CN106632304A - Two-photon RNA (Ribonucleic Acid) fluorescent probe and application thereof in living cell imaging - Google Patents
Two-photon RNA (Ribonucleic Acid) fluorescent probe and application thereof in living cell imaging Download PDFInfo
- Publication number
- CN106632304A CN106632304A CN201611159739.2A CN201611159739A CN106632304A CN 106632304 A CN106632304 A CN 106632304A CN 201611159739 A CN201611159739 A CN 201611159739A CN 106632304 A CN106632304 A CN 106632304A
- Authority
- CN
- China
- Prior art keywords
- rna
- photon
- iodine
- fluorescence probes
- salt compounded
- 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.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
- C09K2211/1037—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom with sulfur
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Materials Engineering (AREA)
- Biotechnology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to a two-photon RNA (Ribonucleic Acid) fluorescent probe and application thereof in living cell imaging. A structural general formula is shown as (I), wherein R is alkyl, hydroxyalkyl or ether radical; X is bromine or iodine. The two-photon RNA fluorescent probe is a novel RNA selectively-recognizable two-photon fluorescent probe and can be applied to mark or display application of RNA in living cell distribution. The probe has the characteristics of good membrane permeability, low cell toxicity and high color rendering property, and can be developed into a biological detection reagent for RNA-related physiological and pathological studies.
Description
Technical field
The present invention relates to a kind of fluorescence probe, more particularly to a kind of two-photon fluorescence spy for RNA imagings in living cells
Pin.
Background technology
In various living matters in the cell, ribonucleic acid (RNA) is important large biological molecule, and it is in catalysis biological
Extremely important role is play in the biological processes such as reaction, gene expression regulation.RNA is primarily present in nuclear kernel
In area and cytoplasm, its positioning, activity, number, form etc. include important life science information, and these information are examined with medical science
Break and treat closely bound up.Therefore, the RNA imagings in kernel and cytoplasm have to biochemistry, biological medicine, life science etc.
It is of great significance.Current Imaging-PAM, especially two-photon fluorescence imaging technology, in real-time monitoring living cells
The aspects such as form, the number of middle biomolecule are widely applied.Two-photon fluorescence imaging technology have high detection sensitivity,
The advantages such as big penetration depth, image high-fidelity, low phototoxicity and photobleaching, obtain the subjects such as biology, life, medical science
The favor of researchers.To adapt to existing situation, the various two-photon fluorescence probes that can be imaged different targets in living cells are
Become study hotspot.
Compared with numerous commercialization probes (such as DNA probe), rna probe is considerably less.There was only molecular phycobiliprotein complexes at present
(Molecular Probes Co.) provides the probe " SYTO RNA-Select " of an available RNA imaging, but the change of the probe
Learn structural formula externally not announce yet.Patent CN103265947A, CN103275699A each provide it is a kind of for living cells into
The RNA fluorescence probes of picture, but the probe fails to realize two-photon fluorescence imaging.Therefore, in living cells RNA fluorescence imagings detection side
Face, still lacks excellent two-photon fluorescence probe, and this present situation is urgently to be resolved hurrily.
The content of the invention
The purpose of the present invention is to be directed to above-mentioned deficiency, and provides a kind of two-photon RNA fluorescence probes, and there is the probe film to lead to
The characteristics of permeability is good, cytotoxicity is little, colour developing is strong.
It is a further object of the present invention to provide two-photon RNA fluorescence probes are being marked or are showing that RNA divides in living cells
The application of cloth.
The technical scheme that the present invention takes is：
A kind of two-photon RNA fluorescence probes, general structure is as shown in (I)：
Wherein：R is alkyl, hydroxyalkyl or ether；X is bromine or iodine.
In above-mentioned two-photon RNA fluorescence probes：The preferred C of described R1-12Alkyl, C1-12Hydroxyalkyl or ether base；Enter
The preferred methyl of one step, ethyl, butyl, dodecyl, ethoxy or ether base.
Described two-photon RNA fluorescence probe most preferably 2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys)
Benzothiazole salt compounded of iodine (referred to as IE).
The preparation method of above-mentioned two-photon RNA fluorescence probes：
First halogenating reaction introduces R bases on 2- methylbenzothiazole nitrogen, and obtains benzothiazolium salt, then using benzo thiophene
Azoles salt obtains final product with 3- formyl indoles by knoevenagel reactions.
