CN111285836A - Preparation and application of near-infrared fluorescent probe - Google Patents
Preparation and application of near-infrared fluorescent probe Download PDFInfo
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Abstract
The invention belongs to the field of fluorescence analysis and detection of biological small molecules, and discloses preparation and application of a near-infrared fluorescent probe for detecting cysteine. The structure of the near-infrared fluorescent probe CN is shown as follows, the preparation method is simple, the fluorescent probe CN has strong fluorescent response and selectivity on cysteine, and when the fluorescence emission wavelength is 655nm, the fluorescence intensity is in linear positive correlation with the concentration of cysteine, so that the near-infrared fluorescent probe CN can be directly applied to the rapid detection and biological imaging of cysteine in a biological environment.
Description
Technical Field
The invention belongs to the field of fluorescence analysis and detection of biological small molecules, and particularly relates to preparation and application of a near-infrared fluorescent probe for detecting cysteine.
Background
Cysteine is an important extracellular reducing agent and is an important substance in protein synthesis. During protein folding, cysteine residues in proteins and peptides stabilize molecular conformation via disulfide bonds, and some enzyme activities are also dependent on the activity of thiols in cysteine residues. In addition, cysteine prevents alcohol-induced damage to the gastric mucosa and inhibits nitric oxide-induced vasodilation. The change of cysteine concentration in plasma and urine samples opens up an effective way for the research of relevant clinical diseases by biomedicines. Increased levels of cysteine in plasma can lead to vascular disease and neurotoxicity. Conversely, cysteine deficiency also leads to a number of serious diseases such as decreased hematopoiesis, leukopenia, psoriasis, and the like. Therefore, the detection of the content of cysteine in the cells has important significance for the research of cell functions.
Compared with other traditional fluorescent probes, the near-infrared fluorescent probe has the advantages of low energy, less biological absorption, small cytotoxicity to cells, strong tissue penetrability and the like, so that the interference of self-absorption and self-fluorescence of a biological sample can be more effectively avoided. Therefore, designing and synthesizing a novel fast, sensitive and highly selective near-infrared fluorescent probe is significant for improving the existing fluorescence detection method.
Disclosure of Invention
In order to overcome the disadvantages and shortcomings of the prior art, the invention provides a near-infrared fluorescent probe molecule.
The invention also aims to provide a synthesis method of the near-infrared fluorescent probe molecule.
The invention further aims to provide application of the near-infrared fluorescent probe molecule in detecting the content of cysteine.
The purpose of the invention is realized by the following scheme:
a near-infrared fluorescent probe molecule CN has the following structure:
the synthesis method of the near-infrared fluorescent probe molecule CN comprises the following steps:
(1) in an organic solvent 1, 2, 4-dihydroxy benzaldehyde, pyridinium p-toluenesulfonate and 3, 4-dihydro-2H-pyran react to generate a compound 1;
(2) PBr is prepared from3Adding into organic solvent 2, adding cyclopentanone, and reactingTo produce compound 2;
(3) in organic solvent 3, compound 1, compound 2 and CS2CO3Reacting to generate a compound 3;
(4) adding the compound 3 into an organic solvent 4, uniformly mixing, adding malononitrile, tetrahydropyrrole and acetic acid for reaction, tracking the reaction by TCL, adding trifluoroacetic acid for continuous reaction after the raw materials disappear, and obtaining a compound 4 after the trifluoroacetic acid disappears;
(5) adding the compound 4 and diisopropylethylamine into an organic solvent 5, and adding methacryloyl chloride for reaction to obtain a target product, namely the near-infrared fluorescent probe molecule CN.
The structural formulas of compound 1, compound 2, compound 3 and compound 4 are respectively as follows:
the organic solvent 1 in the step (1) is dichloromethane;
the molar ratio of the 2, 4-dihydroxy benzaldehyde to the 3, 4-dihydro-2H-pyran in the step (1) is 1: 2-3, and preferably 1: 2.2; the dosage of the pyridinium p-toluenesulfonate in the step (1) meets the condition that the molar ratio of the 2, 4-dihydroxybenzaldehyde to the pyridinium p-toluenesulfonate is 1: 0.01-0.1, preferably 1: 0.04;
the reaction in the step (1) is a stirring reaction at room temperature for 3-5 h, preferably a stirring reaction at room temperature for 4 h;
the method also comprises a purification step after the reaction in the step (1), wherein the purification step comprises the following steps: and distilling the obtained reaction solution under reduced pressure to remove the solvent 1, and purifying by silica gel column chromatography.
The organic solvent 2 in the step (2) is at least one of N, N-Dimethylformamide (DMF) and dichloromethane;
the cyclopentanone in the step (2) is preferably added in a dichloromethane solution of cyclopentanone, wherein the concentration of the dichloromethane solution of cyclopentanone is 10-20 mol/L, and is preferably 11.3 mmol/L;
the step (2) is as describedPBr of3And cyclopentanone in a molar ratio of 2-3: 1, preferably 2.5: 1;
the reaction in the step (2) is carried out at 25 ℃ for 12-20 h, preferably at 25 ℃ for 16 h;
the method also comprises a purification step after the reaction in the step (2), wherein the purification step comprises the following steps: the resulting red solution was poured into ice water, the pH was adjusted to neutral with base, then extracted with dichloromethane, and the organic layer was washed with anhydrous Na2SO4Dried and concentrated.
