CN108126522B - Separation chip, separation device and method for separating target particles in liquid sample - Google Patents
Separation chip, separation device and method for separating target particles in liquid sample Download PDFInfo
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Abstract
The present invention provides a separation chip, which comprises: a cuvette comprising a first side and a second side, the first side being opposite the second side, the first side being provided with a first filter membrane, the second side being provided with a second filter membrane; the first chamber is communicated with the sample pool through the first filtering membrane, and is provided with a first opening used for communicating the first chamber with the outside; and the second chamber is communicated with the sample pool through the second filtering membrane, and is provided with a second opening which is used for communicating the second chamber with the outside. The invention also provides a separation device and a method for separating target particles in the liquid sample.
Description
Technical Field
The invention relates to the field of biotechnology, in particular to a separation chip, a separation device and a separation method for separating target particles in a liquid sample.
Background
Liquid biopsy (liquid biopsy) is a sampling and analysis method that can reflect biological information of tumor cells or tissues comprehensively and in real time. The liquid biopsy has the advantage of non-invasiveness, and can guide medical workers to screen, diagnose, judge prognosis, select a treatment scheme, monitor relapse and the like of tumors by separating and analyzing specific study subjects in blood or other body fluids (urine, saliva, pleural effusion, cerebrospinal fluid and the like) to dynamically observe the tumors. Specific study objects in the liquid biopsy include circulating tumor DNA (ctDNA), Circulating Tumor Cells (CTC), microvesicles (also called exosomes), and the like.
In the prior art, methods such as centrifugation, immunocapture or filtration are generally adopted to separate and purify circulating tumor cells and/or exosomes in blood or body fluid. The centrifugal separation method can cause mechanical damage to the membrane structure of circulating tumor cells (or exosomes) to a certain extent, so that the subsequent analysis and research are influenced, and the complicated centrifugal operation limits the flux of liquid biopsy. The separation method of immunocapture needs to utilize antibody, greatly increases the sample processing cost, and the elution condition after immunocapture may affect the activity of circulating tumor cells (or exosomes). The separation method utilizing the filtration membrane filtration can effectively separate components with different sizes in body fluid, has the characteristics of low cost and high flux, and the target component obtained after filtration can keep high biological activity. However, in practical procedures, the filtration membrane tends to be enriched with a fraction of the components in the biological sample, wherein components larger than the pore size of the membrane tend to clog the pores of the membrane. The clogging of the membrane pores prevents components smaller than the size of the membrane pores from effectively permeating the filtration membrane, and affects the purity of the separated target component. The clogging of the membrane pores can also cause local overpressure and even rupture of the filter membrane.
Disclosure of Invention
In view of the above, it is desirable to provide a separation chip, a separation apparatus and a separation method that can reduce the occurrence of clogging of a filtration membrane during filtration and separation.
The present invention first provides a separation chip, which comprises:
a cuvette comprising a first side and a second side, the first side being opposite the second side, the first side being provided with a first filter membrane, the second side being provided with a second filter membrane;
the first chamber is communicated with the sample pool through the first filtering membrane, and is provided with a first opening used for communicating the first chamber with the outside;
and the second chamber is communicated with the sample pool through the second filtering membrane, and is provided with a second opening which is used for communicating the second chamber with the outside.
Further, the first filter membrane and the second filter membrane are respectively selected from one of porous ceramic materials, porous plastic materials and porous metal materials.
Optionally, the pore size of the first filter membrane and the second filter membrane is 2-20 microns.
Optionally, the pore size of the first filter membrane and the second filter membrane is 5-200 nm.
Further, when the first chamber is subjected to suction through the first opening, a negative pressure is generated in the first chamber; when the second chamber is subjected to suction through the second opening, a negative pressure is created in the second chamber.
Optionally, the sample cell, the first chamber, and the second chamber are made of a polymethylmethacrylate material.