Described two-photon RNA fluorescence probes (IE) to prepare reaction equation as shown in Figure 1.
Above-mentioned two-photon RNA fluorescence probes are in the application for marking or showing RNA to be distributed in living cells.
Described cell is HeLa cells.
Two-photon RNA fluorescence probes of the present invention are applied into the dyeing in HeLa cells, the two-photon fluorescence image after mark
Show, have obvious fluorescence distribution in the cytoplasm and nucleolar zone of cell, clearly point out the probe can be in living cells special secondary school
RNA in one property imaging cells matter and kernel, and it is real to carry out contrast in the cell Jing after ribalgilase (RNase) digestion experiment
It is further characterized by after testing.On HeLa cells the cytotoxicity of the probe is studied with mtt assay, tested HeLa
Cell survival rate of the cell after the probe is incubated 2,4,8,12,24 hours, within dyeing 12 hours, HeLa cells are deposited
Also more than 90%, this shows that two-photon fluorescence probe cytotoxicity of the present invention is little to motility rate, with preferable membrane permeability,
The RNA of cytoplasm and nucleolar zone in living cells can be imaged.
Two-photon RNA fluorescence probes of the present invention are the two-photon fluorescence probes of the new RNA Selective recognitions of a class, the spy
Needle set has the characteristics of membrane permeability is good, cytotoxicity is little, colour developing is strong, can be developed into the related physiology of RNA, pathological research life
Quality testing test agent.
Description of the drawings
Fig. 1 prepares reaction equation for 2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys) benzothiazole salt compounded of iodine；
Fig. 2 is the double light of RNA of 2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys) benzothiazole salt compounded of iodine (IE)
Sub- fluorescence titration figure；A is two-photon fluorescence spectrum；B is maximum fluorescence intensity with RNA and the variation diagram of concentration and probe concentration ratio；
Fig. 3 is 2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys) benzothiazole salt compounded of iodine (IE) to activity
Hela cells are dyeed the two-photon fluorescence photo obtained under 800 nm laser irradiations；A is two-photon fluorescence photo；B is
The differential interference microphoto of light field laser scanning；C is the merging figure (common location figure) of the figures of ab two；
Fig. 4 is HeLa cells Jing after ribalgilase (RNase) before processing, with 2- [2- (1H- indol-3-yls) ethene
Base] -3- (2- ethoxys) benzothiazole salt compounded of iodine (IE) dyeing cell photo comparison figure；A) it is ribalgilase (RNase) place
Before reason, shone with 2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys) benzothiazole salt compounded of iodine (IE) dyeing HeLa cells
Piece；B) it is ribalgilase (RNase) before processing, the differential interference microphoto of light field laser scanning；C) it is ribalgilase
(RNase) after processing, dyeed with 2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys) benzothiazole salt compounded of iodine (IE)
HeLa cell photos；D) after for ribalgilase (RNase) process, the differential interference microphoto of light field laser scanning；
Fig. 5 is that 2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys) benzothiazole salt compounded of iodine (IE) is thin in HeLa
On born of the same parents toxicity (MTT) experiment in cell survival rate with incubation time variation diagram；Incubation time：2nd, 4,8,12,24 hours, incubate
Educate 5 μM of concentration.
Specific embodiment
Further illustrate with reference to specific embodiment.
Embodiment 1：The synthesis of 2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys) benzothiazole salt compounded of iodine (IE)
2- methylbenzothiazoles (0.05mol, 7.46g) and ethylene iodohydrin (0.05mol, 7.81g) are dissolved in into toluene
(50ml) in, stir 4 hours, subsequent back flow reaction 30 minutes is cooled to room temperature, filter, ether washing obtains N- ethanol -2-
Methylbenzothiazole salt compounded of iodine.By N- ethanol -2- methylbenzothiazole salt compounded of iodine (0.01mol, 0.32g) and 3- formyl indoles
(0.01mol, 0.145g) is dissolved in methyl alcohol (20ml), and a few drop piperidines are added dropwise, and back flow reaction 4h, absolute ethanol washing obtains brown
Color powder, yield 77%.
