CN108126522A - The method of target particles in separating chips, separator and separation liquid sample - Google Patents
The method of target particles in separating chips, separator and separation liquid sample Download PDFInfo
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- CN108126522A CN108126522A CN201711396703.0A CN201711396703A CN108126522A CN 108126522 A CN108126522 A CN 108126522A CN 201711396703 A CN201711396703 A CN 201711396703A CN 108126522 A CN108126522 A CN 108126522A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
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Abstract
The present invention provides a kind of separating chips, including：Sample pool, the sample pool include first side and second side, and the first side is opposite with the second side, and the first side is provided with the first filter membrane, and the second side is provided with the second filter membrane；First chamber, the first chamber are connected with the sample pool by first filter membrane, and the first chamber is provided with the first opening, and described first is open that the first chamber is made to be in communication with the outside；Second chamber, the second chamber are connected with the sample pool by second filter membrane, and the second chamber is provided with the second opening, and described second is open that the second chamber is made to be in communication with the outside.The present invention also provides a kind of separators and method for detaching target particles in liquid sample.
Description
Technical field
The present invention relates to biotechnologies, and in particular, to a kind of point for being used to detach target particles in liquid sample
Off-chip piece, separator and separation method.
Background technology
Liquid biopsy (liquid biopsy or fluid biopsy) is that one kind can reflect that tumour is thin comprehensively, in real time
The sampling of born of the same parents or tissue biological's information and analysis method.Liquid biopsy has the advantages that Noninvasive, by separating, analyzing blood
Or specific research object carries out dynamic observation to tumour in other body fluid (urine, saliva, pleural effusion, cerebrospinal fluid etc.), it can
Medical staff to be instructed to carry out screening, diagnosis, judging prognosis, selection therapeutic scheme and monitoring recurrence etc. to tumour.Liquid biopsy
In particular studies object include Circulating tumor DNA (circulating tumor DNA, ctDNA), circulating tumor cell
(circulating tumor cell, CTC), microvesicle (also known as excretion body, exosome) etc..
It is general using following in separation, purifying blood or body fluid the methods of centrifugation, immunocapture or filtering in the prior art
Ring tumour cell and/or excretion body.The separation method of centrifugation can cause one to the membrane structure of circulating tumor cell (or excretion body)
Determine the mechanical damage of degree, influence subsequent analysis and research, and centrifuge the affected cumbersome flux to liquid biopsy and cause to limit.Exempt from
The separation method of epidemic disease capture needs, using antibody, to increase substantially sample process cost, and the elution requirement after immunocapture can
The activity of circulating tumor cell (or excretion body) can be had an impact.It can be effectively using the separation method for filtering membrane filtration
Various sizes of component in body fluid is detached, there is low cost, high throughput, the target component obtained after filtering can be kept very
High bioactivity.The constituent part being easy to but in actual mechanical process, on filter membrane in enriched biological sample, wherein
Component more than membrane pore size can block fenestra.Pore Blocking can prevent the component less than membrane pore size from effectively through filtering
Film influences the purity of target component after separation.It is excessive that Pore Blocking will also result in local pressure, even results in filter membrane rupture.
Invention content
In view of the foregoing, it is necessary to provide it is a kind of can reduce be separated by filtration during filter membrane plug-hole phenomenon occurs
Separating chips, separator and separation method.
Present invention firstly provides a kind of separating chips, including：
Sample pool, the sample pool include first side and second side, the first side and the second side phase
Right, the first side is provided with the first filter membrane, and the second side is provided with the second filter membrane；
First chamber, the first chamber are connected with the sample pool by first filter membrane, first chamber
Room is provided with the first opening, and described first is open that the first chamber is made to be in communication with the outside；
Second chamber, the second chamber are connected with the sample pool by second filter membrane, second chamber
Room is provided with the second opening, and described second is open that the second chamber is made to be in communication with the outside.
Further, first filter membrane is respectively selected from porous ceramic film material, porous plastics with second filter membrane
One kind in material, porous metal material.
Optionally, the aperture of first filter membrane and second filter membrane is 2-20 microns.
Optionally, the aperture of first filter membrane and second filter membrane is 5-200 nanometers.
Further, when the first chamber is open by described first by swabbing action, in the first chamber
Generate negative pressure；When the second chamber is open by described second by swabbing action, negative pressure is generated in the second chamber.
