CN112146956B - Glass needle tip nano-hole filling device based on vacuum method and using method thereof - Google Patents
Glass needle tip nano-hole filling device based on vacuum method and using method thereof Download PDFInfo
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- CN112146956B CN112146956B CN202011017897.0A CN202011017897A CN112146956B CN 112146956 B CN112146956 B CN 112146956B CN 202011017897 A CN202011017897 A CN 202011017897A CN 112146956 B CN112146956 B CN 112146956B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
Abstract
The invention belongs to the technical field of life science and biomolecule detection, and relates to a filling device for a needle tip nanopore, which comprises a centrifuge tube and a clamp holder clamped on the centrifuge tube in a sealing way, wherein a liquid containing groove is integrally formed at the top of the clamp holder, the clamp holder is vertically through, a glass tube with a needle tip nanopore structure is inserted into a through hole in the middle part of the clamp holder, and the problem that the whole glass needle tip nanopore is difficult to fill and the needle tip bubble is difficult to discharge due to the fact that a microinjector is adopted to inject filling liquid into a microtube in the prior filling method for the glass needle tip nanopore is solved.
Description
Technical Field
The invention belongs to the technical fields of life science and biomolecule detection, relates to a filling device for a glass needle tip nanopore, and particularly relates to a filling device for a glass needle tip nanopore based on a vacuum method and a using method of the filling device.
Background
Nanopore technology-based single molecule sensors have been used today for biological, chemical sensor preparation, detection screening of biomolecules, and DNA sequencing. The needle tip nanopore technology is one of solid nanopores, and has the remarkable advantages of easiness in manufacturing, high signal-to-noise ratio, low cost and the like compared with the traditional solid film nanopores. At present, the filling method of the glass needle point nano hole mainly adopts a micro-injector to inject into the micro-tube, and then flicks the needle point to remove bubbles, but the filling of the whole glass needle point nano hole cannot be realized in the mode. And because the opening diameter of the nano hole of the glass needle point is small, bubbles easily generated at the tip of the nano hole are difficult to remove.
It is reported that the method can rapidly fill the needle point by adopting a centrifugal method after the glass tube is filled with liquid through the micro-injector, but a new solution breakpoint can occur under the action of centrifugal force due to the hydrophilic and hydrophobic property of the surface of the glass needle point, so that the filling success rate of the solution is low; it has also been reported that the vacuum pumping method is adopted after the glass tube is injected by the micro-injector, the method consumes more solution and has lower success rate; recently, a microwave method is adopted to improve the success rate of injecting the solution into the small-aperture glass needle tip nano-hole after a vacuum pumping method, however, the concentration property of the solution is easy to change under the microwave heating condition, and the solution is also easy to be polluted.
Disclosure of Invention
In view of the above, the invention provides a glass needle point nano hole filling device based on a vacuum method and a using method thereof, which aims to solve the problems that the whole glass needle point nano hole is difficult to fill and the needle point bubbles are difficult to discharge in the existing glass needle point nano hole filling method for injecting filling liquid into a micro tube by adopting a micro syringe.
In order to achieve the purpose, the invention provides a glass needle point nano hole filling device based on a vacuum method, which comprises a centrifuge tube and a clamp holder clamped on the centrifuge tube in a sealing way, wherein a liquid containing groove is integrally formed at the top of the clamp holder, the clamp holder is in a vertically through shape, and a glass tube with a needle point nano hole structure is inserted into a through hole in the middle of the clamp holder.
The beneficial effect of this basic scheme lies in: the microinjector fills liquid into the centrifuge tube through the glass tube and then is connected with the clamp holder in a sealing way, then the centrifuge tube is centrifuged to remove bubbles, and finally the centrifuge tube is vacuumized.