1H NMR(400MHZ):12.478 (s, 1H), 8.448 (t, J=7.66,2H), 8.342 (d, J=7.96,1H),
8.205 (d, J=4.96,1H), 8.16 (t, J=9.72,1H), 7.798 (t, J=7.76,1H), 7.713 (q, J=9.29,
1H), 7.604 (t, J=5.4,2H), 7.357 (d, J=4.36,2H), 5.261 (t, J=5.68,1H), 4.959 (t, J=
4.34,2H), 3.965 (q, J=4.88,2H).13C NMR(400MHz,DMSO-d6),δ(ppm):173.40,144.44,
142.06,138.38,137.87,129.23,127.81,127.27,,125.17,124.37,124.31,122.89,
121.31,116.72,114.68,113.62,107.11。
Embodiment 2：The synthesis of 2- [2- (1H- indol-3-yls) vinyl] -3- methylbenzothiazole salt compounded of iodine
(10mmol, 1.49g) 2- methylbenzothiazoles and (20mmol, 2.84g) iodomethane are dissolved in 50ml toluene,
Stirring 4 hours, subsequent back flow reaction 30 minutes is cooled to room temperature, filters, ether washing, obtains N- methyl -2- methyl benzo thiophenes
Azoles salt compounded of iodine.(1mmol, 0.291g) N- methyl -2- methylbenzothiazoles salt compounded of iodine is molten with 0.145g (1mmol) 3- formyl indoles
In 20ml methyl alcohol, it is added dropwise 3 and drips piperidines, back flow reaction 6h, absolute ethanol washing obtains brown powder, yield 70%.
IMT-M:1H NMR(400MHZ):12.497 (s, 1H), 8.466 (t, J=10.6,2H), 8.335 (d, J=8,
1H), 8.275 (t, J=4.42,1H), 8.149 (d, J=8.36,1H), 7.815 (t, J=7.72,1H), 7.712 (t, J=
7.64,1H), 7.595 (t, J=4.5,1H), 7.525 (d, J=15.76,1H), 7.525 (d, J=15.76,1H), 7.370-
7.324(m,2H),4.277(s,3H).13C NMR(400MHz,DMSO-d6),δ(ppm):172.45,144.73,142.41,
138.37,138.04,129.31,127.09,125.20,124.40,124.30,122.90,121.48,116.40,114.70,
113.62,106.22。
Embodiment 3：The synthesis of 2- [2- (1H- indol-3-yls) vinyl] -3- ethyl diethyldithiocarbamate ether benzothiazole bromides
(10mmol, 1.49g) 2- methylbenzothiazoles and (20mmol, 6.75g) 2- bromoethyl ethylethers are dissolved in
In 50ml toluene, stir 6 hours, subsequent back flow reaction 30 minutes is cooled to room temperature, filter, ether washing obtains N- ethyl second
Base ether -2- methylbenzothiazole bromides.By (1mmol, 0.617g) N- ethyl diethyldithiocarbamate ether -2- methylbenzothiazoles bromides with
(1mmol, 0.145g) 3- formyl indoles are dissolved in 20ml methyl alcohol, and 3 drop piperidines are added dropwise, and back flow reaction 7h, absolute ethyl alcohol is washed
Wash, obtain brown powder, yield 65%.
Measure of merit
(1) the double light of the RNA of 2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys) benzothiazole salt compounded of iodine (IE)
Sub- fluorescence titration：
Fixed 2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys) benzothiazole salt compounded of iodine (IE) is in Tris-HCl
Concentration in cushioning liquid (pH=7.2, Tris and KCl 100mmol/L), is gradually added into RNA, in 800 nm laser irradiation light
Under obtain 2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys) benzothiazole salt compounded of iodine (IE) two-photon fluorescence titration
Spectrogram, as shown in Figure 2.As a result show, with being continuously added for RNA, the two-photon fluorescence intensity of probe of the present invention was before this
It is increased dramatically, then increases amplitude and slow down.This shows that the probe of the present invention has obvious fluorescence response to RNA.
(2) HeLa cell culture
HeLa cell lines adhere-wall culture is in the nutrient solution for including 10% (v/v) hyclone and a small amount of penicillin/streptomycin
In, in 37 DEG C, 5%CO2(v/v) cultivate in saturated humidity incubator.Treat cell growth to logarithmic phase, contact pin culture：By lid glass
Piece soaks 30 minutes in chromic acid lotion, and clear water, ultra-pure water are cleaned, and drying is put into disposable 35mm sterile petri dish after sterilizing
In；Culture medium in 100mL blake bottles is carefully poured out, cell is washed 2 times with PBS (phosphate buffer, pH=7.4), uses l
ML 0.25% (mass fraction) trypsin solution digests 2～3 minutes, outwells trypsin solution, adds a small amount of fresh culture to blow
Beat uniformly into cell suspending liquid, after cell count, leave the cell of proper density, culture medium is added to into volume required piping and druming equal
It is even, during cell to be then seeded to the culture dish for including cover glass, it is put in incubator and cultivates, grow cell climbing sheet.24～
After 48h, cell density is grown to up to 50%～70%, it is to be dyed.
