CN208604119U - Micro-fluidic chip for circulating tumor cell sorting - Google Patents
Micro-fluidic chip for circulating tumor cell sorting Download PDFInfo
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- CN208604119U CN208604119U CN201821163750.0U CN201821163750U CN208604119U CN 208604119 U CN208604119 U CN 208604119U CN 201821163750 U CN201821163750 U CN 201821163750U CN 208604119 U CN208604119 U CN 208604119U
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Abstract
The utility model discloses a kind of micro-fluidic chips for circulating tumor cell sorting, including sample introduction portion, at least one micro-pillar array area and sample portion out;Wherein, micro-pillar array area includes at least one microtrabeculae matrix and aim cell collection channel for being made of several microtrabeculaes, and aim cell collection channel is located at the side of microtrabeculae matrix along blood flow direction setting and is connected to aim cell collection port;Microtrabeculae matrix is obliquely installed in 1.5~2.5 degree of angles to aim cell collection channel side according to horizontal direction;The cross section of microtrabeculae is that oval, oval diameter is 28~33 μm, and oval symmetrical diameter is 20~25 μm;The ovum heart of adjacent two microtrabeculae of blood flow direction away from being 40~45 μm, perpendicular to blood flow direction adjacent two microtrabeculae the ovum heart away from being 60~65 μm.Leucocyte and circulating tumor cell structural integrity that the utility model obtains, activity are high, are convenient for studying in next step.
Description
Technical field
The utility model relates to a kind of micro-fluidic chip, specifically a kind of miniflow for circulating tumor cell screening
Control chip.
Background technique
Correlative study has confirmed: cancer patient's row tumor radical cure is postoperative, and the circulating tumor cell in blood is in cancer return
And play an important role in transfer, therefore research circulating tumor cell helps thoroughly to block cancer metastasis and recurrence approach,
With important clinical value and meaning.But due to circulating tumor cell rareness, biological stability difference etc., how high efficiency extraction
Circulating tumor becomes hot issue.Currently, circulating tumor cell extraction is broadly divided into positive concentration method and negative concentration method.It is positive
Concentration method identifies circulating tumor cell surface specific antigen (usually EpCAM) by means of different, to reach extraction
The purpose of cell.But circulating tumor cell surface antigen is not continuous expression, is lost when Epithelial and stromal sample change Shi Changhui occurs in it
EpCAM is lost, so as to cause the failure of positive concentration method.Feminine gender enrichment rule reaches enrichment cycles tumour by excluding unrelated cell
The purpose of cell, and existing negative concentration method, predominantly physics crossing sieve method or density gradient centrifugation cooperate traditional immunization magnetic bead
Method, aim cell lose serious, inefficiency.
With microflow control technique to the continuous intensification of cell-biological impact and its in integrated sample pretreatment and blood
Advantage in constituent analysis allows to milder, quickly, consistently manipulates and sort living cells, is conducive to more acurrate efficient
Ground extracts the information in the humoral samples such as blood, is highly suitable for the detection of the body cells such as haemocyte, cancer cell.In recent years, state
Inside and outside scholar achieves a series of achievement in the micro-fluidic sorting field of tumour cell, microfluidic chip technology just gradually at
For the main tool of cell sorting, it is expected to realize the circulating tumor cell detection of high efficiency, high-purity using micro-fluidic chip.
The structure of micro-fluidic chip directly influences the screening efficiency of circulating tumor cell, and micro-fluidic in utilization at present
During cDNA microarray tumour cell, since the designs such as chip structure easily cause tumour cell to be corrupted such that enrichment effect
Fruit is deteriorated, and can not carry out next step culture and drug sensitive test, keeps research limited.For example, a kind of now widely used positivity is rich
Collect fishbone micro-fluidic chip, captures circulating tumor cell using antigen-antibody combined techniques, after cell is in conjunction with chip, need to destroy
Cell is separated after chip, and since cell and antibody Irreversible binding not can avoid when collecting individual cells and tear cell
It splits, to destroy cell activity, losing downstream tests may.
Above-mentioned the problems of the prior art how are solved, are still the hot spot of those skilled in the art's research.
Summary of the invention
The technical problem to be solved by the utility model is to provide one kind can efficiently fast enriching screening circulating tumor it is thin
Born of the same parents, and the tumour cell of enrichment can be kept not to be destroyed, keep the micro-fluidic chip of maximum activity.
