CN105505772B - The device cracked to single excretion body - Google Patents
The device cracked to single excretion body Download PDFInfo
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- CN105505772B CN105505772B CN201510889504.8A CN201510889504A CN105505772B CN 105505772 B CN105505772 B CN 105505772B CN 201510889504 A CN201510889504 A CN 201510889504A CN 105505772 B CN105505772 B CN 105505772B
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
This case is related to the device cracked to single excretion body, including：Cracking room for perforation tubular structure, includes the opposite serrated end set up and open end, and the end of serrated end is equipped with nano-pore；Circuit is cracked, including first electrode, second electrode and power supply；First electrode is located at open end, and second electrode is located at the outside of nano-pore；The vent surface of nano-pore is equipped with first coating, and first coating includes：100 parts by weight of silica；40~42 parts by weight of titanium oxide；10~12 parts by weight of scandium oxide；10~12 parts by weight of silver oxide；3~5 parts by weight of ytterbium oxide.This case is externally secreted body using forceful electric power pressure difference and is cracked, and combines the structure of nano-pore so that excretion body single can only be cleaved by nano-pore, and more accurate detection target is provided for subsequent miRNA detections.By the improvement to nanoporous surface coating and strainer coating, improve the filtering of excretion body and enter the efficiency of nano-pore.
Description
Technical field
The present invention relates to a kind of cracking apparatus of excretion body, more particularly to a kind of dress cracked to single excretion body
It puts.
Background technology
Recent years, the vesicles that one kind is called excretion body (Exosome) are just widely paid close attention to by everybody.Excretion body is
Diameter is about 30-150nm, and density is in the vesicles of 1.13-1.21g/m1.Excretion body is naturally occurring in body fluid, including blood,
Saliva, urine and breast milk, excretion body are the film property capsules from late endosome (also referred to as more vesica bodies) of living cells secretion
Bubble.Excretion body was found before 30 years by people.The research of early stage thinks that excretion body performs albumen transportation function, specifically targets
Recipient cell, exchanger and lipid cause down-stream signaling events.Until 2007, researcher had found that excretion body also transports
Nucleic acid participates in cell-cell communication.In short, its albumen, RNA and fat constituent are special, and carry some important signaling molecules,
It is expected to play a role in the early diagnosis of a variety of diseases, this causes the market of excretion body also in Quick Extended, and as hot topic
Research object.It has now been found that excretion body in contain with the relevant protein rRNA and microRNA of cell derived, and
Excretion body can be by biological barrier, in intercellular trafficking functional nucleic acid, so as to play various biological functions.
In the prior art, the research for RNA in excretion body and detection means mainly all split a large amount of excretion bodies
Solution, then detection mix the RNA in sample：Excretion body specifically is cracked with chemical cracking liquid, RNA's then is performed to mixture
Extraction.But this can have a problem that, the RNA albumen that different excretion bodies carry is different, and corresponding cellularity is not yet
Together, using conventional method obtain excretion body in RNA in often there are more distracter, therefore how more it is careful more accurately
The RNA in single excretion body is obtained, just seems very significant.
Invention content
In view of the deficiencies of the prior art, the present invention intends to provide a kind of how to obtain in single excretion body
How the device of RNA or miRNA realizes the device cracked to single excretion body.
To achieve the above object, the invention is realized by the following technical scheme：
A kind of device cracked to single excretion body, including：
Cracking room for perforation tubular structure, includes the opposite serrated end set up and open end, the end of the serrated end
Portion is equipped with the nano-pore that aperture is 60~100nm；
Circuit is cracked, including first electrode, second electrode and power supply；The first electrode is located at the cracking room
Open end, the second electrode are located at the outside of the nano-pore；
Wherein, the vent surface of the nano-pore is equipped with first coating, and the first coating includes following parts by weight
Material：
Preferably, the device cracked to single excretion body, wherein, it is additionally provided on the outside of the nano-pore
Porous ceramic layer, the porous ceramic layer is between the nano-pore and second electrode.
Preferably, the device cracked to single excretion body, wherein, the aperture of the porous ceramic layer is
50~100nm.
Preferably, the device cracked to single excretion body, wherein, it is additionally provided on the outside of the nano-pore
Strainer, the strainer is between the porous ceramic layer and second electrode.
Preferably, the device cracked to single excretion body, wherein, the aperture of the strainer for 60~
100nm。
Preferably, the device cracked to single excretion body, wherein, the power supply is AC/DC integrated
Power supply.
Preferably, the device cracked to single excretion body, wherein, the thickness of the first coating is 5
~10nm.
Preferably, the device cracked to single excretion body, wherein, the strainer is coated with second coating,
The second coating includes the material of following parts by weight：
Preferably, the device cracked to single excretion body, wherein, the thickness of the second coating is 5
~10nm.
Preferably, the device cracked to single excretion body, wherein, the first coating further includes 2~3
The ferroso-ferric oxide of parts by weight.