The present invention also provides a separation device comprising: the above-mentioned separation chip; a vacuum system connected to the first opening and the second opening of the separation chip, respectively; the frequency conversion module is electrically connected with the vacuum system, and the first cavity and the second cavity alternately generate negative pressure under the action of the frequency conversion module and the vacuum module.
Optionally, the vacuum system includes a first vacuum pump and a second vacuum pump, the first vacuum pump is connected to the first opening of the separation chip, the second vacuum pump is connected to the second opening of the separation chip, and the frequency conversion module is configured to control the first vacuum pump and the second vacuum pump to alternately operate.
Further, the separation device further comprises: a liquid supply unit for automatically supplying a liquid sample to the sample cell of the separation chip; and the sample collection unit is used for automatically collecting the separated liquid sample from the sample pool.
The present invention also provides a method of separating target particles in a liquid sample, comprising the steps of:
providing the separation chip;
providing a liquid sample to the sample cell;
drawing the first chamber through the first opening to create a negative pressure in the first chamber;
stopping pumping the first chamber;
drawing the second chamber through the second opening to create a negative pressure in the second chamber;
stopping pumping the second chamber.
Compared with the prior art, the separation chip provided by the invention can control the gas-liquid flowing direction in the separation chip by alternately changing the negative pressure in the first cavity and the second cavity, so that a liquid sample to be separated can more effectively permeate the filter membrane, and the phenomenon that the filter membrane hole is blocked in the filtering and separating process is reduced. The separation device provided by the invention alternately provides negative pressure for the first cavity and the second cavity of the separation chip through the vacuum system, and the vacuum system is controlled by the frequency conversion module to realize controllable and automatic separation operation. The separation chip, the separation device and the separation method provided by the invention can be used for quickly and high-flux separation and extraction of circulating tumor cells, exosomes and the like in the liquid biopsy process, so that the workload of experimenters is reduced, and the detection cost of liquid biopsy is reduced.
Drawings
FIG. 1 is a schematic structural diagram of a discrete chip provided by the present invention.
Fig. 2 is a disassembled schematic view of a structure of an embodiment of the separation chip provided in the present invention.
Fig. 3 is a functional block diagram of the separation device provided by the present invention.
Fig. 4 is a schematic view of a liquid path of an embodiment of the separation device provided in the present invention.
FIG. 5 is a graph comparing absorbance curves before and after separation of a liquid sample using the separation method provided by the present invention.
Description of the main elements
The following detailed description will further illustrate the invention in conjunction with the above-described figures.
Detailed Description
The technical solution of the present invention will be described below with reference to the preferred embodiments and examples of the present invention. It will be understood that when an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. When an element is referred to as being "disposed on" another element, it can be directly on the other element or intervening elements may also be present. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The names of elements or devices used in the description of the present invention are for the purpose of describing particular embodiments only and are not intended to limit the present invention.
The invention firstly provides a separation chip which utilizes a filter membrane to separate particles with different sizes in a liquid sample so as to obtain target particles with specific sizes. FIG. 1 is a schematic structural diagram of a discrete chip provided by the present invention. As shown in fig. 1, the separation chip 10 includes a sample cell 13, a first chamber 15, and a second chamber 17.
The cuvette 13 comprises a first side 132 and a second side 134, the first side 132 being opposite the second side 134. The first filter membrane 14 is disposed on the first side 132, and the second filter membrane 16 is disposed on the second side 134. The first chamber 15 communicates with the sample cell 13 via the first filter membrane 14. The first chamber 15 is provided with a first opening 152, and the first opening 152 is used for communicating the first chamber 15 with the outside. The second chamber 17 is communicated with the sample cell 13 through the second filter membrane 16, and the second chamber 17 is provided with a second opening 172, wherein the second opening 172 is used for communicating the second chamber 17 with the outside. It will be appreciated that the first chamber 15 and the second chamber 17 are located on opposite sides of the sample cell 13.