Optionally, the sample pool, the first chamber, the second chamber are by polymethyl methacrylate materials system
Into.
The present invention also provides a kind of separator, including：Above-mentioned separating chips；Vacuum system, the vacuum system with
First opening of the separating chips and the described second opening are respectively connected with；Frequency-variable module, the frequency-variable module and institute
State the effect of vacuum system electrical connection, the first chamber and the second chamber in the frequency-variable module and the vacuum module
Under be alternately produced negative pressure.
Optionally, the vacuum system include the first vacuum pump and the second vacuum pump, first vacuum pump with described point
First opening of off-chip piece is connected, and second vacuum pump is connected with the second opening of the separating chips, the frequency conversion
Module is arranged to that first vacuum pump and second vacuum pump is controlled to work alternatively.
Further, the separator further includes：Liquid supplying unit, for the sample of the separating chips
Pond automatically provides liquid sample；Sample collection unit, for automatically collecting the liquid after separation from the sample pool
Sample.
The present invention also provides a kind of methods for detaching target particles in liquid sample, include the following steps：
Above-mentioned separating chips are provided；
Liquid sample is provided to the sample pool；
Aspirating the first chamber by the described first opening makes the first chamber generate negative pressure；
Stop aspirating the first chamber；
Aspirating the second chamber by the described second opening makes the second chamber generate negative pressure；
Stop aspirating the second chamber.
Compared with the prior art, separating chips provided by the present invention pass through in checker first chamber and second chamber
Negative pressure can control the direction of Liquid Flow in separating chips, and liquid sample to be separated is allow more effectively to penetrate filter membrane,
Filtering fenestra during being separated by filtration is reduced simultaneously to be blocked the generation of phenomenon.Separator provided by the present invention passes through vacuum system
It unites and alternately provides negative pressure, and vacuum system is controlled to realize by frequency-variable module for the first chamber and second chamber of separating chips
Controllably, the lock out operation of automation.Separating chips provided by the present invention, separator can be used in liquid biopsy procedure
And the separation and Extraction that separation method is quick, high-throughput to progress such as circulating tumor cell, excretion bodies, mitigate the work of experimenter
Amount reduces the testing cost of liquid biopsy.
Description of the drawings
Fig. 1 is the structure diagram of separating chips provided by the present invention.
Fig. 2 is that the structure of one embodiment of separating chips provided by the present invention disassembles schematic diagram.
Fig. 3 is the high-level schematic functional block diagram of separator provided by the present invention.
Fig. 4 is the fluid path schematic diagram of one embodiment of separator provided by the present invention.
Fig. 5 be using separation method provided by the present invention liquid sample before separation after absorbance curve compare
Figure.
Main element symbol description
Following specific embodiment will be further illustrated the present invention with reference to above-mentioned attached drawing.
Specific embodiment
Technical scheme of the present invention is described below in conjunction with the preferred embodiment of the present invention and embodiment.It needs
Illustrate, when a unit is described as " connecting " in another unit, it can be directly to another unit or
Person may be simultaneously present unit placed in the middle.When a unit is described as " being set to " another unit, it can directly be set
Put on another unit or may be simultaneously present unit placed in the middle.Unless otherwise defined, all technologies used herein
It is had the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs with scientific terminology.In the specification of the present invention
Used in element or the title of equipment be intended merely to the purpose of description specific embodiment, it is not intended that in limiting this hair
It is bright.
Present invention firstly provides a kind of separating chips, the separating chips are using filter membrane to various sizes of in liquid sample
Particle is detached, to obtain the target particles of specific dimensions.Fig. 1 is the structural representation of separating chips provided by the present invention
Figure.As shown in Figure 1, separating chips 10 include sample pool 13, first chamber 15 and second chamber 17.
The sample pool 13 includes first side 132 and second side 134, the first side 132 and 134 phase of second side
It is right.The first filter membrane 14 is provided in the first side 132, the second side 134 is provided with the second filter membrane 16.This
One chamber 15 is connected with the sample pool 13 by first filter membrane 14.The first chamber 15 is provided with the first opening 152, should
First opening 152 is used to that the first chamber 15 to be made to be in communication with the outside.The second chamber 17 passes through second mistake with the sample pool 13
Filter membrane 16 is connected, which is provided with the second opening 172, this second opening 172 for make the second chamber 17 with
External world's connection.It is understood that the first chamber 15 and the second chamber 17 are located at the opposite both sides of the sample pool 13 respectively.
