CN106841139A - A kind of method based on monomolecular detection hydrogen peroxide - Google Patents
A kind of method based on monomolecular detection hydrogen peroxide Download PDFInfo
- Publication number
- CN106841139A CN106841139A CN201710040860.1A CN201710040860A CN106841139A CN 106841139 A CN106841139 A CN 106841139A CN 201710040860 A CN201710040860 A CN 201710040860A CN 106841139 A CN106841139 A CN 106841139A
- Authority
- CN
- China
- Prior art keywords
- hydrogen peroxide
- time
- area
- detecting hydrogen
- slide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2201/00—Features of devices classified in G01N21/00
- G01N2201/06—Illumination; Optics
- G01N2201/061—Sources
- G01N2201/06113—Coherent sources; lasers
Landscapes
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Analytical Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Optics & Photonics (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
Generation, ribozyme the present invention relates to a kind of method based on monomolecular detection hydrogen peroxide, including ribozyme construct are fixed on slide and utilize single molecular fluorescence microscope tracking monitor process.The method is fixed on modification interface by by the compound of G tetrads and hemin, the reaction of fluorescence-causing substance is generated using complex catalysts unstressed configuration substrate, with reference to single molecular fluorescence microscope, real-time monitoring detection process.The advantage of the invention is that using single molecular fluorescence microscopy is highly sensitive, high time resolution the characteristics of, detection process real-time monitored can be imaged, signal to noise ratio is high.Can obtain the information of the single detection unit catalysis single fluorescence-causing substance of generation.
Description
Technical field
The present invention relates to a kind of method that utilization single molecular fluorescence microscopy detects content of hydrogen peroxide.The invention belongs to life
Thing sensing detection field.
Background technology
Traditional optical imagery is different from, imaging mode is pushed away new to the new stage by the appearance of molecule detection：Take off
Show the inhomogeneity and its unique dynamic process for being hidden in molecule under ensemble average level.Using single molecular fluorescence microscopy,
It can be found that the interaction of intermolecular difference and molecule and its residing micro-nano environment, and by the real-time of individual molecule
Follow the trail of, the real-time dynamic change of individual molecule can be obtained.Based on the method, various enzyme molecules are urged with inorganic nano catalyst
Change characteristic to be resolved（Science 1998, Nature Materials 2008 etc.）.Single molecule fluorescence detection method has simultaneously
Splendid signal to noise ratio, detection sensitivity and high temporal resolution, thus in terms of being also applied to detection of nucleic acids（Nature
Communication 2015）.
Hydrogen peroxide（H2O2）Many important physiology enter in regulating cell, in life system, host defense and cell
Signal transduction plays dual parts of, thus " necessary demon " is known as in nearest report.Due to H2O2Important work(
Can, titrating, fluoremetry, the method such as electrochemical method is used for the new H of exploitation2O2The strategy of detection.Wherein, electrochemical method
Have the advantages that direct and detect in real time, however, traditional electrochemical sensing method is difficult to provide intracellular space letter
Breath and image-forming information.Some are to H2O2Very sensitive fluorescence molecule is also used as the probe of intracellular or extracellular detection.
But these fluorescent chromophores are easy to be bleached, therefore it is difficult in physiological environment to H2O2The dynamic change of content is long
Time observation.Therefore at present in the urgent need to a kind of sensitive, imaging clearly, real-time detection method.
The content of the invention
The problem that the present invention exists for existing detection scheme sensitivity, signal to noise ratio, imaging aspect, proposes using using single
Molecular fluorescence microscopy combination ribozyme（DNAzyme）Develop a kind of method of sensitive detection hydrogen peroxide.The method can be to inspection
Survey process carries out real time imagery tracking, and can capture single product generating process.
In the inventive solutions, using ribozyme G- tetrads（G-quadruplex）It is fixed on slide, follows the trail of
The reaction of its catalysis Amplex Red generation fluorescence-causing substance resorufin carries out signal acquisition.