Further, the holder from bottom to top includes bottom, packing ring, upper portion and flourishing cistern in proper order, and the through-hole that runs through has all been seted up at the holder middle part, and the recess is seted up at the base middle part and the external screw thread has been seted up to the outer wall, and the ring channel inner wall has been seted up for seting up ring channel and ring channel inner wall, and flourishing cistern is seted up in the outside on holder upper portion, and the cooperation of internal screw thread and external screw thread is with the packing ring centre gripping between bottom and upper portion.
The beneficial effects are that: the gasket between the bottom and the upper part is used for clamping the glass tube, the bottom and the upper part are in threaded fit with the extrusion gasket to fix the glass tube, and the bottom of the clamp can be just inserted into the opening of the centrifuge tube to form a sealed clamp.
The application method of the glass needle point nano hole filling device based on the vacuum method comprises the following steps:
A. placing a gasket in a groove at the bottom of the clamp holder, mutually biting and fixing the bottom and the upper part through threads, and extruding and fixing the gasket to a glass tube penetrating through the clamp holder;
B. adding a required solution into the centrifuge tube through the tail part of the glass tube by adopting a microinjector, and plugging the glass tube into the centrifuge tube by adopting the clamp holder, wherein the liquid level in the centrifuge tube needs to be beyond the neck part of the needle point of the glass tube;
C. placing the assembled filling device on a centrifugal machine, and prefilling by a centrifugal method;
D. sealing the joint of the clamp holder and the centrifuge tube by using a sealing film, placing the sealing film on a centrifuge tube rack, placing the centrifuge tube rack into a vacuum dryer, standing for more than 10min under a vacuum state after vacuum extraction, and opening the vacuum dryer after filling to finish filling of the needle point nano holes.
Further, the top end of the glass tube in the step A does not exceed the top of the liquid containing tank.
Further, the vacuum in step D is greater than 0.98 atmospheres.
The invention has the beneficial effects that:
1. according to the glass needle point nano hole filling device based on the vacuum method, the micro injector is used for sealing and connecting the centrifuge tube with the clamp holder after filling liquid into the centrifuge tube through the glass tube, then the centrifuge tube is centrifuged to remove bubbles, and finally the centrifuge tube is vacuumized.
2. The glass needle tip nano-pore filling device based on the vacuum method combines the centrifugal method and the vacuum method, and ensures the filling success rate on the premise of ensuring that the consumption of the solution is less and the property of the solution is not changed. Successful application of the device and the technology can effectively improve the filling rate and the use rate of the needle tip nano-holes. Therefore, the filling device of the glass needle point nano hole based on the vacuum method and the using method thereof bring technical innovation to the filling of the glass needle point nano Kong Chong, and have high popularization value.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention. The objects and other advantages of the invention may be realized and obtained by means of the instrumentalities and combinations particularly pointed out in the specification.
Drawings
For the purpose of making the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in the following preferred detail with reference to the accompanying drawings, in which:
FIG. 1 is a schematic diagram of a glass needle tip nanopore filling device based on a vacuum method;
FIG. 2 is a schematic diagram of the structure of a holder in the glass needle tip nanopore filling device based on the vacuum method;
FIG. 3 is a schematic diagram of a vacuum filling of a glass needle tip nanopore filling device based on a vacuum method of the present invention;
FIG. 4 is a microscopic view of a needle tip of a drawn glass tube in a vacuum-based glass needle tip nanopore filling device of the present invention;
FIG. 5 is a microscope image of the tip of a glass tube after the solution is centrifugally filled in example 1 of the present invention;
FIG. 6 is a microscopic view of the tip of a glass tube after filling by the solution vacuum method in example 1 of the present invention.
Reference numerals: a holder 101, centrifuge tube 102, glass tube 103, solution 104, bottom 201, gasket 202, upper 203, and liquid reservoir 204.
Detailed Description
Other advantages and effects of the present invention will become apparent to those skilled in the art from the following disclosure, which describes the embodiments of the present invention with reference to specific examples. The invention may be practiced or carried out in other embodiments that depart from the specific details, and the details of the present description may be modified or varied from the spirit and scope of the present invention. It should be noted that the illustrations provided in the following embodiments merely illustrate the basic idea of the present invention by way of illustration, and the following embodiments and features in the embodiments may be combined with each other without conflict.
