CN105505772A - Pyrolysis device for single exosomes - Google Patents
Pyrolysis device for single exosomes Download PDFInfo
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- CN105505772A CN105505772A CN201510889504.8A CN201510889504A CN105505772A CN 105505772 A CN105505772 A CN 105505772A CN 201510889504 A CN201510889504 A CN 201510889504A CN 105505772 A CN105505772 A CN 105505772A
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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Abstract
The invention relates to a pyrolysis device for single exosomes. The pyrolysis device comprises a pyrolysis chamber and a pyrolysis circuit, wherein the pyrolysis chamber adopts a feed-through tubular structure and comprises a contraction end and a broad end which are mutually opposite, and a nano-pore is formed in the end part of the contraction end; the pyrolysis circuit comprises a first electrode, a second electrode and a power supply; the first electrode is located at the broad end, and the second electrode is located at the outer side of the nano-pore; a first coating is arranged on the orifice surface of the nano-pore and comprises the following components in parts by weight: 100 parts of silicon oxide, 40-42 parts of titanium oxide, 10-12 parts of scandium oxide, 10-12 parts of silver oxide and 3-5 parts of ytterbium oxide. The pyrolysis device utilizes strong voltage difference to pyrolyze exosomes, together with the nano-pore structure, ensures that only single exosomes can be pyrolyzed by the nano-pore, and therefore provides a more accurate detection target for follow-up miRNA detection. As the coating on the surface of the nano-pore and a coating of a filter screen are improved, the exosome filtration efficiency and the efficiency of the exosomes in entering the nano-pore are improved.
Description
Technical field
The present invention relates to a kind of outer cracking apparatus secreting body, particularly a kind ofly secrete the device that body carries out cracking outward to single.
Background technology
Recent years, the vesicles secreting body (Exosome) outside one is called just is being subject to everybody and is paying close attention to widely.Secreting body is outward that diameter is about 30-150nm, and density is at the vesicles of 1.13-1.21g/m1.Secrete that body is natural to be present in body fluid outward, comprise blood, saliva, urine and breast milk, secrete the film vesica deriving from late endosome (also referred to as many vesicas body) that body is viable cell secretion outward.Secrete outward body before 30 years find by people.Early stage research is thought, secrete body outward and perform albumen transportation function, specific target determine recipient cell, exchanger and lipid or initiation down-stream signaling events.Until 2007, secrete body outside researchist finds and also transport nucleic acid, participate in cell-cell communication.In a word, its albumen, RNA and fatty ingredient are special, and carry some important signaling molecules, are expected to play a role in the early diagnosis of various diseases, and this secretes the market of body also at Quick Extended outside making, and become popular research object.Current research finds, secretes outward containing protein rRNA and microRNA relevant to cell derived in body, and secretes body outward and by biological barrier, at intercellular trafficking functional nucleic acid, thus can play various biological function.
In the prior art, mainly outside a large amount of, secrete the whole cracking of body for the research and detection means of secreting RNA in body outward, then detect the RNA in biased sample: specifically secrete body with outside the cracking of chemical cracking liquid, then mixture is performed to the extraction of RNA.But a problem can be there is in this, it is different outer that to secrete the RNA albumen that body carries be different, the cellularity of its correspondence is also different, more distracter is often there is in the outer RNA secreted in body adopting traditional method to obtain, therefore how more careful obtain the single outer RNA secreted in body more accurately, what just seem very is meaningful.
Summary of the invention
For the deficiency that prior art exists, the object of the present invention is to provide a kind ofly how to obtain the single outer device secreting RNA or miRNA in body, namely how to realize secreting the device that body carries out cracking outward to single.
For achieving the above object, the present invention is achieved through the following technical solutions:
Outside single, secrete the device that body carries out cracking, comprise:
Cracking room, it is through tubular structure, includes the serrated end and open end relatively set up, and the end of described serrated end is provided with the nanoporous that aperture is 60 ~ 100nm;
Cracking circuit, it comprises the first electrode, the second electrode and power supply; Described first electrode is positioned at the open end of described cracking room, and described second electrode is positioned at the outside of described nanoporous;
Wherein, the vent surface of described nanoporous is provided with the first coating, and described first coating includes the material of following weight part:
Preferably, described secretes the device that body carries out cracking outside single, and wherein, the outside of described nanoporous is also provided with porous ceramic layer, and described porous ceramic layer is between described nanoporous and the second electrode.
Preferably, described secretes the device that body carries out cracking outside single, and wherein, the aperture of described porous ceramic layer is 50 ~ 100nm.