During using the separating chips 10, liquid sample is added in into sample pool 13, by first opening 152 and second opening
172 are connected respectively with air-extractor.When air-extractor is aspirated the first chamber 15 by first opening 152,
Negative pressure is generated in the first chamber 15.Under the suction function of the first chamber 15, size in the liquid sample in sample pool 13
Ingredient more than the pore size filter of the first filter membrane 14 flows into the first chamber 15 via the first filter membrane 14.When air-extractor leads to
When crossing second opening 172 is aspirated the second chamber 17, negative pressure is generated in the second chamber 17.In the second chamber 17
Suction function under, size is more than the ingredient of the pore size filter of the second filter membrane 16 via the in the liquid sample in sample pool 13
Two filter membranes 16 flow into the second chamber 17.
Alternately and repeatedly make to generate negative pressure in the first chamber 15 and the second chamber 17, can effectively make liquid sample anti-
It reestablishes diplomatic relations and alternately flows through the first filter membrane 14 and the second filter membrane 16, size in liquid sample is made to be more than the first filter membrane 14 and second
The ingredient in 16 aperture of filter membrane is stayed in sample pool 13.The structure design of the separating chips 10 makes to be adsorbed in 14 He of the first filter membrane
The ingredient on 16 surface of the second filter membrane is easy to come off from filter membrane surface in negative pressure variation alternately and repeatedly, can be effectively prevented
The fenestra of filter membrane is blocked.
The sample pool 13 of the separating chips 10, first chamber 15, the main part of second chamber 17 can be by plastics, glass
Glass, metal or composite material are made.In one embodiment, the sample pool 13 of the separating chips 10, first chamber 15, second chamber
17 main part is made of polymethyl methacrylate (PMMA) material.
First filter membrane 14 and second filter membrane 16 can be made of identical membrane material, can also be by different films
Material is made.First filter membrane 14 and second filter membrane 16 can have identical average filtration membrane aperture and/or aperture
Distribution, it is possible to have different average filtration membrane apertures and/or pore-size distribution.First filter membrane 14 (or second filtering
Film 16) it can be formed made of a kind of membrane material or by multiple film Material cladding.First filter membrane, 14 He
Second filter membrane 16 can be porous material, include but are not limited to porous ceramic film material, porous plastic materials and porous gold
Belong to material.Specifically, first filter membrane 14 and second filter membrane 16 can be respectively selected from anodic alumina films, acetate fiber
One or more of film, polyethylene film, polypropylene screen and polystyrene film.
In one embodiment, the aperture of first filter membrane 14 and second filter membrane 16 is between 2-20 microns；Compared with
Goodly, between 5-10 microns.In one embodiment, the aperture of first filter membrane 14 and second filter membrane 16 is 8 microns,
It can be used for the circulating tumor cell in separated plasma sample.
In another embodiment, the aperture of first filter membrane 14 and second filter membrane 16 is between 5-200 nanometers；
Preferably, between 10-100 nanometers.In one embodiment, the aperture of first filter membrane 14 and second filter membrane 16 is 20
Nanometer can be used for separation by the excretion body in the plasma sample of 200 nano-filtration membranes.
It is understood that when first filter membrane 14 and 16 surface of the second filter membrane are not further embellished, it should
Separating chips 10 mainly include according only to the various composition in the aperture screening liquid sample of filter membrane after separation in the sample of gained
Target particles (such as circulating tumor cell, excretion body), it is also possible to containing other with close or large-size particle.Ability
Field technique personnel are appreciated that the absorption to reduce porous material to albumen or gene in fluid to be measured sample, first mistake
The surface of filter membrane 14 and second filter membrane 16 can be modified by sulphation；Specifically to detach target particles, this first
The surface of filter membrane 14 and second filter membrane 16 can be modified by specific biological macromolecular, which divides greatly
Son can be specific one or more of antibody, antigen, polypeptide or base sequence.It should be pointed out that mesh described herein
Mark particle can have the cell of biological significance or component or other kinds of particle, such as liposome of synthesis,
Nanoparticle, nanoparticle etc..