Wherein the drawings are for illustrative purposes only and are shown in schematic, non-physical, and not intended to limit the invention; for the purpose of better illustrating embodiments of the invention, certain elements of the drawings may be omitted, enlarged or reduced and do not represent the size of the actual product; it will be appreciated by those skilled in the art that certain well-known structures in the drawings and descriptions thereof may be omitted.
The same or similar reference numbers in the drawings of embodiments of the invention correspond to the same or similar components; in the description of the present invention, it should be understood that, if there are terms such as "upper", "lower", "left", "right", "front", "rear", etc., that indicate an azimuth or a positional relationship based on the azimuth or the positional relationship shown in the drawings, it is only for convenience of describing the present invention and simplifying the description, but not for indicating or suggesting that the referred device or element must have a specific azimuth, be constructed and operated in a specific azimuth, so that the terms describing the positional relationship in the drawings are merely for exemplary illustration and should not be construed as limiting the present invention, and that the specific meaning of the above terms may be understood by those of ordinary skill in the art according to the specific circumstances.
The glass needle tip nano-hole filling device based on the vacuum method as shown in fig. 1-3 comprises a centrifuge tube 102 and a clamp holder 101 clamped on the centrifuge tube 102, wherein the clamp holder 101 is in a vertical penetrating shape, the clamp holder 101 sequentially comprises a bottom 201, a gasket 202, an upper part 203 and a liquid containing groove 204 from bottom to top, a groove is formed in the middle of a base, an external thread is formed in the outer wall of the groove, an internal thread is formed in the inner wall of the groove, the liquid containing groove 204 is formed in the outer side of the upper part 203 of the clamp holder 101, the gasket 202 is clamped between the bottom 201 and the upper part 203 through the matching of the internal thread and the external thread, the bottom 201 of the clamp holder 101 can be just inserted into an opening of the centrifuge tube 102 to form a sealed clamp holder 101, and a glass tube 103 in a needle tip nano-hole structure is inserted into a through hole in the middle of the clamp holder 101.
When the glass needle tip nano-pore filling device based on the vacuum method is used, firstly, a gasket 202 is placed in a groove at the bottom 201 of a clamp 101, the bottom 201 and the upper 203 are mutually meshed and fixed through threads, the gasket 202 is extruded and fixed to a glass tube 103 penetrating through the clamp 101, and the top end of the glass tube 103 does not exceed the top of a liquid containing groove 204; the required solution 104 is added into the centrifuge tube 102 through the tail of the glass tube 103 by adopting a micro-injector, and the liquid level of the glass tube 103 in the clamp holder 101 in the centrifuge tube 102 is required to be beyond the neck of the needle point. Then placing the assembled filling device on a centrifugal machine, and prefilling by a centrifugal method; after centrifugation, the interface between the holder 101 and the centrifuge tube 102 is sealed by a sealing film, the sealing film is placed on a centrifuge tube 102 frame, the centrifuge tube is placed in a vacuum dryer, and after vacuum extraction, the centrifuge tube is kept stand for more than 10 minutes under a vacuum state (the vacuum degree is more than 0.98 atm), and is filled, the solution 104 in the glass tube 103 overflows upwards in the process of removing bubbles in vacuum, the liquid containing groove 204 on the holder 101 is used for containing the overflowed solution 104, the solution 104 is prevented from overflowing out of the centrifuge tube 102, and meanwhile, after the vacuum dryer is opened, the solution 104 in the liquid containing groove 204 can flow back into the glass tube 103, so that the filling of the needle tip nano holes is completed.
