CN114875378A - Preparation method of cell screening device with vertical structure tungsten diselenide array thin film structure - Google Patents
Preparation method of cell screening device with vertical structure tungsten diselenide array thin film structure Download PDFInfo
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- ROUIDRHELGULJS-UHFFFAOYSA-N bis(selanylidene)tungsten Chemical compound [Se]=[W]=[Se] ROUIDRHELGULJS-UHFFFAOYSA-N 0.000 title claims abstract description 30
- 238000012216 screening Methods 0.000 title claims abstract description 24
- 239000010409 thin film Substances 0.000 title claims description 17
- 238000002360 preparation method Methods 0.000 title description 10
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 claims abstract description 26
- 238000000034 method Methods 0.000 claims abstract description 22
- 239000000758 substrate Substances 0.000 claims abstract description 20
- 238000004519 manufacturing process Methods 0.000 claims abstract description 15
- 239000004408 titanium dioxide Substances 0.000 claims abstract description 13
- 230000005684 electric field Effects 0.000 claims abstract description 7
- 230000008901 benefit Effects 0.000 claims abstract description 6
- 238000005229 chemical vapour deposition Methods 0.000 claims abstract description 4
- 238000001962 electrophoresis Methods 0.000 claims abstract 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 20
- 239000010453 quartz Substances 0.000 claims description 18
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 18
- 238000007873 sieving Methods 0.000 claims description 15
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 claims description 14
- 239000011261 inert gas Substances 0.000 claims description 12
- 238000010438 heat treatment Methods 0.000 claims description 11
- 239000001257 hydrogen Substances 0.000 claims description 11
- 229910052739 hydrogen Inorganic materials 0.000 claims description 11
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims description 10
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 10
- 239000010408 film Substances 0.000 claims description 10
- 239000011780 sodium chloride Substances 0.000 claims description 10
- QGLKJKCYBOYXKC-UHFFFAOYSA-N nonaoxidotritungsten Chemical compound O=[W]1(=O)O[W](=O)(=O)O[W](=O)(=O)O1 QGLKJKCYBOYXKC-UHFFFAOYSA-N 0.000 claims description 9
- 229910001930 tungsten oxide Inorganic materials 0.000 claims description 9
- 239000011810 insulating material Substances 0.000 claims description 7
- 230000007704 transition Effects 0.000 claims description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- 229910052786 argon Inorganic materials 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 238000005086 pumping Methods 0.000 claims description 2
- 238000004544 sputter deposition Methods 0.000 abstract description 6
- 230000001575 pathological effect Effects 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 3
- 210000004027 cell Anatomy 0.000 description 53
- 239000010410 layer Substances 0.000 description 15
- 238000000926 separation method Methods 0.000 description 3
- 230000009182 swimming Effects 0.000 description 3
- 229910010413 TiO 2 Inorganic materials 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 239000003292 glue Substances 0.000 description 2
- 125000006850 spacer group Chemical group 0.000 description 2
- ZNOKGRXACCSDPY-UHFFFAOYSA-N tungsten trioxide Chemical compound O=[W](=O)=O ZNOKGRXACCSDPY-UHFFFAOYSA-N 0.000 description 2
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000002900 effect on cell Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- 239000011229 interlayer Substances 0.000 description 1
- 238000001755 magnetron sputter deposition Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002135 nanosheet Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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- C23—COATING METALLIC MATERIAL; COATING MATERIAL WITH METALLIC MATERIAL; CHEMICAL SURFACE TREATMENT; DIFFUSION TREATMENT OF METALLIC MATERIAL; COATING BY VACUUM EVAPORATION, BY SPUTTERING, BY ION IMPLANTATION OR BY CHEMICAL VAPOUR DEPOSITION, IN GENERAL; INHIBITING CORROSION OF METALLIC MATERIAL OR INCRUSTATION IN GENERAL
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- C23C16/00—Chemical coating by decomposition of gaseous compounds, without leaving reaction products of surface material in the coating, i.e. chemical vapour deposition [CVD] processes
