CN108786941B - Preparation method of novel magnetic self-assembly three-dimensional paper chip - Google Patents
Preparation method of novel magnetic self-assembly three-dimensional paper chip Download PDFInfo
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- CN108786941B CN108786941B CN201810567189.0A CN201810567189A CN108786941B CN 108786941 B CN108786941 B CN 108786941B CN 201810567189 A CN201810567189 A CN 201810567189A CN 108786941 B CN108786941 B CN 108786941B
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
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Abstract
The invention relates to a preparation method of a three-dimensional paper chip, in particular to a preparation method of a novel magnetic self-assembly three-dimensional paper chip, which comprises the following steps: hydrophobic magnetic ink is adopted, and a channel layer, a pretreatment reagent layer and a detection layer of the paper chip are respectively obtained through screen printing; the pretreatment agent is screen printed on the pretreatment agent layer by a patterned printing agent screen by a scraper. Printing a detection medicament on a detection area on the detection layer through a patterned medicament screen by using a scraper; and stacking the channel layer, the pretreatment reagent layer and the detection layer on a gasket made of a magnetic material in sequence to obtain the three-dimensional paper chip. The preparation method of the novel magnetic self-assembly three-dimensional paper chip provided by the invention adopts a screen printing technology, and by improving the printing material, no additional adhesive layer is adopted, and no glue spraying or printing step is required, so that the magnetic material and the hydrophobic material are printed simultaneously, the preparation step is simplified, and the risk of blocking a hydrophilic channel by glue solution permeation is eliminated.
Description
Technical Field
The invention relates to a preparation method of a three-dimensional paper chip, in particular to a preparation method of a novel magnetic self-assembly three-dimensional paper chip.
Background
The microfluidic paper chip is a novel analytical device which is processed into a fluid channel network on a paper substrate or a paper-based material by water conveying or hydrophilic modification and can be used for biological and chemical detection and analysis, and is also called a micro total analysis laboratory on paper. The micro-fluidic chip has the advantages of small volume, low reagent consumption, easy carrying and the like of the traditional micro-fluidic chip, has the advantages of simple preparation, low cost, good biocompatibility and the like, is a novel micro-analysis technology platform which is developed in recent years, and has good application prospect in the fields of disease diagnosis, pathogen detection, food safety, environmental quality monitoring and the like.
The 3D paper chip is obtained by superposing 2D paper chips. The 3D paper chip has a three-dimensional network channel structure, so that the 3D paper chip has certain characteristics superior to those of the 2D paper chip. 1) Rapid transport of fluid because the path in the Z-direction is much shorter than the path of the channel in the x, y-plane; 2) high throughput assay in a single chip device, such as 16 assays performed simultaneously in a 2cm x 1.2mm device; 3) it is easy to design functional parts, such as using the upper paper sheet for filtration.
The sealing technology of the three-dimensional paper chip is one of the key technologies in the preparation of the three-dimensional paper chip. At present, the three-dimensional paper chip processing technology reported in the literature mainly comprises a 2D paper chip bonding method and a paper folding method. In 2011, Crooks group reported a new method for making three-dimensional paper chips by using the traditional Chinese manual paper folding technology. The specific manufacturing process is as follows: firstly, patterning analytical elements such as a channel, a liquid storage tank, a crease and the like on a layer of paper chip by using a simple SU 8-ultraviolet photoetching technology; then folding the layer of paper chip into a multilayer three-dimensional device according to a crease and a certain folding method; finally, four corners are subtracted and put between the pin clamping plates to wait for sample injection analysis. The paper folding method has the disadvantages that the paper layers are difficult to align, gaps exist always, the vertical flow of fluid is influenced, and an additional pin clamping plate needs to be arranged. The Whitesides group utilizes waterproof double-sided adhesive tape which is provided with holes and filled with cellulose powder to naturally and superpose two or more layers of two-dimensional paper chips prepared by SU 8-ultraviolet lithography to prepare three-dimensional paper chips for the first time. Although the paper layers can be tightly combined, the manufacturing process is too difficult and tedious. In 2012, the Phillips group reported a new method for quickly manufacturing three-dimensional paper chips in batches by combining a wax printing method with a glue spraying technology. The glue spraying method has the advantages that the paper layers are tightly combined, the operation is simple, but the glue spraying amount is difficult to control, the patterning is difficult to realize, and the hydrophilic performance of the channel can be influenced by the penetration of glue solution in the filter paper. Zheng et al in 2016 reported a sealing method by screen printing a paste. The glue printing method can realize patterning and better control glue amount, and reduces the influence of glue solution permeation on the channel. However, the printing step needs precise overprinting, and the process is still complicated.
