TWI374266B - - Google Patents

Description

</ RTI> </ RTI> </ RTI> </ RTI> </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; [Prior Art] Biochip inspection technology is an important biotechnology in recent years... It uses molecular biology, genetic information, analytical chemistry and other principles to design, to match wafer, glass or polymer wire Or other processing techniques, 7 produced rain ship components, such as semiconductor crystal #--------------------------------------------------------------------------- Multiple pathogens can be detected simultaneously on a single wafer.

The biochip of the JJ development can be roughly divided into two types: the gene chip (10) e-material rDNA P) "Processjjjgchip or Lab_on_a_chip". The gene chip is the fastest growing of all kinds of different types of biochips. Refers to the number of square centimeters of the φ product with a shuttle or a number of nucleic acid probes, (four) - a test, you can provide a large number of gene phase fine # message, also provide _ (test noisy) gene crystal moon. The microarray type (]VTicroarray) gene wafer is a neatly fixed, arrayed biological probe [deoxyribonucleic acid (DNa)] on the wafer substrate, through dna_DNA hetero-parent reaction or protein-health binding, using a simple sample Analysis or inspection. 1374266 According to the classification of target objects detected by microarray wafers, it can be divided into DNA wafers and egg white matter wafers. DNA wafers are labeled with nucleotides as probes. The corresponding nucleic acid fragment, and the DNA wafer is divided into a complementary deoxyribonucleic acid (cDNA) wafer and an oligonucleic acid wafer (QllgGnudec) tide chiP according to the length of the probe placed on the wafer. The cDNA wafer is most often use Study of a large number of gene expression, the nucleotide chip coupons in addition to gene expression analysis can be done, and more importantly, &quot; X Yau to diagnose things and genotyping of pathogens (Gen〇 ^ yp jjjg).

• DNA Crystal is a feature of her complementary DNA hybridization in the New Zealand, which is complementary to the nuclear nucleus (DNA). It can be used to detect diseases, shorten the development time of new drugs, etc. It is a powerful DNA analysis tool. For example, by using a luminescence-emitting phosphor (also known as a fluorescent-blocking molecule) as a probe (note: a general DNA wafer probe is a single-stranded DNA), reacting with DNA, and before and after the reaction The fluorescence of the needle changes, and the use of changes in the camp light can detect specific DNA fragments. Biochips are widely used, and can be applied to the following fields (Gene expression proflling) (2) Toxic analysis (T〇xic〇bgy Analysis) (3) Gene sequencing ( Gene Sequendn (4) SNP Identificati〇n (5) Forensic medicine orensics) (6) Immunological assay (Immunoassays) (7) Protein chip (8) Detection of biological weapons (Combat Biowarfare (9) drug screening (Drug screening) (1〇) on the phone hardware application (hard drives and microprocessoi^) 〇 However, 'ship biocrystal#' low profile and; ^ side, therefore It is necessary to use ang 1^74266 expensive detection equipment and special imaging technology. In view of this, it is still necessary to develop a new biochip manufacturing method to meet the needs of easy detection and high sensitivity. 4b [Invention content] In the above background of the invention, in order to overcome the shortcomings of the conventional technology, the present invention provides a novel bio-film having a three-dimensional stereoscopic combination and a method of forming the same. # The two-dimensional structure is a hole material having a nano-scale hole Reticulated polymer 'for example, aerogel material. Porosity can be as high as 95%. It has many characteristics due to high porosity: high specific surface area, low density, low thermal conductivity, low-speed transmission, low dielectric Constants, etc., so the application range is very large, and can be used for heat insulating materials, catalysts, adsorbents, electrodes, electronic 'detectors, etc. One of the objects of the present invention is to use a sol-gel technique on a lithographic substrate. A biochip having a three-dimensional structure is synthesized thereon. Another object of the present invention is to transform the surface of a planar substrate into a porous structure of a porous material in a bulk three-dimensional structure, and utilize its large internal specific surface area to capture the calibrated DNA and protein. In this way, the biochip having the three-dimensional structure provided by the present invention has the characteristics of sensitivity quotient and easy detection, and can simplify the detection instrument, for example, only need to set up data. The type of camera (CCD) does not require the use of special imaging technology. According to the present invention, the economic benefits and industrial applicability of the invention are based on the above. For the purpose of the present invention, the 1374266 biochip having a three-dimensional structure has a structure comprising a substrate and a structure formed on the surface of the substrate, wherein the t, three-dimensional continuous perforation wire ^ = After r_定定, _, 雌, 嶋子, and y inventions are also not disclosed - a method for forming a bio-wafer having a layer of a hole material in a three-dimensional structure, which includes - a program of mixing one-year-old, a program of standing, - Drying procedure, - processing procedure, - changing program and coating procedure. • [Embodiment] The present invention is directed to a biocrystal chip having a three-dimensional structure and its green color. In order to understand the present invention in a thorough manner, in the following description: It is apparent that the practice of the invention is not limited to the specific details familiar to those skilled in the art. On the other hand, well-known components or steps are not described in detail to avoid unnecessarily limiting the invention. The preferred embodiments of the present invention will be described in detail below, but in addition to these detailed descriptions, the present invention may also be applied to other actual closures bribely, and the scope of the present invention is not limited. The scope shall prevail. An embodiment of the present invention discloses a method for forming a bio-wafer having a three-dimensional structure. First, an aerogel precursor solution is provided, which comprises an ionic melt (with the function of a template and a solvent), a catalytic hydrolysis and And a condensation reaction reagent and at least one alkoxide monomer and/or an aromatic oxy monomer, wherein the catalytic hydrolysis and/or reductive reagent comprises one of the following groups or any combination thereof. , acidic compounds and test compounds. Further, the above-mentioned alkoxy monomer 1374266 and/or a central element in the aromatic oxy monomer includes one of the following groups (1), he, K, Rb, CS, Be, Mg, Ca, Sr, Ba, B. , Te, CrCuErFe,