The method of the present invention is specially：
A kind of method based on monomolecular detection hydrogen peroxide, it is characterised in that comprise the following steps：
（1）The generation of ribozyme construct
The fixed chain of the double modifications of a certain proportion of G- tetrads DNA and Cy 5, biotin is well mixed, 10 are incubated in 95 DEG C
Min, slow cooling makes G- tetrads hybridize and form tetrad structure with fixed chain, obtains complexⅠ；
（2）Ribozyme is fixed on slide
After dilution complexⅠ to finite concentration, with hemin（hemin）A period of time is incubated in 25 DEG C with certain proportion, is obtained
Complexⅱ；After the slide of complexⅱ and Streptavidin modification is incubated into certain hour, wash buffer is totally excessively multiple
Compound；
（3）Using single molecular fluorescence microscope tracking monitor process
100 microlitres of reaction solutions are added dropwise on slide, reaction solution includes certain density hydrogen peroxide and Amplex Red；Using list
Molecular fluorescence microscope real-time monitored complexⅱ catalysis unstressed configuration Amplex Red generation fluorescence-causing substance resorufins
（resorufin）Process.
The buffer solution for being used is this area routine cushioning liquid, and it is to contain pH=7.4,100 mM K preferably to buffer molten
Cl, 10 mM phosphate buffers.
The G- tetrads DNA for being used is the conventional G- tetrad DNA sequence dnas in this area, preferably in DNA sequence dna table
Sequence；The G- tetrads DNA for being used can be the conventional ratio in this area with the ratio of fixed chain, and preferred ratio is 1:1.
The dilution complexⅠ concentration for being used is to 10-11-10-9 M, preferred concentration is 5 × 10-11M；What is used answers
It can be the conventional ratio in this area that compound I is incubated ratio with hemin, and preferred ratio is 2:1.
The complexⅠ for being used can be this area conventional time with hemin incubation time, and the preferred time is 20
min。
The cover glass step of the Streptavidin modification for being used was published in Nature with reference on May 29th, 2008
The entitled A practical guide to single-molecule FRET's of the 507-516 pages of volume 6 of the 5th phases of Methods
The method of modifying described in S6 in paper Supporting Online Information；The complexⅱ and chain for being used
The incubation time of the cover glass of mould Avidin modification can be this area conventional time, and the preferred time is 10 min.
The single molecular fluorescence microscope for being used for being used is this area conventional microscopy, and preferably total internal reflection is glimmering
Light microscope.Described single molecular fluorescence microscope imaging parameter can be the conventional parameter in this area, preferably 647,561
, in 5 % or so, preferably time for exposure control is in 30-50 ms for nm laser controllings；Described single molecular fluorescence microscope imaging bar
Part can be the conventional condition in this area, and at 20 DEG C or so, the humid control of the imaging is 50 for preferably imaging temperature control
Below %.
In 100-200 nM, preferred concentration is 100 nM for the Amplex Red concentration ranges control for being used.
During data acquisition, the position of complexⅱ is determined first with the fluorescence molecules of Cy 5, and the fluorescence-causing substance for producing exists
561 nm laser excitations, when the two can with common location when, the collection curve that changes over time of fluorescence-causing substance.
The H of minimum detectable 100 pM2O2。
The cover glass step of the Streptavidin modification for being used was published in Nature with reference on May 29th, 2008
The entitled A practical guide to single-molecule FRET's of the 507-516 pages of volume 6 of the 5th phases of Methods
The method of modifying described in S6 in paper Supporting Online Information.
The advantage of the invention is that using single molecular fluorescence microscopy is highly sensitive, high time resolution the characteristics of, can be right
Detection process real-time monitored is imaged.Can obtain the information of the single detection unit catalysis single fluorescence-causing substance of generation.
Brief description of the drawings
The fluorescence microscope image of the product that Fig. 1 is generated by case study on implementation 1.
The real-time curve that Fig. 2 is changed over time for the fluorescence intensity of single complexⅱ generation product in case study on implementation 1.Figure
3 real-time curves changed over time for the fluorescence intensity of single complexⅱ generation product in case study on implementation 2.
The real-time curve that Fig. 4 is changed over time for the fluorescence intensity of single complexⅱ generation product in case study on implementation 3.
Specific embodiment
Technical scheme is further described below by way of specific embodiment.Following embodiment is to this
That invents further illustrates, and does not limit the scope of the invention.