(3) 2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys) benzothiazole salt compounded of iodine (IE) is to HeLa cells
Dyeing observation：
2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys) benzothiazole salt compounded of iodine (IE) is made into into 1mM DMSO
Mother liquor, 5 μM are diluted to during dyeing with PBS (phosphate buffer, pH=7.4).By probe molecule of the cell being inoculated with 5 μM
In solution, 30min is incubated under the conditions of 37 DEG C, with PBS unconjugated unnecessary dye liquor is washed away, cell growth faces lower cover in slide
On；Then sample is placed in fluorescence imaging on objective table.As a result Fig. 3 is seen, two-photon fluorescence image is displayed in cytoplasm and kernel
There is obvious fluorescence distribution in area, shows that the probe of the present invention can be in living cells in narrow spectrum imaging cells matter and kernel
RNA。
(4) 2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys) benzothiazole salt compounded of iodine (IE) probe is to HeLa
The dyeing observation of the HeLa cells after cell, RNase digestion process：
It is prepared by fixed cell：The HeLa cells of culture are soaked in into 4% (mass fraction) paraformaldehyde solution 30 minutes, or
Person fixes 15 minutes with -20 DEG C of methyl alcohol, then penetrating 2 minutes with 0.5%Triton X-100 room temperatures, obtains fixation to be dyed thin
Born of the same parents.
Two groups of above-mentioned fixed cells are taken, with 2- [2- (1H- indol-3-yls) vinyl] -3- (the 2- hydroxyl second that concentration is 5 μM
Base) benzothiazole salt compounded of iodine (IE) probe solution is in CO2Hatching dyeing 30 minutes in incubator.2 are rinsed to two groups of cells with PBS
Time, one group of addition 25mg/mL DNase-Free RNase (GE) (ribalgilase) thereto, then two groups of cells 37 DEG C,
5%CO2Under the conditions of be incubated 2 hours.Subsequently, rinsed 2 times with PBS, the fluorescence in observation, two groups of cells of record under wide-field microscope
Distribution and brightness change etc..
As a result see Fig. 4, without the fluorescence of the HeLa cells of RNase digestion process cytoplasm and nucleolar zone are concentrated mainly on
Domain；And the fluorescence of the HeLa cells Jing after RNase digestion process has focused on nuclear area, its cytoplasm and kernel region base
This is without fluorescence.Because RNase is only capable of hydrolyzing the RNA in cell, therefore, the embodiment can confirm the probe energy of the present invention
RNA in enough selective imaging cells.
(5) MTT cytotoxicity experiments
Logarithmic phase HeLa cell is collected, concentration of cell suspension is adjusted, 100ul is added per hole, it is close that bed board adjusts cell to be measured
Spend to 1000-10000 holes (edge hole is filled with aseptic PBS).5%CO2, 37 DEG C are incubated, and to cell monolayer bottom hole (96 holes are paved with
Flat underside) after start respectively 0,12,16,20, the time of 22h add 5 μM of 2- [2- (1H- indol-3-yls) vinyl] -3-
(2- ethoxys) benzothiazole salt compounded of iodine (IE) fluorescence probe.To during adherent 24h per hole add 20ul MTT solution (5mg/ml, i.e.,
0.5%MTT), continue to cultivate 4h.If medicine can react with MTT, can be first centrifuged and nutrient solution discard afterwards, carefully rush 2-3 with PBS
After, the nutrient solution containing MTT is added.Then, then to every hole 150ul dimethyl sulfoxide (DMSO)s are added, put low-speed oscillation on shaking table
10min, makes crystal fully dissolve.The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument OD490nm.
