CN102061164A - Nano structure for enhancing Raman and fluorescence signals and preparation method thereof - Google Patents
Nano structure for enhancing Raman and fluorescence signals and preparation method thereof Download PDFInfo
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- CN102061164A CN102061164A CN2010105533752A CN201010553375A CN102061164A CN 102061164 A CN102061164 A CN 102061164A CN 2010105533752 A CN2010105533752 A CN 2010105533752A CN 201010553375 A CN201010553375 A CN 201010553375A CN 102061164 A CN102061164 A CN 102061164A
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Abstract
The invention discloses a uniform nano aggregate and a preparation method. The nano aggregate comprises a magnetic nano particle modified with sulfydryl, wherein two sides of the magnetic nano particle are respectively connected with a first metal nano particle and a second metal nano particle through the sulfydryl, the first metal nano particle, the magnetic nano particle and the second metal nano particle are arranged in a line, the particle sizes of the first and the second metal nano particles are 20-400nm and the first and the second metal nano particles are same metal nano particles, and the particle size of the magnetic nano particle is 3-20nm. An object to be tested is orientated to a gap between the metal nano particles by using function groups on the surfaces of the nano particles, thus the great enhancement of the Raman and fluorescence signals of a molecular to be tested is realized. The invention can provides a novel rapid and ultra-sensitive detection method for experiment research, clinical diagnosis and large sample screening.
Description
Technical field
The present invention relates to a kind of function nano material technology field, more specifically, a kind of nanostructure that strengthens Raman and fluorescent signal and preparation method thereof.
Background technology
In current life science, the understanding of significant problems such as vital movement rule and disease generation and development and the research that solution has entered the unit molecule aspect.The development and application molecular level detects the research focus that novel method has become current Medical Biology field, it has not only promoted the revolutionary variation of healthy dependent diagnostic method, and has realistic meaning in biology and medical research, medicament research and development and disease prevention and treatment.
Existing highly sensitive particular sequence nucleic acid detection method generally all is based on polymerase chain reaction (PCR) and fluorescent mark technology.Round pcr can be amplified to target molecule the quantity of enough detections; Fluorescent mark can make detection method have highly sensitive and ease for operation.But because selectivity and non-linear amplification in the PCR process, be that the relative content of amplification gene or sequence all may not reproduction of reality; Fluorescently-labeled detection technique also has many weak points because of the character of fluorescence.
Raman spectrum can provide the chemical structure and the physical message of analyte, and can carry out quantitative or half-quantitative detection.But the application of Raman spectroscopy is general all based on the reinforcing effect of nano material, though the concrete principle of Raman enhanced still is not very clear, but the particle contact position or the gap that more studies show that the metal nanoparticle aggregate have hot spot-effect, [Jiang J, et.al. J. Phys. Chem. B 2003,107:9964-9972; ] can realize that hypersensitive (unit molecule) Raman spectrum detects.At present the hypersensitive raman study of pattern molecule is many is substrate with the colloidal silver aggregate, and the geometrical shape of the particle shape of this substrate and size, gap shape and size and aggregate and polarized state etc. all are difficult to consistent.And Raman signal is subjected to the influence of environment especially big, and therefore, the Raman active substrate that makes up gap character unanimity is very important to using hypersensitive Raman spectrum detection technique.The clearance control of nanometer particle congery is relatively easy, is the Raman active substrate that a kind of potential has homogeneous structural.Have among the preparation method who has reported by direct etching and prepare knot shape (bowtie) aggregate and nanoparticle is assembled into aggregate etc. with bifunctional reagent (molecule) or complementary nucleic acid molecule.[Guerrini L, et.al.
J. Phys. Chem. C2008,112:7527 – 7530; Lim DK, et.al.
Nat. Mater.2010,9:60-67; Pavel I, et.al.