The beneficial effects of the invention are as follows：This case is externally secreted body using forceful electric power pressure difference and is cracked, and combines the knot of nano-pore
Structure so that excretion body single can only be cleaved by nano-pore, and more accurate detection mesh is provided for subsequent miRNA detections
Mark.By the improvement to nanoporous surface coating and strainer coating, improve the filtering of excretion body and enter the efficiency of nano-pore.
Description of the drawings
Fig. 1 is the structure diagram of device cracked to single excretion body.
Fig. 2 be embodiment 1 in, when power supply be DC power supply when, the curve graph of electric current and time.
Specific embodiment
The present invention is described in further detail below in conjunction with the accompanying drawings, to enable those skilled in the art with reference to specification text
Word can be implemented according to this.
Embodiment 1
As shown in Figure 1, the device cracked to single excretion body of an embodiment is listed in this case, including：
Cracking room 1 for perforation tubular structure, includes the opposite serrated end 2 set up and open end 3, the end of serrated end 2
Portion is equipped with the nano-pore 4 that aperture is 60~100nm；
Circuit is cracked, including first electrode 5, second electrode 6 and power supply 7；First electrode 5 is located at the open of cracking room
End 3, second electrode 6 is located at the outside of nano-pore 4；
Wherein, the vent surface of nano-pore 4 is equipped with first coating, and first coating includes the material of following parts by weight：
Embodiment 2
As another embodiment of this case, wherein, the outside of nano-pore 4 is additionally provided with porous ceramic layer 8, the porous ceramic layer 8
Between nano-pore 4 and 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 nano-pore 4 is additionally provided with strainer 9, which is located at porous ceramics
Between layer 8 and second electrode 6.
In the above-described embodiments, the aperture of strainer 9 is preferably 60~100nm.
Power supply 7 is preferably AC/DC integrated power supply.The power supply belongs to the prior art, is commercial product, its inside collection
Into rectifier filter and inverter, its benefit is can freely to control switching AC and DC electric current, when needing alternating current
When just select AC power supply pattern, just select powered by direct current pattern when needing direct current.
The thickness of first coating is preferably 5~10nm.
The course of work of the cracker of excretion body is：Cracker is placed in solution, the solution near nano-pore 4
It is middle to inject excretion body to be cracked, since the diffusion velocity of excretion body in the solution is very slow, it will not from open end 3 into
Enter into cracking room 1；At this point, power supply 7 is adjusted to DC power supply, since excretion body itself is electrically charged, it can be drawn into
Enter in nano-pore 4, and since the aperture of nano-pore 4 limits, single excretion body can only once be allowed to enter, when outside nano-pore 4
When side is equipped with porous ceramic layer 8 and strainer 9, excretion body enters back into after this 2 layers of filter layers can be initially passed through under the driving of electric field
In nano-pore 4；And as to how judge whether excretion body is had been introduced into cracking room 1, then it may determine that there are many method, example
Such as can excretion body be subjected to fluorescent decoration, be marked by fluorescence, can easily observe the track of excretion body；When excretion body into
After entering nano-pore 4, power supply 7 is adjusted to alternating current, forceful electric power pressure difference is generated near nano-pore using alternating voltage, by excretion body
Cracking.Excretion body after cracking can continue to stay in cracking room 1, carry out the detection of subsequent miRNA.The voltage model of direct current
It is trapped among in the range of positive and negative 10V, for the voltage range of alternating current in positive and negative 1V, frequency is 50~100Hz.
But excretion body has certain viscosity in itself, although the pore size of nano-pore and its volume size phase
Match, but want excretion body is allowed to smoothly enter nano-pore nor easy thing, excretion body can be inhaled when across nano-pore because of electrostatic or physics
The factors such as attached are viscous near nano-pore, and even result in blockage nano-pore when serious, it is therefore desirable in the vent surface of nano-pore
One coating is set, so that excretion body can successfully rhythmically " be come into " in nano-pore, improves the cracking speed of excretion body
The stability of rate, the duration of cracking and cracking.
The formula of first coating is an inseparable organic whole, its effect is to improve the antistatic property of nano-pore
Can, its skin-friction force is reduced, so that excretion body can smoothly enter in nano-pore, it has high integrity, i.e.,
If changing the content of some ingredient in formula or changing the type of some ingredient, the hydraulic performance decline that will all make the formula.Therefore, match
The type and content of each ingredient should be limited in side.
Embodiment 4
As the another embodiment of this case, wherein, the surface of strainer 9 is coated with second coating, which includes following
The material of parts by weight：
In the above-described embodiments, the thickness of second coating is preferably 5~10nm.
The effect of second coating is to prevent strainer from blocking, because of the aperture very little of strainer, general coating construction is thick
Degree is difficult to accomplish very thin, the thickness of 5~10nm can be only realized using the coating of special formulation, and be also equipped with excellent
Hardness, extremely low skin-friction coefficient and corrosion resistance；Second coating must be using zinc oxide as main material, it is light, careful, and raw
Object compatibility is good, is not easy to occur with biomolecule viscous, therefore does not easily cause the blocking of strainer, ensure that the high-efficiency continuous of strainer
Strainability.Second coating is also an inseparable organic whole, it has high integrity, if changing in formula
The content of some ingredient or the type for changing some ingredient, the hydraulic performance decline that will all make the formula.Therefore, each ingredient in formula
Type and content should be limited.