As a result Fig. 5 is seen, HeLa cells are in 5 μM of 2- [2- (1H- indol-3-yls) vinyl] -3- (2- ethoxys) benzo thiophene
Azoles salt compounded of iodine (IE) be incubated 2,4,8,12,24 hours after cell survival rate, dyeing 12 hours within, the survival rate of HeLa cells
Also more than 90%, this shows that two-photon fluorescence probe of the present invention can be imaged the RNA of cytoplasm and nucleolar zone in living cells.
It is more than that the present invention is discussed in detail in conjunction with specific embodiments, protection scope of the present invention not limited to this.
Claims (8)
1. a kind of two-photon RNA fluorescence probes, general structure is as shown in (I)：
Wherein：R is alkyl, hydroxyalkyl or ether；X is bromine or iodine.
2. a kind of two-photon RNA fluorescence probes according to claim 1, is characterized in that, the R selects C1-12Alkyl, C1-12
Hydroxyalkyl or ether base.
3. a kind of two-photon RNA fluorescence probes according to claim 2, is characterized in that, the R is methyl, ethyl, fourth
Base, dodecyl, ethoxy or ether base.
4. a kind of two-photon RNA fluorescence probes according to claim 3, is characterized in that, it is 2- [2- (1H- indoles -3-
Base) vinyl] -3- (2- ethoxys) benzothiazole salt compounded of iodine.
5. a kind of preparation method of the two-photon RNA fluorescence probes described in claim 4, is characterized in that, by 2- methyl benzo thiophenes
Azoles is dissolved in toluene with ethylene iodohydrin, stirs 3-4 hours, subsequent back flow reaction 30-40 minute, is cooled to room temperature, is filtered, second
Ether is washed, and obtains N- ethanol -2- methylbenzothiazole salt compounded of iodine；By N- ethanol -2- methylbenzothiazoles salt compounded of iodine and 3- formoxyl Yin
Diindyl is dissolved in methyl alcohol, and a few drop piperidines are added dropwise, and back flow reaction 3-4h, absolute ethanol washing obtains brown powder.
6. a kind of preparation method of two-photon RNA fluorescence probes according to claim 5, is characterized in that, 2- methyl benzos
Thiazole is 1 with the mol ratio of ethylene iodohydrin:1, N- ethanol -2- methylbenzothiazoles salt compounded of iodine is with the mol ratio of 3- formyl indoles
1:1。
7. a kind of two-photon RNA fluorescence probes described in any one of claim 1-4 are being marked or are showing that RNA divides in living cells
The application of cloth.
8. application according to claim 7, is characterized in that, described cell is HeLa cells.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611159739.2A CN106632304B (en) | 2016-12-15 | 2016-12-15 | A kind of two-photon RNA fluorescence probe and its application in living cells imaging |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611159739.2A CN106632304B (en) | 2016-12-15 | 2016-12-15 | A kind of two-photon RNA fluorescence probe and its application in living cells imaging |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106632304A true CN106632304A (en) | 2017-05-10 |
| CN106632304B CN106632304B (en) | 2019-03-26 |
Family
ID=58823722
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611159739.2A Active CN106632304B (en) | 2016-12-15 | 2016-12-15 | A kind of two-photon RNA fluorescence probe and its application in living cells imaging |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106632304B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108285449A (en) * | 2017-10-24 | 2018-07-17 | 泰山医学院 | A kind of pyrido [1,2-a] benzimidazole by thiazole modification can detect fluorescence probe and the application of hypochlorite ion |
| CN108956563A (en) * | 2018-06-21 | 2018-12-07 | 东南大学 | A method of multi-function metal nano-probe is synthesized with tumour cell biology in situ |
| CN111116573A (en) * | 2019-12-31 | 2020-05-08 | 中山大学 | Near-infrared fluorescent probe for simultaneously detecting DNA and RNA under dual channels and preparation method and application thereof |
| CN111995621A (en) * | 2020-08-26 | 2020-11-27 | 广东工业大学 | Benzoindole derivative for G-quadruplex RNA fluorescent probe and preparation method and application thereof |
| CN112410404A (en) * | 2020-05-27 | 2021-02-26 | 江西省肿瘤医院(江西省癌症中心) | Open type two-photon nucleic acid probe and application thereof in FISH |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101555246A (en) * | 2008-04-11 | 2009-10-14 | 大连理工大学 | Halogen-containing asymmetry phthalocyanines compound, preparation method and application thereof |
-
2016