In use of the separation chip 10, a liquid sample is added to the sample cell 13, and the first opening 152 and the second opening 172 are connected to a suction device, respectively. When the first chamber 15 is drawn through the first opening 152 by a suction device, a negative pressure is created in the first chamber 15. Under the negative pressure of the first chamber 15, components of the liquid sample in the cuvette 13 having a size smaller than the filter pore size of the first filter membrane 14 flow into the first chamber 15 via the first filter membrane 14. When the second chamber 17 is drawn through the second opening 172 by a suction device, a negative pressure is created in the second chamber 17. Under the negative pressure of the second chamber 17, the components with a size smaller than the filter pore size of the second filter membrane 16 in the liquid sample in the sample cell 13 flow into the second chamber 17 through the second filter membrane 16.
The repeated alternation of the first chamber 15 and the second chamber 17 to generate negative pressure is effective to cause the liquid sample to repeatedly and alternately flow through the first filtering membrane 14 and the second filtering membrane 16, so that components of the liquid sample having a size larger than the pore size of the first filtering membrane 14 and the second filtering membrane 16 are retained in the sample cell 13. The separation chip 10 is designed such that the components adsorbed on the surfaces of the first filtration membrane 14 and the second filtration membrane 16 are easily detached from the surfaces of the filtration membranes by repeated and alternate negative pressure changes, and the pores of the filtration membranes can be effectively prevented from being clogged.
The body portions of the sample cell 13, the first chamber 15, and the second chamber 17 of the separation chip 10 may be made of plastic, glass, metal, or composite material. In one embodiment, the sample cell 13, the first chamber 15, and the second chamber 17 of the separation chip 10 are made of polymethyl methacrylate (PMMA) material.
The first filter membrane 14 and the second filter membrane 16 may be made of the same membrane material or may be made of different membrane materials. The first filtration membrane 14 and the second filtration membrane 16 may have the same average filtration membrane pore size and/or pore size distribution, or may have different average filtration membrane pore sizes and/or pore size distributions. The first filter membrane 14 (or the second filter membrane 16) may be made of one membrane material, or may be made of a plurality of membrane materials. The first filter membrane 14 and the second filter membrane 16 may be porous materials including, but not limited to, porous ceramic materials, porous plastic materials, and porous metal materials. Specifically, the first filtering membrane 14 and the second filtering membrane 16 may be respectively selected from one or more of an anodic aluminum oxide membrane, a cellulose acetate membrane, a polyethylene membrane, a polypropylene membrane and a polystyrene membrane.
In one embodiment, the pore size of the first filter membrane 14 and the second filter membrane 16 is between 2-20 microns; preferably between 5-10 microns. In one embodiment, the first filter membrane 14 and the second filter membrane 16 have a pore size of 8 microns and can be used to separate circulating tumor cells from a plasma sample.
In another embodiment, the pore size of the first filter membrane 14 and the second filter membrane 16 is between 5 and 200 nanometers; preferably between 10-100 nm. In one embodiment, the first filter membrane 14 and the second filter membrane 16 have a pore size of 20 nm and can be used to separate exosomes from plasma samples that have passed through a 200 nm filter membrane.
It is understood that, when the surfaces of the first filtering membrane 14 and the second filtering membrane 16 are not further modified, the separation chip 10 only screens various components in the liquid sample according to the pore size of the filtering membranes, and the separated sample mainly includes target particles (such as circulating tumor cells, exosomes, etc.), and may also include other particles with similar or larger size. It will be appreciated by those skilled in the art that to reduce the adsorption of the porous material to the proteins or genes in the fluid sample being tested, the surfaces of the first filter membrane 14 and the second filter membrane 16 may be chemically modified; to specifically separate the target particles, the surfaces of the first filter membrane 14 and the second filter membrane 16 may be modified with specific biological macromolecules, which may be specific antibodies, antigens, polypeptides or base sequences. It is noted that the target particles described herein may be cells or components of biological interest, or may be other types of microparticles, such as synthetic liposomes, nanospheres, nanoparticles, and the like.