To achieve the goals above, the utility model adopts the following technical solution:
It is a kind of for circulating tumor cell sorting micro-fluidic chip, including sample introduction portion, at least one micro-pillar array area and
Sample portion out;Micro-pillar array area one end is connected to sample introduction portion, and the other end is connected to sample portion out;The sample introduction portion includes blood sample
This injection port;The sample portion out includes aim cell collection port and waste collection mouth;Wherein, the micro-pillar array area includes at least
The one microtrabeculae matrix and aim cell collection channel being made of several microtrabeculaes, the aim cell collection channel is along blood flow direction
Side positioned at the microtrabeculae matrix is set and is connected to the aim cell collection port;The microtrabeculae matrix according to blood flow side
It is obliquely installed in 1.5~2.5 degree of angles to aim cell collection channel side;The cross section of the microtrabeculae be it is oval, it is described
Oval diameter is 28~33 μm, and the oval symmetrical diameter is 20~25 μm;The ovum of adjacent two microtrabeculae of the blood flow direction
The heart is away from being 40~45 μm, and the ovum heart of adjacent two microtrabeculae perpendicular to blood flow direction is away from being 60~65 μm.
Wherein more preferably, the quantity in the micro-pillar array area is two.
Wherein more preferably, the both ends in the micro-pillar array area are equipped with grating structure area, and the microtrabeculae arranged in matrix is in two grid
Between shape structural area.
Wherein more preferably, the sample introduction portion further includes buffer inlet.
Wherein more preferably, the blood sample injection port is connected by blood sample sample feeding pipe and micro-pillar array area end
It is logical;The buffer inlet is connected with the blood sample sample feeding pipe and the micro-pillar array area simultaneously by buffer inlet tube
It is logical.
Wherein more preferably, the blood sample sample feeding pipe is camber pipe.
Wherein more preferably, grating structure is equipped in the blood sample sample feeding pipe.
Wherein more preferably, grating structure is equipped in the buffer inlet tube.
Wherein more preferably, the aim cell collection port is connected to by aim cell collecting pipe with micro-pillar array area end;
The waste collection mouth is connected to by waste collection pipe with micro-pillar array area end.
Wherein more preferably, the aim cell collecting pipe is camber pipe.
The structure design of this micro-fluidic chip can be such that cell sorting process realizes by physical method, and cell flows through chip
Respective collection port is only flowed to by preset path with different size of cell after microtrabeculae generation elastic collision, that directly collects is thin
Born of the same parents pass through the visible leucocyte and circulating tumor cell obtained by the utility model of immunofluorescence dyeing without remaining operation damage
Structural integrity, activity are high, are convenient for studying in next step.
Detailed description of the invention
Fig. 1 is the overall structure diagram of the utility model;
Fig. 2 is the sample introduction portion structural schematic diagram of the utility model;
Fig. 3 is the area Tu1Zhong W enlarged drawing, that is, microtrabeculae matrix structure schematic diagram;
Fig. 4 is the sample portion structural schematic diagram out of the utility model;
Fig. 5 is the microtrabeculae enlarged drawing of the utility model;
Fig. 6 is the operating procedure schematic diagram of the utility model;
Fig. 7 is the leucocyte and circulating tumor cell after immunofluorescence dyeing.
Specific embodiment
As shown in Figures 1 to 5, the utility model provides a kind of micro-fluidic chip for circulating tumor cell sorting,
Including sample introduction portion 1, Liang Ge micro-pillar array area 2 and sample portion 3 out;It micro-pillar array area 2 can basis in actual production use process
It needs to be arranged quantity, at least Yao Youyige micro-pillar array area, uses two in the present embodiment.The design in Liang Ge micro-pillar array area is not
The screening efficiency of aim cell only can be improved.
The one end in micro-pillar array area 2 is connected to sample introduction portion 1, and the other end is connected to sample portion 3 out.Sample introduction portion 1 includes blood sample
This injection port 11 and buffer inlet 12.Sample portion 3 includes aim cell collection port 31 and waste collection mouth 32 out.Blood sample
Injection port 11 is connected to by blood sample sample feeding pipe 111 with 2 end of micro-pillar array area;Buffer inlet 12 by buffer into
Liquid pipe 121 is connected to blood sample sample feeding pipe 111 and micro-pillar array area 2 simultaneously.Blood sample sample feeding pipe 111 is camber pipe, this
Design the purpose is to reduce fluid resistances.Foreign matter enters chip in order to prevent, is equipped with grating structure in blood sample sample feeding pipe 111
112.Grating structure 122 is again provided in buffer inlet tube 121, in order to prevent obligation from entering chip.Aim cell
Collection port 31 is connected to by aim cell collecting pipe 311 with 2 end of micro-pillar array area, and aim cell collecting pipe 311 is camber pipe.
Waste collection mouth 32 is connected to by waste collection pipe 321 with 2 end of micro-pillar array area.
