CN110331188A - A kind of detection method of internal pH - Google Patents
A kind of detection method of internal pH Download PDFInfo
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- CN110331188A CN110331188A CN201910642270.5A CN201910642270A CN110331188A CN 110331188 A CN110331188 A CN 110331188A CN 201910642270 A CN201910642270 A CN 201910642270A CN 110331188 A CN110331188 A CN 110331188A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6816—Hybridisation assays characterised by the detection means
- C12Q1/6818—Hybridisation assays characterised by the detection means involving interaction of two or more labels, e.g. resonant energy transfer
Abstract
The present invention relates to a kind of internal pH measuring methods, it is response pH element with i-motif structure, by in a pair of of fluorescence resonance energy transfer (FRET) fluorescence pair of i-motif connection, positive charge fluorescence is quenched in conjunction with group of the lanthanides organic metal framework lamella in i-motif, after subsequent i-motif and proton respond, it is dissociated from lamella, generates fluorescence resonance energy transfer, the measurement of internal pH is realized by the common location and color change of two kinds of fluorescence.The present invention designs FRET pairs of the both ends i-motif label a pair of, and positively charged TAMRA fluorescence is quenched using lanthanide series metal organic frame nanometer layer, retains another electronegative FAM fluorescence.Method of the invention has preferable photostability, anti-interference, and detection has splendid time and spatial resolution, quickly can change quantitative detection to pH in the physiology courses such as cell transfecting intracellular.
Description
Technical field
The present invention relates to a kind of detection methods for measuring internal pH, and in particular to one kind is based on i-motif response pH hair
Raw shape changes, and changes so as to cause fluorescence probe FRET effect, the method for measuring pH by common location.The invention belongs to
Single cell analysis field.
Background technique
Internal pH it is many it is intracellular activity in play important purposes, as cell cycle, cell phagocytosis, Apoptosis,
Multiple drug resistance, ion channel and contraction of muscle etc..PH can regulate and control the structure and function of intracellular protein, some organelles, such as
Lysosome and mitochondria utilize proton channel synthesis ATP to keep normal physiological function.The pH environment of disorder intracellular is not only
It will affect cell internalizing approach, it might even be possible to nervous system is damaged, so as to cause the generation of some diseases, such as myocardial ischemia, cancer
Disease and Alzheimer's disease etc..Therefore disease is found for understanding and quantifying intracellular events to the detection of pH variation in living cells
The origin cause of formation has great importance and researching value.
Traditional internal pH measuring method has: nuclear magnetic resonance method (Chinese patent: a kind of pH measuring method, publication number:
CN108195867A.)；Microelectrode method: (Chinese patent: biological neural chip and its preparation of collection optoelectronic pole and microelectrode one
Method, publication number: CN108389931A.)；Fluorescence probe method: (Chinese patent: a kind of internal pH value detection method, it is open
Number: CN108535169A.).However nuclear-magnetism method minute resolution ratio is poor, not can be carried out unicellular pH value measurement；Micro- electricity grade
Method need to precisely hold electrode insertion position, and operation difficulty is big；And the fluorescence probe quantum yield of pH sensitivity is low, easily drift in the cell
It is high to moor background, and has certain toxic side effect to cell.Disadvantages described above limits the application that these methods measure internal pH,
It is badly in need of researching and developing a kind of detection method of sensitive, stable internal pH.
DNA has unique advantage in molecular recognition field, and specificity and flexibility become building nanoscale essence
The foundation stone of true self-assembled structures.I-motif structure is four sensitive chain helical DNA structures of a kind of pair of proton, it has been designed
For the efficient molecular motor and different kinds of molecules device driven using acid/base.Artificial synthesized nanostructure can use not
Same Complementary hybridization, aptamers and metal ion combines the structure for converting itself, to cope with the stimulation of external environment.DNA nanometers
There is structure excellent procedural and biocompatibility can simulate the intracellular events of organelle in a certain respect.It is rich
DNA nano-component containing cytimidine can be used as proton detection device, and in acid condition, the nucleotide containing cytimidine can be with shape
At i-motif structure, i.e. two parallel C-HC+The hairpin structure of composition is mutually embedding in antiparallel mode.This i-
Motif structure can change quick response to pH, higher stability is able to maintain in several circulations, and will not generate nocuousness
By-product.I-motif can change structure in the range of pH 5-7, be very suitable for detecting the change of internal pH.Therefore
The novel detection method research significance with higher and application of response can be changed to internal pH using the design of i-motif structure
Value.