It is understood that the volume of the sample cell 13 can be designed according to the actual application scenario. For biopsy application scenarios, the volume of the sample reservoir 13 may be between 0.1-10 ml, alternatively between 0.5-2 ml. In one embodiment, the volume of the cuvette 13 is 1 ml. The cuvette 13 may comprise a cuvette opening 138 for adding and/or removing a liquid sample.
In the embodiment shown in fig. 1, the first chamber 15 comprises a first side wall 156 opposite the first filter membrane 14, the first opening 152 being provided on the first side wall 156; the second chamber 17 comprises a second side wall 176 opposite to said second filter membrane 16, the second opening 172 being provided in the second side wall 176. In the present embodiment, the discrete chip 10 has a symmetrical structure. It should be noted that the discrete chip 10 may also be an asymmetric structure or any other structure capable of implementing the inventive concept.
FIG. 2 is an exploded view of an embodiment of a discrete chip provided by the present invention. As shown in fig. 2, the first opening 152 of the discrete chip 10 is a circular hole opened on the first sidewall 156, and the second opening 172 of the discrete chip 10 is a circular hole opened on the second sidewall 176. In this embodiment, the first side wall 156 and the second side wall 176 are two sheets of PMMA cover sheets. The first chamber 15 is formed by heating and pressing the PMMA cover plate of the first side wall 156 and another PMMA substrate. The PMMA substrate of the first chamber 15 is provided with a circular hole, which is sealed by the first filter membrane 14 from the outside of the first chamber 15. It will be appreciated that the second chamber 17 is formed by the heated press-fit of the PMMA cover sheet and a sheet of PMMA base sheet of the second side wall 176. The PMMA substrate of the second chamber 17 is also provided with a circular hole which is sealed by a second filter membrane 16 from the outside of the second chamber 17. The PMMA cover plate of the first chamber 15, a U-shaped substrate and the PMMA cover plate of the second chamber 17 are sequentially attached together to form the sample cell 13. Based on the method, the separation chip according to the spirit of the invention can be prepared at lower cost, and the separation chip is provided with a sample cell, a first chamber and a second chamber which are respectively communicated with the sample cell through a filtering membrane, and two openings which are respectively arranged in the first chamber and the second chamber and are communicated with the outside, so that air can be respectively pumped in two directions of the separation chip, and the filtering separation effect is optimized.
In the embodiment illustrated in fig. 2, the discrete chip 10 may alternatively have a length of about 30 mm, a width of about 23 mm, and a thickness of about 6 mm. The first opening 152 and the second opening 172 have circular holes with a diameter of about 1 mm. The diameter of the circular holes for the filter membrane on the two PMMA substrates of the first chamber 15 and the second chamber 17 is about 13 mm. The sample cell volume of the separation chip 10 is about 1 ml.
The invention further provides a separation device. Fig. 3 shows a functional block diagram of the separating device. The separating apparatus includes the separating chip 10, the vacuum system 20, the frequency conversion module 30, the controller 80 and the power supply module 90 as described above.
The vacuum system 20 is used to generate negative pressure in the first chamber 15 and the second chamber 17 of the separation chip 10, respectively. The vacuum system 20 may be two separate vacuum systems or may be one vacuum system designed. The vacuum system may also include a micro vacuum pump or a micro suction pump. It is understood that the vacuum system 20 and the separation chip 10 may be connected by a pipe having a better air tightness.
The inverter module 30 is electrically connected to the vacuum system 20, and the inverter module 30 is also electrically connected to the power module 90, so that the inverter module 30 can control the power voltage supplied to the vacuum module, thereby alternately generating the negative pressure in the first chamber 15 and the second chamber 17. It is understood that the frequency conversion module 30 may include two frequency converters for controlling the variation of the negative pressure in the first chamber 15 and the second chamber 17 of the separation chip 10, respectively; the frequency conversion module 30 may also include only one frequency converter and a specific switching circuit to achieve similar technical effects.