Preferably, described secretes the device that body carries out cracking outside single, and wherein, the outside of described nanoporous is also provided with filter screen, and described filter screen is between described porous ceramic layer and the second electrode.
Preferably, described secretes the device that body carries out cracking outside single, and wherein, the aperture of described filter screen is 60 ~ 100nm.
Preferably, described secretes the device that body carries out cracking outside single, and wherein, described power supply is AC/DC integrated power supply.
Preferably, described secretes the device that body carries out cracking outside single, and wherein, the thickness of described first coating is 5 ~ 10nm.
Preferably, described secretes the device that body carries out cracking outside single, and wherein, described filter screen is coated with the second coating, and described second coating includes the material of following weight part:
Preferably, described secretes the device that body carries out cracking outside single, and wherein, the thickness of described second coating is 5 ~ 10nm.
Preferably, described secretes the device that body carries out cracking outside single, and wherein, described first coating also comprises the Z 250 of 2 ~ 3 weight parts.
To the invention has the beneficial effects as follows: this case utilizes forceful electric power pressure reduction externally to secrete body to carry out cracking, and the structure in combining nano hole, make outer secrete body can only be single cleaved by nanoporous, detect target more accurately for follow-up miRNA detects to provide.By the improvement to nanoporous surface coating and filter screen coating, improve the outer efficiency of secreting body and filtering and entering nanoporous.
Accompanying drawing explanation
Fig. 1 secretes the structural representation that body carries out the device of cracking outside single.
Fig. 2 is in embodiment 1, when power supply is direct supply, and the graphic representation of electric current and time.
Embodiment
Below in conjunction with accompanying drawing, the present invention is described in further detail, can implement according to this with reference to specification sheets word to make those skilled in the art.
Embodiment 1
As shown in Figure 1, what an embodiment was listed in this case outer secretes the device that body carries out cracking to single, comprising:
Cracking room 1, it is through tubular structure, includes the serrated end 2 and open end 3 relatively set up, and the end of serrated end 2 is provided with the nanoporous 4 that aperture is 60 ~ 100nm;
Cracking circuit, it comprises the first electrode 5, second electrode 6 and power supply 7; Open end 3, second electrode 6 that first electrode 5 is positioned at cracking room is positioned at the outside of nanoporous 4;
Wherein, the vent surface of nanoporous 4 is provided with the first coating, and the first coating includes the material of following weight part:
Embodiment 2
As another embodiment of this case, wherein, the outside of nanoporous 4 is also provided with porous ceramic layer 8, and this porous ceramic layer 8 is between nanoporous 4 and the second electrode 6.
In the above-described embodiments, the aperture of porous ceramic layer 8 is preferably 50 ~ 100nm.
Embodiment 3
As the another embodiment of this case, wherein, the outside of nanoporous 4 is also provided with filter screen 9, and this filter screen 9 is between porous ceramic layer 8 and the second electrode 6.
In the above-described embodiments, the aperture of filter screen 9 is preferably 60 ~ 100nm.
Power supply 7 is preferably AC/DC integrated power supply.This power supply belongs to prior art, be commercially available prod, its inside is integrated with rectifier filer and invertor, and its benefit can freely control to switch AC and DC electric current, just select AC-powered pattern when needs alternating-current, just select powered by direct current pattern when needs direct current.
The thickness of the first coating is preferably 5 ~ 10nm.
The working process of secreting the cracker of body is outward: cracker is placed in solution, inject in solution near nanoporous 4 and treat that the outer of cracking secretes body, very slow owing to secreting body velocity of diffusion in the solution outward, therefore, it can not enter into cracking room 1 from open end 3; Now, power supply 7 is adjusted to direct supply, electrically charged owing to secreting body itself outward, therefore it can be drawn in nanoporous 4, and limit due to the aperture of nanoporous 4, once can only allow singlely outer secrete body and enter, when the outside of nanoporous 4 is provided with porous ceramic layer 8 and filter screen 9, secretes body outward and can first enter in nanoporous 4 again through after these 2 layers of filtering layers under the driving of electric field; And such as, as how judging that whether secrete body outward has entered in cracking room 1, then having multiple method to judge, can carry out fluorescent decoration outward by secreting body, mark by fluorescence, the outer track secreting body can be observed easily; When secreting after body enters nanoporous 4 outward, power supply 7 being adjusted to alternating-current, utilizing alternating voltage near nanoporous, produce forceful electric power pressure reduction, body cracking being secreted outward.Outer body of secreting after cracking can continue to stay in cracking room 1, carries out the detection of follow-up miRNA.Galvanic voltage range is within the scope of positive and negative 10V, and the voltage range of alternating-current is in positive and negative 1V, and frequency is 50 ~ 100Hz.