All current three-dimensional paper chip fabrication involves complex assembly box sealing processes. This process is often a technical bottleneck in achieving chip mass production.
Disclosure of Invention
The invention provides a preparation method of a novel magnetic self-assembly three-dimensional paper chip to solve the technical problems.
In order to achieve the technical effects, the invention comprises the following technical scheme: a preparation method of a novel magnetic self-assembly three-dimensional paper chip comprises the following steps:
(1) hydrophobic magnetic ink is adopted, and a channel layer, a pretreatment reagent layer and a detection layer of the paper chip are respectively obtained through screen printing;
(2) the pretreatment agent is screen printed on the pretreatment agent layer by a patterned printing agent screen by a scraper.
(3) Printing a detection medicament on a detection area on the detection layer through a patterned medicament screen by using a scraper;
(4) and stacking the channel layer, the pretreatment reagent layer and the detection layer on a gasket made of a magnetic material in sequence to obtain the three-dimensional paper chip.
Further, the step (1) includes: uniformly mixing hydrophobic ink and a magnetic powder material to obtain hydrophobic magnetic ink, printing the hydrophobic magnetic ink on paper through a patterned ink screen by using a scraper, penetrating the paper to form a hydrophobic area, wherein the part which is not printed is a hydrophilic area, and respectively obtaining a channel layer, a pretreatment reagent layer and a detection layer of the paper chip.
Furthermore, the hydrophobic ink is prepared from hydrophobic substances and a solvent, and the hydrophobic magnetic ink is prepared from hydrophobic ink and magnetic powder; wherein, the hydrophobic substance comprises but is not limited to one of screen printing ink, paraffin, polydimethylsiloxane and acrylic pigment; the solvent includes but is not limited to one of ethyl acetate, n-heptane, toluene, ethanol; the magnetic powder comprises but is not limited to one of iron-cobalt-nickel alloy, rubidium-boron-iron alloy and ferroferric oxide. When the hydrophobic ink consists of screen printing ink and ethyl acetate, the volume ratio of the screen printing ink to the ethyl acetate is 5-3: 1; when the printing liquid consists of paraffin and n-heptane, the mass ratio of the paraffin to the n-heptane is 1: 150-200; when the printing liquid consists of propylene pigment and ethanol, the volume ratio of the propylene pigment to the ethanol is 1: 1-1.5; when the printing liquid consists of hydrophobic material polydimethylsiloxane and solvent toluene, the printing liquid also comprises cross-linking agent ethyl orthosilicate, and the volume ratio of the polydimethylsiloxane to the ethyl orthosilicate to the toluene is 10: 1: 2-3.
Preferably, the ink printing screen and the medicine printing screen are both nylon screens, the ink printing screen is 50-300 meshes, and the medicine printing screen is 200-400 meshes. The tension of the ink printing silk screen and the tension of the medicine printing silk screen are both 12-18N/cm. The angle of the scraper in contact with the silk screen during printing is 40-50 degrees, and the printing speed is 0.2-0.6 m/s. The paper includes, but is not limited to, one or more of filter paper, cellulose acetate film, cellulose nitrate film or mixed cellulose film.
Preferably, the mass ratio of the hydrophobic ink to the magnetic powder is 1: 0.3-1.5.
Further, the channel layer has three layers including a first channel layer, a second channel layer and a third channel layer; the three-dimensional paper chip is formed by sequentially stacking a first channel layer, a second channel layer, a pretreatment reagent layer, a third channel layer, a detection layer and a gasket from top to bottom in sequence.
Preferably, the detection agent includes, but is not limited to, a chemical masking agent, a buffer, an enzyme, an antibody, a fluorescent label or an enzyme-labeled antibody, a chemical chromogenic agent, a fluorescent probe, a DNA molecule, a bacterium, a cell, or a substrate. The chemical color developing agent and the fluorescent probe are respectively prepared by distilled water or corresponding organic solvents; the DNA, bacteria, cells or substrate may or may not be coated with cellulose or silica microspheres.
By adopting the technical scheme, the method has the following beneficial effects: the preparation method of the novel magnetic self-assembly three-dimensional paper chip provided by the invention adopts a screen printing technology, and by improving a printing material, an additional adhesive layer is not adopted, and a step of spraying or printing glue is not needed, so that the magnetic material and the hydrophobic material are printed simultaneously, the preparation step is simplified, and the risk of blocking a hydrophilic channel by glue solution permeation is eliminated;
the magnetic material is adopted for self-assembly, and on the premise of ensuring tight bonding of the paper chip layer, complicated chip packaging steps are reduced, so that the industrialization of the preparation of the three-dimensional paper chip is easy to realize, and the production cost is obviously reduced;
the magnetic material is adopted for reversible self-assembly and easy disassembly, so that the color reaction of the chip layer in the three-dimensional paper chip can be observed, and the application range of the three-dimensional paper chip is expanded.