Example 1
First step, preparation in early stage of filling
Quartz capillaries with the outer diameter of 1.0mm and the inner diameter of 0.7mm are sequentially treated for 30min under the heating state of piranha washing liquid (volume ratio of 98% H2SO4 to 30% H2O2=3:1), then are ultrasonically cleaned and flushed by deionized water, dried by nitrogen for 30min at 60 ℃, cooled to room temperature and dried and stored for later use; the glass tube 103 with the glass nano-pore needle tip and the needle tip diameter of which is about 30nm is drawn by a P-2000 laser drawing instrument, the microscopic image of the needle tip of the drawn glass tube 103 is shown in fig. 4, and a micro-injector is adopted to inject electrolyte solution 104 of LiCl (pH=8) with the required concentration of 1mol/L into a centrifuge tube 102; then, the glass tube 103 thereof was fixed to the holder 101, the upper tip 203 of the needle was not beyond the bottom 201 of the liquid container 204, the holder 101 was inserted into the mating 1.5ml microcentrifuge tube 102, and the needle tip was immersed in the electrolyte solution 104.
Second step, glass needle tip nanopore prefilling
Placing the prepared filling device on a centrifuge, pre-filling the solution 104 on a high-speed centrifuge under the condition of 2500g and 5min, and performing centrifugal filling on the solution 104, wherein a microscopic view of the needle tip of the glass tube 103 is shown in FIG. 5;
third step, vacuum filling of the glass needle tip nano holes
Sealing the interface between the holder 101 and the centrifuge tube 102 by using a sealing film, placing the sealing film on a centrifuge tube 102 frame, placing the centrifuge tube 102 frame into a vacuum dryer, extracting vacuum, observing that the solution 104 in the glass tube 103 overflows upwards into a liquid containing groove 204 on the holder 101 in the process of removing bubbles in vacuum, standing for 30min for filling after the vacuum reaches 1.01 atm, slowly opening the vacuum dryer, refluxing the solution 104 in the liquid containing groove 204, and completing filling of the needle tip nano holes, wherein the needle tip microscopic view of the glass tube 103 after the filling of the solution 104 by a vacuum method is shown in fig. 6.
Example 2
Example 2 differs from example 1 in that the solution 104 was replaced in the first step, the quartz capillary having a gauge of 1.0mm in outside diameter and 0.5mm in inside diameter. The glass tip nano Kong Jian filled with the solution 104 had a diameter of about 25nM, and its tip was fixed on the holder 101 with the tip upper portion 203 not exceeding the bottom 201 of the liquid container 204, the holder 101 was inserted into a mating 1.5ml microcentrifuge tube 102, and the tip was immersed in the electrolyte solution 104 (1 nM. Lambda. DNA,1mol/L LiCl, pH=8) containing the DNA sample.
Thirdly, sealing the interface between the holder 101 and the centrifuge tube 102 by using a sealing film, then placing the sealed interface on the centrifuge tube 102 frame into a vacuum dryer, vacuumizing, observing the upward overflow amount of the solution 104 in the glass tube 103 in the process of removing bubbles in vacuum, allowing the overflowed solution 104 to enter a liquid containing groove 204 on the holder 101, standing for 1h after the vacuum degree reaches 1.01 atm for filling, and slowly opening the vacuum dryer to finish the filling of the solution 104 with the needle tip nano holes.
Finally, it is noted that the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications and equivalents may be made thereto without departing from the spirit and scope of the present invention, which is intended to be covered by the claims of the present invention.