The controller 80 is electrically connected to the frequency conversion module 30 and the power module 90. The controller 80 may be a collection of logical relationships embedded in hardware or firmware (firmware) or may be a series of programs written in a programming language stored in memory or other firmware. The controller 80 can control the operation of the frequency conversion module 30, so that the vacuum system 20 can automatically and alternately suck the first chamber 15 and the second chamber 17 of the separation chip 10 to separate target particles in the liquid sample.
In one embodiment of the detaching apparatus provided in the present invention, the vacuum system 20 includes a first vacuum pump 210 and a second vacuum pump 220, the first vacuum pump 210 is connected to the first opening 152 of the detaching chip 10, and the second vacuum pump 220 is connected to the second opening 172 of the detaching chip 10. The frequency conversion module 30 includes a first frequency converter electrically connected to the first vacuum pump 210 and a second frequency converter electrically connected to the second vacuum pump 220. The two frequency converters can repeatedly and alternately operate the first vacuum pump 210 and the second vacuum pump 220 under the control of the controller 80. For example, the first frequency converter controls the first vacuum pump 210 to operate, so as to pump air through the first opening 152 to generate a negative pressure in the first chamber 15; then, the first frequency converter controls the first vacuum pump 210 to stop running; then, the second frequency converter controls the second vacuum pump 220 to operate, so as to pump air through the second opening 172, so that negative pressure is generated in the second chamber 17; then, the second frequency converter controls the second vacuum pump 220 to stop running; after repeating the above steps for a plurality of times, the vacuum system 20 is controlled to stop running, and the separated liquid sample is collected.
Fig. 4 shows a schematic liquid path of the separation device. It will be appreciated by those skilled in the art that the separation device may further comprise a liquid supply unit 50 and a liquid collection unit 60 to achieve an automated separation process of target particles in the liquid sample.
The liquid supply unit 50 is used to automatically inject liquid into the sample cell 13 of the separation chip 10. In the embodiment shown in fig. 4, the liquid supply unit 50 includes a sample chamber 510 to be tested, a washing liquid chamber 530, and a control valve 550. The control valve 550 may be a fluid circuit switch, including but not limited to a solenoid valve, a rotary valve. The control valve 550 is connected to the sample chamber 510 to be tested and the sample cell 13, so that the liquid sample in the sample chamber 510 to be tested can be provided to the sample cell 13 of the separation chip 10 for separating the target particles; it is also possible to supply the washing liquid in the washing liquid chamber 530 to the sample cell 13 of the separation chip 10 for washing the separation chip 10 by changing the setting of the control valve 550 so that the control valve 550 communicates the washing liquid chamber 530 and the sample cell 13. It will be appreciated that the fluid supply unit 50 may also include a power component, such as a power pump or a suction pump, to provide power to the fluid flow. In some embodiments, the liquid supply unit 50 may not include a power component, and the liquid is supplied by the suction action of the vacuum system 20.
The liquid collection unit 60 is used for automatically collecting the separated liquid sample from the sample cell 13 of the separation chip 10. In one embodiment, the liquid collection unit 60 includes a sampling needle, and the controller 80 controls the sampling needle to move and extend into the sample cell 13 of the separation chip 10 to draw the separated liquid sample.
Further, the separation device may also include a first liquid storage chamber 410 and a second liquid storage chamber 420. The first liquid storage chamber 410 is disposed between the first vacuum pump 210 and the first opening 152 of the separation chip 10, and the first liquid storage chamber 410 is respectively communicated with the first vacuum pump 210 and the first chamber 15 of the separation chip 10. Similarly, the second liquid storage chamber 420 is disposed between the second vacuum pump 220 and the second opening 172 of the separation chip 10, and the second liquid storage chamber 420 is respectively communicated with the second vacuum pump 220 and the second chamber 17 of the separation chip 10. The first liquid storage chamber 410 and the second liquid storage chamber 420 can be used as safety bottles to prevent the liquid in the separation chip 10 from entering the vacuum pump, and can also be used as a waste liquid bottle to collect the liquid or cleaning liquid remaining in the separation chip 10 after each separation.