- 2016-12-15 CN CN201611159739.2A patent/CN106632304B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101555246A (en) * | 2008-04-11 | 2009-10-14 | 大连理工大学 | Halogen-containing asymmetry phthalocyanines compound, preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| PANKAJ GAUR，ET AL.: ""Selenium Incorporated Cationic Organochalcogen: Live Cell Compatible and Highly Photostable Molecular Stain for Imaging and Localization of Intracellular DNA"", 《ACS APPLIED MATERIALS & INTERFACES》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108285449A (en) * | 2017-10-24 | 2018-07-17 | 泰山医学院 | A kind of pyrido [1,2-a] benzimidazole by thiazole modification can detect fluorescence probe and the application of hypochlorite ion |
| CN108956563A (en) * | 2018-06-21 | 2018-12-07 | 东南大学 | A method of multi-function metal nano-probe is synthesized with tumour cell biology in situ |
| CN111116573A (en) * | 2019-12-31 | 2020-05-08 | 中山大学 | Near-infrared fluorescent probe for simultaneously detecting DNA and RNA under dual channels and preparation method and application thereof |
| CN112410404A (en) * | 2020-05-27 | 2021-02-26 | 江西省肿瘤医院(江西省癌症中心) | Open type two-photon nucleic acid probe and application thereof in FISH |
| CN111995621A (en) * | 2020-08-26 | 2020-11-27 | 广东工业大学 | Benzoindole derivative for G-quadruplex RNA fluorescent probe and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106632304B (en) | 2019-03-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106632304B (en) | A kind of two-photon RNA fluorescence probe and its application in living cells imaging | |
| CN103265947B (en) | A kind of indolepyridinium salt fluorescent probe for RNA in viable cell and kernel imaging | |
| Trampe et al. | Functionalized bioink with optical sensor nanoparticles for O2 imaging in 3D‐bioprinted constructs | |
| CN102127055B (en) | Single-photon and two-photon homocysteine fluorescent probes and use thereof | |
| CN106905309B (en) | A kind of very high selectivity and the amphiphilic fat drips fluorescence probe of tool and its application | |
| CN106946869A (en) | The fluorescence probe of fat drips in a kind of specific marker cell | |
| CN109293633A (en) | A kind of non-reactive mitochondria tracking fluorescence probe IVPI-12 and its application | |
| CN108844931A (en) | LZQ fluorescence probe detects SO at the same time2With the application in HSA | |
| CN103275699B (en) | Pyrrole pyridine salt fluorescent probe used for RNA (ribonucleic acid) and nucleolus imaging in living cell | |
| CN106590630B (en) | A kind of mitochondrial membrane potential fluorescence probe of space dependent form and its application | |
| CN105566207A (en) | Two-photon deep red emission fluorescent probe for imaging cell membranes in tissues based on molecular rotors | |
| WO2021109862A1 (en) | Light-sensitive living cell ribonucleic acid fluorescent probe | |
| CN107325814A (en) | A kind of fluorescence silicon nano dots and preparation method and application | |
| CN110057804A (en) | Application of the fluorescent carbon point based on N- methyl-o-phenylenediamine hydrochloride in lysosome targeting | |
| CN105062467A (en) | Rotor-type two-photon mitochondrion fluorescence probe and application thereof | |
| CN108373447A (en) | A kind of fluorescence probe that distinguishing dead/living cells and its synthetic method and application | |
| CN103820104B (en) | Near infrared fluorescent probe, its method for making and the application of one class taking Nile blue as parent | |
| CN108822031A (en) | A kind of red transmitting fluorescence probe of two-photon detecting mitochondria | |
| CN103896928A (en) | pH fluorescent chemical sensor as well as synthetic method and application thereof | |
| CN105153733B (en) | Membrane permeability dye with large two-photon fluorescence active cross section and application of membrane permeability dye | |
| CN103710021B (en) | Fluorescent dye with nitrobenzimidazole as RNA (ribonucleic acid) recognition group as well as preparation method and application of fluorescent dye | |
| CN106496126B (en) | A kind of two-photon fluorescence probe that RNA is imaged and its application in living cells | |
| CN109422758B (en) | Near-infrared fluorescence ratio fluorescence probe and preparation method and application thereof | |
| CN109293632A (en) | A kind of non-reactive mitochondria tracking fluorescence probe and its application containing 12 carbon alkyl chains | |
| CN112239455B (en) | RNA fluorescent probe for rapidly distinguishing cancer tissues from normal tissues by using morphological change of nucleolus |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |