CN114395463A - CTC enrichment and release system based on micro-fluidic and low-light-level tweezers array and preparation method - Google Patents
CTC enrichment and release system based on micro-fluidic and low-light-level tweezers array and preparation method Download PDFInfo
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Abstract
The invention discloses a CTC enrichment release system based on a micro-fluidic and micro-optical tweezers array and a preparation method thereof, wherein the system comprises a micro-fluidic chip provided with a micro-fluidic vortex system and a planar superlens optical tweezers array; the microfluidic vortex system comprises a plurality of microfluidic channels with a plurality of isosceles triangles, and when blood flows through the sudden expansion and sudden contraction areas, large-size cells are captured in the vortex of the triangular structure; the planar super-lens optical tweezer array comprises a plurality of rows of planar super-lenses with different focal lengths, and the optical tweezer array is formed to capture cells at different heights in the micro-channel. The invention can carry out multi-stage separation and capture on the circulating tumor cells in the whole blood, and realizes nondestructive controllable release, thereby realizing the enrichment and release of the circulating tumor cells with high purity, high activity and no damage. The invention also discloses a preparation method of the system, which is integrated and low in cost and can be applied to the fields of industry, medicine, biology and the like in a large scale.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a CTC enrichment and release system based on a micro-fluidic and low-light-level tweezer array and a preparation method thereof.
Background
Human blood contains abundant information. In early cancer, cells called Circulating Tumor Cells (CTCs) are removed from the original tumor tissue and circulated through the body's blood to various sites in the body. If the circulating tumor cells can be screened and accurately captured in the early stage of cancer, the method has more pertinence to the later-stage treatment of patients and improves the survival rate of the patients.
To date, there are several main methods for capturing circulating tumor cells: the method comprises a filtering technology based on different cell size differences, a capturing technology based on a microfluidic system, an antibody marking recognition technology and a cell separation technology based on a material surface micro-nano structure. However, the above methods have many disadvantages such as easy clogging of the channel by the cells, expensive apparatus, poor operability, low capturing efficiency, and complicated preparation process. In particular, only the drug screening aiming at the high-activity undamaged circulating tumor cells has clinical guidance significance, and the protein fragments obtained by the research of cell molecule typing are more complete, but the contact and the damage of the circulating tumor cells are inevitable in the traditional method. Therefore, the current capture of circulating tumor cells not only needs to meet the requirements of high purity and high efficiency, but also needs to meet the requirements of high activity and non-destructive release capture function.
The microfiltration technology based on cell size difference has strong operability and does not damage cell protein, but has low extraction purity due to similar physical characteristics of partial cells and tumor cells, and a microchannel filtering structure is easy to block, so that the capture efficiency and the capture precision are low. When the capture technology based on the microfluidic system captures the circulating tumor cells, nonspecific adsorption with other cells can occur, so that the capture purity is low, and the accuracy of subsequent cancer analysis is influenced. The cell separation technology based on the material surface wiener structure needs to use a pump as a driving force for filtration, has long detection time, low capture efficiency under high blood flow rate and complicated preparation process, and the method inevitably directly contacts circulating tumor cells, destroys the original form of the cells and greatly influences the analysis of the tumor cells. In contrast to physical property enrichment methods, biochemical marker affinity enrichment methods isolate target cells by protein biomarkers specifically expressed on the cell surface. The method has good biochemical specificity, can realize high-purity enrichment, but needs to modify the sample environment in advance, often only identifies specific cells, is difficult to capture circulating tumor cells with different characteristics, and simultaneously, the used instrument is high in price and is not suitable for large-scale community screening.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a CTC enrichment and release system based on a micro-fluidic and low-light-level tweezer array, which is used for performing multi-stage separation and capture on circulating tumor cells in whole blood and realizing nondestructive controllable release, thereby realizing the enrichment of the circulating tumor cells with high purity, high activity and no damage.
The invention also aims to provide a preparation method of the CTC enrichment and release system based on the micro-fluidic and the micro-optical tweezers array.