Ta, V, Zn, Zr, A1, Si, Ge, s_b. Secondly, the aerogel precursor solution is subjected to a uniform mixing procedure to facilitate hydrolysis and polymerization of at least one activating oxygen monomer and/or aromatic oxy monomer, and the specificity of the precursor solution is specific. Viscosity is greater than or equal to 150 Cps. Next, the aerogel precursor solution is allowed to stand so that at least one of the alkoxy monomers and/or the aryloxy monomers are continuously hydrolyzed and polymerized, thereby forming a gas condensing. The lining gas is transferred to extract the ionic melt n contained in the aerogel (10) t to carry out the solvent contained in the pores of the aerogel. Then, an addition engineering and processing procedure is carried out to grind the gas condensing shaft-aerogel powder, and the recording number is from coffee to gamma. . After the process is completed, the procedure is carried out by mixing-changing_and gas-condensing=end' to guide at least the "functional" based on the inner surface and the outer surface of the aerogel powder. In the present invention, the aerogel powder has a plurality of pores inside, and the inner surface of all the pores and the wall is called the "inner surface"; relatively, the surface of the pore between the gas particles is called the "outer surface". The total surface area of the two gels is also high in volume due to the high silkiness of the gas. The side of the _ _ light borrowing domain to super =: rr gel powder inside and outside the surface, thereby making = ^ 曰 片 film. Finally, a coating process is carried out, the purpose of which is to use the old-and-two gas-condensing 7-nuclear substrate on the substrate, and to form a three-dimensional image of the material, wherein the substrate material contains the following groups. '·································································································· The material (_) is used for the purpose of using the solvent in the '7 stock solution, and adding a line W having a three-dimensional 颂 structure to the two axes of the modified gas gel powder and the substrate. The above-mentioned series is 80 to 12 inches. The helium gel precursor solution may contain an acidic compound or an organic compound to catalyze the hydrolysis and polymerization of the sulphur-based monomer and/or the aryloxy monomer. In addition, the gas condensing solution for the solution includes: Money, age recording base monomer or aromatic oxy-early body and ion test and form - the first mixture. Secondly, add human - acidic compound to the first: mixed wealth and formed - second mixture 4, add people - alkaline Compound to the first mixture to improve the hydrolysis of silk-based monomers and / or aromatic oxy monomers The degree of polymerization. The common component of the above aerogel comprises one of the following groups or any combination thereof. SiO2, Ti〇2, ν2〇5 and A1A are replaced by a solvent to extract a gas condensate. The ionic melt contained in the gas _ hole shot. The above solution _ is preferably selected as a low boiling point solvent (having a boiling point of less than or equal to 2 〇〇〇c;), and includes one of the following groups: nitrile, alcohol Classes, ketones and water. After drying, the grinding process is carried out to form aerogel powder. The average pore diameter of the aerogel powder is about 2 to 5 ,, and the specific surface area is greater than or equal to l〇〇m2/ g, the porosity ranges from 5〇% to 99〇/〇. The aerogel powder is then subjected to a modification procedure, and the modified 1374266 agent used in the modification procedure has at least one specific group 'the specific group contains the following groups One of them: an amine group, a hydroxyl group, a carboxylic acid group and an epoxy group. Commonly used modifiers include: N-[3-(trimethoxysilyl)propyl]-l,2-ethanediamine (DAMO), 3-Glycidoxypropyl-trimethoxysilane ( GLYMO),

3 - Aminopropyltriethoxy silane (APTS), N-(2-Aminoethyl) 3-aminopropyltriethoxysilane (TMsen), and the like. After the modification, the aerogel powder is subjected to a coating process, and the modified aerogel powder is applied to a specific region on a substrate to thereby form a bio-wafer having a three-dimensional structure. In a preferred embodiment of the present embodiment, a conversion process can be performed after completion of the coating process. First, a converter is provided which includes a first group and at least a second group. Then, a specific functional group on the inner and outer surfaces of the aerogel powder and a first group of the converter are bonded to form a biochip having a second group on the surface. For example, when the modifier is N-[3-(trimethoxysilyl)propyl]-l,2-ethanediamine (DAMO), a disk can be used as a conversion to form a mesopores with acid groups on the surface. Floor. The above-described transformant comprises one of the following groups: an antigen, a monoclonal antibody, a primary antibody, a multi-drug antibody (p〇iyCi〇nai ^tibodies), a nucleic acid (nucleic acids) [including a monomeric state ( Monomeric) and oligomeric], pr〇teins, enzymes, lipids, polysaccharides, sugars, peptides, and more Polypeptides, drugs, viruses, microorganisms and bioligands ° 12 1374266 In this embodiment, another method is preferred. After the completion of the procedure, the process can be carried out. First, a blocking agent is provided which has a -third group. 'Then' then links the specific functional groups on the outer surface of the gum powder handle with the blocker's third base®' to determine no on the biochip. Residual unreacted specific functional groups. In another preferred embodiment of the present embodiment, after the blocking process is completed, a -specific matching procedure can be performed. First, a pairing is provided, which includes a Four groups and - fifth group. Then, special The second group on the sheet and the fourth group on the pair to form a biochip having a fifth group on the surface, wherein the pair includes the group of the next group: antigen , monoclonal antibodies, polyclonal antibodies, polyclonal antibodies, nucleic acids, nucleic acids, nucleic acid, nucleic acid, nucleic acid, nucleic acid, nucleic acid, nucleic acid, nucleic acid, nucleic acid, (Proteins), enzymes, lipids, polysaccharides, sugars, ureas, peptides, peptides, viruses, microorganisms Bioligands. In the re-preferred example of the present embodiment, after the completion of the specific pairing procedure, the 'can be performed-calibration procedure' first provides a calibration carrier comprising at least a sixth a group and a -seventh group, wherein the seventh group is bonded to the - calibrator, and the fifth group of the post-ligand is paired with the sixth group of the calibration carrier to form a target-treated organism The calibration of the upper ball is one of the following or Hexian County, Dawning cold light enzyme, radioactive substance. In addition, the above calibration vector contains the following groups: monoclonal antibody, primary antibody, 13 1374266 secondary antibody, labeled primary antibody, multiple antibodies (Polyclonal such as his heart ^), Nucleic acids [including monomeric and oligomeric], proteins, enzymes, lipids, poly- ( Polysaccharides), sugars, peptides, peptides, drugs, viruses, microorganisms, and bioligands