Embodiment 1
Streptavidin modifies slide：Slide is sequentially placed into 10 % Alconox solution, acetone, piranha solution（The concentrated sulfuric acid：
30% H2O2=3:1）, need ultra-pure water to clean after each step ultrasound after ultrasonic 30 min in 1 M K OH solution, nitrogen drying.Flame
Lamp calcination slide（Several seconds）, to remove the fluorescent organic molecule remained on slide.Contain 1 by what slide was put into fresh configuration
% silylating reagents（APTES）Ethanol solution in, concussion be incubated 1 hr, subsequent slide ethanol thoroughly wash and with surpass
Pure water rinsing, N2 Drying is standby.The polyethylene glycol of 0.2 mg biotin modifications（biotin-PEG）And 8 mg polyethylene glycol
（mPEG）It is dissolved in 0.1 M sodium bicarbonate of fresh configuration（Na HCO3, pH 8.5）In solution, then mixed liquor is incubated
In slips over night（In wet box）.By slide ultra-pure water thoroughly cleaning, N2Drying.By company's enzyme Avidin drop of 0.2 mg/mL
It is added on the slide of biotin modification, clean with ultra-pure water cleaning down after 10 min are incubated in wet box, N2Drying is stand-by.
By the fixed chain DNA of 1 μM of G- tetrads DNA and double modifications with 1:1 ratio is well mixed, in 95 DEG C of water-baths 10
Min, slow cooling makes G- tetrads hybridize and form tetrad structure with fixed chain, obtains complexⅠ.
50 μ L complexⅠs are diluted to 100 pM, with 50 pM hemins（hemin）With 2:1 ratio is well mixed, and 25
DEG C be incubated 20 min, obtain complexⅱ.After sticking fence on the slide of Streptavidin modification, build sample cell, by 100
μ L complexⅱs are added dropwise in sample cell, after 25 DEG C are incubated 10 min（Note lucifuge）, wash buffer is totally excessively combined
Thing.
Slide is placed on microscope work platform, 100 μ L reaction solutions are added dropwise（100 nM Amplex Red, 1 μM
H2O2）, single molecular fluorescence microscope parameter is adjusted, 5% or so, the time for exposure controls in 30 ms 647,561 nm laser controllings,
Real-time monitored complexⅱ catalysis unstressed configuration Amplex Red generation fluorescence-causing substance resorufins（resorufin）Process.
The fluorescence microscope image of the product that Fig. 1 is generated by case study on implementation 1.Can be obtained by figure, when containing H2O2It is anti-
After answering liquid to add sample cell, can observe on originally clean interface and multiple fluorescence bright spots occur, show there is product to generate.Specifically
Collecting method is shown in the content of the invention.Fig. 2 is the fluorescence intensity anaplasia at any time of single complexⅱ generation product in case study on implementation 1
The real-time curve of change.Can be obtained by figure, with this understanding, the Mean Speed of single complexⅱ generation product is 1.13 s-1。
Embodiment 2
Streptavidin modifies slide：Slide is sequentially placed into 10 % Alconox solution, acetone, piranha solution（The concentrated sulfuric acid：
30% H2O2=3:1）, need ultra-pure water to clean after each step ultrasound after ultrasonic 30 min in 1 M K OH solution, nitrogen drying.Flame
Lamp calcination slide（Several seconds）, to remove the fluorescent organic molecule remained on slide.Contain 1 by what slide was put into fresh configuration
% silylating reagents（APTES）Ethanol solution in, concussion be incubated 1 hr, subsequent slide ethanol thoroughly wash and with surpass
Pure water rinsing, N2Drying is standby.The polyethylene glycol of 0.2 mg biotin modifications（biotin-PEG）And 8 mg polyethylene glycol
（mPEG）It is dissolved in 0.1 M sodium bicarbonate of fresh configuration（Na HCO3, pH 8.5）In solution, then mixed liquor is incubated
In slips over night（In wet box）.By slide ultra-pure water thoroughly cleaning, N2Drying.By company's enzyme Avidin drop of 0.2 mg/mL
It is added on the slide of biotin modification, clean with ultra-pure water cleaning down after 10 min are incubated in wet box, N2Drying is stand-by.
By the fixed chain DNA of 1 μM of G- tetrads DNA and double modifications with 1:1 ratio is well mixed, in 95 DEG C of water-baths 10
Min, slow cooling makes G- tetrads hybridize and form tetrad structure with fixed chain, obtains complexⅠ.
50 μ L complexⅠs are diluted to 100 pM, with 50 pM hemins（hemin）With 2:1 ratio is well mixed, and 25
DEG C be incubated 20 min, obtain complexⅱ.After sticking fence on the slide of Streptavidin modification, build sample cell, by 100
μ L complexⅱs are added dropwise in sample cell, after 25 DEG C are incubated 10 min（Note lucifuge）, wash buffer is totally excessively combined
Thing.