J. Phys. Chem. C2008,112:4880-4883] but this class formation can not satisfy actual biomolecule detection.Because of the bifunctional reagent molecule is nonrigid, the gap of aggregate can be in a kind of fluctuation status because of the pedesis of nanoparticle; More difficult is how with (signal) to be measured molecular orientation to gap area and can limit the orientation of molecule.The research of Nam etc. has realized the gap location of signaling molecule (dyestuff), but and carry out deposition of silver again after the target molecule hybridization, grope silver-colored thickness of the shell and could measure Raman signal, can not satisfy actual sample amplifying nucleic acid fast super sensitivity detection demand.[Lim DK, et.al.
Nat. Mater.2010,9:60-67] make up the metal nanoparticle aggregate that nucleic acid molecule can be directed to the homogeneous of hot spot region and be still a challenging problem so far.
The present invention is a bridge with the magnetic nano-particle of radical functinos such as sulfydryl, amino, aldehyde radical, carboxyl and hydroxyl, connects two metal nanoparticles by sulfydryl; Groups such as amino, aldehyde radical, carboxyl and hydroxyl can be directed to magnetic nano particle sub-surface (hot spot region) for probe molecules such as antibody, aglucon and nucleotide sequences, obtain enhanced Raman or fluorescent signal.
Summary of the invention
Technical problem: the purpose of this invention is to provide a kind of nanostructure that strengthens Raman and fluorescent signal and preparation method thereof, have the incident that to discern mutually based on molecule to occur in the advantage of the magnetic nano particle sub-surface of nanostructure.
Technical scheme: the present invention adopts following technical scheme:
The nanostructure of enhancing Raman of the present invention and fluorescent signal is the magnetic nano-particle that is modified with sulfydryl by, and the both sides of magnetic nano-particle connect first metal nanoparticle and second metal nanoparticle by described sulfydryl respectively, described first metal nanoparticle, the arrangement that is in line of the magnetic nano-particle and second metal nanoparticle, the particle diameter of described first metal nanoparticle and second metal nanoparticle is that 20-400 nm and first metal nanoparticle and second metal nanoparticle are metal nanoparticle of the same race, and the particle diameter of described magnetic nano-particle is 3-20 nm.
The nanostructure preparation method of enhancing Raman of the present invention and fluorescent signal:
Step 1) gets functionalized magnetic nano particle at the finishing sulfydryl of magnetic nano-particle, and method therefor is known in this area;
Described magnetic nano-particle is the spheric nanoparticle, can be metallicl magnetic material such as electrical steel, nickel-base alloy and rare earth alloy etc., can be Ferrite Material;
Step 2) functionalized magnetic nano particle being added concentration is 10
6-10
18Individual particle/liter metal nanoparticle suspension in stir or concussion after 2-48 hour, carry out magnetic field separation, the nanostructure of be enhanced Raman and fluorescent signal, wherein, the number of functionalized magnetic nano particle and metal nanoparticle is than being 1:2-50.Separating the magnetic nano-particle method therefor is known in this area.
Described metal nanoparticle is the spheric nanoparticle, can be that single metal nanoparticle comprises gold and silver, copper, platinum; Can be that compound spheric nanoparticle comprises gold and silver composite nanoparticle and gold, copper and platinum composite nanoparticle; The nanoparticle that can be the compound spherical nucleocapsid comprises galactic nucleus gold core/shell nanoparticles, the compound core/shell nanoparticles of copper nuclear gold and silver, copper nuclear gold and silver double shells nanoparticle; Can be nonmetal SiO
2Examine golden shell, silver-colored shell, copper shell, platinum shell or compound core/shell nanoparticles, described compound core/shell nanoparticles is the core/shell nanoparticles of two or more metal composite in gold and silver, copper and the platinum.
Beneficial effect: compared with prior art, the present invention has following advantage:
(1) the present invention utilizes the nanoparticle of structure homogeneous to make up the nanostructure that strengthens Raman and fluorescent signal, helps obtaining the nanostructure of enhancement factor unanimity, thereby can be used for the equimolecular quantitative molecular of nucleic acid-protein; The inflexible magnetic nano-particle connects two metal nanoparticles, not only is convenient to isolate the purpose nanostructure but also guarantee that the gap is stable, is convenient to quantitative analysis.
