CN111154620A - Micro-fluidic chip for magnetic quantity analysis carried by magnetic labeled cells - Google Patents
Micro-fluidic chip for magnetic quantity analysis carried by magnetic labeled cells Download PDFInfo
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Abstract
The invention provides a micro-fluidic chip for analyzing magnetic quantity carried by magnetically labeled cells, which is characterized in that cell suspensions in a magnetic separation area are respectively conveyed to different outlet channels through an external magnetic field to realize magnetic separation, and in combination with an electrode pair arranged on a glass substrate layer, when passing through an impedance detection area, the magnetically labeled cells can change the impedance of the impedance detection area, so that a pulse amplitude value can be generated, and the size and the quantity of the magnetically labeled cells can be reflected based on the pulse amplitude value. That is to say, the microfluidic chip can continuously and efficiently sort the magnetic labeled cells, analyze and evaluate the magnetic carrying quantity and the number of the magnetic labeled cells, analyze and sort the high magnetic carrying cells, can be used for magnetic resonance imaging, and has good imaging effect.
Description
Technical Field
The invention relates to the technical field of microfluidics, in particular to a microfluidic chip for magnetic quantity analysis carried by magnetically-labeled cells.
Background
Magnetic labeling of cells plays a very important role in vivo cell tracking and cell therapy, and current methods for magnetic labeling of cells include: direct incubation, electroporation, and sonoporation. However, the magnetic loading of magnetically labeled cells is an important indicator of the impact on both tracking and therapy, regardless of the labeling method used, and therefore, there is a need for effective methods for analyzing and sorting cells of different magnetic loading.
It is important to provide a microfluidic chip that can simultaneously perform analysis and sorting of magnetically labeled cells.
Disclosure of Invention
In view of the above, in order to solve the above problems, the present invention provides a microfluidic chip for magnetic quantity analysis carried on magnetically labeled cells, which has the following technical scheme:
a magnetically labeled cell-borne magnetic flux analysis microfluidic chip, the microfluidic chip comprising: a microfluidic chip layer and a glass substrate layer;
the microfluidic chip layer is provided with a cell suspension inlet, a buffer solution inlet, a cell suspension inlet channel, a buffer solution inlet channel, a magnetic separation area, a plurality of outlet channels and a plurality of outlets;
wherein a cell suspension is delivered to the magnetic sorting region through the cell suspension inlet and the cell suspension inlet channel; a buffer is delivered to the magnetic sorting zone through the buffer inlet and the buffer inlet channel;
the external magnetic field is used for conveying the cell suspension liquid in the magnetic separation area to different outlet channels respectively;
each of the outlet channels has an impedance detection zone;
a plurality of groups of electrode pairs are arranged on the glass substrate layer, and each group of electrode pairs is provided with a first electrode end, a second electrode end, a third electrode end and a fourth electrode end;
the first electrode end is connected with the second electrode end through an electrode lead, and the third electrode end is connected with the fourth electrode end through another electrode lead;
the first electrode end and the fourth electrode end form an external electrode, a gap exists between the second electrode end and the third electrode end, and the gap corresponds to the impedance detection area.
Preferably, in the above microfluidic chip, the microfluidic chip layer is a polydimethylsiloxane microfluidic chip layer.
Preferably, in the microfluidic chip, the thickness of the microfluidic chip layer is 5mm to 8 mm;
the thickness of the glass substrate layer is 1mm-2 mm.
Preferably, in the microfluidic chip, the length of the microfluidic chip layer is 6cm-8cm, and the width of the microfluidic chip layer is 4cm-6 cm;
the length of the glass substrate layer is 9cm-11cm, and the width of the glass substrate layer is 6cm-8 cm.
Preferably, in the above microfluidic chip, the heights of the cell suspension inlet channel, the buffer inlet channel and the magnetic separation region are 70 μm to 100 μm.
Preferably, in the above microfluidic chip, the height of the outlet channel and the impedance detection region is 15 μm to 20 μm.