Drawings
Fig. 1 is a schematic structural view of a three-dimensional paper chip prepared in example 1 of the present invention;
fig. 2 is a schematic structural view of a three-dimensional paper chip prepared in embodiment 2 of the present invention.
In the figure, the position of the upper end of the main shaft,
1. a first channel layer; 2. a second channel layer; 3. pretreating a reagent layer; 4. a third channel layer; 5. a detection layer; 6. a gasket; 7. an absorbent pad.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the present invention, the terms "upper", "lower", "left", "right", "front", "rear", "top", "bottom", "inner", "outer", "center", "vertical", "horizontal", "lateral", "longitudinal", and the like indicate an orientation or positional relationship based on the orientation or positional relationship shown in the drawings. These terms are used primarily to better describe the invention and its embodiments and are not intended to limit the indicated devices, elements or components to a particular orientation or to be constructed and operated in a particular orientation.
Moreover, some of the above terms may be used to indicate other meanings besides the orientation or positional relationship, for example, the term "on" may also be used to indicate some kind of attachment or connection relationship in some cases. The specific meanings of these terms in the present invention can be understood by those skilled in the art as appropriate.
Furthermore, the terms "mounted," "disposed," "provided," "connected," and "coupled" are to be construed broadly. For example, it may be a fixed connection, a removable connection, or a unitary construction; can be a mechanical connection, or an electrical connection; may be directly connected, or indirectly connected through intervening media, or may be in internal communication between two devices, elements or components. The specific meanings of the above terms in the present invention can be understood by those of ordinary skill in the art according to specific situations.
"plurality" means two or more unless otherwise specified.
The present invention will be described in further detail below with reference to specific embodiments and with reference to the attached drawings.
Example 1:
the embodiment provides a preparation method of a novel magnetic self-assembly three-dimensional paper chip, which comprises the following steps:
s101, adopting hydrophobic magnetic ink, and respectively obtaining a channel layer, a pretreatment reagent layer 3 and a detection layer 5 of the paper chip through screen printing;
uniformly mixing hydrophobic ink and a magnetic powder material to obtain hydrophobic magnetic ink, printing the hydrophobic magnetic ink on paper through a patterned ink screen by using a scraper, penetrating the paper to form a hydrophobic area, wherein the part which is not printed is a hydrophilic area, and respectively obtaining a channel layer, a pretreatment reagent layer and a detection layer of the paper chip.
Specifically, a 120-mesh patterned nylon ink printing screen is selected, the tension of the screen is 15N/cm, the screen is used for printing a designed pattern on filter paper by using hydrophobic magnetic ink, and the filter paper is dried. The magnetic ink in the printing process consists of hydrophobic ink and magnetic powder. Wherein the hydrophobic ink comprises screen printing ink and ethyl acetate at a volume ratio of 4: 1, the magnetic powder is 150-rubidium ferromagnetic powder of 250 meshes, and the weight ratio of the hydrophobic ink to the magnetic powder is 1: 0.7. The distance between the screen and the filter paper is 1.5mm during printing, the contact angle between the scraper and the screen is 45 degrees, the printing speed is 0.2m/s, and the paper bearing pressure is 12N.
S102, printing the pretreatment agent on the pretreatment agent layer through a patterned chemical printing screen by using a scraper.
The pretreatment agent in this example is a chemical masking agent, i.e., a 20% sodium malonate and 15% sodium fluoride aqueous solution by mass ratio.
S103, printing a detection medicament on a detection area on the detection layer through a patterned medicament screen by using a scraper;
the detection medicament is prepared by the following method: preparing a color developing agent sodium diethyldithiocarbamate into a solution of 1.0mg ml-1 by using a mixed solution of EDTA and hydrogen fluoride (the concentration of the EDTA is 0.05mol1-1, and the concentration of the hydrogen fluoride is 0.05mol1-1), and adjusting the pH value to 8.5 by using 2% (v/v) ammonia water to obtain the detection medicament.