In order to achieve the purpose, the invention adopts the following technical scheme:
in one aspect of the invention, a CTC enrichment and release system based on a micro-fluidic and micro-optical tweezers array is provided, which comprises a micro-fluidic chip provided with a micro-fluidic vortex system and a planar super-lens optical tweezers array, and a light source device;
the microfluidic vortex system comprises a plurality of microfluidic channels connected with inlets of microfluidic chips, and each microfluidic channel comprises a plurality of sudden-expansion and sudden-shrinkage structures; the sudden expansion and sudden contraction structure is an isosceles triangle, a vertical bisector on the bottom side of the sudden expansion and sudden contraction structure is superposed with the microfluidic channel, and liquid is introduced from the bottom side to the top point; the outlets of all the micro-flow channels are converged and connected with the inlet of the micro-flow channel where the planar superlens optical tweezers array is positioned;
the planar super-lens optical tweezers array comprises a plurality of rows of planar super lenses, each row comprises a plurality of planar super lenses, and the row direction is vertical to the fluid flowing direction; the planar super lens comprises a plurality of circles of nano cylinders arranged in a concentric ring mode; the outlet of the microfluidic channel where the planar superlens optical tweezers array is positioned is connected with the outlet of the microfluidic chip;
the light source device provides parallel laser forming an array, the parallel laser corresponds to the planar superlens optical tweezers array one by one, and the parallel laser is normally incident to the planar superlens optical tweezers array.
As a preferred technical scheme, the microfluidic chip is made of SiO2The glass slide and the PDMS structure are bonded by a plasma bonding method.
As a preferable technical scheme, the diameter of the inlet and the outlet of the microfluidic chip is 300 μm.
As a preferred technical scheme, the diameter of a microfluidic channel of the microfluidic vortex system is 100 microns.
As a preferable technical scheme, the isosceles triangle of the sudden expansion and sudden contraction structure has the bottom side of 500 microns and the height of 500 microns, and the distance between two adjacent sudden expansion and sudden contraction structures in the same microfluidic channel is 700 microns.
As a preferred technical scheme, the sudden expansion and sudden shrinkage structures are arranged in a plurality of stages in the same microfluidic channel and are distributed in a staggered manner on different microfluidic channels.
As a preferable technical solution, the planar superlens optical tweezer array is periodically arranged in a group of three lines along the fluid flow direction, the focal lengths of the planar superlenses in each line are the same, and the focal lengths of the planar superlenses in each group of three lines are sequentially increased along the fluid flow direction, and are respectively 20 μm, 30 μm and 40 μm.
As a preferred technical scheme, the nano cylinder material is titanium dioxide; the height of the nano cylinder is 600 nm; the diameters of the nano cylinders in different circles are different, and the diameters are between 45nm and 280 nm; the distance between the ring and the next ring is 450 μm.
As a preferable technical solution, the light source device includes a laser light source and a spatial light modulator, the power of the laser light source is 50mW, the wavelength of the emitted laser light is 650nm, and the emitted laser light forms parallel laser light of an array after passing through the spatial light modulator.
In another aspect of the present invention, a method for preparing a CTC enrichment and release system based on a micro-fluidic and micro-optical tweezers array is provided, which is applied to the CTC enrichment and release system based on a micro-fluidic and micro-optical tweezers array, and comprises the following steps:
spin-coating an SU8 photoresist layer with the thickness of 50 microns on a silicon-based wafer, and forming a micro-channel SU8 male die through photoetching treatment;
preparing 10:1 of PDMS elastomer and curing agent mixed solution, uniformly stirring, removing bubbles in vacuum, pouring the mixture onto an SU8 male mold, heating and curing, peeling PDMS, and forming holes with the size of 300 mu m at the inlet and outlet respectively as the outlet and inlet of a microfluidic chip to obtain a PDMS layer having a flow channel cavity corresponding to the microfluidic vortex system and the planar superlens optical tweezers array structure;
in the clean SiO2Spin-coating an electron beam photoresist PMMA layer with the thickness of 1 mu m on the glass slide, and forming a plane figure with a required nano structure by electron beam lithography;
depositing TiO with the thickness of 800nm on the PMMA layer by an atomic layer deposition technology2Layer, removal of surface TiO2Layer and PMMA layer, keeping TiO with thickness of 600nm2A layer forming a planar superlens nanostructure;
adopting a plasma bonding method to bond the PDMS layer with the SiO carrying the planar super lens array2And (5) carrying out glass slide bonding.