In the present embodiment, the above ionic melt is a room temperature ionic liquid which is formed by mixing an organic base with a Lewis acid. When the Lewis acid used is a functionalized metal acid, it is also possible to form a normal temperature ion refining liquid, but such an ionic liquid reacts with water to produce a hydrohalic acid, and thus is not suitable for use in the present invention. Therefore, the Lewis acid used in the present invention does not contain a metal acid, thereby preparing an ionic melt which can be used in the seed. In the preferred example of the present invention, the general formula of the cationic group in the above organic test is as follows.

The choices are as follows: R1 --^ 2 CH; Η' 53⁄4 ---JJ ζϊζ~~Η --- ch3ch2c h2ch2 Η

R3 - R 4 CH; · H CH3 H ~ 1 -~--- H ch3 -- H 1374266 (ch3)2ch ch2 H ch3 H CH3CH2C h2ch2 H C2H5 H ch3 H CH3OCH2CH2 H ch3 H cf3ch2 H ch3 c H3 c2h5 H Ch3 c H3 CH3CH2CH2 H c6h6ch2 c H3 ch3ch2ch2 H c6h6ch2 c H3 ch3ch2ch2c . h2 H c6h6ch2 c H3 (CH3CH2)(CH3 )CH H c6h6ch2 c H3 CH3CH2CH2C h2ch2 H ch3 H c2h5 C h3 c2h5 H c2h5 c H3 For example, more common The organic base comprises one of the following groups: ln-butyl-3-methylimidazolium chloride (BMIC), 15 1374266 1 -octanyl-3-methylimidazolium bromide ( ΟΜΓΒ ) , • 1 -dodecanyl-3-methylimidazolium bromide ( DMIB ) and . 1 -hexadecanyl-3-methylimidazolium bromide (HDMEB). Further, the anionic group of the Lewis acid_ described above includes one of the following groups: BF4·, .· PF6·, AsF6_, SbF6_, F(HF)n·, CF3S03·, CF3CF2CF2CF2S03-, (CF3S02) 2N-, (CF3S02)3C·, CF3CO〇- and CF3CF2CF2COCT. When the type of the organic base used above is fixed, the hydrophilicity of the ionic solution can be controlled by adjusting the anionic group in the Lewis acid, for example, the hydrophilic ion solution bmic_bf4 and the hydrophobic ion melt BMC-CTFSI; Taking a decyloxy monomer as an example, it hydrolyzes to form a hydrophilic sterol (_Si_0H), so that the 'hydrophilic ionic melt is more attractive to sterols and stabilizes the formation of dioxide-dream and 纟. 'Get a more stable two-dimensional three-dimensional junction; ^ hole material. In the present embodiment, the above ionic melt has a weight ranging from about -70 to the weight of at least one alkoxy monomer. /. Preferably, the range is about 2% to 5%. When the amount of addition of the + solution is greater than the upper limit of addition, the concentration of the sol decreases and the rate of gel reaction slows, causing structural instability. In the present invention, the method for forming a three-dimensional structure of a biochip includes the following methods: direct immunization method, indirect immunization method, Gu-binding immunoassay, Sanming treatment immunotherapy, and the like.

Example, the actual _ dew - the formation method of the organism (9) with the three-gel layer (the steps) as shown in the first figure: 1374266 (1 2) cleaning the glass: * immersing the slide in the sodium hydroxide In the middle of the aqueous solution, as for the ultrasonic shock absorber. Shock for 30 minutes, and then change the water for 30 minutes. Finally, the acetone is shaken for 30 minutes, and taken out and dried in an oven. (2) Aerogel production: BMIC-BF3 is mixed with citric acid solution, and then rapidly mixed with butyl hydrazine 5, and left to stand for hydrolysis and heating. The sample is aged after aging, then lyophilized, and finally coarsely ground into a powder. The Lu gas gel reaction formula is as follows:

OH

OH

I

4H20 —»&gt; HO— Si — OH + 4ROH

I

OH

〇H OH OH

I I I

HO— Si — OH --- HO— Si — 〇 — Si — OH

I - H20 I I

〇H OH OH

OH

OH n H〇- Si - 〇_

PH in

〇— Si —〇H I OH

HO - Si _ 0 - Si - OH I - I - ► Ο O OH η H20 I I I HO - Si _ 0 - Si - 0 - Si - 〇 H 0

HO— Si ～〇H

OH (4) aerogel amine-based upgrading: continuous stirring for a small amount of coarsely ground aerogel mixed with 2% DAMO. solution after 'drying' cleaning - day, placed in the drying oven. (4) Synthetic aerogel on the surface of the slab: 17 1 2