Taking a 400-mesh patterned nylon screen for printing the medicine, wherein the tension of the screen is 18N/cm; printing the pretreatment agent on the pretreatment agent layer shown in figure 1, printing the obtained detection agent on the detection agent layer shown in figure 1, drying, wherein the silk screen patterns are respectively consistent with the second layer and the fifth layer, the distance between the silk screen and the filter paper is 1.5mm, the contact angle between the scraper and the medicine printing silk screen is 45 degrees, the printing speed is 0.5m/s, and the paper bearing pressure is 8N.
S104, stacking the channel layer, the pretreatment reagent layer and the detection layer on a gasket 6 made of a magnetic material in sequence to obtain the three-dimensional paper chip.
In this embodiment, the channel layer has three layers, including a first channel layer 1, a second channel layer 2, and a third channel layer 4; the three-dimensional paper chip is formed by sequentially stacking a first channel layer 1, a second channel layer 2, a pretreatment reagent layer 3, a third channel layer 4, a detection layer 5 and a gasket 6 from top to bottom.
The chips are sequentially stacked on a gasket 6 made of magnetic materials according to the structure shown in figure 1, and the magnetic self-assembly chip can be obtained. The paper penetration rate of the hydrophobic material of the chip is 100 percent, and the channel resolution is 0.3 mm. The structure schematic diagram of the three-dimensional paper chip prepared according to the steps is shown in fig. 1. By the chip, a single sample can be parallelly distributed into 16 parallel samples, and the detection flux is improved.
Example 2:
the embodiment provides a preparation method of a novel magnetic self-assembly three-dimensional paper chip, which comprises the following steps:
s201, hydrophobic magnetic ink is adopted, and a channel layer, a pretreatment reagent layer 3 and a detection layer 5 of the paper chip are respectively obtained through screen printing;
and selecting a 100-mesh patterned ink-printing nylon screen, and printing the magnetic hydrophobic printing liquid on the nitrocellulose membrane. The tension of the silk screen is 18N/cm during printing, the components of the hydrophobic ink are composed of the silk screen printing ink and ethyl acetate in the volume ratio of 3: 1, the magnetic powder is 150-mesh ferroferric oxide magnetic powder with the particle size of 250 meshes, and the weight ratio of the hydrophobic ink to the magnetic powder is 1: 0.8. And during printing, the mesh distance between the ink screen and the nitrocellulose membrane is 1.0mm, the contact angle between the scraper and the ink screen is 50 degrees, the printing speed is 0.2m/s, and the pressure borne by the nitrocellulose membrane is 12N, so that each channel layer, the pretreatment reagent layer and the detection layer of the paper chip are obtained.
S202, printing the pretreatment agent on the pretreatment agent layer through a patterned chemical printing screen by using a scraper.
The pretreatment agent adopts monodisperse silica microspheres (5.0 muL, 2.5% (w/V)) mixed with gold-labeled rabbit anti-HAV antibody 1 or monodisperse silica microspheres (5.0 muL, 2.5% (w/V)) mixed with gold-labeled rabbit anti-HBs (HBV s antigen) antibody 1; the detection reagent used was monodisperse silica microspheres (5.0. mu.L, 2.5% (w/V)) mixed with rabbit anti-HAV antibody 2 or monodisperse silica microspheres (5.0. mu.L, 2.5% (w/V)) mixed with rabbit anti-HBs (HBV s antigen) antibody 2.
S203, printing a detection medicament on a detection area on the detection layer through a patterned medicament screen by using a scraper;
selecting a 400-mesh patterned nylon screen for printing the chemicals, wherein the tension of the screen is 16N/cm; printing the pretreatment agent on the pretreatment agent layer shown in figure 2, printing the detection agent on the detection layer shown in figure 2, and drying, wherein the mesh distance between the drug printing screen and the nitrocellulose membrane is 1.0mm during printing, the angle between the scraper and the drug printing screen is 50 degrees when the scraper is in contact with the drug printing screen, the printing speed is 0.6m/s, and the pressure borne by the cellulose acetate membrane is 6N.
S204, stacking the channel layer, the pretreatment reagent layer and the detection layer on a gasket made of a magnetic material in sequence to obtain the three-dimensional paper chip.
The channel layer has three layers including a first channel layer 1, a second channel layer 2 and a third channel layer 4; the three-dimensional paper chip is formed by sequentially stacking a first channel layer 1, a second channel layer 2, a pretreatment reagent layer 3, a third channel layer 4, a detection layer 5 and a gasket 6 from top to bottom.
Further comprising the steps of: the water absorption pad is arranged on the detection layer and the gasket made of the magnetic material, and can promote liquid to flow downwards, so that the antigen or antibody to be detected combined with the colloidal gold can quickly reach the detection area.