It is understood that the volume of the sample pool 13 can be designed according to practical application scene.For biological biopsy
Application scenarios, the volume of the sample pool 13 can be between 0.1-10 milliliters, optionally, between 0.5-2 milliliters.Implement one
In example, the volume of the sample pool 13 is 1 milliliter.The sample pool 13 can include sample pool opening 138, for adding in and/or taking
Go out liquid sample.
In the embodiment shown in figure 1, which includes the first side wall opposite with first filter membrane 14
156, which is arranged on the first side wall 156；The second chamber 17 includes and 16 phase of the second filter membrane
To second sidewall 176, which 172 is arranged in the second sidewall 176.In the present embodiment, the separation
Chip 10 has symmetrical structure.It should be noted that separating chips 10 can also be dissymmetrical structure or other it is any can be real
The structure of existing present inventive concept.
Fig. 2 shows the decomposition diagrams of an embodiment of separating chips provided by the present invention.As shown in Fig. 2, separation
First opening 152 of chip 10 is the circular hole for being opened in the first side wall 156, and the second opening 172 of the separating chips 10 is opens up
In the circular hole of second sidewall 176.In the present embodiment, the first side wall 156 and the second sidewall 176 are two panels PMMA cover plates.
First chamber 15 is formed by the PMMA cover plates of the first side wall 156 with another PMMA substrates heating pressing.The first chamber 15
PMMA substrates on offer circular hole, which is sealed by the first filter membrane 14 from the outside of the first chamber 15.It is appreciated that
Ground, second chamber 17 are formed by the PMMA cover plates of the second sidewall 176 with the heating pressing of a piece of PMMA substrates.The second chamber 17
PMMA substrates on also offer circular hole, which is sealed by the second filter membrane 16 from the outside of the second chamber 17.By this
The PMMA cover plates of one chamber 15, a U-shaped substrate, the second chamber 17 PMMA cover boards fit together to form sample pool successively
13.Based on above method, the separating chips for meeting spirit of the present invention can be made with relatively low cost, this separating chips have
One sample pool, the first chamber connected respectively with the sample pool by filter membrane and second chamber and being set to is set respectively
Two openings being in communication with the outside of first chamber and second chamber are placed in, so as to divide in the both direction of separating chips
It is not evacuated, optimization is separated by filtration effect.
In Fig. 2 the embodiment described, optionally, the length of the separating chips 10 is about 30 millimeters, and width is about 23 millis
Rice, thickness is about 6 millimeters.The Circularhole diameter of first opening 152 and second opening 172 is about 1 millimeter.The first chamber 15
With to be used to set the Circularhole diameter of filter membrane on the two panels PMMA substrates of the second chamber 17 be about 13 millimeters.The separating chips 10
Sample pool volume be about 1 milliliter.
Invention further provides a kind of separators.Fig. 3 shows the high-level schematic functional block diagram of the separator.It should
Separator includes separating chips 10, vacuum system 20, frequency-variable module 30, controller 80 and power module as described above
90。
The vacuum system 20 is used for the first chamber 15 for making the separating chips 10 respectively and second chamber 17 generates negative pressure.It should
Vacuum system 20 can be two independent vacuum systems or a vacuum system by designing.The vacuum system
It can also include the equipment such as minipump or micro-suction pump.It is understood that the vacuum system 20 and the separating chips
It can be connected between 10 by the preferable pipeline of air-tightness.
The frequency-variable module 30 is electrically connected with the vacuum system 20, which is also electrically connected with the power module 90,
The supply voltage of the vacuum module is supplied to so as to control the frequency-variable module 30, so as to make 15 and second chamber of first chamber
Room 17 is alternately produced negative pressure.It is understood that the frequency-variable module 30 can include two frequency converters, for controlling this point respectively
Negative pressure variation in the first chamber 15 and second chamber 17 of off-chip piece 10；The frequency-variable module 30 can also only include a frequency conversion
Device and specific switching circuit, to realize similar technique effect.
