CN113621516A - Tumor cell morphology dynamic observation microfluidic chip device based on vortex cell rotation - Google Patents
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- 210000004027 cell Anatomy 0.000 title claims abstract description 45
- 210000004881 tumor cell Anatomy 0.000 title claims abstract description 9
- 208000005443 Circulating Neoplastic Cells Diseases 0.000 claims abstract description 39
- 210000004369 blood Anatomy 0.000 claims abstract description 10
- 239000008280 blood Substances 0.000 claims abstract description 10
- 210000005266 circulating tumour cell Anatomy 0.000 claims abstract 11
- 210000005259 peripheral blood Anatomy 0.000 claims description 8
- 239000011886 peripheral blood Substances 0.000 claims description 8
- 230000001276 controlling effect Effects 0.000 claims description 7
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- 239000004205 dimethyl polysiloxane Substances 0.000 claims description 4
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- 230000003993 interaction Effects 0.000 claims description 3
- 239000001301 oxygen Substances 0.000 claims description 2
- 229910052760 oxygen Inorganic materials 0.000 claims description 2
- 235000013870 dimethyl polysiloxane Nutrition 0.000 claims 2
- 230000006399 behavior Effects 0.000 claims 1
- 239000007788 liquid Substances 0.000 claims 1
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 claims 1
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- 201000011510 cancer Diseases 0.000 description 3
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- 239000002245 particle Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 206010027476 Metastases Diseases 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
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- 238000011528 liquid biopsy Methods 0.000 description 1
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Abstract
The invention discloses a microfluidic chip device for dynamically observing tumor cell morphology based on vortex cell rotation, which consists of a chip main body structure, an inlet, a main channel, symmetrically distributed round and rectangular micro-groove structures, an outlet and a lower bottom plate. The vortex cell flow field generated by the micro-groove is utilized to realize the accurate control of the three-dimensional space rotation motion behavior of the captured circulating tumor cells, so that the dynamic tracking observation of the three-dimensional morphology is carried out, and the defect that the traditional single-cell observation device can only observe the unilateral morphology of the static cells is overcome. The chip device is simple in structure, the three-dimensional vortex cell structure generated by the micro groove is fully utilized, the three-dimensional space rotation observation of the cell to be detected is realized, the three-dimensional shape reconstruction of the CTCs cell can be further realized, and the integrated operation requirement of the sorting and the three-dimensional shape observation of the CTCs cell in blood is met. The invention is beneficial to the dynamic observation of the morphology of the CTCs, lays a foundation for further single cell analysis, and has important significance for scientific research on human tumor diseases.
Description
Technical Field
The invention relates to a novel microfluidic chip channel device for dynamically observing the three-dimensional morphology of Circulating Tumor Cells (CTCs). The invention belongs to the research fields of microfluidic technology, experimental hydromechanics and cell biology.
Background
Malignant tumor has become one of the most serious diseases in our country, and tumor metastasis is the main cause of death of patients. Detection of Circulating Tumor Cells (CTCs) with low concentration in peripheral blood of a patient has become one of emerging liquid biopsy technologies, and is of great importance for early diagnosis and treatment of tumors. Under the single cell observation condition, the three-dimensional space morphology observation of the internal tissue of the captured CTCs is carried out, which is helpful for further understanding the single cell structure and behavior characteristics.
Microfluidic technology (Microfluidics) is a new scientific technology for controlling micro-volume fluid (nanoliter to microliter) by using a microchannel, and is applied to the fields of new-generation medical diagnosis, drug detection and the like due to the subversive characteristics of precision, miniaturization, integration and the like. The micro-fluidic technology based on the micro-groove structure utilizes the inertial separation theory to enable the content of blood to be extremely low (10)-9) The CTCs of (1) are sorted from a large number of blood cells. Meanwhile, a three-dimensional vortex cell flow field structure formed by the micro-groove structure can realize the accurate control of the CTCs and the three-dimensional dynamic tracking and morphology observation of single cells/particles.