Compared with the prior art, the invention has the following advantages and beneficial effects:
(1) according to the dual high-purity capture mode, cells with different sizes are separated through the multi-stage micro-scale vortex, circulating tumor cells are enriched and captured in the structural vortex, finally, secondary purification is carried out on the circulating tumor cells in the downstream section through the optical tweezer array, the circulating tumor cells in whole blood are subjected to multi-stage separation and capture, and capture efficiency can be improved;
(2) in the two-step combined capturing mode, the capturing and enriching modes of the upstream section according to the size difference and the downstream section according to the light absorption difference of the cells are both physical unmarked capturing modes, the releasing process only needs to turn off a light source, is more convenient than the capturing mode of the traditional filter screen, has no damage to the activity of the cells, does not need hydrolysis protease to separate the captured circulating tumor cells, avoids the contact damage in the capturing process and the change of protein indexes in the releasing process, is beneficial to the analysis and the detection of a late cell whole genome, and realizes the damage-free controllable release, thereby realizing the enrichment of the circulating tumor cells with high purity, high activity and no damage;
(3) the invention adopts the superlens array with high numerical aperture to realize the cell capture of the optical tweezers and integrates the superlens array with high numerical aperture into the microfluidic chip, thereby forming the technology of dual capture of CTCs.
Drawings
FIG. 1 is a schematic diagram of the overall structure of a CTC enrichment and release system based on microfluidics and a micro-optical tweezers array according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of a sudden-expansion and sudden-contraction microfluidic channel structure and its specific dimensions according to an embodiment of the present invention;
fig. 3 is a schematic diagram of a surface structure of a planar superlens optical tweezers array in a microfluidic channel according to an embodiment of the present invention, where fig. 3(a) is an overall top view of the planar superlens optical tweezers array in the microfluidic channel; FIG. 3(b) is an overall side view of a planar superlens optical tweezer array in a microfluidic channel; FIG. 3(c) is a top plan view of a planar superlens; FIG. 3(d) is a schematic perspective view of a single nanocylinder;
FIG. 4 is a flow chart of multi-stage separation and capture of circulating tumor cells in whole blood according to an embodiment of the present invention, wherein FIG. 4(a) is a schematic diagram of primary separation and enrichment of circulating tumor cells in a microfluidic channel; fig. 4(b) is a schematic diagram of releasing circulating tumor cells in a microfluidic vortex system into downstream for capture by optical tweezers; FIG. 4(c) is a schematic representation of further purification of circulating tumor cells;
fig. 5 is a schematic diagram of a preparation method of a CTC enrichment release system based on microfluidics and a micro-optical tweezers array in an embodiment of the invention.
The reference numbers illustrate: 1. a microfluidic chip inlet; 2. a microfluidic channel; 3. a sudden expansion and contraction structure; 4. a planar superlens; 5. a microfluidic chip outlet; 6. small size leukocytes; 7. red blood cells; 8. large size leukocytes; 9. circulating tumor cells; 10. parallel laser; 11. SiO 22A glass slide; 12. a PMMA photoresist layer; 13. TiO 22A layer; 14. a silicon-based wafer; 15. SU8 photoresist layer; 16. a PDMS layer.
Detailed Description
In order to make the technical solutions better understood by those skilled in the art, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. It is to be understood that the embodiments described are only a few embodiments of the present application and not all 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 application.
Examples
Optical tweezers are a device in which focused light can exert a force on particles near a focal point, and the force is comprehensively expressed by attracting and stabilizing the particles at the focal point as if the light is used as tweezers to capture the particles. It features no mechanical contact and low absorption of light to complete the non-damage capture of cell. When the light source is turned off, the particles or cells can be released again freely. The conventional optical tweezers trapping is generally based on a high-cost, large-volume optical lens and a complex control system, and the multi-focus trapping function is difficult to realize.
The planar superlens is a micro-nano structure array prepared on a planar substrate (such as a glass slide) by a micro-nano photoetching process on the planar substrate, each micro-nano structure is smaller than the wavelength of incident light for research, and the light field at the point can be modulated. The combined array can flexibly change the transmission or reflection characteristics of light according to the artificial design of the micro-nano structure, wherein the light is converged into one point, and the wave front of the light can be flexibly regulated and controlled (such as single-point or multi-point focusing). Unlike conventional lenses that rely on an arcuate thickness, planar superlenses macroscopically behave as planar structures.
As shown in fig. 1, the present embodiment provides a CTC enrichment and release system based on microfluidics and micro-optical tweezers array, including a microfluidic chip provided with a microfluidic vortex system and a planar superlens optical tweezers array, and a light source device;
the micro-fluidic chip is divided into two-stage structures, the first-stage structure is a micro-fluidic vortex system, and the second-stage structure is a planar superlens optical tweezers array;
further, the microfluidic chip is made of SiO2The glass slide is bonded with the PDMS structure.