- C23C16/22—Chemical coating by decomposition of gaseous compounds, without leaving reaction products of surface material in the coating, i.e. chemical vapour deposition [CVD] processes characterised by the deposition of inorganic material, other than metallic material
- C23C16/30—Deposition of compounds, mixtures or solid solutions, e.g. borides, carbides, nitrides
- C23C16/305—Sulfides, selenides, or tellurides
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- C23C16/00—Chemical coating by decomposition of gaseous compounds, without leaving reaction products of surface material in the coating, i.e. chemical vapour deposition [CVD] processes
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Abstract
The invention provides a method for manufacturing a cell screening device with a vertical structure tungsten diselenide array film structure, which comprises the steps of sputtering a layer of titanium dioxide on a customized conductive substrate, and preparing a vertically-grown tungsten diselenide film by using a chemical vapor deposition method; then symmetrically fixing the two conductive substrates at a certain interval; and then, the structural advantages of the vertical tungsten diselenide three-dimensional sheet are utilized, and the electrophoresis phenomenon of the cells caused by an external electric field is combined, so that the cells are effectively screened and separated. The cell screening device prepared by the method can screen different pathological states of different types of cells and the same type of cells, does not need to mark the cells, and ensures the activity of the cells.
Description
Technical Field
The invention belongs to the technical field of biomedicine, and particularly relates to a preparation method of a cell screening device with a vertical structure tungsten diselenide array thin film structure.
Background
Cells are the basic building blocks of each living body structure, and are the research foundation and the focus in the fields of life science and biomedicine. In the research process, the first thing to do is to screen different kinds of cells and different pathological states of the same kind of cells. Therefore, cell sorting technology is crucial in this field of research.
The cell screening and separation through the cell screen is a simple and effective screening mode, and a plurality of cell screens with different pore sizes and different materials are produced on the market at present so as to screen cells with different sizes to obtain single cell suspension. However, the cell can be screened only from the aspect of cell size by simply passing through a cell screen. It is difficult to further screen the same kind of cells in different pathological states and meet some special screening requirements.
Flow cytometry is another technique for rapid and accurate screening of cells. It is irradiated by laser beam, and utilizes sensor to detect scattered light and fluorescence excited by cell, if the target cell signal is detected, the cell can be transferred into storage pool by means of controlling cell flow path. This technique can be used for screening single cells with high accuracy and has a high separation purity, but has a problem. First, laser irradiation of fluorescent labels has a negative effect on cell viability. Second, some markers are difficult to match into cells and have false positive errors. In addition, the technical equipment is complex, high in cost, time-consuming and labor-consuming.
Disclosure of Invention
Based on the problems in the prior art, the invention provides a preparation method of a cell screening device with a vertical structure tungsten diselenide array thin film structure.
According to the technical scheme of the invention, the preparation method of the cell screening device with the vertical structure tungsten diselenide array thin film structure is providedMethod for preparing vertically grown WSe by chemical vapor deposition by sputtering a layer of titanium dioxide on a customized conductive substrate 2 A film; then symmetrically fixing the two conductive substrates at a certain interval; it utilizes a vertical WSe 2 The three-dimensional sheet structure advantage combines the swimming phenomenon that outside electric field made the cell take place, can effectually screen the separation to the cell.
Further, the manufacturing method of the cell sieving device with the vertical structure tungsten diselenide array thin film structure comprises the following steps:
step S1, pre-sputtering a layer of TiO on the conductive substrate 2 As a growth transition layer.
