum-Jin, platinum-Yin or platinum-copper to have the metal material of big thermopower difference right at normal temperatures, or use copper nickel-copper, platinum Pt Rh, the commercial alloy material of nickel chromium triangle-copper nickel right.
Said insulation course adopts SiO
2Or MgO, realize through the magnetron sputtering insulating material.
Beneficial effect: the unicellular detector probe through the method obtains, when realizing traditional ion channel, film potential measurement function, also can realize the measurement of cell internal and external temperature, and intracellular ph value and ion concentration.The Temperature Feedback that wherein temperature survey function provides has also further improved the precision that the pH value is measured.
Description of drawings
Fig. 1 is the utility model multiprobe structural representation.
Fig. 2 is second step of the utility model, uses the PH sensing glass and uses PH sensitive material liquid film in a microtubule therein.
Fig. 3 is the 3rd step of the utility model, uses ion-sensitive glass and ion sensitive polymer film in a microtubule therein.
Fig. 4 is the 4th step of the utility model, the film thermocouple of making in the glass electrode outside.
More than comprise among the figure: temperature-measuring thin film thermopair 11, film potential are measured and ion channel detects microelectrode 12, and pH value is measured microelectrode 13, and ion concentration detects microelectrode 14; Glass microelectrode 21, PH sensing glass 22, the most advanced and sophisticated sealing-in 23 of PH sensing glass, PH sensing glass and glass microelectrode inwall sealing-in 24, the responsive liquid film 25 of PH; Glass microelectrode 31, ion-sensitive glass 32, the most advanced and sophisticated sealing-in 33 of ion-sensitive glass, ion-sensitive glass and glass microelectrode inwall sealing-in 34, ion concentration sensitive polymer film 35; Glass microelectrode 41, film thermocouple 42; Metallic film 1, insulation course 44, metallic film 2 45.
Embodiment
The utility model is used for the multiprobe of unicellular detection; Comprise temperature-measuring thin film thermopair 11 and three microelectrodes in the cell; Described three microelectrodes are: film potential is measured and ion channel detects microelectrode 12, and pH value is measured microelectrode 13, and ion concentration detects microelectrode 14; Said three microelectrodes all are conical, and three microelectrodes abut against together, and the epiconus of three microelectrodes is positioned at same direction; Said temperature-measuring thin film thermopair 11 is made up of two kinds of metallic films and insulation course, and wherein two kinds of metallic films are at the most advanced and sophisticated formation in the outside of three microelectrodes thermometric knot, remainder mutually insulated.Wherein three tube cores are realized patch-clamp basic function (film potential measurement, ion channel, microinjection), pH measurement function, ion concentration measurement function respectively.
The preparation method is:
The first step draws appearance through glass electrode and draws three core glass electrodes, through hot polishing, makes eletrode tip become Paint Gloss, thereby can improve the sealing-in rate with cell membrane.
In second step, integrated PH sensitive sensor in microtubule can be used the PH sensing glass therein, adopts the method for heat sealing, perhaps adopts PH sensitive material liquid-film method.
The 3rd step, integrated ion concentration sensitive sensor in another root microtubule therein.Can use ion-sensitive glass and ion-sensitive material polymers film.
In the 4th step,, use the method for sputter or vapor deposition, preparation metallic film and insulation course in the glass electrode outside.
Instance:
At first select the pyrex pipe that contains pipe in three for use, 10cm is long, single tube external diameter 1mm, internal diameter 0.75mm, no filament.On glass electrode drawing appearance, draw out the glass microelectrode that tip diameter is 1um, make eletrode tip become Paint Gloss through hot polishing, thereby can improve the sealing-in rate.
Next selects PH sensing glass pipe and potassium ion sensing glass pipe for use, and long 10cm, external diameter of pipe are 0.7mm, and internal diameter is 0.35mm, no filament.Draw the glass electrode that appearance draws out tip diameter 0.2um through glass electrode, handle, make most advanced and sophisticated sealing-in closed through annular heat.Control two glass microelectrodes and insert wherein two in the glass electrode that contains pipe in three, and slowly be inserted into most advanced.End at two electrodes feeds nitrogen and keeps certain air pressure then, utilizes thermal treatment that it is heated simultaneously, makes outer wall of inner tube and outer tube wall sealing-in.