The controller 80 is electrically connected with the frequency-variable module 30, the power module 90.The controller 80 can be embedded in firmly
Part or logical relation set on firmware (firmware) or a series of it is stored in storage with what programming language was write
Program in device or other firmwares.The controller 80 can control the working method of the frequency-variable module 30, so as to make vacuum system
20 automatically alternately aspirate the first chamber 15 of the separating chips 10 and second chamber 17, detach target in liquid sample
Grain.
In an embodiment of separator provided by the present invention, which includes the first vacuum pump 210
With the second vacuum pump 220, which is connected with the first opening 152 of the separating chips 10, second vacuum pump
220 are connected with the second opening 172 of the separating chips 10.The frequency-variable module 30 includes the first frequency converter and the second frequency converter,
First frequency converter is electrically connected with the first vacuum pump 210, which is electrically connected with the second vacuum pump 220.The two become
Frequency device can make the first vacuum pump 210 and the second vacuum pump 220 alternately and repeatedly work under the control of controller 80.It for example, should
First Frequency Converter Control, first vacuum pump 210 is run, and makes to generate negative pressure in first chamber 15 by 152 pumping of the first opening；
Then, first Frequency Converter Control, first vacuum pump 210 is out of service；Later, second Frequency Converter Control, second vacuum
220 operation of pump makes to generate negative pressure in second chamber 17 by 172 pumping of the second opening；Followed by, which should
Second vacuum pump 220 is out of service；It after above-mentioned steps are multiple repeatedly, controls the vacuum system 20 out of service, collects separation
Liquid sample afterwards.
Fig. 4 shows the fluid path schematic diagram of the separator.It will be understood by those skilled in the art that the separator may be used also
To further comprise liquid supplying unit 50 and liquid collecting unit 60, so as to fulfill the automation of target particles in liquid sample
Separation process.
The liquid supplying unit 50 is used to automatically inject liquid into the sample pool 13 of separating chips 10.In Fig. 4 institutes
In the embodiment shown, which includes sample to be tested room 510, cleaning liquid chamber 530 and control valve 550.The control
Valve 550 processed can be fluid path converter, include but are not limited to solenoid valve, rotary valve.The control valve 550 is connected into sample to be tested
Room 510 and sample pool 13, the sample pool 13 that the liquid sample in sample to be tested room 510 can be provided to separating chips 10 are used for
Detach target particles；The control valve 550 can also be connected into cleaning liquid chamber 530 and sample by changing the setting of the control valve 550
The cleaning solution cleaned in liquid chamber 530 is provided to the sample pool 13 of separating chips 10 for cleaning separating chips 10 by this pond 13.It can
With understanding, which can also include a power part, such as kinetic pump or aspiration pump, be provided for liquid stream dynamic
Power.In certain embodiments, which may not also include power part, be made by the suction of vacuum system 20
With realization feed liquor.
The liquid collecting unit 60 is for automatically from the liquid-like after the collection separation of the sample pool 13 of separating chips 10
This.In one embodiment, which includes sampling needle, and controller 80 controls sampling needle mobile and stretches into separating core
The liquid sample after separation is drawn in the sample pool 13 of piece 10.
Further, which can also include the first liquid locker room 410 and second liquid locker room 420.It should
First liquid locker room 410 is arranged between first vacuum pump 210 and the first opening 152 of separating chips 10, this first
First chamber 15 of the liquid locker room 410 respectively with the first vacuum pump 210 and separating chips 10 is connected.Similarly, this second
Liquid locker room 420 is arranged between second vacuum pump 220 and the second opening 172 of separating chips 10, the second liquid
Second chamber 17 of the locker room 420 respectively with the second vacuum pump 220 and separating chips 10 is connected.First liquid locker room 410
Safety flack can be used as with second liquid locker room 420, the liquid in separating chips 10 is avoided to enter vacuum pump, can also be used as
Waste liquid bottle remains in liquid or cleaning solution in separating chips 10 after collecting separation every time.
Automatically the target particles in liquid sample can be detached using separator provided by the present invention,
It will can not be separated by the component of filter membrane in sample pool, while changed by the negative pressure variation in the cavity of sample pool both sides
Liquid Flow direction in sample pool reduces the component for being adhered to filter membrane surface, avoids filter membrane quilt during being separated by filtration
What is blocked happens.The separator cost is relatively low, easy to use, significantly reduces the workload of experimenter.