Step S2, mixing tungsten oxide and sodium chloride according to the ratio of 5:1, putting the mixture into a quartz boat, and putting the substrate TiO 2 The layer is downwards arranged at the position above the powder on the quartz boat;
step S3, placing the selenium powder and the quartz boat in different temperature areas of the tube furnace, firstly vacuumizing the furnace chamber, preferably to a vacuum degree within 1mTorr, then introducing inert gas to atmospheric pressure and keeping a flow of 200 sccm;
step S4, heating the temperature zone of selenium powder to 150 ℃, heating the temperature zone of quartz boat to 1100 ℃, and introducing hydrogen to grow vertical WSe 2 。
Step S5, after the growth is finished, the hydrogen is closed, and after the furnace chamber is cooled to room temperature in the inert gas environment, the substrate sample is taken out;
step S6, WSe of two-piece sample 2 The layers were facing each other, and the two samples were separated by a gap of 25 μm on both sides by an insulating material.
Wherein, TiO in step S1 2 The thickness of the transition layer is 5nm-20 nm. In the step S2, the mass of tungsten oxide is 10mg-200mg, the mass of sodium chloride is 0.5mg-200mg, and the mass ratio of tungsten oxide to sodium chloride is 1: 1-20: 1. In step S4, the heating temperature of the temperature zone where the selenium powder is located is 100-400 ℃. The heating temperature of the temperature zone of the quartz boat is 900-1100 ℃.
Further, WSe in step S4 2 The growth time is 1-30 minutes. In step S6, the gap between the two samples is 20-30 μm.
Preferably, the hydrogen flow rate is 1sccm to 200 sccm.
More preferably, the mass of the selenium powder in step S3 is 10mg-300mg, the inert gas refers to one or more of argon, nitrogen or other inert gases, and the flow rate is maintained at 1 sccm-300 sccm.
Compared with the prior art, the preparation method of the cell screening device with the vertical structure tungsten diselenide array thin film structure has the beneficial effects that:
1. the cell screening device prepared by the method can screen different pathological states of different types of cells and the same type of cells, does not need to mark the cells, and ensures the activity of the cells.
2. The cell screening device prepared by the method has high flux, high screening speed and wide application range.
3. The cell screening device prepared by the method has the advantages of simple structure, convenient operation and low cost.
Drawings
Fig. 1 is a schematic cross-sectional structure view of a cell sieving device with a vertical structure tungsten diselenide array thin film structure according to the present invention.
The reference numbers in the drawings are: conductive substrate 1, vertical structure WSe 2 2, an insulating material spacer 3.
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. It is to be understood that the described embodiments are merely a few embodiments of the invention, 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 invention. Additionally, the scope of the present invention should not be limited to the particular structures or components or the particular parameters described below.
In the description of the present invention, it is to be understood that the terms "upper", "lower", "front", "rear", "left", "right", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, are merely for convenience in describing the present invention and simplifying the description, and do not indicate or imply that the device or component being referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, should not be construed as limiting the present invention.
The invention discloses tungsten diselenide (WSe) with a vertical structure 2 ) The preparation method of the cell sieving device with the array film structure comprises the step of sputtering a layer of titanium dioxide (TiO) on a customized conductive substrate 2 ) Preparation of vertically grown WSe by chemical vapor deposition 2 A film; then, the two conductive substrates are symmetrically fixed at a certain interval. The present invention utilizes a vertical WSe 2 The three-dimensional sheet structure advantage is combined with the external electric field to enable the swimming phenomenon of the cells to occur, so that the cells can be effectively screened and separated; the invention has the obvious advantages of effective method, simple structure, lower cost, wide application range and the like.
The invention relates to tungsten diselenide (WSe) with a vertical structure 2 ) The preparation method of the cell screening device with the array film structure is based on the following working principle: setting the cell sieving device at certain intervals, firstly applying proper voltage on the cell sieving device, and generating a response electric field at the middle interval; then, a cell suspension with a certain pH value passes through the cell screening device at a constant speed, and cells can generate a swimming phenomenon under the action of an external electric field. The invention utilizes the fact that the cell surface has net positive or negative charges under a certain PH value, and different cells or the same cell have certain difference of the positive or negative charges and the charge quantity under different physiological states and pathological states; this results in differential deflection of different cell types under the influence of the electric field of the sieving device, the present invention re-uses WSe 2 The vertical structure of the nanosheets fixes the cells, so that the screening effect on the cells is realized.