HO — Si — OH 3

OH 4

I 1374266 Disperse the modified aerogel in a second degree of water and prepare a dispersion of 1% by weight to continue to disturb the magnet. Pipette 2 of the dispersion with a micropipette and drop onto the slide - the planned position. · (5) Quantum dot modification: ·- Prepare SBB buffer, add 3.07 g boric acid to 19.07 g sodium borate, adjust pH=9 with NaOH or HC1. The number of grams required for ED.EDC, PEG, sulfo-NHS is required to calculate the drug. Scale medicine, • Store in a microcentrifuge tube and record the actual number of grams. The amount of buffer required for each drug is then calculated based on the actual number of grams. Mix the desired quantum dots with the buffer. Mix the PEG and buffer first, and shake until the PEG is dissolved in the buffer. Rapid mixing of the roc with the buffer was immediately added to the quantum dot reaction. Mix sulfo-NHS with buffer. Finally, sulfo-NHS was added to the quantum dots and PEG was added immediately. The mixture was shaken in a refrigerator at 4 ° C for two hours. The solution after completion of the reaction through the molecular sieve is filtered to leave the remaining quantum dots. The above quantum dot modification _ reaction is shown in the following scheme: 1374266

(6) Grafting of quantum dots with antibodies: A PBS buffer was prepared, and 〇.24 g of KH2P04 was dissolved in dihydrated water, followed by addition of 1.44 g of Na2HP〇4, 0.2 g of KC1 and 8 g of Naa, and pH = 7.5 with NaOH or HC1. Prepare the wash buffer, add 580 mL PBS to 20.5 //L Tween 20, and mix evenly. Calculate the desired antibody and quantum dots, mix the antibody with the quantum dots, and place them in a refrigerator at 4 °C overnight. The quantum dot and antibody graft reaction scheme is as follows:

(7) Antigen and antibody on the crystal: Dilute different concentrations of antibody (Mouse anti 19 1374266 human, IL_6) at the planned position on the slide. The fine amount is i at 37t_, the reaction is for two hours, after removal Wash with a small amount of washing buffer, one minute at a time, three times. The blocking reaction was carried out, and a 1% solution of BSA and PBS was spread over the entire slide, and the slide was placed in a water-filled box and placed in 4. Hey; the refrigerator is overnight. After taking out, wash it with an appropriate amount of washing buffer, one minute at a time, three times in total. Dilute the diluted antigen (Human IL-6) at the same point, each drop is 1 "L, placed in an oven at 30 °C for - hour, remove and wash with a small amount of washing buffer, one minute at a time. A total of three times. At the same point, the antibody that has been bound to the quantum dot (Rabbitantihu_, IL-6) '1 per drop, placed in an oven at 30 °C, reacted for one hour, taken out, and washed in the buffer; Shock cleaning, one minute at a time, three times in total. Finally, 1 / L pure quantum dots are dropped on the positive control point. The glass sheet having the aerogel layer formed by the above steps (1)-(7) The surface is white, and the formed area is the sample-measuring area. As shown in the second figure, it is the result of infrared absorption spectrum analysis of the aerogel structure. The fruit is displayed at 3200~3700 cm1 and 1620~1640 cm-1. Where the absorption peak of the -〇·Η bond is present; the absorption peak at 3200~3700 cm·1 is mainly 0-H bonding vibration of =Si-〇H, and the wave number is 93〇~950 cm·1 The absorption peak is the bond vibration of ^Si-〇H. The wave number is 1000~1200 cm·!, 78〇~82〇(10)-丨 and ~46〇^ is 3Si-0-S _ bond stretching vibration. Any atom with spin angular momentum, that is, atomic order or mass arbitrarily, will have NMR absorption, common liquid nuclear magnetic 20 1374266 _) regular money machine ride The vertical, but in the _ conditions often • Ships in the choice of D solvent. With the advancement of _, nuclear magnetic resonance spectrum analyzer, the solid state sample can be directly side, and step-by-step on chemical structure, bond, 纟_联Sex and motivation - learning can have a deeper study. The use of solid, difficult R can analyze the bond distribution along the aerogel, in order to understand the mesh formed by TE〇S in the sol-condensation Cross-linking · Bonding in the structure. Analysis of the dihydrated Weigel by 29Si solid-state NMR image reveals that 'when Silica is disubstituted', there is a characteristic absorption peak at ·99~_1〇2ppm; Φ When it is tetrasubstituted, there is a characteristic absorption peak at _107~-110PPm. From the 29Si solid-state NMR spectrum analysis in the second figure, it can be found that the main bond appears at -111 ppm for Si tetra-substitution (Q4). 'Secondary bond in -1〇2ppn^Si three substitution (Q3) 'Show two Oxidation &gt;5 aerogel can form Silica network cross-linking and stable structure. • As shown in the fourth and fifth figures, the microscopic shape of the aerogel surface is observed by SEM' by a magnification of 30k. In the fourth graph, the surface of the cerium oxide aerogel can be observed to have a spherical shape, and then observed in the fifth graph with a magnification of 300k times, then the sphere can be found; the shape is more and more Small ruthenium dioxide is composed; the size of the oxidized stone is about 20~30 nm, which proves that the material is a nano-aerogel pore material. · Nitrogen adsorption/desorption analyzer measures the surface properties of materials. Its working principle is to use inert gas to measure the surface physical properties of the material such as pore size, specific surface area and hole structure. The degree of adsorption is closely related to the nature of the sample to be tested and the adsorbed gas used, and is a function of pressure (or concentration) and temperature. Usually we take the P/IVi map for the adsorption amount per gram of sample at constant temperature, and P〇 is the saturated vapor pressure of the analyzed gas at the experimental temperature. This curve is called the adsorption isotherm curve 21 1374266 (MS〇_〇n Is 〇the_). From the sixth figure, the nitrogen absorption/desorption desorption curve of the dioxide dioxide aerogel is prepared. The specific surface area is 8i2m2/g, the pore volume is DW/g, and the average pore size is 94nm. . Further, it can be seen from the seventh figure that the needle-like distribution _ ' indicates that the prepared dihydrated Wei gel has a very concentrated pore distribution range.