Embodiment 5
As the another embodiment of this case, wherein, first coating further includes the ferroso-ferric oxide of 2~3 parts by weight.We know
Road, the shape of excretion body is irregular, and its own is with charge, under the electric field, when excretion body is passed through containing four oxidations three
During the first coating of iron, ferroso-ferric oxide can generate a faint induction field, which can cause electrically charged outer
It secretes body to spin, the benefit of spin is that excretion body can be made to be quickly found out best angle to enter in nano-pore, such as can
So that excretion body, so as to improve the smooth degree that excretion body enters nano-pore, improves out of most elongated one end drilling nano-pore
The lysis efficiency of excretion body.But it should be noted that the content of ferroso-ferric oxide must be limited, the range beyond setting will be led
Cause can not generate best induction field, so as to have a negative impact instead, reduce the efficiency that excretion body enters nano-pore.
Fig. 2 be embodiment 1 in, when power supply 7 be DC power supply when, the curve graph of electric current and time, when an excretion body is worn
When crossing nano-pore, electric current, which can jump, generates a trough.From figure 2 it can be seen that the interval of trough is uneven, this shows
Excretion body enters uneven continuous.In the current versus time curve figure obtained in embodiment 4, the interval of trough becomes more
It is even, and in the current versus time curve figure obtained in embodiment 5, the interval of trough just becomes visibly homogeneous.This illustrates coating
It improves with obvious effects into improved efficiency to excretion body.
Although the embodiments of the present invention have been disclosed as above, but its be not restricted in specification and embodiment it is listed
With it can be fully applied to various fields suitable for the present invention, for those skilled in the art, can be easily
Realize other modification, therefore without departing from the general concept defined in the claims and the equivalent scope, it is of the invention and unlimited
In specific details and legend shown and described herein.
Claims (7)
1. a kind of device cracked to single excretion body, which is characterized in that including：
Cracking room for perforation tubular structure, includes the opposite serrated end set up and open end, the end of the serrated end is set
There is the nano-pore that aperture is 60~100nm；
Circuit is cracked, including first electrode, second electrode and power supply；The first electrode is located at the open of the cracking room
End, the second electrode are located at the outside of the nano-pore；
Wherein, the vent surface of the nano-pore is equipped with first coating, and the first coating includes the material of following parts by weight：
Be additionally provided with porous ceramic layer on the outside of the nano-pore, the porous ceramic layer be located at the nano-pore and second electrode it
Between；
Strainer is additionally provided on the outside of the nano-pore, the strainer is between the porous ceramic layer and second electrode；
The strainer is coated with second coating, and the second coating includes the material of following parts by weight：
2. the device cracked as described in claim 1 to single excretion body, which is characterized in that the porous ceramic layer
Aperture is 50~100nm.
3. the device cracked as described in claim 1 to single excretion body, which is characterized in that the aperture of the strainer is
60~100nm.
4. the device cracked as described in claim 1 to single excretion body, which is characterized in that the power supply is alternating current-direct current
Integrated power supply.
5. the device cracked as described in claim 1 to single excretion body, which is characterized in that the thickness of the first coating
It spends for 5~10nm.
6. the device cracked as described in claim 1 to single excretion body, which is characterized in that the thickness of the second coating
It spends for 5~10nm.
7. the device cracked as described in claim 1 to single excretion body, which is characterized in that the first coating is also wrapped
Include the ferroso-ferric oxide of 2~3 parts by weight.
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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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|---|---|---|---|
| CN201510889504.8A CN105505772B (en) | 2015-12-07 | 2015-12-07 | The device cracked to single excretion body |
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| CN105505772B true CN105505772B (en) | 2018-07-10 |
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| US20210285952A1 (en) * | 2017-12-01 | 2021-09-16 | Cornell University | Nanoparticles and distinct exosome subsets for detection and treatment of cancer |
| 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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| EP1726959A3 (en) * | 1996-04-25 | 2007-07-11 | BioArray Solutions Ltd. | Light-controlled electrokinetic Assembly of particles near surfaces |
| US8501668B2 (en) * | 2005-04-29 | 2013-08-06 | University Of Rochester | Drug screening via nanopore silicon filters |
| ITMI20061063A1 (en) * | 2006-05-31 | 2007-12-01 | Mindseeds Lab S R L | METRODO AND PE SYSTEM RLA SELECTION AND MODIFICATION OF SINGLE CELLS AND THEIR SMALL AGGREGATES |
| EP3238782A3 (en) * | 2010-04-06 | 2018-01-03 | ExoCyte Therapeutics Pte, LTD | Methods of treating cancer |
| KR20150014925A (en) * | 2012-04-16 | 2015-02-09 | 바이오로지컬 다이나믹스, 인크. | Nucleic acid sample preparation |
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