Preferably, in the above microfluidic chip, the microfluidic chip further includes:
a plurality of support cylinders disposed within the magnetic separation region.
Preferably, in the microfluidic chip, the diameter of the bottom surface of the support cylinder is 100 μm to 150 μm.
Preferably, in the above microfluidic chip, the microfluidic chip further includes:
a PDMS mixed toluene film arranged between the microfluidic chip layer and the glass substrate layer;
the volume ratio of PDMS to toluene in the PDMS mixed toluene film is 1: 3.
Preferably, in the above microfluidic chip, a width of a gap existing between the second electrode terminal and the third electrode terminal is 1mm to 2 mm.
Compared with the prior art, the invention has the following beneficial effects:
according to the micro-fluidic chip for analyzing the magnetic quantity carried by the magnetically labeled cells, provided by the invention, the cell suspension in the magnetic separation area is respectively conveyed to different outlet channels through an external magnetic field, so that the magnetic separation is realized, and in combination with the electrode pairs arranged on the glass substrate layer, when passing through the impedance detection area, the magnetically labeled cells can change the impedance of the impedance detection area, so that a pulse amplitude value can be generated, and the size and the number of the magnetically labeled cells can be reflected on the basis of the pulse amplitude value.
That is to say, the microfluidic chip can continuously and efficiently sort the magnetic labeled cells, analyze and evaluate the magnetic carrying quantity and the number of the magnetic labeled cells, analyze and sort the high magnetic carrying cells, can be used for magnetic resonance imaging, and has good imaging effect.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
Fig. 1 is a schematic structural diagram of a microfluidic chip for magnetic quantity analysis carried by magnetically labeled cells according to an embodiment of the present invention;
fig. 2 is a schematic structural diagram of a microfluidic chip layer according to an embodiment of the present invention;
FIG. 3 is a schematic cross-sectional view of a microfluidic chip layer according to an embodiment of the present invention;
fig. 4 is a schematic structural diagram of a glass substrate layer according to an embodiment of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in further detail below.
Referring to fig. 1, fig. 1 is a schematic structural diagram of a microfluidic chip for magnetic quantity analysis on magnetically labeled cells according to an embodiment of the present invention.
The microfluidic chip includes: a microfluidic chip layer 1 and a glass substrate layer 3.
Referring to fig. 2, fig. 2 is a schematic structural diagram of a microfluidic chip layer according to an embodiment of the present invention.
Referring to fig. 3, fig. 3 is a schematic cross-sectional view of a microfluidic chip layer according to an embodiment of the present invention.
The microfluidic chip layer 1 is provided with a cell suspension inlet 4, a buffer solution inlet 5, a cell suspension inlet channel 6, a buffer solution inlet channel 7, a magnetic sorting area 8, a plurality of outlet channels 11 and a plurality of outlets 12;
wherein a cell suspension is delivered to the magnetic sorting zone 8 through the cell suspension inlet 4 and the cell suspension inlet channel 6; buffer is delivered to the magnetic sorting zone through the buffer inlet 5 and the buffer inlet channel 7;
the external magnetic field is used for conveying the cell suspension liquid in the magnetic sorting area 8 to different outlet channels 11 respectively;
each of the outlet channels 11 has an impedance detection zone 10.
Wherein 19 in FIG. 3 shows a schematic cross-sectional view of the impedance detection zone 10 and the outlet channel 11; a schematic cross-sectional view of the magnetic sorting region 8 and the inlet channel is shown at 20.
Referring to fig. 4, fig. 4 is a schematic structural diagram of a glass substrate layer according to an embodiment of the present invention.
A plurality of groups of electrode pairs are arranged on the glass substrate layer 3, and each group of electrode pairs is provided with a first electrode end 13, a second electrode end 17, a third electrode end 18 and a fourth electrode end 14;
the first electrode end 13 and the second electrode end 17 are connected through an electrode lead, and the third electrode end 18 and the fourth electrode end 14 are connected through another electrode lead;
the first electrode end 13 and the fourth electrode end 14 form an external electrode, and a gap exists between the second electrode end 17 and the third electrode end 18, as shown in fig. 1, where the gap corresponds to the impedance detection region 10.