Micro-pillar array area 2 includes the microtrabeculae matrix 22 that at least one is made of several microtrabeculaes 21.The one of the utility model
In a embodiment, each micro-pillar array area 2 is equipped with 13 microtrabeculae matrixes 22;Each microtrabeculae matrix 22 includes that 57 column are micro-
Column, each column microtrabeculae are 13 rows, amount to 741 microtrabeculaes.Microtrabeculae quantity in microtrabeculae matrix 22 can be according to actual needs
Setting.In the utility model, aim cell can successfully be imported cell collection channel and flow into mesh by the setting of microtrabeculae matrix
Cell collection port.Micro-pillar array area 2 further includes an aim cell collection channel 23, and the aim cell collection channel 23 is along blood
The setting of stream direction is located at the side of microtrabeculae matrix 22 and is connected to the aim cell collection port 31.Microtrabeculae matrix 22 according to blood
Stream direction is obliquely installed in 1.5~2.5 angles to 23 side of aim cell collection channel.
In one embodiment of the utility model, microtrabeculae matrix 22 (is denoted as with horizontal direction in 1.7 degree of angles in figure
F).The both ends in micro-pillar array area 2 are additionally provided with grating structure area 24, several settings of microtrabeculae matrixes 22 two grating structure areas 24 it
Between.The purpose of setting in grating structure area 24 is to guarantee that fluid enters chip according to certain orientation, and prevent foreign matter from entering chip.
The cross section of microtrabeculae 21 is that oval, oval diameter B recommended range is 28~33 μm, and oval symmetrical diameter A is pushed away
Recommend is 20~25 μm;The ovum heart of adjacent two microtrabeculae of blood flow direction is recommended as 40~45 μm away from C, perpendicular to adjacent the two of blood flow direction
The ovum heart of microtrabeculae 21 is recommended as 60~65 μm away from D.Above-mentioned numberical range is feasible and preferable numberical range, in the utility model
One embodiment in, the preference data of use are as follows: oval diameter B recommended range is 31 μm, and oval symmetrical diameter A recommends
It is 23 μm;The ovum heart of adjacent two microtrabeculae of blood flow direction is recommended as 41 μm away from C, perpendicular to the ovum of adjacent two microtrabeculae 21 of blood flow direction
The heart is recommended as 62 μm away from D.
As shown in fig. 6, the application method of the utility model are as follows:
(1) patient whole blood is mixed into incubation (in figure shown in a) with novel whole blood CD45 magnetic bead, by leucocyte in whole blood and magnetic
Pearl combines.
(2) whole blood after step (1) is in conjunction with magnetic bead enters chip by blood sample injection port 11, simultaneous buffering liquid b by
Buffer inlet 12 enters chip.In the micro-pillar array area 2 of micro-fluidic chip be greater than sorting critical radius big substance (with
Magnetic bead combine leucocyte and circulating tumor cell) with 21 array of microtrabeculae collision after lateral displacement occurs, can be received to aim cell
Collect the convergence of channel side, and be less than the small substance (red blood cell and blood platelet etc. interfere cell) of sorting critical radius with microtrabeculae 21
Sidesway does not occur after array collision, flows through array according to its original;The structure and size and microtrabeculae of the utility model microtrabeculae it
Between distance determine the critical deflection radius of the array of the microtrabeculae matrix be 4~5 μm.I.e. greater than 5 μm cell can along array to
Upper deflection, the cell less than 5 μm will be flowed along original direction, will not change flow direction.Aim cell collection port collects mesh
Cell c (leucocyte and circulating tumor cell in conjunction with magnetic bead), waste liquid d by waste collection mouth discharge (red blood cell and blood are small
Plate etc. interferes cell).
(3) the aim cell c for obtaining step (2) removes the leucocyte in conjunction with magnetic bead by specific magnetic fields e, thus
Obtain high-purity circulating tumor cell f.Gained circulating tumor cell is efficiently quick in screening enrichment process, and whole operation process
In do not destroy circulating tumor cell, it is high to make to collect obtained circulating tumor cell activity, is conducive to the detection in later period.
Using this micro-fluidic chip, cell is sorted by physical method, cell flows through chip and elastic collision only occurs with microtrabeculae
Hit, after can directly collect, it is no remaining operation damage, Fig. 7 be immunofluorescence dyeing after leucocyte and circulating tumor cell, it is seen that
The eucaryotic cell structure obtained using this micro-fluidic chip is complete, active high, is convenient for studying in next step.