Summary of the invention
For the demand of existing detection, it is an object of that present invention to provide a kind of detection methods of internal pH, and one kind is with i-
Motif structure is to respond the internal pH measuring method of pH element.
The object of the invention is achieved through the following technical solutions: a kind of detection method of internal pH is with i-motif structure
PH element is responded, by connecting a pair of of fluorescence resonance energy transfer (FRET) fluorescence pair, i-motif in i-motif structural DNA
Positive charge fluorescence is quenched in structural DNA in conjunction with group of the lanthanides organic metal framework lamella, and subsequent i-motif structural DNA and proton respond
Afterwards, it is dissociated from lamella, generates fluorescence resonance energy transfer, pass through the common location and color change realization internal pH of two kinds of fluorescence
Measurement, comprising the following steps:
(1) i-motif structural DNA and lanthanide series metal organic frame nanoscale twins are incubated for:
2 15 ~ 30 μM of the μ L linear ss-DNA of i-motif, including nucleic acid sequence table Seq. No.1-4 DNA 1-4 and 0.2
Mg/mL lanthanide series metal organic frame nanoscale twins (MOF-La) are in 1 mL, 10 mM phosphate buffer I(PBS I) mixing is
It is even, in being incubated at room temperature 10 min, obtain i-motif+MOF-La compound, wherein PBS I is 100 mM Na Cl, 4 mM Mg
Cl2；
(2) internal pH measures:
Cell after six orifice plates, culture 1-2 days, is discarded culture medium, takes 20 μ L i-motif+MOF-La with suitable density kind
After compound is completely dissolved in fresh culture, with 4 hr of cell incubation, keep i-motif+MOF-La compound abundant by cell
Intake, then, cell are flushed three times using 10 mM buffers (PBS II), and the PBS II is 10 mM Na2 HPO4、2 mM
K H2PO4, 137 mM Na Cl, 2.7 mM K Cl, 250 mM glucose, pH 7.4；Cell, which uses, contains 10 μM of Ni Li
The 20 mM HEPES buffer solutions (20 mM HEPES) of sub- rhzomorph concentration handle 30 min, the 20 mM HEPES are as follows: 20
mM Na Cl、125 mM K Cl、0.5 mM Ca Cl2、0.5 mM Mg Cl2, 5 mM glucose, pH 5.0 ~ 7.5；Then,
Cell is flushed three times using 10 mM PBS II buffers, using Laser Scanning Confocal Microscope to cell imaging.
The method of the present invention by a pair of of fluorescence resonance energy transfer (FRET) fluorescence pair of i-motif connection, i-motif with
Group of the lanthanides organic metal framework lamella, which combines, is quenched positive charge fluorescence, and subsequent i-motif is dissociated from lamella, produced with after proton response
Raw fluorescence resonance energy transfer realizes the measurement of internal pH by the common location and color change of two kinds of fluorescence.This method tool
There are preferable photostability, anti-interference, detection has splendid time and spatial resolution, can be quickly to cell transfecting intracellular
Etc. in physiology courses pH change quantitative detection.
In the inventive solutions, the 5 of i-motif structural DNA is designed,Terminal modified carboxyl tetramethylrhodamine
(TAMRA), 3,Fluorescein (FAM) modification is held, is incubated for using lanthanide series metal organic frame nanoscale twins and i-motif structural DNA,
TAMRA fluorescence is quenched, and FAM fluorescence can retain.Further, by lamella and i-motif compound and cell incubation, to it
Into after cell, under the conditions of comparatively acidic pH, pattern falls off from linearly tetramer structure is become from lamella i-motif, FAM
FRET effect is generated with TAMRA, restores TAMRA fluorescence, according to the difference of internal pH, FAM and TAMRA fluorescence intensity occur
Corresponding change can be measured internal pH by the variation of the common location color of FAM and TAMRA and common location parameter.
Preferably, the lanthanide series metal organic frame nanoscale twins (MOF-La) are to be published on March 10th, 2017NPG Asia MaterialsThe entitled Lanthanide-based metal-organic framework of 9th phase e354
nanosheets with unique fluorescence quenching properties for two-color
MOF-Lns documented by page 2 in the paper of intracellular adenosine imaging in living cells
Synthetic method.