In this embodiment, the cell suspension in the magnetic separation region 8 is delivered to different outlet channels 11 by an external magnetic field, so as to realize magnetic separation, and in combination with the electrode pairs disposed on the glass substrate layer 3, when passing through the impedance detection region 10, the magnetically labeled cells change the impedance of the impedance detection region 10, so as to generate a pulse amplitude, based on which the size and number of the magnetically labeled cells can be reflected. That is, the cell impedance analysis is an important precondition for realizing the magnetic quantity analysis chip of the magnetically labeled cell, when the cell passes through the micro-channel of the impedance detection area, the cell can replace the original medium, so as to cause the change of the impedance, the changed impedance corresponds to the corresponding pulse amplitude, the number and the volume of the cell can be analyzed according to the pulse amplitude, the micro-fluidic cell impedance analysis can be applied to the fields of cell counting, cell subtype analysis, cell performance evaluation and the like.
That is to say, the microfluidic chip can continuously and efficiently sort the magnetic labeled cells, analyze and evaluate the magnetic carrying quantity and the number of the magnetic labeled cells, analyze and sort the high magnetic carrying cells, can be used for magnetic resonance imaging, and has good imaging effect.
The micro-fluidic chip is a functional integrated micro-fluidic chip which can simultaneously realize the analysis and the sorting of magnetic labeling cells, can continuously operate, can classify the cells with different magnetic carrying quantities through the magnetic sorting function, the cells with different classes have corresponding magnetic carrying quantities, the classified cells can carry out continuity statistics on the number of the cells in an impedance detection area, the number is counted, the density is calculated according to the flow, and the cells with high magnetic carrying quantity with known cell density have better nuclear magnetic resonance imaging effect.
Further, based on the above embodiment of the present invention, as shown in fig. 1, the microfluidic chip further includes:
a PDMS mixed toluene film 2 arranged between the microfluidic chip layer 1 and the glass substrate layer 3;
in the PDMS mixed toluene film 2, the volume ratio of PDMS (polydimethylsiloxane) to toluene is 1: 3.
In this embodiment, the PDMS mixed toluene film 2 can effectively insulate the electrode and the cell solution, and the collected signal is affected when the film thickness is larger than 10 μm, so the film thickness is set to be 5 μm-10 μm.
Further, based on the above embodiments of the present invention, the microfluidic chip layer 1 is a polydimethylsiloxane microfluidic chip layer.
The glass substrate layer 3 can be a common glass substrate.
Further, based on the above embodiment of the present invention, the thickness of the microfluidic chip layer 1 is 5mm to 8 mm;
the thickness of the glass substrate layer is 1mm-2 mm.
In this embodiment, when the thickness of the microfluidic chip layer 1 is less than 5mm, the connection stability of the components disposed on the microfluidic chip layer 1 is reduced, and when the thickness is greater than 8mm, the material is wasted.
When the thickness of the glass substrate layer 3 is less than 1mm, damage may be easily caused, and when the thickness is more than 2mm, observation with a microscope is not performed.
Further, based on the above embodiment of the present invention, the length of the microfluidic chip layer 1 is 6cm to 8cm, and the width thereof is 4cm to 6 cm;
the length of the glass substrate layer 3 is 9cm-11cm, and the width is 6cm-8 cm.
In this embodiment, the length and width of the microfluidic chip layer 1 are determined by the channel size, less than 4cm results in inconvenient processing, and more than 6cm wastes material; the length and the width of the glass substrate layer 3 are optimally larger than the microfluidic chip layer by 2-3 cm according to the length and the width of the microfluidic chip layer 1, the small glass substrate layer is not suitable for electrode routing arrangement, and materials are wasted due to the large glass substrate layer.