The present invention further provides a kind of methods for detaching target particles in liquid sample, include the following steps：
Step S200 provides separating chips 10 of the present invention；
Step S220 provides liquid sample into the sample pool 13 of the separating chips 10；
Step S230 by the 152 suction first chamber 15 of the first opening of the separating chips 10, makes to produce in first chamber 15
Raw negative pressure；
Step S250 stops suction first chamber 15；
Step S270 makes to produce in second chamber 17 by the 172 suction second chamber 17 of the second opening of the separating chips 10
Raw negative pressure；
Step S290 stops suction second chamber 17.
Wherein, step S230-S290 is recycled repeatedly, to realize better separating effect.It is understood that above-mentioned point
Method from target particles in liquid sample can be manual, semi-automatic or full-automatic realization.When the liquid sample
During for cleaning solution (such as buffer solution), separating chips 10 can be cleaned by the step S220-S290 in this method.
Step S220-S290 is described in detail below in conjunction with the embodiment of separator shown in Fig. 4.
In step S220, liquid sample (or cleaning solution) is added in sample pool 13.Optionally, which can pass through
The liquid supplying unit 50 of separator performs, and liquid sample can also be added in sample pool either manually or by liquid-transfering gun or syringe
13。
Before step S230, by the first of separating chips 10 the 152, second opening 172 of opening respectively with vacuum system (such as
Two vacuum pumps) it is connected.
In step S230, vacuum system 20 makes to produce in first chamber 15 by 152 suction first chamber 15 of the first opening
Raw negative pressure.Liquid in liquid sample (such as blood sample, body fluid sample) and size in sample pool 13 are less than the first filtering
The component in 14 aperture of film under suction function by the first filter membrane 14, into first chamber 15.In some cases, such as first
The volume of chamber 15 is relatively small, and also or, the negative pressure variation in first chamber 15 is too fast, and liquid and size are less than the first filtering
The component in 14 aperture of film may also be further by 152 outflow of the first opening, into the first liquid locker room 410.
In step s 250, vacuum system 20 stops suction first chamber 15.
In step S270, vacuum system 20 makes to produce in second chamber 17 by 172 suction second chamber 17 of the second opening
Raw negative pressure.In the implementation procedure of step S270, the component for being adhered to 14 surface of the first filter membrane can be with air-flow and/or liquid
It flows in sample pool 13, the liquid in liquid sample (such as blood sample, body fluid sample) and size in sample pool 13 are less than the
The component in two filter membranes, 16 aperture under suction function by the second filter membrane 16, into second chamber 17.In some cases,
Volume such as second chamber 17 is relatively small, and also or, the negative pressure variation in second chamber 17 is too fast, and liquid and size are less than the
The component in two filter membranes, 16 aperture may also be further by 172 outflow of the second opening, into second liquid locker room 420.It can
To understand ground, step S250 and step S270 can be performed successively, can also be performed simultaneously successively.
In step S290, vacuum system 20 stops suction second chamber 17.
Later, step S230-S290 repeated filtration liquid samples can be repeated several times.
For example, performing step S230 again after step S290,152 suction first of the first opening is passed through by vacuum system 20
Chamber 15, in the process, the component for being adhered to 16 surface of the second filter membrane can be with air-flow and/or liquid stream sample pool 13
In, and liquid in the liquid sample (such as blood sample, body fluid sample) in sample pool 13 and size are less than the first filter membrane
The component in 14 apertures under suction function by the first filter membrane 14, into first chamber 15.Then step S250 is performed again,
Vacuum system 20 stops suction first chamber 15.And step S270 is performed, vacuum system 20 passes through 172 suction second of the second opening
Chamber 17.
Can be removed the component in liquid sample less than filter membrane aperture by performing step S230-S290 repeatedly,
Component more than filtering membrane aperture is trapped in sample pool 13.Changed by the alternating of negative pressure in first chamber, second chamber
The permeability of filter membrane in filter process can be improved, reduce filter membrane plug-hole, improve filter effect.
It is understood that in step S220, cleaning solution can also be added in sample pool 13, by performing step repeatedly
Rapid S230-S290 cleans separating chips 10.It can be buffer solution in the cleaning solution, can also be containing specific raw
The solution of object bioactive molecule (such as nuclease, protease), for remove 10 each Adsorption on Surface of separating chips biology into
Point.