The eighth_dioxotomy red is similar to the rotation (under the condition of nitrogen, D = 3 〇 ~ _ ΐ, heating rate = 1 () to (4). The results from the solid line show that the dioxide is a low temperature Yu Qing.c, there is a weight loss of 2.6 Qing, the loss may come from the second silk surface suction _ a little read; # temperature rise to the brain when the 'weight loss of 6.2 wt%, the loss can be silk dioxide On the surface, there are some hydrogen peroxides that are not completely hydrolyzed and condensed. In addition, the part of the dotted line is incompletely aerogel-like in the form of sulfur dioxide. If the temperature rises, the template will start to crack when the temperature rises. Come out, it can also identify whether the cleaning is complete. Different sizes of nano-scale quantum dots can see different glory under the illumination of UV light. The ninth picture shows the image taken by the digital camera after the UV light is irradiated. It can be seen from the figure that the excitation light of the quantum dot used here is in the green light range. The quantum dot modification is to modify the amine group on the surface of the hydrophilic quantum dot, and is prepared for reaction with the carboxyl group on the antibody as an experimental calibration. It can be seen from the tenth figure that the modified quantum dots are measured by a fluorescence spectrometer. The excitation wavelength drifts to about 540 nm, which is caused by the amine group modified to the surface of the deficient point. That is, the modification is the work. The same voltage and time are given in the electrophoresis, and the molecular weight is large. The material with a small molecular weight is slow, and the modified quantum dot has a larger molecular weight than the unmodified quantum dot. Therefore, 22 can be determined by electrophoresis to determine whether the modification is successful. In the eleventh figure, the modified quantum The point moves slower than the unmodified quantum dot, so it can be seen that the modification is successful. In the process of upgrading, because of the addition of buffer solution, the concentration is different from that before the modification. The concentration of the modified quantum dot (4) is measured at a single point of 527 nm, the absorbance is measured, and the concentration is calculated. The formula for calculating the concentration: C = A7Γ Calculate the concentration: where A is the absorbance, ε is the molar absorption coefficient (M example-!, L is the right-handed quantum dot in the particle facet (4) Nine®, _ above formula, A = 0.04 L = 0.512 £ = 77793.9851 Brought into the formula 0.04/(77793.9851 χ 〇.512)==1〇〇χΐ〇,