The separation device provided by the invention can automatically separate target particles in a liquid sample, components which cannot pass through the filtering membrane in the sample pool are separated, and meanwhile, the gas-liquid flow direction in the sample pool is changed through the negative pressure change in the cavities at two sides of the sample pool, so that the components adhered to the surface of the filtering membrane are reduced, and the situation that the filtering membrane is blocked in the filtering and separating process is avoided. The separating device has the advantages of low cost and convenient use, and greatly reduces the workload of experimenters.
The present invention further provides a method of separating target particles in a liquid sample, comprising the steps of:
step S200, providing the separation chip 10 of the invention;
step S220, providing a liquid sample to the sample cell 13 of the separation chip 10;
step S230, sucking the first chamber 15 through the first opening 152 of the separation chip 10 to generate a negative pressure in the first chamber 15;
step S250, stopping the suction of the first chamber 15;
step S270, sucking the second chamber 17 through the second opening 172 of the separation chip 10 to generate a negative pressure in the second chamber 17;
in step S290, the suction of the second chamber 17 is stopped.
Wherein, steps S230-S290 can be cycled for a plurality of times to achieve a better separation effect. It will be appreciated that the above-described method of separating target particles from a liquid sample may be performed manually, semi-automatically or fully automatically. When the liquid sample is a cleaning solution (e.g., a buffer solution), the separation chip 10 may be cleaned through steps S220-S290 in the method.
The steps S220-S290 will be described in detail below with reference to an embodiment of the separation device shown in fig. 4.
In step S220, a liquid sample (or a washing liquid) is added to the sample cell 13. Alternatively, this step may be performed by the liquid supply unit 50 of the separation device, or the liquid sample may be manually added to the cuvette 13 by a pipette or syringe.
Before step S230, the first opening 152 and the second opening 172 of the separation chip 10 are respectively connected to a vacuum system (e.g., two vacuum pumps).
In step S230, the vacuum system 20 sucks the first chamber 15 through the first opening 152, so as to generate a negative pressure in the first chamber 15. The liquid in the liquid sample (e.g., blood sample, body fluid sample, etc.) in the sample cell 13 and the components having a size smaller than the pore size of the first filter membrane 14 pass through the first filter membrane 14 under the negative pressure and enter the first chamber 15. In some cases, if the volume of the first chamber 15 is relatively small, or if the negative pressure in the first chamber 15 changes too rapidly, the liquid and components having a size smaller than the pore size of the first filter membrane 14 may further flow out through the first opening 152 into the first liquid storage chamber 410.
In step S250, the vacuum system 20 stops pumping the first chamber 15.
In step S270, the vacuum system 20 sucks the second chamber 17 through the second opening 172, so that a negative pressure is generated in the second chamber 17. During the execution of step S270, the components adhered to the surface of the first filtering membrane 14 can pass through the second filtering membrane 16 under the negative pressure in the sample cell 13 along with the air flow and/or the liquid flow, and the liquid in the liquid sample (such as blood sample, body fluid sample, etc.) in the sample cell 13 and the components with the size smaller than the pore size of the second filtering membrane 16 enter the second chamber 17. In some cases, if the volume of the second chamber 17 is relatively small, or if the negative pressure in the second chamber 17 changes too quickly, the liquid and the components with the size smaller than the pore size of the second filter membrane 16 may further flow out through the second opening 172 and enter the second liquid storage chamber 420. It is understood that step S250 and step S270 may be executed sequentially or simultaneously.
In step S290, the vacuum system 20 stops pumping the second chamber 17.
Thereafter, steps S230-S290 may be repeated a plurality of times to repeatedly filter the liquid sample.