The chips are sequentially stacked on the magnet gasket according to the illustration in fig. 2, and the magnetic self-assembly chip can be obtained. The paper penetration rate of the hydrophobic material of the chip is 100 percent, and the channel resolution is 0.25 mm. By the chip, detection of HAV and HBV virus antigens of two samples can be completed. By simply repeating the detection unit, a higher throughput of detection paper chips can be obtained.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (6)
1. A preparation method of a magnetic self-assembly three-dimensional paper chip is characterized by comprising the following steps:
(1) hydrophobic magnetic ink is adopted, and a channel layer, a pretreatment reagent layer and a detection layer of the paper chip are respectively obtained through screen printing;
(2) printing the pretreatment agent on the pretreatment agent layer through a patterned drug screen by using a scraper;
(3) printing a detection medicament on a detection area on the detection layer through a patterned medicament screen by using a scraper;
(4) stacking the channel layer, the pretreatment reagent layer and the detection layer on a gasket made of a magnetic material in sequence to obtain a three-dimensional paper chip;
the step (1) comprises the following steps: uniformly mixing hydrophobic ink and a magnetic powder material to obtain hydrophobic magnetic ink, printing the hydrophobic magnetic ink on paper through a patterned ink screen by using a scraper, penetrating the paper to form a hydrophobic area, wherein the part which is not printed is a hydrophilic area, and respectively obtaining a channel layer, a pretreatment reagent layer and a detection layer of the paper chip.
2. The preparation method according to claim 1, wherein the hydrophobic ink is prepared from a hydrophobic substance and a solvent, and the hydrophobic magnetic ink is prepared from a hydrophobic ink and magnetic powder; wherein, the hydrophobic substance comprises but is not limited to one of screen printing ink, paraffin, polydimethylsiloxane and acrylic pigment; the solvent includes but is not limited to one of ethyl acetate, n-heptane, toluene, ethanol; the magnetic powder comprises but is not limited to one of iron-cobalt-nickel alloy, rubidium-boron-iron alloy and ferroferric oxide.
3. The production method according to claim 1 or 2, wherein the mass ratio of the hydrophobic ink to the magnetic powder is 1: (0.3-1.5).
4. The production method according to claim 1, wherein the channel layer has three layers including a first channel layer, a second channel layer, and a third channel layer; the three-dimensional paper chip is formed by sequentially stacking a first channel layer, a second channel layer, a pretreatment reagent layer, a third channel layer, a detection layer and a gasket from top to bottom in sequence.
5. The method of claim 1, wherein the detection agent includes, but is not limited to, chemical masking agents, buffers, enzymes, fluorescent or enzyme-labeled antibodies, chemo-chromogenic agents, fluorescent probes, DNA molecules, bacteria, cells, or substrates.
6. The method for preparing according to claim 1, wherein the step (4) further comprises the steps of: and a water absorption pad is arranged on the detection layer and the gasket made of the magnetic material.
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|---|---|---|---|---|
| US3160093A (en) * | 1962-11-05 | 1964-12-08 | Warren S D Co | Planographic printing plate receptive to carbon ribbon imaging |
| CN1448723A (en) * | 2002-03-29 | 2003-10-15 | 成都夸常科技有限公司 | Biological chip capable of being disassembled and assembled |
| CN102016595A (en) * | 2008-03-27 | 2011-04-13 | 哈佛学院院长等 | Three-dimensional microfluidic devices |
| CN106111219A (en) * | 2016-06-05 | 2016-11-16 | 大连大学 | Three-dimensional paper chip preparation methoies based on many editions patterning silk screen associating printing technologies |
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| US7303860B1 (en) * | 2001-12-06 | 2007-12-04 | Massachusetts Institute Of Technology | System and method for performing multi-resolution lithography |
| CN1432655A (en) * | 2002-11-04 | 2003-07-30 | 成都夸常科技有限公司 | Multilateral biochip |
| US20100227382A1 (en) * | 2005-05-25 | 2010-09-09 | President And Fellows Of Harvard College | Nanoscale sensors |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3160093A (en) * | 1962-11-05 | 1964-12-08 | Warren S D Co | Planographic printing plate receptive to carbon ribbon imaging |
| CN1448723A (en) * | 2002-03-29 | 2003-10-15 | 成都夸常科技有限公司 | Biological chip capable of being disassembled and assembled |
| CN102016595A (en) * | 2008-03-27 | 2011-04-13 | 哈佛学院院长等 | Three-dimensional microfluidic devices |
| CN106111219A (en) * | 2016-06-05 | 2016-11-16 | 大连大学 | Three-dimensional paper chip preparation methoies based on many editions patterning silk screen associating printing technologies |
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