Excretion body can be detached from blood sample using separating chips provided by the present invention and separation method.In Fig. 5
Curve a be blood sample before separation in wavelength be the absorbance curve between 220-340 nanometers, wherein, the blood sample
For obtained liquid sample after the membrane filtration by 200 nano apertures.Curve b in Fig. 5 is that the blood sample after separation exists
Wavelength is the absorbance curve between 220-340 nanometers.Absorbance curve a is compared with b it is found that the sample after detaching is in wavelength
Absorbance near 280 nanometers is substantially reduced, and illustrates that the separating chips and separation method can be effectively in liquid samples
Component is detached and is collected.
Above-described embodiment is the preferable embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, embodiment of above are only for interpreting the claims.Right protection scope of the present invention is not limited to specification.Appoint
What those familiar with the art is in the technical scope of present disclosure, the variation or replacement that can readily occur in,
It is included within protection scope of the present invention.
Claims (10)
1. a kind of separating chips, which is characterized in that including：
Sample pool, the sample pool include first side and second side, and the first side is opposite with the second side, institute
It states first side and is provided with the first filter membrane, the second side is provided with the second filter membrane；
First chamber, the first chamber are connected with the sample pool by first filter membrane, and the first chamber is set
The first opening is equipped with, described first is open that the first chamber is made to be in communication with the outside；
Second chamber, the second chamber are connected with the sample pool by second filter membrane, and the second chamber is set
The second opening is equipped with, described second is open that the second chamber is made to be in communication with the outside.
2. separating chips as described in claim 1, which is characterized in that first filter membrane is distinguished with second filter membrane
One kind in porous ceramic film material, porous plastic materials, porous metal material.
3. separating chips as claimed in claim 2, which is characterized in that the hole of first filter membrane and second filter membrane
Diameter is 2-20 microns.
4. separating chips as claimed in claim 2, which is characterized in that the hole of first filter membrane and second filter membrane
Diameter is 5-200 nanometers.
5. separating chips as described in claim 1, which is characterized in that when the first chamber by described first opening by
During swabbing action, negative pressure is generated in the first chamber；When the second chamber by the described second opening by swabbing action
When, negative pressure is generated in the second chamber.
6. separating chips as claimed in claim 5, which is characterized in that the sample pool, the first chamber, second chamber
Room is made of polymethyl methacrylate materials.
7. a kind of separator, including：
Separating chips as described in any one of claim 1-6；
Vacuum system, the vacuum system are respectively connected with first opening of the separating chips and second opening
It connects；
Frequency-variable module, the frequency-variable module are electrically connected with the vacuum system, and the first chamber and the second chamber are in institute
It states and is alternately produced negative pressure under the action of frequency-variable module and the vacuum module.
8. separator as claimed in claim 7, which is characterized in that the vacuum system includes the first vacuum pump and second very
Sky pump, first vacuum pump are connected with the first opening of the separating chips, second vacuum pump and the separating core
Second opening of piece is connected, and the frequency-variable module is arranged to that first vacuum pump and second vacuum pump is controlled to replace
Work.
9. separator as claimed in claim 7, which is characterized in that further comprise：
Liquid supplying unit, for automatically providing liquid sample to the sample pool of the separating chips；
Sample collection unit, for automatically collecting the liquid sample after separation from the sample pool.
10. a kind of method for detaching target particles in liquid sample, includes the following steps：
Separating chips as described in any one of claim 1-6 are provided；
Liquid sample is provided to the sample pool；
Aspirating the first chamber by the described first opening makes the first chamber generate negative pressure；
Stop aspirating the first chamber；
Aspirating the second chamber by the described second opening makes the second chamber generate negative pressure；
Stop aspirating the second chamber.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711396703.0A CN108126522B (en) | 2017-12-21 | 2017-12-21 | Separation chip, separation device and method for separating target particles in liquid sample |
| US16/203,652 US10758867B2 (en) | 2017-11-30 | 2018-11-29 | Isolation device and isolation method |
| US16/203,649 US10758866B2 (en) | 2017-11-30 | 2018-11-29 | Isolation chip and manufacturing method of the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711396703.0A CN108126522B (en) | 2017-12-21 | 2017-12-21 | Separation chip, separation device and method for separating target particles in liquid sample |
Publications (2)
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