Furthermore, the diameter of the inlet and the outlet of the microfluidic chip is 300 μm.
Further, the diameter of the microfluidic channel 2 of the microfluidic vortex system is 100 μm, as shown in fig. 2.
(1) The microfluidic vortex system comprises a plurality of microfluidic channels 2 connected with a microfluidic chip inlet 1, and each microfluidic channel 2 comprises a plurality of sudden-expansion and sudden-shrinkage structures 3; the sudden expansion and sudden contraction structure 3 is an isosceles triangle, a vertical bisector on the bottom side of the structure is superposed with the microfluidic channel 2, and liquid is introduced along the direction from the bottom side to the top point; the outlets of all the micro-flow channels 2 are converged and connected with the inlet of the planar superlens optical tweezers array;
furthermore, the isosceles triangle of the sudden-expansion and sudden-contraction structure 3 has a bottom side of 500 μm and a height of 500 μm, and the distance between two adjacent sudden-expansion and sudden-contraction structures 3 in the same microfluidic channel 2 is 700 μm, as shown in fig. 2.
Furthermore, the sudden expansion and sudden contraction structures 3 are arranged in a plurality of stages in the same microfluidic channel 2 and are distributed in a staggered manner on different microfluidic channels 2.
(2) As shown in fig. 3, the planar superlens optical tweezer array includes a plurality of rows of planar superlenses 4, each row includes a plurality of planar superlenses 4, and the row direction is perpendicular to the fluid flow direction; the planar superlens 4 comprises a plurality of circles of concentric nano cylinders; the outlet of the planar superlens optical tweezers array is connected with the outlet 5 of the microfluidic chip;
further, as shown in fig. 3(a) and (b), the planar superlens optical tweezer array is periodically arranged in three rows along the fluid flow direction, the focal lengths of the planar superlenses 4 in each row are the same, and the focal lengths of the planar superlenses 5 in each group and three rows are sequentially increased along the fluid flow direction, and are respectively 20 μm, 30 μm and 40 μm.
Further, in the planar superlens optical tweezers array, the nano cylinder is made of titanium dioxide, the height of the nano cylinder is 600nm, the diameters of the nano cylinders in different circles are different and are shown in fig. 3(c) and (d), the diameter di is 45nm-280nm, and the specific parameters are shown in table 1(a), table 1(b) and table 1 (c); the concentric circles are spaced 450 μm apart.
TABLE 1(a) diameter parameters of each turn of nanocylinder of a planar superlens with a focal length of 20 μm
TABLE 1(b) diameter parameters of each turn of nanocylinder of a planar superlens with a focal length of 30 μm
TABLE 1(c) diameter parameters of each turn of nanocylinder of a planar superlens with a focal length of 40 μm
Furthermore, based on the idea of the invention, the size of the micro-nano structure can be changed, and the planar superlens can be designed according to the requirements of different focal lengths.
(3) The light source device provides parallel lasers 10 forming an array, the parallel lasers correspond to the planar superlens optical tweezers array one by one, and the parallel lasers are normally incident to the planar superlens optical tweezers array.
Further, the light source device comprises a laser light source and a spatial light modulator, the power of the laser light source is 50mW, the wavelength of the emitted laser light is 650nm, and the emitted laser light forms parallel laser light 10 of an array after passing through the spatial light modulator.
In order to make persons skilled in the art better understand the scheme of the present application, in the following, a CTC enrichment and release system based on microfluidics and micro-optical tweezers array in the embodiment of the present application is combined to perform multistage separation capture on circulating tumor cells 9 in whole blood and achieve nondestructive controllable release, as shown in fig. 4, including the following steps:
s1, preparation of blood samples: approximately 200 and 700 unequal circulating tumor cells 9 were added to 1mL of blood, and the blood was diluted 5X fold with PBS.
S2, separating the circulating tumor cells 9 in the microfluidic channel: a blood sample is first introduced into the microfluidic channel 2 at a high flow rate of 500. mu.l/hr. When the cells do Poiseuille shear flow in the straight channel along with the blood, the cells are moved laterally to the stress balance position by the inertial lift force and the wall effect, and when the blood flows through the triangular area of the sudden expansion and sudden contraction structure 3, based on the research mechanism that the fluid can induce the secondary vortex in the curved channel with high curvature and low Reynolds number, the circulating tumor cells 9 with larger size and a small part of white blood cells 8 with larger size can be captured in the triangular area of the sudden expansion and sudden contraction structure 3, while the red blood cells 7 with smaller size and the white blood cells 6 with most small size flow out of the channel along with the blood, as shown in FIG. 4 (a). After completion, the effect of the separation of the circulating tumor cells 9 in the region of the knob 3 can be observed under a microscope.