But, secrete body itself outward and there is certain viscosity, although the pore size of nanoporous and its volume size match, but wanting to allow outward, secreting body enters nanoporous also non-easy thing smoothly, secrete body outward when through nanoporous, can because the factor such as electrostatic or physical adsorption viscous be near nanoporous, even result in blockage time serious nanoporous, therefore need to arrange a coating in the vent surface of nanoporous, can successfully rhythmically " come into " nanoporous in make secreting body outward, secretes the heating rate of body, the persistence of cracking and splits stability of solution outside improving.
The formula of the first coating is an inseparable organic whole, its effect is the antistatic property improving nanoporous, reduce its skin friction force, can smoothly enter nanoporous in make secreting body outward, it has the integrity of height, even change the content of certain composition in formula or change the kind of certain composition, all will make the degradation of this formula.Therefore, in formula, the kind of each composition and content should be limited.
Embodiment 4
As the another embodiment of this case, wherein, the surface of filter screen 9 is coated with the second coating, and this second coating includes the material of following weight part:
In the above-described embodiments, the thickness of the second coating is preferably 5 ~ 10nm.
The effect of the second coating prevents filter screen from blocking, because the aperture of filter screen is very little, general coating construction thickness is difficult to accomplish very thin, only have the thickness adopting the coating of special formulation can realize 5 ~ 10nm, and possess excellent hardness, extremely low skin friction coefficient and corrosion resistance nature; Second coating must with zinc oxide for main material, its light weight, careful, and good biocompatibility, not easily with biomolecules generation viscous, therefore not easily cause the blocking of filter screen, ensure that the high-efficiency continuous strainability of filter screen.Second coating is also an inseparable organic whole, and it has the integrity of height, if change the content of certain composition in formula or change the kind of certain composition, all will make the degradation of this formula.Therefore, in formula, the kind of each composition and content should be limited.
Embodiment 5
As the another embodiment of this case, wherein, the first coating also comprises the Z 250 of 2 ~ 3 weight parts.We know, the shape of secreting body is outward irregularity, and himself is with electric charge, under the electric field, when secreting body outward through the first coating containing Z 250, Z 250 can produce a faint induction field, this induction field can make chargedly outer secrete body and spin, the benefit of spin to make outer body of secreting find best angle to enter into nanoporous fast, such as, secreting body outside can making creeps in nanoporous from the most elongated one end, thus secrete the smooth degree that body enters nanoporous outside improve, improve the outer lysis efficiency secreting body.But it should be noted that the content of Z 250 must be limited, the scope exceeding setting cannot produce best induction field by causing, thus can have a negative impact on the contrary, outside making, secrete the efficiency reduction that body enters nanoporous.
Fig. 2 is in embodiment 1, and when power supply 7 is direct supply, the graphic representation of electric current and time, when secreting body outward through nanoporous for one, electric current can jump and produce a trough.As can be seen from Figure 2, the interval of trough is uneven, this show outer secrete that body enters continuously uneven.In the current versus time curve figure that embodiment 4 obtains, the interval of trough becomes comparatively even, and in the current versus time curve figure obtained in embodiment 5, the interval of trough just becomes very even.What this illustrated that the improvement of coating externally secretes body enters improved efficiency successful.
Although embodiment of the present invention are open as above, but it is not restricted to listed in specification sheets and embodiment utilization, it can be applied to various applicable the field of the invention completely, for those skilled in the art, can easily realize other amendment, therefore do not deviating under the universal that claim and equivalency range limit, the present invention is not limited to specific details and illustrates here and the legend described.
Claims (10)
1. outside single, secrete the device that body carries out cracking, it is characterized in that, comprising:
Cracking room, it is through tubular structure, includes the serrated end and open end relatively set up, and the end of described serrated end is provided with the nanoporous that aperture is 60 ~ 100nm;
Cracking circuit, it comprises the first electrode, the second electrode and power supply; Described first electrode is positioned at the open end of described cracking room, and described second electrode is positioned at the outside of described nanoporous;
Wherein, the vent surface of described nanoporous is provided with the first coating, and described first coating includes the material of following weight part:
2. outside single, secrete the device that body carries out cracking as claimed in claim 1, it is characterized in that, the outside of described nanoporous is also provided with porous ceramic layer, and described porous ceramic layer is between described nanoporous and the second electrode.