Slide is placed on microscope work platform, 100 μ L reaction solutions are added dropwise（100 nM Amplex Red, 10 nM
H2O2）, single molecular fluorescence microscope parameter is adjusted, 5% or so, the time for exposure controls in 30 ms 647,561 nm laser controllings,
Real-time monitored complexⅱ catalysis unstressed configuration Amplex Red generation fluorescence-causing substance resorufins（resorufin）Process.
The real-time curve that Fig. 2 is changed over time for the fluorescence intensity of single complexⅱ generation product in case study on implementation 1.By
Can be obtained in figure, with this understanding, the Mean Speed of single complexⅱ generation product is 0.14 s-1。
Embodiment 3
Streptavidin modifies slide：Slide is sequentially placed into 10 % Alconox solution, acetone, piranha solution（The concentrated sulfuric acid：
30% H2O2=3:1）, need ultra-pure water to clean after each step ultrasound after ultrasonic 30 min in 1 M K OH solution, nitrogen drying.Flame
Lamp calcination slide（Several seconds）, to remove the fluorescent organic molecule remained on slide.Contain 1 by what slide was put into fresh configuration
% silylating reagents（APTES）Ethanol solution in, concussion be incubated 1 hr, subsequent slide ethanol thoroughly wash and with surpass
Pure water rinsing, N2Drying is standby.The polyethylene glycol of 0.2 mg biotin modifications（biotin-PEG）And 8 mg polyethylene glycol
（mPEG）It is dissolved in 0.1 M sodium bicarbonate of fresh configuration（Na HCO3, pH 8.5）In solution, then mixed liquor is incubated
In slips over night（In wet box）.By slide ultra-pure water thoroughly cleaning, N2Drying.By company's enzyme Avidin drop of 0.2 mg/mL
It is added on the slide of biotin modification, clean with ultra-pure water cleaning down after 10 min are incubated in wet box, N2Drying is stand-by.
By the fixed chain DNA of 1 μM of G- tetrads DNA and double modifications with 1:1 ratio is well mixed, in 95 DEG C of water-baths 10
Min, slow cooling makes G- tetrads hybridize and form tetrad structure with fixed chain, obtains complexⅠ.
50 μ L complexⅠs are diluted to 100 pM, with 50 pM hemins（hemin）With 2:1 ratio is well mixed, and 25
DEG C be incubated 20 min, obtain complexⅱ.After sticking fence on the slide of Streptavidin modification, build sample cell, by 100
μ L complexⅱs are added dropwise in sample cell, after 25 DEG C are incubated 10 min（Note lucifuge）, wash buffer is totally excessively combined
Thing.
Slide is placed on microscope work platform, 100 μ L reaction solutions are added dropwise（100 nM Amplex Red, 1 nM
H2O2）, single molecular fluorescence microscope parameter is adjusted, 5% or so, the time for exposure controls in 30 ms 647,561 nm laser controllings,
Real-time monitored complexⅱ catalysis unstressed configuration Amplex Red generation fluorescence-causing substance resorufins（resorufin）Process.
The real-time curve that Fig. 2 is changed over time for the fluorescence intensity of single complexⅱ generation product in case study on implementation 1.By
Can be obtained in figure, with this understanding, the Mean Speed of single complexⅱ generation product is 0.05 s-1。
<110>Shanghai National Engineering Research Center for Nanotechnology Co., Ltd
<120>A kind of method based on monomolecular detection hydrogen peroxide
<140>
<141>
<160>2
<210>1
<211>45
<212>DNA
<213>Artificial sequence
<400>
aataataata ataataataa taattttggg tagggcgggt tggga
<210>2
<211>28
<212>DNA
<213>Artificial sequence
<220>
<221>misc feature
<222>(1)
<223>3' ends biotin modification, 5' ends cy 5 is modified
<400>
aataataata ataataataa taattttt
Claims (10)
1. it is a kind of based on the monomolecular method for detecting hydrogen peroxide, it is characterised in that to comprise the following steps：
（1）The generation of ribozyme construct
The fixed chain of the double modifications of a certain proportion of G- tetrads DNA and Cy 5, biotin is well mixed, 10 are incubated in 95 DEG C
Min, slow cooling makes G- tetrads hybridize and form tetrad structure with fixed chain, obtains complexⅠ；
（2）Ribozyme is fixed on slide
After dilution complexⅠ to finite concentration, with hemin（hemin）A period of time is incubated in 25 DEG C with certain proportion, is obtained
Complexⅱ；After the slide of complexⅱ and Streptavidin modification is incubated into certain hour, wash buffer is totally excessively multiple
Compound；
（3）Using single molecular fluorescence microscope tracking monitor process
100 microlitres of reaction solutions are added dropwise on slide, reaction solution includes certain density hydrogen peroxide and Amplex Red；Using list
Molecular fluorescence microscope real-time monitored complexⅱ catalysis unstressed configuration Amplex Red generation fluorescence-causing substance resorufins
（resorufin）Process.