Further, according to the above embodiment of the present invention, the height of the cell suspension inlet channel 6, the buffer inlet channel 7 and the magnetic separation region 8 is 70 μm to 100 μm.
In this embodiment, the microchannel height of the magnetic sorting region 8 affects the flow velocity of the sorting region, and when the flow rate is fixed, the microchannel height of the magnetic sorting region 8 is less than 70 μm or more than 100 μm, which has a relatively large influence on the sorting efficiency of the chip.
Further, according to the above embodiment of the present invention, the height of the outlet channel 11 and the impedance detection section 10 is 15 μm to 20 μm.
In this embodiment, the height of the impedance detection area 10 and each outlet channel 11 is matched with the cell, most of the immune cells are distributed between 10-20 μm in diameter, the accuracy of impedance detection is affected when the height of the channel is greater than 20 μm, and blockage is easily caused when the height of the channel is less than 15 μm.
Further, based on the above embodiment of the present invention, the microfluidic chip further includes:
a plurality of support cylinders 9 disposed within the magnetic sorting zone 8.
Optionally, the diameter of the bottom surface of the support cylinder 9 is 100 μm to 150 μm.
In this embodiment, the bottom surface of the support cylinder 9 is directly less than 100 μm and does not support very well, and more than 150 μm has an excessive effect on the fluid in the channel.
Further, according to the above embodiment of the present invention, each of the electrode tips has a rectangular shape, a length of 2mm to 3.5mm and a width of 1.5mm to 2.5mm, and a gap existing between the second electrode tip and the third electrode tip has a width of 1mm to 2 mm.
Based on all the above-mentioned embodiments of the invention, in another embodiment of the invention, a preferred embodiment is provided,
the whole microfluidic chip layer 1 is 6cm long, 4cm wide and 5mm thick; wherein the microchannel comprises a cell and buffer solution inlet, and the height of the magnetic sorting area is 100 μm, so as to ensure that the flow is improved as much as possible under the condition of constant flow velocity; the width of the cell and buffer solution inlet channel is 300 μm, the length of the magnetic sorting area is 13mm, and the width is 3.3 mm; the height of 6 outlet channels and impedance detection area is 20 μm to ensure the accuracy of impedance detection, the side length of the impedance detection area is 20 μm, the width of the outlet channel is 250 μm.
The glass substrate layer 3 is formed by using glass as a substrate and forming an indium tin oxide electrode by a laser etching technique, as shown in fig. 3. The length of the whole glass substrate layer was 9cm, the width was 6cm, and the thickness was 1 mm. The indium tin oxide electrode etched by laser has 6 pairs, which correspond to 6 impedance detection areas, and each electrode has a length of 2mm and a width of 1.5 mm. The distance between the two electrodes of each electrode pair was 1.5 mm. The width of the electrode line conducting with the electrode is 250 μm.
The magnetic labeled cells and the buffer solution respectively enter the magnetic sorting area from the cell inlet and the buffer solution inlet, and under the action of the magnetic field generated by the permanent magnet, the cells with different magnetic carrying quantities generate different deflections to realize sorting. When the magnetic sorting cells enter the impedance detection areas of different outlet channels, the media of the impedance detection areas can be changed, so that impedance changes are caused, and the number and the size of the cells can be analyzed through the processing of the acquisition circuit, the acquisition board card and computer software.
The present invention provides a magnetic labeled cell magnetic quantity analysis microfluidic chip, which is described in detail above, and the specific examples are applied herein to illustrate the principles and embodiments of the present invention, and the description of the examples is only used to help understand the method and the core concept of the present invention; meanwhile, for a person skilled in the art, according to the idea of the present invention, there may be variations in the specific embodiments and the application scope, and in summary, the content of the present specification should not be construed as a limitation to the present invention.
It should be noted that, in the present specification, the embodiments are all described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments may be referred to each other. The device disclosed by the embodiment corresponds to the method disclosed by the embodiment, so that the description is simple, and the relevant points can be referred to the method part for description.