(2) the present invention is the magnetic nano-particle functionalization, guarantees that testing molecule navigates to the magnetic nano particle sub-surface and the nanostructure Raman that is in this patent strengthens the strongest position, can improve detection signal to the full extent.Nano structural material with this function is not studied report both at home and abroad.
(3) the present invention is based on the Raman reinforcing effect of nanometer aggregate, set up the detection method of fast super sensitivity, do not need to sample concentrate, processing such as amplification, can reduce the consumption of reagent and shorten the time of detecting.
Description of drawings
The structural representation of Fig. 1 linear pattern nanostructure (wherein the intermediary nanoparticle is a magnetic nano-particle, both sides be metal nanoparticle).
Fig. 2 linear pattern gold nano aggregate Electronic Speculum figure.
The synoptic diagram that Fig. 3 gold nanoshell nanostructure is used in detection of nucleic acids.
Embodiment:
Embodiment 1: the preparation method who strengthens the gold nano structure of Raman and fluorescent signal
Regulate the golden nanometer particle suspension of particle diameter 50 nm, making its concentration is 1 * 10
6-18Individual/liter, the ion of surfaces of metal nanoparticles absorption can form stable double electrical layers, and particle repels mutually because of the electrostatic interaction of its stable double electrical layers.Again in golden nanometer particle suspension with the 1:2-50(number) ratio slowly drip the particle diameter 8 nm ferrite magnetic nano particles (the used method of functionalization is known in this area) of the functionalization of dilution.A spot of magnetic nano-particle in excessive golden nanometer particle suspension, thereby around guaranteeing to have around the magnetic nano-particle a plurality of golden nanometer particles rather than a golden nanometer particle a plurality of magnetic nano-particles are arranged.Magnetic nano-particle is owing to the chemical b ` of its surperficial sulfydryl and gold is adsorbed onto the golden nanometer particle surface, and because the space steric effect that the size difference of two kinds of particles causes, magnetic nano-particle and golden nanometer particle in the aggregate that forms mostly the ratio with 1:2 exist, again because the Coulomb repulsion effect of golden nanometer particle, the gold nano structure that forms is to be the linear pattern enhancing Raman at center and the gold nano structure (as attached Fig. 1 and 2) of fluorescent signal with the magnetic nano-particle, is separating by the action of a magnetic field; Method therefor is known in this area, and can be used.
Embodiment 2: the preparation method who strengthens the golden shell nanostructure of Raman and fluorescent signal
Regulate the nanoparticle suspension of the silica core gold shell structure of particle diameter 180 nm, making its concentration is 1 * 10
10Individual/liter, the ion of golden core/shell nanoparticles surface adsorption can form stable double electrical layers, and particle repels mutually because of the electrostatic interaction of its stable double electrical layers.Again in golden nanometer particle suspension with the 1:10(number) ratio slowly drip the particle diameter 10 nm ferrite magnetic nano particles (the used method of functionalization is known in this area) of the functionalization of dilution.A spot of magnetic nano-particle in excessive golden nanometer particle suspension, thereby around guaranteeing to have around the magnetic nano-particle a plurality of golden core/shell nanoparticles rather than a golden core/shell nanoparticles a plurality of magnetic nano-particles are arranged.Magnetic nano-particle is owing to the chemical b ` of its surperficial sulfydryl and gold is adsorbed onto golden core/shell nanoparticles surface, and because the space steric effect that the size difference of two kinds of particles causes, magnetic nano-particle and golden core/shell nanoparticles in the aggregate that forms mostly the ratio with 1:2 exist, again because the Coulomb repulsion effect of golden core/shell nanoparticles, the golden shell nanostructure that forms is to be the linear pattern enhancing Raman at center and the gold nano structure of fluorescent signal with the magnetic nano-particle, is separating by the action of a magnetic field; Method therefor is known in this area, and can be used.