2. it is according to claim 1 a kind of based on the monomolecular method for detecting hydrogen peroxide, it is characterised in that what is used is slow
Fliud flushing is this area routine cushioning liquid, and it is to delay containing pH=7.4,100 mM K Cl, 10 mM phosphate preferably to buffer molten
Fliud flushing.
3. it is according to claim 1 a kind of based on the monomolecular method for detecting hydrogen peroxide, it is characterised in that the G- for being used
Tetrad DNA is the conventional G- tetrad DNA sequence dnas in this area, preferably sequence in DNA sequence dna table；The G- tetrads for being used
Body DNA can be the conventional ratio in this area with the ratio of fixed chain, and preferred ratio is 1:1.
4. it is according to claim 1 a kind of based on the monomolecular method for detecting hydrogen peroxide, it is characterised in that to be used
Dilute complexⅠ concentration to 10-11-10-9 M, preferred concentration is 5 × 10-11M；The complexⅠ for being used is incubated with hemin
Ratio can be the conventional ratio in this area, and preferred ratio is 2:1.
5. it is according to claim 1 a kind of based on the monomolecular method for detecting hydrogen peroxide, it is characterised in that to be used
ComplexⅠ can be this area conventional time with hemin incubation time, and the preferred time is 20 min.
6. it is according to claim 1 a kind of based on the monomolecular method for detecting hydrogen peroxide, it is characterised in that what is used answers
Compound II can be this area conventional time with the incubation time of the cover glass of Streptavidin modification, and the preferred time is 10
min。
7. it is according to claim 1 a kind of based on the monomolecular method for detecting hydrogen peroxide, it is characterised in that the institute for being used
The single molecular fluorescence microscope for using is this area conventional microscopy, preferably utilizing total internal reflection fluorescence microscope, described list
Molecular fluorescence microscope imaging parameter can be the conventional parameter in this area, and preferably 647,561 nm laser controllings are left in 5 %
The right side, preferably time for exposure are controlled in 30-50 ms；Described single molecular fluorescence microscope imaging condition can be normal for this area
The condition of rule, preferably imaging temperature are controlled at 20 DEG C or so, and the humid control of the imaging is in 50 below %.
8. it is according to claim 1 a kind of based on the monomolecular method for detecting hydrogen peroxide, it is characterised in that to be used
In 100-200 nM, preferred concentration is 100 nM to the control of Amplex Red concentration ranges.
It is 9. according to claim 1 a kind of based on the monomolecular method for detecting hydrogen peroxide, it is characterised in that during data acquisition,
The position of complexⅱ, and the fluorescence-causing substance for producing are determined first with the fluorescence molecules of Cy 5 in 561 nm laser excitations, when two
Person can with common location when, the collection curve that changes over time of fluorescence-causing substance.