Claims (2)
1. The application method of the glass needle point nano hole filling device based on the vacuum method is characterized in that the filling device comprises a centrifuge tube and a clamp holder clamped on the centrifuge tube in a sealing way, a liquid containing groove is integrally formed at the top of the clamp holder, the clamp holder is vertically through, and a glass tube with a needle point nano hole structure is inserted into a through hole in the middle of the clamp holder;
the clamp comprises a bottom, a gasket, an upper part and a liquid containing groove from bottom to top in sequence, wherein a through hole is formed in the middle of the clamp, a groove is formed in the middle of the bottom, an external thread is formed in the outer wall of the bottom, an annular groove is formed in the upper part, an internal thread is formed in the inner wall of the annular groove, the liquid containing groove is formed in the outer side of the upper part of the clamp, and the gasket is clamped between the bottom and the upper part by matching the internal thread and the external thread; the bottom of the clamp is just inserted into the opening of the centrifuge tube to form a sealed clamp;
the using method comprises the following steps:
A. placing a gasket in a groove at the bottom of the clamp holder, mutually biting and fixing the bottom and the upper part through threads, and extruding and fixing the gasket to a glass tube penetrating through the clamp holder;
B. adding a required solution into the centrifuge tube through the tail part of the glass tube by adopting a microinjector, and plugging the glass tube into the centrifuge tube by adopting the clamp holder, wherein the liquid level in the centrifuge tube needs to be beyond the neck part of the needle point of the glass tube;
C. placing the assembled filling device on a centrifugal machine, and prefilling by a centrifugal method;
D. sealing the joint of the clamp holder and the centrifuge tube by using a sealing film after centrifugation is completed, placing the joint on a centrifuge tube rack, placing the centrifuge tube rack into a vacuum dryer, standing for more than 10 minutes under a vacuum state after vacuum extraction for filling, wherein a liquid containing groove on the clamp holder is used for containing solution overflowed from a glass tube in the process of removing bubbles in vacuum, opening the vacuum dryer after filling, and refluxing the solution in the liquid containing groove into the glass tube to finish filling of the needle tip nano holes;
wherein, the top end of the glass tube in the step A does not exceed the top of the liquid containing tank.
2. The method of claim 1, wherein the vacuum level in step D is greater than 0.98 atmospheres.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011041671A1 (en) * | 2009-10-02 | 2011-04-07 | Simpore, Inc. | Methods for facilitating fluid flow through nanoporous membranes |
| JP2012016875A (en) * | 2010-07-07 | 2012-01-26 | National Institute Of Advanced Industrial Science & Technology | Transparent incombustible material and method for manufacturing the same |
| CN104034833A (en) * | 2014-06-30 | 2014-09-10 | 厦门大学 | Preparing device and method for nanofiller capillary chromatography microcolumn |
| CN104195527A (en) * | 2014-09-16 | 2014-12-10 | 山东省科学院激光研究所 | Device and technology for plating surface of glass capillary tube with carbon film |
| CN106602153A (en) * | 2016-12-27 | 2017-04-26 | 中科泰能高铭科技发展有限公司 | Electrolyte injection method |
| CN209018801U (en) * | 2018-08-01 | 2019-06-25 | 上海市肺科医院 | A kind of new medical Exhaled breath condensate carbon dioxide depassing unit |
| CN110669654A (en) * | 2019-09-26 | 2020-01-10 | 苏州科技大学 | Preparation method of methane microbial sensor |
-
2020
- 2020-09-24 CN CN202011017897.0A patent/CN112146956B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011041671A1 (en) * | 2009-10-02 | 2011-04-07 | Simpore, Inc. | Methods for facilitating fluid flow through nanoporous membranes |
| JP2012016875A (en) * | 2010-07-07 | 2012-01-26 | National Institute Of Advanced Industrial Science & Technology | Transparent incombustible material and method for manufacturing the same |
| CN104034833A (en) * | 2014-06-30 | 2014-09-10 | 厦门大学 | Preparing device and method for nanofiller capillary chromatography microcolumn |
| CN104195527A (en) * | 2014-09-16 | 2014-12-10 | 山东省科学院激光研究所 | Device and technology for plating surface of glass capillary tube with carbon film |
| CN106602153A (en) * | 2016-12-27 | 2017-04-26 | 中科泰能高铭科技发展有限公司 | Electrolyte injection method |
| CN209018801U (en) * | 2018-08-01 | 2019-06-25 | 上海市肺科医院 | A kind of new medical Exhaled breath condensate carbon dioxide depassing unit |
| CN110669654A (en) * | 2019-09-26 | 2020-01-10 | 苏州科技大学 | Preparation method of methane microbial sensor |
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