The size of the lanthanide series metal organic frame nanoscale twins is 100 ~ 200 nm, more preferably 100 nm.
The cell is cancer cell, including Hela cell and MCF-7 cell.
The pH measurement range is 5.0-7.5.
The present invention has the advantages that
(1) present invention design both ends i-motif label is FRET pairs a pair of of, and band is being quenched just using lanthanide series metal organic frame nanometer layer
The TAMRA fluorescence of electricity retains another electronegative FAM fluorescence.After nanoscale twins and i-motif compound enter cell,
The variation of pH makes i-motif deformation and falls off from nanoscale twins, and FRET effect occurs for FAM and TAMRA, promotes TAMRA to generate glimmering
Light.And pH is lower, then i-motif falls off more, and TAMRA fluorescence is stronger, can be to thin by the common location of FAM and TAMRA fluorescence
PH intracellular is measured.It is this according to FRET ratio carry out signal detection mode can the influence by environment to detection process be down to
It is minimum, to realize the accurate detection of internal pH.
(2) method of the invention has preferable photostability, anti-interference, and detection has splendid time and space point
Resolution quickly can change quantitative detection to pH in the physiology courses such as cell transfecting intracellular.
Specific embodiment
Below by way of specific embodiment, the technical scheme of the present invention will be further described.Embodiment below is to this
The further explanation of invention, and do not limit the scope of the invention.
Embodiment 1
A kind of detection method of internal pH is response pH element with i-motif structure, by connecting in i-motif structural DNA
A pair of of fluorescence resonance energy transfer (FRET) fluorescence pair is connect, i-motif structural DNA is quenched in conjunction with group of the lanthanides organic metal framework lamella
Go out positive charge fluorescence, after subsequent i-motif structural DNA and proton response, dissociates from lamella, generates fluorescence resonance energy transfer,
The measurement of internal pH is realized by the common location and color change of two kinds of fluorescence, according to the following steps:
(1) i-motif and lanthanide series metal organic frame nanoscale twins are incubated for:
The 2 linear ss-DNA(of 25 μM of μ L i-motif, that is, nucleic acid sequence table DNA 1-4 sequence) and 0.2 mg/mL lanthanide series metal
Organic frame nanoscale twins (MOF-La) are in 1 mL 10 mM phosphate buffer I(PBS I, 100 mM Na Cl, 4 mM Mg
Cl2) be uniformly mixed, in being incubated at room temperature 10 min, obtain i-motif+MOF-La compound.
(2) internal pH measures:
Hela cell inoculation is based on 37 DEG C of incubator cultures in six orifice plates, using the culture of RPMI 1640 containing 20% inactivation BSA
After 1 day, culture medium is discarded.After taking 20 μ L i-motif+MOF-La compounds to be completely dissolved in fresh culture, incubated with cell
4 hr are educated, absorb i-motif+MOF-La compound sufficiently by cell.Subsequent cell uses 10 mM PBS II buffers (10
mM Na2 HPO4, 2 mM K H2PO4, 137 mM Na Cl, 2.7 mM K Cl, 250 mM glucose, pH 7.4) and rinse three
It is secondary.Cell uses 20 mM HEPES buffer solutions (the 20 mM HEPES, 20 mM Na containing 10 μM of nigericin concentration
Cl, 125 mM K Cl, 0.5 mM Ca Cl2, 0.5 mM Mg Cl2, 5 mM glucose, pH 7.0) and 30 min of processing.Then
Cell is flushed three times using 10 mM PBS II buffers, is carried out using Laser Scanning Confocal Microscope to intracellular FAM and TAMRA fluorescence
Navy blue is presented in common location, cell imaging color.
Embodiment 2
A kind of detection method of internal pH, according to the following steps:
(1) i-motif and lanthanide series metal organic frame nanoscale twins are incubated for:
The 2 linear ss-DNA(of 25 μM of μ L i-motif, that is, nucleic acid sequence table DNA 1-4 sequence) and 0.2 mg/mL lanthanide series metal
Organic frame nanoscale twins (MOF-La) are in 1 mL 10 mM phosphate buffer I(PBS I, 100 mM Na Cl, 4 mM Mg
Cl2) be uniformly mixed, in being incubated at room temperature 10 min, obtain i-motif+MOF-La compound.