The experimental development is based on the microfluidic technology, the micro-groove CTCs are sorted and captured, a set of symmetrical micro-groove CTCs are used for sorting the microfluidic chip device, a special three-dimensional vortex cell flow field structure in the micro-groove is utilized, the three-dimensional dynamic appearance observation is carried out on the captured tumor cells, and the three-dimensional reconstruction is carried out on the cell appearance. Has important application prospect in the fields of cell biology, clinical medicine and the like, and has important significance for the research of single cell biology.
The invention content is as follows:
the invention aims to realize a microfluidic chip device for dynamically observing the three-dimensional morphology of Circulating Tumor Cells (CTCs) based on vortex cell rotation, which integrates the sorting and three-dimensional morphology observation of CTCs in blood, has simple operation, wide application prospect range and higher scientific research application value, and adopts the following technical scheme:
a micro-fluidic chip device for realizing the sorting and three-dimensional appearance observation of circulating tumor cells CTCs. The channel comprises a chip main body structure 1, an inlet 2, a main channel 3, a circular and rectangular micro-groove structure 4, an outlet 5 and a lower bottom plate 6 which are symmetrically distributed.
Specifically, the inlet 2, the main channel 3, the symmetrically distributed circular and rectangular micro-groove structures 4 and the outlet 5 are groove or hole structures on the chip main body structure 1, and each structure is a fluid flowing area when the chip works; the width range of the inlets of the symmetrically distributed round and rectangular micro-groove structures 4 is set to be 600 micrometers, the height of the rectangular groove and the radius of the symmetrically distributed round groove are both 400 micrometers, so that the three-dimensional vortex cell structures in the groove are fully utilized to realize the capture and morphology dynamic tracking of the CTCs, and the three-dimensional vortex cell structures in the groove for capturing and observing the CTCs are shown in FIG. 2; circular and rectangular micro-groove structures 4 which are symmetrically distributed are positioned at two sides of the main channel, and capture the circulating tumor cells into the grooves by utilizing the inertia lift force action and the collision capture mechanism, and meanwhile, the accurate control and morphology observation of the captured cells are realized by utilizing the three-dimensional flow field characteristics in the micro-grooves and the interaction force of fluid and the cells through the regulation and control of the Reynolds number at the inlet; the peripheral blood extraction device is a medical injector of 5-30ml, and is connected with an experimental device through a micro-flow pump, so that the peripheral blood extraction device enters the symmetrically distributed round and rectangular micro-groove structures 4 through the inlet 2.
The main body structure 1 and the lower bottom plate 6 are both made of polydimethylsiloxane and are fixed in an up-and-down bonding mode through oxygen ions, and the lower bottom plate 6 is arranged at the bottom of the main body structure 1 to support the main body structure of the chip and provide a flowing space;
the overall working process of the invention is as follows:
peripheral blood from a cancer patient enters the main channel structure 3 from the inlet 2, acquires inertial lift at the groove inlet due to the size difference of CTCs and other cells in the blood, and is captured in the symmetrically distributed circular and rectangular micro-groove structures 4. Meanwhile, by using different three-dimensional vortex cell structures generated by two micro grooves, the captured CTCs perform stable orbital motion and three-dimensional rotation motion in the grooves, and the experimental phenomenon is shown in FIG. 3. Accurate control of cell orbital motion behavior is achieved by regulating and controlling the Reynolds number of the inlet, so that three-dimensional observation of CTCs can be achieved, three-dimensional reconstruction is carried out on the appearance of the CTCs, and the observation effect is shown in figure 4. The modified device meets the operation requirements of the integration of sorting and three-dimensional shape observation of the CTCs in blood, and the CTCs captured by the grooves and observed and recorded can be discharged by reducing the Reynolds number of the inlet so as to carry out subsequent researches on identification, culture, enrichment and the like.
Drawings
FIG. 1 is a schematic diagram of a three-dimensional structure of a chip according to the present invention.
FIG. 2 is a schematic diagram of a three-dimensional vortex flow field structure and a cross-sectional particle velocity gradient in a micro-groove.
Fig. 3 is a diagram showing the trajectory of a particle to be labeled and the three-dimensional spin effect.
FIG. 4 is a topographical view of circulating tumor cells observed in the microgrooves.
FIG. 5 is a schematic diagram of the structure of the chip of the present invention.