3. outside single, secrete the device that body carries out cracking as claimed in claim 2, it is characterized in that, the aperture of described porous ceramic layer is 50 ~ 100nm.
4. outside single, secrete the device that body carries out cracking as claimed in claim 2, it is characterized in that, the outside of described nanoporous is also provided with filter screen, and described filter screen is between described porous ceramic layer and the second electrode.
5. outside single, secrete the device that body carries out cracking as claimed in claim 4, it is characterized in that, the aperture of described filter screen is 60 ~ 100nm.
6. outside single, secrete the device that body carries out cracking as claimed in claim 1, it is characterized in that, described power supply is AC/DC integrated power supply.
7. outside single, secrete the device that body carries out cracking as claimed in claim 1, it is characterized in that, the thickness of described first coating is 5 ~ 10nm.
8. outside single, secrete the device that body carries out cracking as claimed in claim 4, it is characterized in that, described filter screen is coated with the second coating, and described second coating includes the material of following weight part:
9. outside single, secrete the device that body carries out cracking as claimed in claim 8, it is characterized in that, the thickness of described second coating is 5 ~ 10nm.
10. outside single, secrete the device that body carries out cracking as claimed in claim 1, it is characterized in that, described first coating also comprises the Z 250 of 2 ~ 3 weight parts.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510889504.8A CN105505772B (en) | 2015-12-07 | 2015-12-07 | The device cracked to single excretion body |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510889504.8A CN105505772B (en) | 2015-12-07 | 2015-12-07 | The device cracked to single excretion body |
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| CN105505772A true CN105505772A (en) | 2016-04-20 |
| CN105505772B CN105505772B (en) | 2018-07-10 |
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| CN201510889504.8A Active CN105505772B (en) | 2015-12-07 | 2015-12-07 | The device cracked to single excretion body |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109628277A (en) * | 2019-01-23 | 2019-04-16 | 东南大学 | The separation of excretion in-vivo tumour mark miRNA a kind of and detection system and method |
| CN111971560A (en) * | 2017-12-01 | 2020-11-20 | 康奈尔大学 | Nanoparticles and different exosome subsets for detecting and treating cancer |
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| EP1726958A2 (en) * | 1996-04-25 | 2006-11-29 | BioArray Solutions Ltd. | Light-controlled electrokinetic assembly of particles near surfaces |
| CN101460253A (en) * | 2006-05-31 | 2009-06-17 | 明达西德实验室有限公司 | Method and apparatus for manipulating single cells and small aggregates thereof |
| US20110152107A1 (en) * | 2005-04-29 | 2011-06-23 | University Of Rochester | Drug Screening Via Nanpore Silicon Filters |
| CN102905534A (en) * | 2010-04-06 | 2013-01-30 | 约翰·W·霍拉迪 | Methods of treating cancer |
| CN104583420A (en) * | 2012-04-16 | 2015-04-29 | 生物动力学公司 | Nucleic acid sample preparation |
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2015
- 2015-12-07 CN CN201510889504.8A patent/CN105505772B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| EP1726958A2 (en) * | 1996-04-25 | 2006-11-29 | BioArray Solutions Ltd. | Light-controlled electrokinetic assembly of particles near surfaces |
| US20110152107A1 (en) * | 2005-04-29 | 2011-06-23 | University Of Rochester | Drug Screening Via Nanpore Silicon Filters |
| CN101460253A (en) * | 2006-05-31 | 2009-06-17 | 明达西德实验室有限公司 | Method and apparatus for manipulating single cells and small aggregates thereof |
| CN102905534A (en) * | 2010-04-06 | 2013-01-30 | 约翰·W·霍拉迪 | Methods of treating cancer |
| CN104583420A (en) * | 2012-04-16 | 2015-04-29 | 生物动力学公司 | Nucleic acid sample preparation |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111971560A (en) * | 2017-12-01 | 2020-11-20 | 康奈尔大学 | Nanoparticles and different exosome subsets for detecting and treating cancer |
| CN111971560B (en) * | 2017-12-01 | 2024-06-11 | 康奈尔大学 | Nanoparticles and different exosome subsets for detection and treatment of cancer |
| CN109628277A (en) * | 2019-01-23 | 2019-04-16 | 东南大学 | The separation of excretion in-vivo tumour mark miRNA a kind of and detection system and method |
| CN109628277B (en) * | 2019-01-23 | 2022-02-01 | 东南大学 | System and method for separating and detecting tumor marker miRNA in exosome |
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| CN105505772B (en) | 2018-07-10 |
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