(2) internal pH measures:
Hela cell inoculation is based on 37 DEG C of incubator cultures in six orifice plates, using the culture of RPMI 1640 containing 20% inactivation BSA
After 1 day, culture medium is discarded.After taking 20 μ L i-motif+MOF-La compounds to be completely dissolved in fresh culture, incubated with cell
4 hr are educated, absorb i-motif+MOF-La compound sufficiently by cell.Subsequent cell uses 10 mM PBS II buffers (10
mM Na2 HPO4, 2 mM K H2PO4, 137 mM Na Cl, 2.7 mM K Cl, 250 mM glucose, pH 7.4) and rinse three
It is secondary.Cell uses 20 mM HEPES buffer solutions (the 20 mM HEPES, 20 mM Na containing 10 μM of nigericin concentration
Cl, 125 mM K Cl, 0.5 mM Ca Cl2, 0.5 mM Mg Cl2, 5 mM glucose, pH 6.5) and 30 min of processing.Then
Cell is flushed three times using 10 mM PBS II buffers, is carried out using Laser Scanning Confocal Microscope to intracellular FAM and TAMRA fluorescence
Indigo color is presented in common location, cell imaging color.
Embodiment 3
A kind of detection method of internal pH, according to the following steps:
(1) i-motif and lanthanide series metal organic frame nanoscale twins are incubated for:
The 2 linear ss-DNA(of 25 μM of μ L i-motif, that is, nucleic acid sequence table DNA 1-4 sequence) and 0.2 mg/mL lanthanide series metal
Organic frame nanoscale twins (MOF-La) are in 1 mL 10 mM phosphate buffer I(PBS I, 100 mM Na Cl, 4 mM Mg
Cl2) be uniformly mixed, in being incubated at room temperature 10 min, obtain i-motif+MOF-La compound.
(2) internal pH measures:
Hela cell inoculation is based on 37 DEG C of incubator cultures in six orifice plates, using the culture of RPMI 1640 containing 20% inactivation BSA
After 1 day, culture medium is discarded.After taking 20 μ L i-motif+MOF-La compounds to be completely dissolved in fresh culture, incubated with cell
4 hr are educated, absorb i-motif+MOF-La compound sufficiently by cell.Subsequent cell uses 10 mM PBS II buffers (10
mM Na2 HPO4, 2 mM K H2PO4, 137 mM Na Cl, 2.7 mM K Cl, 250 mM glucose, pH 7.4) and rinse three
It is secondary.Cell uses 20 mM HEPES buffer solutions (the 20 mM HEPES, 20 mM Na containing 10 μM of nigericin concentration
Cl, 125 mM K Cl, 0.5 mM Ca Cl2, 0.5 mM Mg Cl2, 5 mM glucose, pH 6.0) and 30 min of processing.Then
Cell is flushed three times using 10 mM PBS II buffers, is carried out using Laser Scanning Confocal Microscope to intracellular FAM and TAMRA fluorescence
Yellow green is presented in common location, cell imaging color.
Embodiment 4
A kind of detection method of internal pH, according to the following steps:
(1) i-motif and lanthanide series metal organic frame nanoscale twins are incubated for:
The 2 linear ss-DNA(of 25 μM of μ L i-motif, that is, nucleic acid sequence table DNA 1-4 sequence) and 0.2 mg/mL lanthanide series metal
Organic frame nanoscale twins (MOF-La) are in 1 mL 10 mM phosphate buffer I(PBS I, 100 mM Na Cl, 4 mM Mg
Cl2) be uniformly mixed, in being incubated at room temperature 10 min, obtain i-motif+MOF-La compound.
(2) internal pH measures:
Hela cell inoculation is based on 37 DEG C of incubator cultures in six orifice plates, using the culture of RPMI 1640 containing 20% inactivation BSA
After 1 day, culture medium is discarded.After taking 20 μ L i-motif+MOF-La compounds to be completely dissolved in fresh culture, incubated with cell
4 hr are educated, absorb i-motif+MOF-La compound sufficiently by cell.Subsequent cell uses 10 mM PBS II buffers (10
mM Na2 HPO4, 2 mM K H2PO4, 137 mM Na Cl, 2.7 mM K Cl, 250 mM glucose, pH 7.4) and rinse three
It is secondary.Cell uses 20 mM HEPES buffer solutions (the 20 mM HEPES, 20 mM Na containing 10 μM of nigericin concentration
Cl, 125 mM K Cl, 0.5 mM Ca Cl2, 0.5 mM Mg Cl2, 5 mM glucose, pH 5.5) and 30 min of processing.Then
Cell is flushed three times using 10 mM PBS II buffers, is carried out using Laser Scanning Confocal Microscope to intracellular FAM and TAMRA fluorescence
Common location, cell imaging color present orange red.