Further, the invention relates to tungsten diselenide (WSe) with a vertical structure 2 ) The manufacturing method of the cell sieving device with the array film structure comprises the following steps:
step S1, pre-sputtering a layer of TiO on the conductive substrate 2 As a growth transition layer.
Step S2, mixing tungsten oxide and sodium chloride according to the ratio of 10:1, putting the mixture into a quartz boat, and putting the substrate TiO 2 The layer is downwards arranged at the position above the powder on the quartz boat;
step S3, placing the selenium powder and the quartz boat in different temperature areas of the tube furnace, firstly, vacuumizing the furnace chamber, wherein the vacuum degree is preferably within 1mTorr, then introducing inert gas to the atmospheric pressure, and keeping the flow of 100sccm to stabilize the growth environment;
step S4, heating the temperature zone of selenium powder to 100 ℃, heating the temperature zone of the quartz boat to 1000 ℃, and introducing hydrogen for 10 minutes to grow vertical WSe 2 。
Step S5, after the growth is finished, the hydrogen is closed, and after the furnace chamber is cooled to room temperature in the inert gas environment, the substrate sample is taken out;
step S6, WSe of two-piece sample 2 The layers were opposed, and the two samples were separated by a gap of 20 μm by an insulating material on both sides so that the cell fluid flowed therebetween.
Wherein, TiO in step S1 2 The thickness of the transition layer is 5nm-20 nm; in the step S2, the mass of tungsten oxide is 10mg-200mg, the mass of sodium chloride is 0.5mg-200mg, and the mass ratio of tungsten oxide to sodium chloride is 1: 1-20: 1. In the step S3, the selenium powder has a mass of 10-300 mg, the inert gas refers to one or more of argon, nitrogen or other inert gases, and the flow rate is kept at 1-300 sccm.
Further, the temperature zone of the selenium powder in the step S4 is heated to 100-400 ℃. The heating temperature of the temperature zone of the quartz boat is 900-1100 ℃. The hydrogen flow rate is 1sccm to 200 sccm. WSe in step S4 2 The growth time is 1-30 minutes. The insulating material in step S6 includes but is not limited to insulating glue, heat sealing film, etc.; the gap between the two samples is 20-30 μm.
The preparation method of the invention is explained in detail below with reference to the attached figure 1:
vertical Structure WSe in embodiment 1 2 The invention relates to a method for manufacturing a cell screening device with an array film structure, in particular to a vertical structure WSe 2 Array film structure cell screening deviceThe further refinement of the manufacturing method comprises the following steps:
step W1, pre-sputtering a layer of TiO with the thickness of 20nm on the conductive substrate 1 in a magnetron sputtering mode 2 A transition layer;
step W2, weighing 150mg of tungsten trioxide and 15mg of sodium chloride powder, uniformly mixing, and putting into a quartz boat; taking a silicon chip as a conductive substrate to be buckled upside down above the powder;
and step W3, placing the selenium powder and the quartz boat into two different temperature zones of the tube furnace. Firstly, pumping a tube furnace to a vacuum state, wherein the vacuum degree is preferably within 1mTorr, then introducing argon to the conventional atmospheric pressure, and keeping the flow of 120 sccm;
step W4, heating temperature zones of selenium powder to 70 ℃, heating temperature zones of quartz boat to 850 ℃, introducing hydrogen with the flow of 40sccm, and growing for 5 minutes; after the reaction is finished, closing the hydrogen, and waiting for the furnace chamber to be cooled to the room temperature;
step W5, argon is closed, and the well grown WSe is taken out after the furnace chamber is cooled to room temperature 2 A sample;
step W6, vertical structures WSe of two samples 2 The layers were facing each other, and the two samples were separated by a 20 μm gap by an insulating material spacer 3 on both sides and fixed with a jig. The insulating material interlayer is preferably insulating glue.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. It will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the present invention as defined by the following claims.