For example, step S230 is performed again after step S290, and the first chamber 15 is sucked by the vacuum system 20 through the first opening 152, during which process, the components adhered to the surface of the second filtering membrane 16 can flow into the sample cell 13 along with the air flow and/or the liquid flow, and the liquid in the liquid sample (such as blood sample, body fluid sample, etc.) in the sample cell 13 and the components with the size smaller than the pore size of the first filtering membrane 14 pass through the first filtering membrane 14 under the negative pressure and enter the first chamber 15. Then, the step S250 is performed again, and the vacuum system 20 stops sucking the first chamber 15. And step S270 is performed, the vacuum system 20 sucks the second chamber 17 through the second opening 172.
By repeatedly performing steps S230 to S290, components smaller than the pore size of the filtration membrane in the liquid sample can be removed, and components larger than the pore size of the filtration membrane can be trapped in the sample cell 13. The permeability of the filtering membrane in the filtering process can be improved through the alternate change of negative pressure in the first cavity and the second cavity, the blocking hole of the filtering membrane is reduced, and the filtering effect is improved.
It is understood that, in step S220, a washing solution may be added to the cuvette 13, and the separation chip 10 may be washed by repeatedly performing steps S230 to S290. The cleaning solution may be a buffer solution or a solution containing specific bioactive molecules (e.g., nuclease, protease, etc.) for removing biological components adsorbed on the respective surfaces of the separation chip 10.
The separation chip and the separation method provided by the invention can be used for separating exosomes from blood samples. Curve a in FIG. 5 is the absorbance curve between the wavelengths of 220 and 340 nm of a blood sample before separation, wherein the blood sample is a liquid sample obtained after filtration through a filter with a pore size of 200 nm. Curve b in FIG. 5 is the absorbance curve between the wavelengths of 220 and 340 nm for the separated blood sample. Comparing the absorbance curves a and b shows that the absorbance of the separated sample at a wavelength of about 280 nm is obviously reduced, which indicates that the separation chip and the separation method can effectively separate and collect the components in the liquid sample.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and the above embodiments are only used for explaining the claims. The scope of the invention is not limited by the description. Any changes or substitutions that can be easily made by those skilled in the art within the technical scope of the present disclosure are included in the scope of the present invention.
Claims (8)
1. A separation device, comprising:
a separation chip, the separation chip comprising: a cuvette comprising a first side and a second side, the first side being opposite the second side, the first side being provided with a first filter membrane, the second side being provided with a second filter membrane;
the first chamber is communicated with the sample pool through the first filtering membrane, and is provided with a first opening used for communicating the first chamber with the outside;
the second chamber is communicated with the sample pool through the second filtering membrane, and is provided with a second opening which is used for communicating the second chamber with the outside;
a vacuum system connected to the first opening and the second opening of the separation chip, respectively;
the frequency conversion module is electrically connected with the vacuum system, and the first cavity and the second cavity alternately generate negative pressure under the action of the frequency conversion module and the vacuum system.
2. The separation device of claim 1, wherein the vacuum system comprises a first vacuum pump and a second vacuum pump, the first vacuum pump is connected to the first opening of the separation chip, the second vacuum pump is connected to the second opening of the separation chip, and the frequency conversion module is configured to control the first vacuum pump and the second vacuum pump to alternately operate.
3. The separation device of claim 1, further comprising:
a liquid supply unit for automatically supplying a liquid sample to the sample cell of the separation chip;
and the sample collection unit is used for automatically collecting the separated liquid sample from the sample pool.
4. The separation device according to claim 1, wherein the first filter membrane and the second filter membrane are each selected from one of a porous ceramic material, a porous plastic material, and a porous metal material.
5. The separation device of claim 4, wherein the pore size of the first filter membrane and the second filter membrane is 2-20 microns.
6. The separation device of claim 4, wherein the pore size of the first filtration membrane and the second filtration membrane is between 5 and 200 nanometers.
7. The separation device of claim 1, wherein the sample cell, the first chamber, and the second chamber are formed from a polymethylmethacrylate material.