Embodiment 5
A kind of detection method of internal pH, according to the following steps:
(1) i-motif and lanthanide series metal organic frame nanoscale twins are incubated for:
The 2 linear ss-DNA(of 25 μM of μ L i-motif, that is, nucleic acid sequence table DNA 1-4 sequence) and 0.2 mg/mL lanthanide series metal
Organic frame nanoscale twins (MOF-La) are in 1 mL 10 mM phosphate buffer I(PBS I, 100 mM Na Cl, 4 mM Mg
Cl2) be uniformly mixed, in being incubated at room temperature 10 min, obtain i-motif+MOF-La compound.
(2) internal pH measures:
Hela cell inoculation is based on 37 DEG C of incubator cultures in six orifice plates, using the culture of RPMI 1640 containing 20% inactivation BSA
After 1 day, culture medium is discarded.After taking 20 μ L i-motif+MOF-La compounds to be completely dissolved in fresh culture, incubated with cell
4 hr are educated, absorb i-motif+MOF-La compound sufficiently by cell.Subsequent cell uses 10 mM PBS II buffers (10
mM Na2 HPO4, 2 mM K H2PO4, 137 mM Na Cl, 2.7 mM K Cl, 250 mM glucose, pH 7.4) and rinse three
It is secondary.Cell uses 20 mM HEPES buffer solutions (the 20 mM HEPES, 20 mM Na containing 10 μM of nigericin concentration
Cl, 125 mM K Cl, 0.5 mM Ca Cl2, 0.5 mM Mg Cl2, 5 mM glucose, pH 5.0) and 30 min of processing.Then
Cell is flushed three times using 10 mM PBS II buffers, is carried out using Laser Scanning Confocal Microscope to intracellular FAM and TAMRA fluorescence
Common location, cell imaging color present red.
<110>Shanghai National Engineering Research Center for Nanotechnology Co., Ltd
<120>a kind of detection method of internal pH
<160>4
<210>1
<211>26
<212>DNA
<213>artificial sequence
<221>misc feature
<222>(1)
<223>end 5' TRAMA is modified, the end 3' FAM label
<400>atccctaacc ctaaccctaa cccata
<210>2
<211>20
<212>DNA
<213>artificial sequence
<221>misc feature
<222>(1)
<223>end 5' TRAMA is modified, the end 3' FAM label
<400>ccccgccccg ccccgcccca
<210>3
<211>27
<212>DNA
<213>artificial sequence
<221>misc feature
<222>(1)
<223>end 5' TRAMA is modified, the end 3' FAM label
<400>cccctaaccc ctaaccccta accccca
<210>4
<211>24
<212>DNA
<213>artificial sequence
<221>misc feature
<222>(1)
<223>end 5' TRAMA is modified, the end 3' FAM label
<400>ccccaacccc aaccccaacc ccaa
Claims (5)
1. a kind of detection method of internal pH, which is characterized in that with i-motif structure be response pH element, by i-
Motif structural DNA connects a pair of of fluorescence resonance energy transfer (FRET) fluorescence pair, i-motif structural DNA and group of the lanthanides organic metal
Frame lamella, which combines, is quenched positive charge fluorescence, after subsequent i-motif structural DNA and proton response, dissociates from lamella, generates fluorescence
Resonance energy transfer realizes the measurement of internal pH by the common location and color change of two kinds of fluorescence, comprising the following steps:
(1) i-motif structural DNA and lanthanide series metal organic frame nanoscale twins are incubated for:
2 15 ~ 30 μM of μ the L linear ss-DNA of i-motif, the DNA 1-4 and 0.2 including nucleic acid sequence table Seq. No.1-4
Mg/mL lanthanide series metal organic frame nanoscale twins (MOF-La), in 1 mL, 10 mM phosphate buffer I(PBS I) mixing is
It is even, in being incubated at room temperature 10 min, obtain i-motif+MOF-La compound, wherein PBS I is 100 mM Na Cl, 4 mM Mg
Cl2；
(2) internal pH measures:
Cell after six orifice plates, culture 1-2 days, is discarded culture medium, takes 20 μ L i-motif+MOF-La with suitable density kind
After compound is completely dissolved in fresh culture, with 4 hr of cell incubation, keep i-motif+MOF-La compound abundant by cell
Intake, then, cell are flushed three times using 10 mM buffers (PBS II), and the PBS II is 10 mM Na2 HPO4、2 mM
K H2PO4, 137 mM Na Cl, 2.7 mM K Cl, 250 mM glucose, pH 7.4；Cell, which uses, contains 10 μM of Ni Li
The 20 mM HEPES buffer solutions (20 mM HEPES) of sub- rhzomorph concentration handle 30 min, the 20 mM HEPES are as follows: 20
mM Na Cl、125 mM K Cl、0.5 mM Ca Cl2、0.5 mM Mg Cl2, 5 mM glucose, pH 5.0 ~ 7.5；Then,
Cell is flushed three times using 10 mM PBS II buffers, using Laser Scanning Confocal Microscope to cell imaging.