Claims (10)
1. A method for manufacturing a cell screening device with a vertical structure tungsten diselenide array film structure is characterized in that a layer of titanium dioxide is sputtered on a customized conductive substrate, and a vertically-grown tungsten diselenide film is prepared by a chemical vapor deposition method; then symmetrically fixing the two conductive substrates at a certain interval; and then, the structural advantages of the vertical tungsten diselenide three-dimensional sheet are utilized, and the electrophoresis phenomenon of the cells caused by an external electric field is combined, so that the cells are effectively screened and separated.
2. The method for manufacturing the cell sieving device with the vertical structure of the tungsten diselenide array thin film structure as claimed in claim 1, wherein the method comprises the following steps:
step S1, a layer of titanium dioxide is pre-sputtered on the conductive substrate to be used as a growth transition layer.
Step S2, mixing tungsten oxide and sodium chloride according to a ratio of 15:1, putting the mixture into a quartz boat, and putting a substrate titanium dioxide layer downwards at a position above powder on the quartz boat;
step S3, placing the selenium powder and the quartz boat in different temperature areas of the tube furnace respectively, firstly pumping the furnace chamber to vacuum with the vacuum degree within 1mTorr, then introducing inert gas to atmospheric pressure and keeping 150 sccm;
and step S4, heating the temperature zone of the selenium powder to 200 ℃, heating the temperature zone of the quartz boat to 1200 ℃, and introducing hydrogen to grow the vertical tungsten diselenide.
Step S5, after the growth is finished, the hydrogen is closed, and after the furnace chamber is cooled to room temperature in the inert gas environment, the substrate sample is taken out;
step S6, the tungsten diselenide layer surfaces of the two samples are opposite, and the two samples are separated by a gap of 30 μm on two sides through insulating materials.
3. The method for manufacturing a cell sieving device with a vertical structure tungsten diselenide array thin film structure as claimed in claim 2, wherein the thickness of the titanium dioxide transition layer in the step S1 is 5nm to 20 nm.
4. The manufacturing method of the cell screening device with the vertical structure tungsten diselenide array thin film structure as claimed in claim 2, wherein in the step S2, the mass of tungsten oxide is 10mg-200mg, the mass of sodium chloride is 0.5mg-200mg, and the mass ratio of tungsten oxide to sodium chloride is 1: 1-20: 1.
5. The method for manufacturing the cell sieving device with the vertical structure tungsten diselenide array thin film structure according to claim 3, wherein the temperature zone where the selenium powder is located in the step S4 is heated to 100-400 ℃.
6. The method for manufacturing a cell sieving device with a vertical structure of tungsten diselenide array thin film structure as claimed in claim 3, wherein the quartz boat is heated at 900-1100 ℃ in the temperature zone.
7. The method for manufacturing a cell sieving device with a vertical structure tungsten diselenide array thin film structure as claimed in claim 2, wherein the growth time of tungsten diselenide in step S4 is 1 minute to 30 minutes.
8. The method for manufacturing a cell sieving device with a vertical structure of tungsten diselenide array thin film structure as claimed in claim 2, wherein the gap between two tungsten diselenide samples is 20 μm to 30 μm.
9. The method for manufacturing a cell sieving device with vertical structure tungsten diselenide array thin film structure as claimed in claim 4, wherein the hydrogen flow rate is 1sccm to 200 sccm.
10. The method for manufacturing a cell sieving device with a vertical structure tungsten diselenide array thin film structure as claimed in claim 3, wherein the mass of the selenium powder in step S3 is 10mg-300mg, the inert gas refers to one or more of argon, nitrogen or other inert gases, and the flow rate is maintained at 1 sccm-300 sccm.
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