8. A method of separating target particles in a liquid sample, comprising the steps of:
providing a separation device according to any one of claims 1 to 7;
providing a liquid sample to the sample cell;
drawing the first chamber through the first opening to create a negative pressure in the first chamber;
stopping pumping the first chamber;
drawing the second chamber through the second opening to create a negative pressure in the second chamber;
stopping pumping the second chamber.
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| CN201711396703.0A CN108126522B (en) | 2017-12-21 | 2017-12-21 | Separation chip, separation device and method for separating target particles in liquid sample |
| US16/203,649 US10758866B2 (en) | 2017-11-30 | 2018-11-29 | Isolation chip and manufacturing method of the same |
| US16/203,652 US10758867B2 (en) | 2017-11-30 | 2018-11-29 | Isolation device and isolation method |
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| CN109628277B (en) * | 2019-01-23 | 2022-02-01 | 东南大学 | System and method for separating and detecting tumor marker miRNA in exosome |
| CN112601959A (en) * | 2019-07-17 | 2021-04-02 | 深圳汇芯生物医疗科技有限公司 | Separation device and method for separating target particles in liquid sample |
| CN112694969A (en) * | 2020-09-22 | 2021-04-23 | 深圳汇芯生物医疗科技有限公司 | Separation detection chip, separation detection device and separation detection method |
| KR20230096955A (en) * | 2020-11-11 | 2023-06-30 | 센젠 후이신 라이프 테크놀로지스 컴퍼니 리미티드 | Separation device and separation method |
| JP7395212B2 (en) * | 2020-11-11 | 2023-12-11 | 深▲せん▼匯芯生物医療科技有限公司 | separation chip assembly |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102803467A (en) * | 2009-12-22 | 2012-11-28 | 液体活体检视公司 | Device and method for biological sample purification and enrichment |
| CN104711188A (en) * | 2015-04-01 | 2015-06-17 | 刘韬 | Device for separating tumor cells in blood fluid |
| CN105536898A (en) * | 2015-12-14 | 2016-05-04 | 清华大学 | Microfluidic chip, hemocyte separation method and system, and preparation method of system |
| CN107073397A (en) * | 2014-05-21 | 2017-08-18 | 非链实验室 | The system and method exchanged for cushioning liquid |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5227290A (en) * | 1990-08-29 | 1993-07-13 | Pocock Douglas A | Method for conducting diagnostic assays |
| AU703488B2 (en) * | 1995-06-30 | 1999-03-25 | Pall Corporation | Separation systems and methods |
| CN2836814Y (en) * | 2004-12-01 | 2006-11-15 | 上海日科贸易有限公司 | Blood purification device having bidirectional circulation |
| US8247191B2 (en) * | 2008-11-13 | 2012-08-21 | Ritzen Kalle | Disposable cassette and method of use for blood analysis on blood analyzer |
| CA2896047C (en) * | 2012-12-21 | 2021-04-13 | Porifera, Inc. | Separation systems and elements utilizing laterally offset membranes |
| CN106794295B (en) * | 2014-08-21 | 2020-10-23 | 查尔斯斯塔克布料实验室公司 | Systems and methods for increasing convective clearance of undesired particles in a microfluidic device |
| CN206396221U (en) * | 2016-10-09 | 2017-08-11 | 陈静 | The equipment that a kind of high flux fast Acquisition purifies circulating tumor cell |
-
2017
- 2017-12-21 CN CN201711396703.0A patent/CN108126522B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102803467A (en) * | 2009-12-22 | 2012-11-28 | 液体活体检视公司 | Device and method for biological sample purification and enrichment |
| CN107073397A (en) * | 2014-05-21 | 2017-08-18 | 非链实验室 | The system and method exchanged for cushioning liquid |
| CN104711188A (en) * | 2015-04-01 | 2015-06-17 | 刘韬 | Device for separating tumor cells in blood fluid |
| CN105536898A (en) * | 2015-12-14 | 2016-05-04 | 清华大学 | Microfluidic chip, hemocyte separation method and system, and preparation method of system |
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