S3, capture of circulating tumor cells 9 in planar superlens array: and starting incident laser of the planar superlens array, wherein the light source is 650nm laser, the power is 50mW, the laser forms parallel laser 10 of the array after passing through the spatial light modulator, the parallel laser is normally incident to the corresponding planar superlens 4 substrate one by one, and the parallel laser penetrates through the planar superlens 4 substrate to form the optical tweezers potential well array. The planar super lens array is divided into a plurality of groups, each group has a plurality of planar super lenses 4 with three different focal lengths, the focal lengths are respectively 20, 30 and 40 mu m, and the planar super lens array is convenient for capturing circulating tumor cells with different heights in a flow channel. After determining the capture of the circulating tumor cells 9 in the sudden expansion and contraction structure 3 of the microfluidic channel 2 at S2, the flow rate of the introduced PBS solution is reduced to 100 μ l/hr, and the circulating tumor cells 9 and a small amount of participating large-sized white blood cells 8 are discharged from the region of the sudden expansion and contraction structure 3, enter the region of the planar superlens 4 and are captured by the optical tweezers, as shown in fig. 4 (b).
S4, observing and distinguishing the large-sized white blood cells 8 from the circulating tumor cells 9 under the microscope, by adjusting the spatial light modulator, the incident light of the super-surface optical tweezers capturing the large-sized white blood cells 8 is turned off, thereby releasing the large-sized white blood cells 8, leaving only the circulating tumor cells 9, and realizing further purification of the circulating tumor cells 9, as shown in fig. 4 (c).
And S5, after the large-size white blood cells 8 are completely released, the laser light source is turned off, and the circulating tumor cells 9 separated twice are released.
In another aspect of this embodiment, a method for preparing a CTC enrichment and release system based on microfluidics and micro-optical tweezers array is also provided, as shown in fig. 5, including the following steps:
(1) preparing a micro channel: the micro-channel preparation material adopts PDMS (polydimethylsiloxane) with good biocompatibility and light transmittance, and the substrate adopts SiO which can be bonded with the PDMS and can be used for planar superlens growth2 A slide glass 11;
a. casting: an SU8 photoresist layer 15(MicroChem, USA) with the thickness of 50 microns is spin-coated on the silicon-based wafer 14, and a micro-channel SU8 photoresist layer 15 is formed as a male die after photoetching;
b. molding: preparing 10:1 of PDMS elastomer and curing agent mixed solution, uniformly stirring, removing bubbles in vacuum, pouring the mixture onto a male mold formed by an SU8 photoresist layer 15, heating and curing, peeling off the PDMS layer 16, and forming holes with the size of 300 mu m at an inlet and an outlet respectively as an outlet and an inlet of a microfluidic chip to obtain the PDMS layer 16 with a flow channel cavity corresponding to a microfluidic vortex system and a planar superlens optical tweezers array structure;
(2) preparing a planar superlens: in the clean SiO2A PMMA photoresist layer 12 (polymethyl methacrylate) with the thickness of 1 mu m is coated on the glass slide 11 in a spin mode, and a plane figure with a required nano structure is formed through electron beam lithography; deposition of TiO on a substrate to a thickness of 800nm by atomic layer deposition2Layer 13, surface layer of TiO abraded2 Layer 13 and PMMA photoresist layer 12, with retention of TiO 600nm thick2Layer 13 forming a planar superlens nanostructure;
3) integration of microfluidic-metamaterial: bonding the PDMS layer 16 with the SiO carrying the planar superlens array by plasma bonding2The slide 11 is bonded.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. The CTC enrichment and release system based on the micro-fluidic and micro-optical tweezers array is characterized by comprising a micro-fluidic chip provided with a micro-fluidic vortex system and a planar super-lens optical tweezers array, and a light source device;
the microfluidic vortex system comprises a plurality of microfluidic channels connected with inlets of microfluidic chips, and each microfluidic channel comprises a plurality of sudden-expansion and sudden-shrinkage structures; the sudden expansion and sudden contraction structure is an isosceles triangle, a vertical bisector on the bottom side of the sudden expansion and sudden contraction structure is superposed with the microfluidic channel, and liquid is introduced from the bottom side to the top point; the outlets of all the micro-flow channels are converged and connected with the inlet of the micro-flow channel where the planar superlens optical tweezers array is positioned;
the planar super-lens optical tweezers array comprises a plurality of rows of planar super lenses, each row comprises a plurality of planar super lenses, and the row direction is vertical to the fluid flowing direction; the planar super lens comprises a plurality of circles of nano cylinders arranged in a concentric ring mode; the outlet of the microfluidic channel where the planar superlens optical tweezers array is positioned is connected with the outlet of the microfluidic chip;
the light source device provides parallel laser forming an array, the parallel laser corresponds to the planar superlens optical tweezers array one by one, and the parallel laser is normally incident to the planar superlens optical tweezers array.