Note that:
1. chip main structure, 2 inlet, 3 main channel, 4 symmetrically distributed round and rectangular micro-groove structure, 5 outlet, 6 lower bottom plate
Detailed Description
The working process and effect of the invention will be further explained with reference to the structure drawings.
Fig. 1 is a schematic structural diagram of a microfluidic chip device for dynamically observing the three-dimensional morphology of circulating tumor cells CTCs based on vortex cell rotation.
A micro-fluidic chip device for realizing the sorting of tumor CTCs cells and the observation of three-dimensional appearance. The channel comprises a chip main body structure 1, an inlet 2, a main channel 3, a circular and rectangular micro-groove structure 4, an outlet 5 and a lower bottom plate 6 which are symmetrically distributed. The inlet 2, the main channel 3, the symmetrically distributed round and rectangular micro-groove structures 4, the outlet 5 and the lower bottom plate 6 are groove or hole structures on the chip main body structure 1, and each structure is a fluid flowing area when the chip works; circular and rectangular micro-groove structures 4 which are symmetrically distributed are positioned at two sides of the main channel, the circulating tumor cells are captured into the grooves by utilizing the inertia lift force action and the collision capture mechanism, and meanwhile, the captured cells are accurately controlled and observed in shape by utilizing the three-dimensional flow field characteristics in the micro-grooves and the interaction force of fluid and the cells and regulating and controlling the Reynolds number of an inlet. The width range of the inlets of the symmetrically distributed round and rectangular micro-groove structures 4 is set to be 600 micrometers, the height of the rectangular groove and the radius of the symmetrically distributed round groove are both 400 micrometers, so that the three-dimensional vortex cell structures in the groove are fully utilized to realize the capture and morphology dynamic tracking of CTCs;
the chip main body structure 1 and the lower base plate 6 are made of polydimethylsiloxane.
The blood inlet is a hole structure which is arranged on the main body structure 1 and is communicated up and down.
The working process of the device is as follows: peripheral blood from a cancer patient enters the main channel structure 3 from the inlet 2, acquires inertial lift at the groove inlet due to the size difference of CTCs and other cells in the blood, and is captured in the symmetrically distributed circular and rectangular micro-groove structures 4. Meanwhile, by using different three-dimensional vortex cell structures generated by two micro grooves, the captured CTCs perform stable orbital motion and three-dimensional rotation motion in the grooves, and the experimental phenomenon is shown in FIG. 3. Accurate control of cell orbital motion behavior is achieved by regulating and controlling the Reynolds number of the inlet, so that three-dimensional observation of CTCs can be achieved, three-dimensional reconstruction is carried out on the appearance of the CTCs, and the observation effect is shown in figure 4. The modified device meets the operation requirements of the integration of sorting and three-dimensional shape observation of the CTCs in blood, and the CTCs captured by the grooves and observed and recorded can be discharged by reducing the Reynolds number of the inlet so as to carry out subsequent researches on identification, culture, enrichment and the like.
Claims (5)
1. A microfluidic chip device for dynamically observing the appearance of tumor cells based on vortex rotation comprises a chip main body structure (1), an inlet (2), a main channel (3), symmetrically distributed round and rectangular micro-groove structures (4), an outlet (5) and a lower bottom plate (6);
the inlet (2), the main channel (3), the symmetrically distributed round and rectangular micro-groove structures (4) and the outlet (5) are groove or hole structures on the chip main body structure 1, and each structure is a fluid flowing area when the chip works;
the width range of inlets of the symmetrically distributed round and rectangular micro-groove structures (4) is set to be 600 micrometers, the height of the rectangular groove and the radius of the symmetrically distributed round micro-groove structures are both 400 micrometers, so that the three-dimensional vortex cell structures in the groove are fully utilized to realize the capture and morphology dynamic tracking of CTCs;
circular and rectangular groove structures (4) which are symmetrically distributed are positioned at two sides of the main channel, the circulating tumor cells are captured into the grooves by utilizing the inertia lift force action and the collision capture mechanism, and meanwhile, the captured cells are accurately controlled and observed in appearance by utilizing the three-dimensional flow field characteristics in the micro-grooves and the interaction force of fluid and the cells and regulating and controlling the Reynolds number of an inlet;
the peripheral blood extraction device is a medical injector of 5-30ml, and is connected with an experimental device through a micro-flow pump, so that the peripheral blood extraction device enters a circular and rectangular micro-groove structure (4) which are symmetrically distributed through an inlet (2); the chip main body structure (1) and the lower base plate (6) are fixed in an up-and-down bonding mode through oxygen ions, and the lower base plate (6) is arranged at the bottom of the chip main body structure (1) to support the chip main body structure and provide a flowing space.