Embodiment 3: the application of gold nano structure in detection---the detection of nucleic acids (Raman signal) that strengthens Raman and fluorescent signal
The nucleic acid probe molecules of 10 microlitre activated carboxylics (10 nmoles/liter) is added to the gold nano structure (1 * 10 of the graceful and fluorescent signal of haling of the above-mentioned preparation of 50 microlitres
8Individual/liter) in, 4 ℃ are spent the night, the unreacted nucleic acid probe molecules of flush away.Solution to be measured is added in the gold nanoshell nanostructure suspension of stationary probe, hybridized 5 minutes for 72 ℃, 3 times (60 ℃/4 ℃) are handled in thermal cycling again.37 ℃ add the single-chain nucleic acid lytic enzyme again and handled washing and magnetic field separation gold nano structure, Raman spectrometer mensuration nucleic acid characteristic signal (experiment flow such as accompanying drawing 3) 5 minutes.Method therefor is known in this area, and can be used.
Embodiment 4: the application of golden shell nanostructure in detection---the Protein Detection (Raman signal) that strengthens Raman and fluorescent signal
The folate molecule of 10 microlitre activated carboxylics (1 nmole/liter) is added to 50 microlitres gold shell nanoshell nanostructure (1 * 10
10Individual/liter) in, 4 ℃ are spent the night, the unreacted folate molecule of flush away.The epicyte protein extracting solution is added in the golden nanometer particle aggregate suspension of fixing folate molecule, 37 ℃ of incubations 30 minutes, washing and magnetic field separation gold shell nanostructure, Raman spectrometer is measured the characteristic signal of folacin receptor protein molecular.Method therefor is known in this area, and can be used.
Embodiment 5: the application of golden shell nanostructure in detection---the detection of nucleic acids (fluorescent signal) that strengthens Raman and fluorescent signal
The nucleic acid probe molecules of the activated carboxylic of 10 microlitre mark fluorescent signaling molecules (10 nmoles/liter) is added to 50 microlitres gold shell nanostructure (1 * 10
14Individual/liter) in, 4 ℃ are spent the night, the unreacted nucleic acid molecule of flush away, the strength of signal of mensuration fluorescence.Solution to be measured is added in the gold nanoshell nanostructure suspension of stationary probe, hybridized 5 minutes for 72 ℃, 3 times (60 ℃/4 ℃) are handled in thermal cycling again.37 ℃ add the single-chain nucleic acid lytic enzyme again and handled 5 minutes, and washing and magnetic field separation gold shell nanostructure are measured the strength of signal of fluorescence once more, and the decay of calculating fluorescent signal obtains the amount of bind nucleic acid.Method therefor is known in this area, and can be used.
Claims (10)
1. nanostructure that strengthens Raman and fluorescent signal, it is characterized in that, comprise: a magnetic nano-particle that is modified with sulfydryl, and the both sides of magnetic nano-particle connect first metal nanoparticle and second metal nanoparticle by described sulfydryl respectively, described first metal nanoparticle, the arrangement that is in line of the magnetic nano-particle and second metal nanoparticle, the particle diameter of described first metal nanoparticle and second metal nanoparticle is that 20-400 nm and first metal nanoparticle and second metal nanoparticle are metal nanoparticle of the same race, and the particle diameter of described magnetic nano-particle is 3-20 nm.
2. the nanostructure of enhancing Raman according to claim 1 and fluorescent signal is characterized in that, is modified with a kind of in amino, aldehyde radical, carboxyl and the hydroxyl on magnetic nano-particle at least.
3. the nanostructure of enhancing Raman according to claim 1 and fluorescent signal is characterized in that, magnetic nano-particle is a metallicl magnetic material, and described metallicl magnetic material comprises electrical steel, nickel-base alloy or rare earth alloy.
4. the nanostructure of enhancing Raman according to claim 1 and fluorescent signal is characterized in that, magnetic nano-particle is a Ferrite Material.
5. the nanostructure of enhancing Raman according to claim 1 and fluorescent signal is characterized in that, first metal nanoparticle and second metal nanoparticle are metal nanoparticle, and described metal nanoparticle comprises gold and silver, copper, platinum.