2. The micro-fluidic and micro-optical tweezers array-based CTC (CTC concentration release system) according to claim 1, wherein the micro-fluidic chip is SiO2The glass slide and the PDMS structure are bonded by a plasma bonding method.
3. The micro-fluidic and micro-optical tweezers array based CTC enrichment release system of claim 1, wherein the micro-fluidic chip inlet and outlet diameters are 300 μm.
4. The micro-fluidic and micro-optical tweezers array based CTC enrichment release system of claim 1, wherein the micro-fluidic channel diameter of the micro-fluidic vortex system is 100 μm.
5. A CTC enrichment release system based on microfluidics and micro tweezers array according to claim 1, wherein the isosceles triangle of the sudden expansion and sudden shrinkage structure has a bottom side length of 500 μm and a height of 500 μm, and the distance between two adjacent sudden expansion and sudden shrinkage structures in the same microfluidic channel is 700 μm.
6. The CTC enrichment release system based on microfluidics and micro-optical tweezers array according to claim 1, wherein the sudden expansion and sudden shrinkage structures are arranged in multiple stages in the same microfluidic channel and are distributed in a staggered manner on different microfluidic channels.
7. A CTC enrichment release system according to claim 1, wherein the planar superlens optical tweezers array is arranged in a group of three rows along the fluid flow direction, the focal length of the planar superlens in each row is the same, and the focal length of the planar superlens in each group of three rows increases sequentially along the fluid flow direction, and is 20 μm, 30 μm, and 40 μm respectively.
8. The micro-fluidic and micro-optical tweezers array based CTC enrichment release system of claim 1, wherein the nano cylinder material is titanium dioxide; the height of the nano cylinder is 600 nm; the diameters of the nano cylinders in different circles are different, and the diameters are between 45nm and 280 nm; the distance between the ring and the next ring is 450 μm.
9. A CTC enrichment release system according to claim 1, wherein the light source device comprises a laser source with 50mW power and 650nm wavelength of outgoing laser, and a spatial light modulator, and the outgoing laser passes through the spatial light modulator to form parallel laser of the array.
10. The preparation method of the CTC enrichment and release system based on the micro-fluidic and micro-optical tweezers array is characterized by being applied to the CTC enrichment and release system based on the micro-fluidic and micro-optical tweezers array in any one of claims 1 to 9, and comprising the following steps:
spin-coating an SU8 photoresist layer with the thickness of 50 microns on a silicon-based wafer, and forming a micro-channel SU8 male die through photoetching treatment;
preparing 10:1 of PDMS elastomer and curing agent mixed solution, uniformly stirring, removing bubbles in vacuum, pouring the mixture onto an SU8 male mold, heating and curing, peeling PDMS, and forming holes with the size of 300 mu m at the inlet and outlet respectively as the outlet and inlet of a microfluidic chip to obtain a PDMS layer having a flow channel cavity corresponding to the microfluidic vortex system and the planar superlens optical tweezers array structure;
in the clean SiO2Spin-coating an electron beam photoresist PMMA layer with the thickness of 1 mu m on the glass slide, and forming a plane figure with a required nano structure by electron beam lithography;
depositing TiO with the thickness of 800nm on the PMMA layer by an atomic layer deposition technology2Layer, removal of surface TiO2Layer and PMMA layer, keeping TiO with thickness of 600nm2A layer forming a planar superlens nanostructure;
adopting a plasma bonding method to bond the PDMS layer with the SiO carrying the planar super lens array2And (5) carrying out glass slide bonding.
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