It is further noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include or include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
1. A magnetic-labeled cell-borne magnetic quantity analysis microfluidic chip, comprising: a microfluidic chip layer and a glass substrate layer;
the microfluidic chip layer is provided with a cell suspension inlet, a buffer solution inlet, a cell suspension inlet channel, a buffer solution inlet channel, a magnetic separation area, a plurality of outlet channels and a plurality of outlets;
wherein a cell suspension is delivered to the magnetic sorting region through the cell suspension inlet and the cell suspension inlet channel; a buffer is delivered to the magnetic sorting zone through the buffer inlet and the buffer inlet channel;
the external magnetic field is used for conveying the cell suspension liquid in the magnetic separation area to different outlet channels respectively;
each of the outlet channels has an impedance detection zone;
a plurality of groups of electrode pairs are arranged on the glass substrate layer, and each group of electrode pairs is provided with a first electrode end, a second electrode end, a third electrode end and a fourth electrode end;
the first electrode end is connected with the second electrode end through an electrode lead, and the third electrode end is connected with the fourth electrode end through another electrode lead;
the first electrode end and the fourth electrode end form an external electrode, a gap exists between the second electrode end and the third electrode end, and the gap corresponds to the impedance detection area.
2. The microfluidic chip according to claim 1, wherein the microfluidic chip layer is a polydimethylsiloxane microfluidic chip layer.
3. The microfluidic chip according to claim 1, wherein the thickness of the microfluidic chip layer is 5mm to 8 mm;
the thickness of the glass substrate layer is 1mm-2 mm.
4. The microfluidic chip according to claim 1, wherein the microfluidic chip layer has a length of 6cm to 8cm and a width of 4cm to 6 cm;
the length of the glass substrate layer is 9cm-11cm, and the width of the glass substrate layer is 6cm-8 cm.
5. The microfluidic chip according to claim 1, wherein the height of the cell suspension inlet channel, the buffer inlet channel and the magnetic separation region is 70 μm to 100 μm.
6. The microfluidic chip according to claim 1, wherein the height of the outlet channel and the impedance detection area is 15 μm to 20 μm.
7. The microfluidic chip according to claim 1, further comprising:
a plurality of support cylinders disposed within the magnetic separation region.
8. The microfluidic chip according to claim 7, wherein the diameter of the bottom surface of the support cylinder is 100 μm to 150 μm.
9. The microfluidic chip according to claim 1, further comprising:
a PDMS mixed toluene film arranged between the microfluidic chip layer and the glass substrate layer;
the volume ratio of PDMS to toluene in the PDMS mixed toluene film is 1: 3.
10. The microfluidic chip according to claim 1, wherein a width of a gap existing between the second electrode terminal and the third electrode terminal is 1mm to 2 mm.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112094839A (en) * | 2020-11-06 | 2020-12-18 | 深圳市赛特罗生物医疗技术有限公司 | Automatic cell magnetic sorting method and device |
| CN112871229A (en) * | 2021-01-21 | 2021-06-01 | 中国科学技术大学 | Chip for water dielectrophoresis bacteria sorting |
| CN112903793A (en) * | 2021-01-21 | 2021-06-04 | 中国科学技术大学 | Multi-parameter rapid detection device and method for water body |
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2020
- 2020-01-17 CN CN202010053102.5A patent/CN111154620A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112094839A (en) * | 2020-11-06 | 2020-12-18 | 深圳市赛特罗生物医疗技术有限公司 | Automatic cell magnetic sorting method and device |
| CN112094839B (en) * | 2020-11-06 | 2021-03-16 | 深圳市赛特罗生物医疗技术有限公司 | Automatic cell magnetic sorting method and device |
| CN112871229A (en) * | 2021-01-21 | 2021-06-01 | 中国科学技术大学 | Chip for water dielectrophoresis bacteria sorting |
| CN112903793A (en) * | 2021-01-21 | 2021-06-04 | 中国科学技术大学 | Multi-parameter rapid detection device and method for water body |
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