10. it is according to claim 1 a kind of based on the monomolecular method for detecting hydrogen peroxide, it is characterised in that minimum to examine
Measure the H of 100 pM2O2。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710040860.1A CN106841139A (en) | 2017-01-20 | 2017-01-20 | A kind of method based on monomolecular detection hydrogen peroxide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710040860.1A CN106841139A (en) | 2017-01-20 | 2017-01-20 | A kind of method based on monomolecular detection hydrogen peroxide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106841139A true CN106841139A (en) | 2017-06-13 |
Family
ID=59119255
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710040860.1A Pending CN106841139A (en) | 2017-01-20 | 2017-01-20 | A kind of method based on monomolecular detection hydrogen peroxide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106841139A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107630066A (en) * | 2017-08-10 | 2018-01-26 | 上海纳米技术及应用国家工程研究中心有限公司 | A kind of method that cascade reaction is studied under the conditions of unimolecule |
| CN107630067A (en) * | 2017-08-10 | 2018-01-26 | 上海纳米技术及应用国家工程研究中心有限公司 | Single molecular fluorescence observation based on DNA paper foldings is lower to determine signal acquisition site |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105018474A (en) * | 2014-08-22 | 2015-11-04 | 江苏省原子医学研究所 | Probe based on G-quadruplex-chlorine heme DNA enzyme and application of probe |
| CN105802963A (en) * | 2016-04-01 | 2016-07-27 | 中国科学院成都生物研究所 | Oligonucleotide probe |
-
2017
- 2017-01-20 CN CN201710040860.1A patent/CN106841139A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105018474A (en) * | 2014-08-22 | 2015-11-04 | 江苏省原子医学研究所 | Probe based on G-quadruplex-chlorine heme DNA enzyme and application of probe |
| CN105802963A (en) * | 2016-04-01 | 2016-07-27 | 中国科学院成都生物研究所 | Oligonucleotide probe |
Non-Patent Citations (3)
| Title |
|---|
| CHI-LIN LI 等: "Fluorescence Detection of Lead(II) Ions Through Their Induced Catalytic Activity of DNAzymes", 《ANALYTICAL CHEMISTRY》 * |
| EYAL GOLUB 等: "Hemin/G-quadruplexes as DNAzymes for the fluorescent detection of DNA,aptamer-thrombin complexes, and probing the activity of glucose oxidase", 《ANALYST》 * |
| 段晓雷: "Pif1解旋酶解旋G4 DNA分子机制的研究", 《中国博士学位论文全文数据库 基础科学辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107630066A (en) * | 2017-08-10 | 2018-01-26 | 上海纳米技术及应用国家工程研究中心有限公司 | A kind of method that cascade reaction is studied under the conditions of unimolecule |
| CN107630067A (en) * | 2017-08-10 | 2018-01-26 | 上海纳米技术及应用国家工程研究中心有限公司 | Single molecular fluorescence observation based on DNA paper foldings is lower to determine signal acquisition site |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20210164039A1 (en) | Method of in situ gene sequencing | |
| Haustein et al. | Single-molecule spectroscopic methods | |
| Zhang et al. | Resolving cadherin interactions and binding cooperativity at the single-molecule level | |
| Orosco et al. | Real-time monitoring of enzyme activity in a mesoporous silicon double layer | |
| Tanaka et al. | Orientation dependence of displacements by a single one-headed myosin relative to the actin filament | |
| Treffer et al. | Recent advances in single-molecule sequencing | |
| Licatalosi et al. | Approaches for measuring the dynamics of RNA–protein interactions | |
| CN107727705A (en) | A kind of enzyme reaction detects nano-pore electric sensor | |
| Ni et al. | A fluorescent assay for alkaline phosphatase activity based on inner filter effect by in-situ formation of fluorescent azamonardine | |
| CN106841139A (en) | A kind of method based on monomolecular detection hydrogen peroxide | |
| US8916343B2 (en) | DNA biochip and methods of use | |
| Ding et al. | Recent advances in single cell analysis by electrochemiluminescence | |
| Bracamonte | Microarrays towards nanoarrays and the future Next Generation of Sequencing methodologies (NGS) | |
| CN107884373A (en) | The method that prostate specific antigen is detected under the conditions of unimolecule | |
| Michelotti et al. | A bird's eye view: tracking slow nanometer-scale movements of single molecular nano-assemblies | |
| US20230109070A1 (en) | Clinical- and industrial-scale intact-tissue sequencing | |
| Thompson et al. | Total internal reflection with fluorescence correlation spectroscopy: Applications to substrate-supported planar membranes | |
| Zhang et al. | Instrumentation and metrology for single RNA counting in biological complexes or nanoparticles by a single-molecule dual-view system | |
| CN103207167B (en) | Preparation method of fluorescence resonance system for rapid detection of ATP in mitochondria | |
| CN106918584B (en) | A method of the single molecular fluorescence Resonance energy transfer based on photoactivation | |
| Wang et al. | A group-targeting biosensor for sensitive and rapid detection of quinolones in water samples | |
| Kahlscheuer et al. | Single-molecule pull-down FRET to dissect the mechanisms of biomolecular machines | |
| CN106337082A (en) | Enzyme process generated random G-quadruplex-based cell apoptosis label-free detection method | |
| CN104991090A (en) | Method for detecting single molecule horizontal intermolecular interaction by use of atomic force microscope (AFM) | |
| Hwang et al. | Single-molecule FRET: methods and biological applications |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170613 |
|
| RJ01 | Rejection of invention patent application after publication |