The concentration to be paid is 1.00 X. The protein is produced here. The quality of the protein, the small molecule of the small molecule, the specificity of the structure, the protein white matter, and the protein are shown in Figure 12. This experiment is applied. The principle of sandwich immune response, using 23 1374266 antibody as a biomaterial, gj is set on (9), and then specifically combined with the corresponding antigen. In addition, antibodies grafted with quantum dots were utilized as calibrations in experiments. If 'specific antigens, antibodies do not bind or antibodies and antigens do not correspond to each other, then 'there is no fluorescence on the scanned image. • The GenePix 4000B, a fluorescent wafer scanner used, uses a simultaneous dual laser (dual_laser) scanning system to produce a ratio image in a timely manner. The proportional image is a composite 24-bit red, green, and blue composite image. The internal reference values of the Kawasaki system are 635 and 532 nm laser light, respectively. The thirteenth picture shows the cleaned blank slide. The background value is very dark blue. Based on this background value, compare the background values of other modified slides to see if the background value is good. As a result of testing the gel wafer, a two-dimensional wafer made of a directly modified slide was used as a comparison group, so the background value of the modified amine-based (2% DAMO) two-dimensional wafer was first discussed. The slide modification procedure was performed by soaking the slide in a dyeing pot containing 2% DAMO solution in ethanol for four hours, washing with a second degree of water, and finally drying in an oven for twenty minutes. In the fourteenth figure, it can be seen that most of the background value of the slide is still dark blue after the amino group modification. Although fine light blue spots appear, the background value is still good. The 10 wt% aerogel dispersion has a three-dimensional structure - a scanned image of the biochip, the dot is a drop of aerogel, and the image is light blue, still falling within the range of low background values. The sixteenth picture shows an amine-modified aerogel, which rises slightly to blue-green. The seventeenth figure shows a three-dimensional "photograph of the appearance of a glass sheet of a gas-gel layer", in which the sample to be tested is The above steps are formed by (1)·(7). Secondly, the positive control 24 points are the epoxy-modified gas condensation directly connected to the amine-modified quantum dots, and the negative control point is the unmodified gas condensation. The purpose of the positive control point is to standardize the biochips produced in different batches, because the quality of different batches of quantum dots may not cause the brightness of the wafer broom to be uncorrelated, resulting in data cannot be compared. Fresh sub-organisms use _ test filaments to make the results more meaningful. In addition, the purpose of the negative control points is to measure the background value of the individual aerogels so that the background value can be subtracted from subsequent data processing. It can be seen that after the probe antibody (M〇use touch gamma positive 6) is dropped, the background value is still _ blue ride. The nineteenth shows that after the sunny reaction, the background value can still be converted to _ color. Twenty-figure can be seen after the sputum reaction and the antigen (HumanIL6) is dropped. The value of the scene is slightly reduced to light blue. The twenty-first picture shows the final-step mark with a good mark &amp; _ city post interest - h Li ^ IL-6), its knot · all light up orange , indicating that the whole sandwich immunoassay succeeded. The twenty-second picture shows the wafer scanning of the two-dimensional protein wafer. This is a two-dimensional protein wafer that will be compared with the three-dimensional aerogel wafer to complete the antibody and antigen specificity in the whole process. The results of the image scanning after the reaction were analyzed by signal analysis on the silk of the DNA chip on the wafer (GenePixPn) 6()). The results are summarized in Tables 1 and 2. Table 1 shows the results of the three-dimensional aerogel wafer analysis, as shown in the following table: Sample position - developer concentration (QD) intensity color 1 " 1.44 χίο'6 '--- 26213 orange yellow 25 2 1.44 xlO·6 28527 3 - ----- 1.44 xlO&quot;6 29041 4 ------- 1.46 xlO'6 ---~ 31609 5 ^---- - 11125 Dial yellow orange yellow orange yellow table 2 for 2D protein wafer analysis results , as shown in the following table: Sample position ^ - Concentration (Μ) Strength ^ Color 1 ------- 1.0 --- 52029 White 2 1.0 X 10'1 One —. 23130 Red 3 -----. 1.0 X 10'2 ~--- 30067 Orange 4 ---- 1.0 X 10*3 22615 Green 5 5.0 X 10'4 16756 Blue According to Table 2, the sample concentration on the 2D protein wafer is 5.0 X 1〇4 At M, the signal strength on the chip is 16750, which is a light blue image close to the background value. Therefore, it is speculated that when the sample is indifferent (1〇_5), the chip scanner cannot detect the signal. value. However, when the three-dimensional aerogel wafer is under a low concentration of L44 X 〇〇-6Μ, the signal intensity is enhanced to 26213~31609 due to the stereostructure relationship of the aerogel, and the three-dimensional aerogel wafer can be seen to increase the surface area while amplifying the signal. strength. Obviously, the invention may have many modifications and differences in accordance with the description in the above embodiments. It is therefore to be understood within the scope of the appended claims, in addition to 26 lines. 1External 'The present invention shoots in the other embodiments to apply for a patent to reproduce the best implementation of the program, and _ give the effect of the invention. #Μ:, 匕 辑 not the spirit of the invention disclosed T The finished red ornaments should be included in the following application. BRIEF DESCRIPTION OF THE DRAWINGS A flow chart of forming a biofilm having a three-dimensional structure and a first embodiment of the present invention is in accordance with a preferred embodiment of the present invention; Three-dimensional structure aerogel layer FTIR spectrum; the first figure is in the preferred embodiment of the invention, the unmodified three-dimensional structure gas condensation wins 2% solid state nuclear magnetic resonance analysis results; the fourth figure is according to the invention A preferred example towel, the surface type of the three-dimensional read structure gas layer is observed by a time-lapse electron microscope (SEM) amplification material; the fifth figure is a scan type according to a preferred example of the present invention. The electron microscope (SEM) magnification is 200,000 times to observe the surface morphology of the three-dimensional structure aerogel layer; 'The sixth figure is a modified example of the three-dimensional structure aerogel layer after QLYMO modification according to the preferred embodiment of the present invention. The measurement data of nitrogen adsorption/desorption; the seventh figure is a pore distribution diagram of the aerogel powder of the three-dimensional structure aerogel layer according to the preferred embodiment of the present invention; the eighth figure is preferred according to the present invention. 'Three-dimensional structure Thermogravimetric analysis (TGA) comparison data before and after gel layer modification; 27 according to the invention, the quantum dot is, the image under excitation; the tenth figure is the preferred detail of the invention, the quality has been changed The sub-point excitation wavelength spectrum; the tenth-picture is an electropherogram of the unmodified and modified mass points according to a preferred embodiment of the present invention; and the twelfth figure is a sandwich immunization according to a preferred example of the present invention Figure 13 is a broom image of a blank glass sheet in accordance with a preferred embodiment of the present invention. Figure 14 is a scanning of a modified amine-based two-dimensional wafer in accordance with a preferred embodiment of the present invention. The fifteenth figure is a biocrystalline image having a three-dimensional structure in the preferred embodiment of the present invention, and the sixteenth embodiment is a preferred example according to the present invention. Wherein the amine-based modified scan image of the bio-wafer having a three-dimensional structure; and the seventeenth embodiment is a photograph of the appearance of the glass sheet having the three-dimensional structure aerogel layer according to a preferred embodiment of the present invention; Eight diagrams are preferred in accordance with the present invention In the example, the amino-based modified bio-wafer having a three-dimensional structure is further immersed in a sweep image of the probe antibody; and the nineteenth aspect is a preferred example according to the present invention. The amine-based modified organism having a three-dimensional structure Crystals&gt;{(4) Knockback image of the broom; the tenth figure is a preferred example of the present invention, wherein the amine-based modification has a three-dimensional standing 28 1374266 body structure and the biofilm is blocked by the reaction. The second scan image is a scan image of a biofilm having a three-dimensional structure and a quantum dot chromogenic antibody on the basis of a preferred embodiment of the present invention; BRIEF DESCRIPTION OF THE DRAWINGS In accordance with a preferred embodiment of the present invention, a two-dimensional protein wafer wafer scan is shown.

29

Claims (1)