6. the nanostructure of enhancing Raman according to claim 1 and fluorescent signal, it is characterized in that, first metal nanoparticle and second metal nanoparticle are compound spheric nanoparticle, and described compound spheric nanoparticle comprises gold and silver composite nanoparticle and gold, copper and platinum composite nanoparticle.
7. the nanostructure of enhancing Raman according to claim 1 and fluorescent signal, it is characterized in that, first metal nanoparticle and second metal nanoparticle are the nanoparticle of compound spherical nucleocapsid, and the nanoparticle of described compound spherical nucleocapsid comprises galactic nucleus gold core/shell nanoparticles, the compound core/shell nanoparticles of copper nuclear gold and silver, copper nuclear gold and silver double shells nanoparticle.
8. the nanostructure of enhancing Raman according to claim 1 and fluorescent signal is characterized in that, first metal nanoparticle and second metal nanoparticle are nonmetallic SiO
2Examine golden shell, silver-colored shell, copper shell, platinum shell or compound core/shell nanoparticles, described compound core/shell nanoparticles is the core/shell nanoparticles of two or more metal composite in gold and silver, copper and the platinum.
9. the preparation method of the nanostructure of described enhancing Raman of claim 1 and fluorescent signal is characterized in that step is as follows:
Step 1) gets functionalized magnetic nano particle at the finishing sulfydryl of magnetic nano-particle;
Step 2) functionalized magnetic nano particle being added concentration is 10
6-10
18Individual particle/liter metal nanoparticle suspension in stir or concussion after 2-48 hour, carry out magnetic field separation, the nanostructure of be enhanced Raman and fluorescent signal, wherein, the number of functionalized magnetic nano particle and metal nanoparticle is than being 1:2-50.
10. preparation method according to claim 9 is characterized in that, modifies amino, aldehyde radical, carboxyl, hydroxyl in the finishing sulfydryl of magnetic nano-particle.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102746848A (en) * | 2011-12-26 | 2012-10-24 | 河北大学 | Preparation method for carboxyl functionalized beta-NaYF4 substrate upconversion fluorescence nanomaterial |
| CN111624186A (en) * | 2020-06-24 | 2020-09-04 | 江南大学 | Enterotoxin spectral analysis method based on fluorescence and Raman double-signal enhancement |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101089615A (en) * | 2007-06-22 | 2007-12-19 | 东南大学 | Preparation method of magnetic surface strength laman scattering mark |
| CN101680900A (en) * | 2007-03-20 | 2010-03-24 | 贝克顿·迪金森公司 | Use the check of Surface enhanced raman spectroscopy (SERS)-active particle |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101680900A (en) * | 2007-03-20 | 2010-03-24 | 贝克顿·迪金森公司 | Use the check of Surface enhanced raman spectroscopy (SERS)-active particle |
| CN101089615A (en) * | 2007-06-22 | 2007-12-19 | 东南大学 | Preparation method of magnetic surface strength laman scattering mark |
Non-Patent Citations (1)
| Title |
|---|
| 蒋彩云; 钱卫平: "纳米金-PNIPAM类智能凝胶", 《化学进展》, vol. 22, no. 8, 31 August 2010 (2010-08-31), pages 1626 - 1632 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102746848A (en) * | 2011-12-26 | 2012-10-24 | 河北大学 | Preparation method for carboxyl functionalized beta-NaYF4 substrate upconversion fluorescence nanomaterial |
| CN102746848B (en) * | 2011-12-26 | 2013-07-31 | 河北大学 | Preparation method for carboxyl functionalized beta-NaYF4 substrate upconversion fluorescence nanomaterial |
| CN111624186A (en) * | 2020-06-24 | 2020-09-04 | 江南大学 | Enterotoxin spectral analysis method based on fluorescence and Raman double-signal enhancement |
| CN111624186B (en) * | 2020-06-24 | 2021-03-16 | 江南大学 | Enterotoxin spectral analysis method based on fluorescence and Raman double-signal enhancement |
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Application publication date: 20110518 |