At the thick platinum film of pyrex pipe outer wall sputter 100nm, the outside sputter of platinum film covers the thick SiO of 20nm through magnetron sputtering
2Insulation course is removed most advanced and sophisticated SiO through ion beam etching
2Layer makes the platinum film expose out in glass tube tip, in the thick gold thin film of outermost layer sputter 100nm.So just formed platinum-Pt/Au thermocouple thermometric knot at pyrex pipe outer wall tip.
Claims (6)
1. multiprobe that is used for unicellular detection; It is characterized in that this probe comprises temperature-measuring thin film thermopair (11) and three microelectrodes in the cell; Described three microelectrodes are: film potential is measured and ion channel detects microelectrode (12); PH value is measured microelectrode (13), and ion concentration detects microelectrode (14); Said three microelectrodes all are conical, and three microelectrodes abut against together, and the epiconus of three microelectrodes is positioned at same direction; Said temperature-measuring thin film thermopair (11) is made up of two kinds of metallic films and insulation course, and wherein two kinds of metallic films are at the most advanced and sophisticated formation in the outside of three microelectrodes thermometric knot, remainder mutually insulated.
2. by the described multiprobe that is used for unicellular detection of claim 1, it is characterized in that three microelectrodes draw through the glass tube thermal treatment that contains tube core in three and form.
3. by the described multiprobe that is used for unicellular detection of claim 1, it is characterized in that said pH value is measured microelectrode (13), adopt the responsive liquid film of PH, PH sensing glass or solid-state PH sensor to realize.
4. by the described multiprobe that is used for unicellular detection of claim 1, it is characterized in that said ion concentration measurement microelectrode (14) adopts ion-sensitive glass or ion sensitive polymer film.
5. by the described multiprobe that is used for unicellular detection of claim 2, it is characterized in that described temperature-measuring thin film thermopair (11), but adopt the target of sputter or vapor deposition; Wherein select platinum-tungsten, platinum-Jin, platinum-Yin or platinum-copper to have the metal material of big thermopower difference right at normal temperatures, or use copper nickel-copper, platinum Pt Rh, the commercial alloy material of nickel chromium triangle-copper nickel right.
6. by the described multiprobe that is used for unicellular detection of claim 2, it is characterized in that said
Insulation course adopts SiO
2Or MgO, realize through the magnetron sputtering insulating material.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011202686436U CN202330238U (en) | 2011-07-27 | 2011-07-27 | Multifunctional probe for single cell detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011202686436U CN202330238U (en) | 2011-07-27 | 2011-07-27 | Multifunctional probe for single cell detection |
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| Publication Number | Publication Date |
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| CN202330238U true CN202330238U (en) | 2012-07-11 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN2011202686436U Withdrawn - After Issue CN202330238U (en) | 2011-07-27 | 2011-07-27 | Multifunctional probe for single cell detection |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102262113A (en) * | 2011-07-27 | 2011-11-30 | 东南大学 | Multifunctional probe for unicellular detection |
| CN104458191A (en) * | 2014-12-10 | 2015-03-25 | 中国航天空气动力技术研究院 | Miniature thin film platinum resistor heat flux transducer and manufacturing method thereof |
-
2011
- 2011-07-27 CN CN2011202686436U patent/CN202330238U/en not_active Withdrawn - After Issue
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102262113A (en) * | 2011-07-27 | 2011-11-30 | 东南大学 | Multifunctional probe for unicellular detection |
| CN102262113B (en) * | 2011-07-27 | 2013-05-01 | 东南大学 | Multifunctional probe for unicellular detection |
| CN104458191A (en) * | 2014-12-10 | 2015-03-25 | 中国航天空气动力技术研究院 | Miniature thin film platinum resistor heat flux transducer and manufacturing method thereof |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| AV01 | Patent right actively abandoned |
Granted publication date: 20120711 Effective date of abandoning: 20130501 |
|
| RGAV | Abandon patent right to avoid regrant |