1374266 X. Patent application scope: Announcement. 丨.—The method for forming a bio-wafer having a three-dimensional structure, the method for forming a three-dimensional three-dimensional structure*, comprising: providing a gas precursor test, The aerogel transfer solution comprises an ion refining solution, a catalytic hydrolysis and/or condensation reaction reagent, and at least one alkoxide monomer and/or an aromatic oxygen monomer (IV). The catalytic water solution and/or condensation reaction reagent comprises the following groups or any combination thereof, an alcohol, an acidic compound and a basic composition; and the gas lifter precursor solution is subjected to a solution a uniform mixing procedure to facilitate the hydrolysis and polymerization of at least one of the alkoxy monomers and/or aromatic oxy monomers until the specific viscosity of the gas phoenix precursors is greater than the scale of 150 cps; The aerogel precursor solution is allowed to stand so as to continuously hydrolyze and polymerize at least a methoxy- and/or aryloxy monomer, thereby forming an aerogel; • Le-solvent-transfer-extraction The ionic melt contained in the pores of the aerogel; the solvent contained in the dry-sequence gas-filled gel pore towel; the processing-processing procedure is performed by grinding the aerogel into a gas-solid powder Carrying-modification procedure. The quality of the process is performed by mixing-changing_ with the end of the gasification county to introduce at least one specific function on the inner surface and the outer surface of the aerogel powder; and performing-coating Procedure, the coating procedure _ Mingding the modified aerogel The powder is applied to a specific area on a substrate, a biochip, thereby forming a method for forming a 1=specific 1 bio-wafer having a three-dimensional structure, wherein the coating procedure described above comprises: :: the modified coagulation The 谬 powder is dissolved in a sword to form a modified liquid; the modified liquid is applied to a specific area on the substrate; and the m2-baking, baking process is used to remove the "flying" The bath agent 'and increases the adhesion of the modified aerogel powder to the substrate' thereby forming the bio-wafer having a three-dimensional structure. 3. The method of forming a bio-wafer having a three-dimensional structure as described in claim 2, wherein the above-described baking process temperature ranges from 12 叱. The invention relates to a method for forming a bio-wafer having a three-dimensional structure as described in Item 1 wherein the aerogel precursor solution further comprises an acidic compound or an inspective compound. 5. The method for forming a bio-wafer having a three-dimensional structure as described in the application of the third aspect of the invention, wherein the method for forming the aerogel precursor solution comprises: the oxy-monomer and the /-silicoxy group Forming a first mixture with the ionic melt; adding an acidic compound to the first mixture and forming a second mixture; and adding a basic compound to the second mixture to form the precursor solution. 6. The method of forming a bio-wafer having a three-dimensional structure as described in the application of the patent specification, wherein the central element 31 of the alkoxy monomer and/or the aryloxy monomer described above comprises one of the following groups: Li, Na, K, Rb, Cs, Be, Mg, Ca, Sr, Ba, Ti, Te, Cr, Cu, Er, Fe, Ta, V, Zn, Zr, A1, Si, Ge, Sn and Pb. 7' The method for forming a bio-wafer having a three-dimensional structure as described in claim 1, wherein the alkoxy monomer and/or the aryloxy monomer comprises the following group: one of the groups: Tetramethyl orthosilicate (TMOS), tetraethoxy orthosilicate (TEOS), bis (triethoxysilyl)ethane (BTSE), bis〇iethoxySilyi) benzene (BTSB) and tetrabutyl titanate (TBOT) 〇 8. 3D as described in claim i The method for forming a biochip of the structure, wherein the ionic melt is a normal temperature ionic liquid. 9. The method for forming a three-dimensionally structured bio-wafer according to claim i, wherein the above-mentioned ion refining liquid is obtained by mixing an organic base with a Lewis acid, and the Lewis acid is not a halogenated metal acid. . 10. The method of forming a biochip having a three-dimensional standing structure according to claim 9 of the invention, wherein the cationic group in the above organic test may be a leuco group or an aromatic group, and the general formula is as follows: g'Rl, R2, R, 5^# is as follows: R3 R1 R2 ch3 H —&quot; c2h5 H R4 HH 32 1374266 C2H5 H c2h5 H ch3ch2ch2ch2 H ch3 H — (CH3)2CHCH2 H ch3 H CH3CH2CH2CH2 H c2h5 H Ch3 H ch3och2ch2 H ch3 H cf3ch2 H — ch3 ch3 C2Hs H — ch3 ch3 ch3ch2ch2 H C6H6CH2 ch3 CH3CH2CH2 H c6h6ch2 ch3 ch3ch2ch2ch2 H C6H6CH2 ch3 (CH3CH2)(CH3)CH H — c6h6ch2 ch3 ch3ch2ch2ch2ch2 H ch3 H C2H5 ch3 c2h5 H c2H5 ch3 — 11. The method of forming a bio-wafer having a three-dimensional structure according to claim 9, wherein the organic base comprises one of the following groups: ln-butyl-3-methylimidazolium chloride (BMIC), 1 -〇ctanyl -3-methylimidazolium bromide (OMIB), 1 -dodecanyl-3-methylimidazolinm bromide (DMIB) and l-hexadecanyl-3, methylimidazolium bromide (HDMIB). 12. The method of forming a bio-wafer having a three-dimensional structure according to claim 9, wherein the anionic group in the Lewis acid comprises one of the following groups: BF4-, PF6, AsF6-, SbF6 ', F(HF)n-, CF3S〇3·, cf3cf2cf2cf2so3·, (CF3S〇2)2N, (cf3so2)3c_, CF3C〇〇· and cf3cf2cf2coo. 13. The method of forming a bio-wafer having a three-dimensional structure as described in the above-mentioned patent application, wherein the ionic melt has a weight range of about at least one alkoxy 33-based monomer and/or an aromatic oxy group. 10% to 90% of the body weight. 14. The method of forming a bio-wafer having a three-dimensional structure according to the above-mentioned claim, wherein the ionic melt has a weight range of about at least one alkoxy monomer and/or an aryloxy monomer. 50% to 7〇% by weight. I5. The method for forming a bio-wafer having a three-dimensional structure as described in the application of the above-mentioned item, wherein the substrate material comprises one of the following groups: a germanium wafer, a glass or a polymer material. For example, the composition of the bio-wafer having the three-dimensional structure described in the patent application Laid-No., the gas-reducing component of the above-mentioned gas-removing composition includes one of the following groups or any combination thereof: Si〇2, Ti02, V205 and Oh! 2〇3.