Claims (10)
1. a kind of micro-fluidic chip for circulating tumor cell sorting, including sample introduction portion (1), at least one micro-pillar array area
(2) and out sample portion (3);Described micro-pillar array area (2) one end is connected to sample introduction portion (1), and the other end is connected to sample portion (3) out;Institute
Stating sample introduction portion (1) includes blood sample injection port (11);The sample portion out includes aim cell collection port (31) and waste collection
Mouth (32);It is characterized by:
The micro-pillar array area (2) includes microtrabeculae matrix (22) and the aim cell receipts that at least one is made of several microtrabeculaes (21)
Collect channel (23), the aim cell collection channel (23) is located at the side of the microtrabeculae matrix (22) simultaneously along blood flow direction setting
It is connected to the aim cell collection port (31);The microtrabeculae matrix (22) according to horizontal direction in 1.5~2.5 degree of angles to
Aim cell collection channel (23) side is obliquely installed;The cross section of the microtrabeculae (21) be it is oval, the oval diameter is
28~33 μm, the oval symmetrical diameter is 20~25 μm;The ovum heart of adjacent two microtrabeculae of the blood flow direction is away from for 40~45 μ
M, the ovum heart of adjacent two microtrabeculae perpendicular to blood flow direction is away from being 60~65 μm.
2. micro-fluidic chip as described in claim 1, it is characterised in that:
The quantity of the micro-pillar array area (2) is two.
3. micro-fluidic chip as described in claim 1, it is characterised in that:
The both ends of the micro-pillar array area (2) are equipped with grating structure area (24), and the microtrabeculae matrix (22) is arranged in two palisade knots
Between structure area (24).
4. micro-fluidic chip as described in claim 1, it is characterised in that:
The sample introduction portion (1) further includes buffer inlet (12).
5. micro-fluidic chip as claimed in claim 4, it is characterised in that:
The blood sample injection port (11) is connected to by blood sample sample feeding pipe (111) with micro-pillar array area (2) end;
The buffer inlet (12) by buffer inlet tube (121) simultaneously with the blood sample sample feeding pipe (111) and described
Micro-pillar array area (2) connection.
6. micro-fluidic chip as claimed in claim 5, it is characterised in that:
The blood sample sample feeding pipe (111) is camber pipe.
7. such as micro-fluidic chip described in claim 5 or 6, it is characterised in that:
Grating structure (112) are equipped in the blood sample sample feeding pipe (111).
8. such as micro-fluidic chip described in claim 5 or 6, it is characterised in that:
Grating structure (122) are equipped in the buffer inlet tube (121).
9. micro-fluidic chip as described in claim 1, it is characterised in that:
The aim cell collection port (31) is connected to by aim cell collecting pipe (311) with micro-pillar array area (2) end;It is described
Waste collection mouth (32) is connected to by waste collection pipe (321) with micro-pillar array area (2) end.
10. micro-fluidic chip as claimed in claim 9, it is characterised in that:
The aim cell collecting pipe (311) is camber pipe.
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Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
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CN111733074A (en) * | 2020-07-30 | 2020-10-02 | 首都医科大学附属北京友谊医院 | Micro-fluidic chip and system for high-flux magnetic sorting of circulating tumor cells |
WO2021013066A1 (en) * | 2019-07-23 | 2021-01-28 | 北京大学 | Microfluidic chip suitable for capturing circulating tumour cells |
CN113652333A (en) * | 2021-08-03 | 2021-11-16 | 中国科学院上海微系统与信息技术研究所 | Micro-column type multi-phase displacement channel for optimizing fluid distribution |
CN115957838A (en) * | 2023-01-13 | 2023-04-14 | 睿思生命(广东)科技有限公司 | Microfluidic chip and application method thereof |
CN115970775A (en) * | 2022-12-08 | 2023-04-18 | 中南大学 | Centrifugal driving micro-fluidic chip, preparation method and application |
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2018
- 2018-07-20 CN CN201821163750.0U patent/CN208604119U/en active Active
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
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WO2021013066A1 (en) * | 2019-07-23 | 2021-01-28 | 北京大学 | Microfluidic chip suitable for capturing circulating tumour cells |
CN111733074A (en) * | 2020-07-30 | 2020-10-02 | 首都医科大学附属北京友谊医院 | Micro-fluidic chip and system for high-flux magnetic sorting of circulating tumor cells |
CN111733074B (en) * | 2020-07-30 | 2020-12-04 | 首都医科大学附属北京友谊医院 | Micro-fluidic chip and system for high-flux magnetic sorting of circulating tumor cells |
CN113652333A (en) * | 2021-08-03 | 2021-11-16 | 中国科学院上海微系统与信息技术研究所 | Micro-column type multi-phase displacement channel for optimizing fluid distribution |
CN115970775A (en) * | 2022-12-08 | 2023-04-18 | 中南大学 | Centrifugal driving micro-fluidic chip, preparation method and application |
CN115957838A (en) * | 2023-01-13 | 2023-04-14 | 睿思生命(广东)科技有限公司 | Microfluidic chip and application method thereof |
CN115957838B (en) * | 2023-01-13 | 2023-09-26 | 睿思生命(广东)科技有限公司 | Microfluidic chip |
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