2. The microfluidic chip device for realizing tumor cell morphology dynamic observation based on vortex cell rotation as claimed in claim 1, wherein: the method comprises the steps of realizing accurate control of capturing CTCs by controlling Reynolds numbers, carrying out accurate control by controlling cell orbital motion behaviors by using different vortex flow fields generated by micro grooves under different Reynolds numbers, realizing three-dimensional observation of circulating tumor cells, and carrying out three-dimensional reconstruction on the appearance of the circulating tumor cells; CTCs captured by the groove and observed and recorded are discharged by reducing the inlet Reynolds number for subsequent identification, culture and enrichment.
3. The microfluidic chip device for realizing tumor cell morphology dynamic observation based on vortex cell rotation as claimed in claim 1, wherein: circular and rectangular micro-groove structures (4) are symmetrically distributed, two micro-groove forms generate three-dimensional vortex cell flow fields with different structures, and the two structures can realize accurate control of the captured CTCs three-dimensional space rotation motion, so that dynamic tracking observation of three-dimensional morphology is carried out.
4. The microfluidic chip device for realizing tumor cell morphology dynamic observation based on vortex cell rotation as claimed in claim 1, wherein: peripheral blood enters the main channel (3) from the inlet (2), and due to the size difference between CTCs and other cells in the blood, the CTCs obtain inertial lift force at the groove inlet and are captured in symmetrically distributed round and rectangular micro-groove structures (4); the captured CTCs perform stable orbital motion in the groove; meanwhile, a three-dimensional vortex cell structure generated by the micro-groove is fully utilized, the three-dimensional space rotation observation of the cells to be detected is realized, and the integrated operation requirements of the sorting and three-dimensional shape observation of the CTCs in the blood are met.
5. The microfluidic chip device for realizing tumor cell morphology dynamic observation based on vortex cell rotation as claimed in claim 1, wherein: the inlet (2), the main channel (3), the symmetrically distributed round and rectangular micro-groove structures (4) and the outlet (5) are groove or hole structures on the chip main body structure (1), each structure is a liquid flowing area when the chip works, and the chip main body structure (1) and the lower base plate (6) are made of polydimethylsiloxane PDMS.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113832005A (en) * | 2021-08-30 | 2021-12-24 | 北京工业大学 | Blood cancer cell filter chip device based on microchannel groove vortex cell flow |
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| CN107974392A (en) * | 2017-12-27 | 2018-05-01 | 深圳市合川医疗科技有限公司 | A kind of method of circulating tumor cell in micro-fluidic chip and separating trap blood |
| CN110257223A (en) * | 2019-07-15 | 2019-09-20 | 北京工业大学 | A kind of cell micro-environment regulation chip apparatus based on groove droplet capture |
| CN110923117A (en) * | 2019-12-11 | 2020-03-27 | 北京工业大学 | Chip device is selected separately to high-efficient circulation tumor cell based on heart type arcuation recess |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107974392A (en) * | 2017-12-27 | 2018-05-01 | 深圳市合川医疗科技有限公司 | A kind of method of circulating tumor cell in micro-fluidic chip and separating trap blood |
| CN110257223A (en) * | 2019-07-15 | 2019-09-20 | 北京工业大学 | A kind of cell micro-environment regulation chip apparatus based on groove droplet capture |
| CN110923117A (en) * | 2019-12-11 | 2020-03-27 | 北京工业大学 | Chip device is selected separately to high-efficient circulation tumor cell based on heart type arcuation recess |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113832005A (en) * | 2021-08-30 | 2021-12-24 | 北京工业大学 | Blood cancer cell filter chip device based on microchannel groove vortex cell flow |
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