2. the detection method of internal pH according to claim 1, it is characterised in that the lanthanide series metal organic frame is received
The size of rice lamella is 100 ~ 200 nm.
3. the detection method of internal pH according to claim 2, it is characterised in that the lanthanide series metal organic frame is received
The size of rice lamella is 100 nm.
4. the detection method of internal pH according to claim 1, which is characterized in that the cell is cancer cell, packet
Include Hela cell and MCF-7 cell.
5. the detection method of internal pH according to claim 1, which is characterized in that the pH measurement range is 5.0-
7.5。
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101434984A (en) * | 2007-11-13 | 2009-05-20 | 中国科学院上海应用物理研究所 | Optical means for detecting i-motif conformation of DNA |
| US20110223676A1 (en) * | 2010-03-10 | 2011-09-15 | Yamuna Krishnan | Dna-based molecular switches and uses thereof |
| CN104086536A (en) * | 2014-06-27 | 2014-10-08 | 中国科学院化学研究所 | Fluorescent probe for detecting pH value and preparation method thereof, and special detection kit |
| CN104391119A (en) * | 2014-11-19 | 2015-03-04 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of pH sensitive element based on DNA molecule configuration change |
| CN106908424A (en) * | 2017-02-09 | 2017-06-30 | 南京大学 | A kind of fluorescent nano probe and preparation method thereof and its application in bio-sensing |
| CN108535169A (en) * | 2018-04-17 | 2018-09-14 | 中国农业科学院特产研究所 | A kind of internal pH value detection method |
| CN109897887A (en) * | 2019-01-04 | 2019-06-18 | 中国科学院苏州生物医学工程技术研究所 | Activity test method of telomerase based on FRET |
-
2019
- 2019-07-16 CN CN201910642270.5A patent/CN110331188A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101434984A (en) * | 2007-11-13 | 2009-05-20 | 中国科学院上海应用物理研究所 | Optical means for detecting i-motif conformation of DNA |
| US20110223676A1 (en) * | 2010-03-10 | 2011-09-15 | Yamuna Krishnan | Dna-based molecular switches and uses thereof |
| CN104086536A (en) * | 2014-06-27 | 2014-10-08 | 中国科学院化学研究所 | Fluorescent probe for detecting pH value and preparation method thereof, and special detection kit |
| CN104391119A (en) * | 2014-11-19 | 2015-03-04 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of pH sensitive element based on DNA molecule configuration change |
| CN106908424A (en) * | 2017-02-09 | 2017-06-30 | 南京大学 | A kind of fluorescent nano probe and preparation method thereof and its application in bio-sensing |
| CN108535169A (en) * | 2018-04-17 | 2018-09-14 | 中国农业科学院特产研究所 | A kind of internal pH value detection method |
| CN109897887A (en) * | 2019-01-04 | 2019-06-18 | 中国科学院苏州生物医学工程技术研究所 | Activity test method of telomerase based on FRET |
Non-Patent Citations (2)
| Title |
|---|
| HUAI-SONG WANG ET AL.: "Lanthanide-based metal-organic framework nanosheets with unique fluorescence quenching properties for two-color intracellular adenosine imaging in living cells", 《NPG ASIA MATERIALS》 * |
| QIMING QIU ET AL.: "Recent advances in the rational synthesis and sensing applications of metal-organic framework biocomposites", 《COORDINATION CHEMISTRY REVIEWS》 * |
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