八17J7Please refine the first item of the biofilm of the Sanshe bribe. The method of the above-mentioned aerogel powder has a particle size range of about (7) coffee to (five) nm 〇 shape to 18:2 patent range item 1 The bio-wafer having a three-dimensional structure, wherein the upper air-guiding powder has an average pore size range of about 2 (10), and the three-dimensional one has a boiling point of less than or equal to the boiling point of the solvent. The shape of the bio-wafer having the three-dimensional structure described in Item 1 of the above-mentioned item 1. The above-mentioned solvent type is included in the following groups: guessing (mtnle), _, _, water, and the like. 1 21. If the patent application method is applied, the temperature axis of the above-mentioned solvent and the dimensional structure of the bio-dimensional structure is about 200 34 ο. 22·= Patent application _ Item 1 The shape of the biochip, the method wherein the modifier used in the above modification procedure has at least one specific group, the specific group comprising one of the following groups: a group and an epoxy group. Amino group, silk, and citric acid 23. The method for forming a biochip having a three-dimensional Lixian structure as described in the above-mentioned application item i after the above-mentioned coating procedure is further included, and further includes a conversion program, the program comprising: Providing a converter 'wherein the converter comprises a first group and at least a second group; a specific functional group linking the inner and outer surfaces of the aerogel powder with the soil and the first of the converter a group to form the biochip having the second group on the surface. 24. The method of forming a bio-wafer having a three-dimensional structure according to claim 23, wherein the transformant comprises one of the following groups: an antigen, a monoclonal antibody, a primary antibody, and a plurality of strains. Polyclonal antibodies, nucleic acids [including monomeric and oligomeric], protein (proteinS), enzymes, lipids, polysaccharides (p〇 iySacchari (ies), sugar (sug s), peptide (p-tides), p〇iypeptides, drugs, viruses, microorganisms and bioligands (bi〇iigands) 0 25 The method for forming a bio-wafer having a three-dimensional structure according to claim 23, further comprising a blocking program after the conversion process is completed, the blocking program comprising: 35 providing - resistance _j 'the towel 'Miscellaneous_comprising-third group; and linking the gum powder (4) to the table Φ without reclining the specific functional group and the third agent of the agent, to determine that the organism has no residue remaining unreacted Functional group. 26. If applied Fill 'material 25 is formed with three groups of biological method for the structure of the wafer, after said blocking process is completed community, further comprising - specific pairing process, the specificity, I1 green pairing procedure comprising: a base-providing pair 'where the pair includes a fourth group and a fifth group; and a fourth group that is sexually bonded to the partner on the biocrystal # to form a surface containing the The biochip of the fifth group. The method for forming a bio-wafer having a three-residue structure according to claim 26, wherein the pairing unit comprises one of the following groups: an antigen, a monoclonal antibody, a primary antibody, and a plurality of Polyclonal antibodies, nucieic acids [including monomeric states φ (m〇n〇meric) and oligomeric states (〇iig〇meric)], proteins (卩(1)(四)阳), enzymes, lipids Ipid, pdysaccharides, sugars, peptides, p〇iypeptides, drugs, viruses, microorganisms, and bioligands. 28. The method for forming a bio-wafer having a three-dimensional structure according to claim 26, further comprising a calibration procedure after the completion of the specific pairing procedure, the calibration procedure comprising: providing a calibration carrier, wherein the calibration The carrier comprises at least a sixth group 36 1J/4Z00 Λ 曰 "seventh group", the seventh substrate is bonded to the calibrator, and the fifth group of the pair is linked to the calibration vector a sixth group to form a biochip for calibration processing. 29. The domain method of a bio-wafer having a three-dimensional structure as described in claim 28, wherein the calibration system is under--or a combination thereof : Fluorescent, light-filling, luminescent, enzymatic, radioactive materials, quantum dots, nano-diamonds. 3〇_The method of forming a bio-wafer of a three-dimensional stereoscopic structure as described in claim 28, wherein the above The calibration vector comprises one of the following groups: monoclonal antibodies (m〇n〇d〇nal antibodies), post-calibration primary antibodies, secondary antibodies, polyclonal antibodies (polyclonaUntibodies), nucleic acids (nucleic acids) [ Including monomeric states (_〇meric) and oligomeric states (〇l_ieriC)], proteins, enzymes, lipids, p〇lysaccharides, sugars, Peptides, polypeptides (p〇lyp immortal (4), drugs (drugs), diseased Qin, microbes and biochemical ligands (bi〇Ugands). 31·3D as described in the patent application The method for forming a three-dimensional structure of a biochip includes one of the following methods: direct immunoassay, indirect immunization, complement-binding immunoassay, and sandwich immunoassay. 32. Formed according to the method of forming the third paragraph of the patent application scope An array type biochip having a three-dimensional structure, the array type biochip having a three-dimensional structure formed according to the method for forming a third aspect of the patent application comprises: a substrate; a three-dimensional formed on the surface of the substrate a three-dimensional structural layer, wherein the three-dimensional structural layer is composed of an aerogel powder, the inner and outer surfaces of the aerogel powder having 37 #53⁄4 id L1% j to a specific g energy group, the specific functional group One of the following groups: an amine group, a hydroxyl group, a carboxylic acid group, and an epoxy group. = An array type organism having a three-dimensional structure formed according to the forming method of item i of claim 32 of claim 32. The wafer, wherein the specific surface area of the gas condensed powder is greater than or equal to 1 〇〇 m 2 /g. 3 "As described in claim 32, according to the forming party of the scope of claim i, the three-dimensional form is formed. A structured array of biochips wherein the aerogel powder has an average pore diameter of less than or equal to 2 〇 nm. 35. An array type biochip having a three-dimensional structure formed according to the forming method of claim section ii according to claim 32, wherein the aerogel powder has a porosity ranging from 5% to 99%. 36. An array type biochip having a three-dimensional structure formed according to the method for forming a patent scope 帛4 according to claim 32, wherein the aerogel powder has a particle size ranging from about 1 nm to about 150! .