The organic solvent 3 in the step (3) is N, N-Dimethylformamide (DMF);
the reaction in the step (3) is stirred and reacted for 16-30 h at 30-40 ℃, preferably for 24h at 35 ℃;
the mol ratio of the compound 1 to the compound 2 in the step (3) is 1: 1-3, preferably 1: 2; CS as described in step (3)2CO3The amount of (A) satisfies CS2CO3The mol ratio of the compound to the compound 1 is 2-4: 1, preferably 3: 1;
the method also comprises a purification step after the reaction in the step (3), wherein the purification step is as follows: filtering the obtained reaction product, washing with ethanol, concentrating the obtained filtrate, dissolving the obtained residue again, and performing silica gel chromatography column chromatography;
the organic solvent 4 in the step (4) is at least one of tetrahydrofuran and ethanol;
the trifluoroacetic acid, the compound 3 and the malononitrile in the step (4) are used in the following amounts: the molar ratio of the trifluoroacetic acid to the compound 3 to the malononitrile is 1: 10-15, preferably 1:12: 13.6; the dosages of the pyrrolidine and the acetic acid in the step (4) meet the following requirements: in the step (4), 1 drop of pyrrolidine and 1 drop of acetic acid are correspondingly dropped every 0.1mmol of trifluoroacetic acid;
the reactions in the step (4) are all reactions carried out at room temperature;
after the reaction in the step (4) is finished, a purification step of distilling the reaction product under reduced pressure to remove the solvent and then carrying out chromatographic separation by using a silica gel column is also included;
the organic solvent 5 in the step (5) is tetrahydrofuran;
the step (5) of adding methacryloyl chloride refers to adding under the ice bath condition;
the reaction in the step (5) is carried out for 0.5-2 h in an ice-water bath, then the reaction is carried out for 8-12 h at room temperature, preferably for 1h in the ice-water bath, and then the reaction is carried out for 10h at room temperature;
the dosage of the compound 4 and the diisopropylethylamine in the step (5) meets the condition that 4-5 mmol of the compound 4 is added to every 4mL of diisopropylethylamine, and preferably 4.2mmol of the compound 4 is added to every 4mL of diisopropylethylamine; the dosage of the methacrylic chloride in the step (5) meets the following requirements: the molar ratio of the compound 4 to the methacryloyl chloride is 2-4: 1, preferably 3: 1;
the method also comprises a purification step after the reaction in the step (5) is finished, wherein the purification step comprises the following steps: adding dichloromethane into the obtained reaction product for dissolving, then removing the solvent under reduced pressure, and then separating by silica gel column chromatography;
the solvent used in steps (1) to (5) functions as a reaction medium, and thus the amount of the solvent used is not particularly limited; the stirring in steps (1) to (5) is performed for the purpose of bringing the raw materials into sufficient contact with each other, and therefore, the stirring speed is not particularly limited.
The temperature is not indicated to be carried out at room temperature, and the room temperature refers to 25 ℃;
the application of the near-infrared fluorescent probe molecule CN in cysteine detection.
Preferably, in the application of the near-infrared fluorescent probe molecule CN in cysteine detection, the required excitation wavelength range is 500-650 nm during detection, and the wavelength range of fluorescence emission light is 620-750 nm; and the fluorescence intensity ratio of the fluorescence emission wavelength at 655nm is in linear positive correlation with the concentration of cysteine.
Compared with the prior art, the invention has the following advantages and beneficial effects:
(1) the synthesis method of the near-infrared fluorescent probe molecule CN is simple;
(2) the near-infrared fluorescent probe molecule CN has strong fluorescent response and selectivity to cysteine;
(3) the near-infrared fluorescent probe CN can be directly applied to the rapid detection and biological imaging of cysteine in a biological environment.
Drawings
FIG. 1 shows the color appearance change of a near-infrared fluorescent probe molecule CN solution before and after adding cysteine; the left image in fig. 1 is under a UV lamp; the right image in FIG. 1 is under sunlight;
FIG. 2 is a fluorescence emission spectrogram of a fluorescent probe molecule CN gradually dropwise added with cysteine (0-110 mol/L);
FIG. 3 is a graph showing the linear relationship between the concentration (0-110 mol/L) of cysteine (Cys) and the intensity of fluorescence emission wavelength at 655nm of a fluorescent probe CN;
FIG. 4 shows the fluorescence intensity changes of the fluorescence probe molecule CN for common amino acids such as cysteine (Cys), proline (Pro), tyrosine (Tyr), lysine (Lys) and inorganic salts;
FIG. 5 is a biological imaging diagram of the fluorescent probe molecule CN in the detection of cysteine in living cells.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the embodiments of the present invention are not limited thereto.
The reagents used in the examples are commercially available without specific reference.
Example 1: synthesis of near-infrared fluorescent probe molecule CN
(1) Synthesis of Compound 1
Stirring 2, 4-dihydroxybenzaldehyde (2.76g,20.00mmol), pyridinium p-toluenesulfonate (0.20g,0.80mmol) and 3, 4-dihydro-2H-pyran (4.01mL,44.00mmol) and 20mL of dichloromethane at room temperature for 4H, and after the reaction is finished, distilling the reaction solution under reduced pressure and purifying the reaction solution by silica gel column chromatography to obtain a compound 1(4.01g, 90%);
the reaction formula is as follows:
(2) synthesis of Compound 2
PBr was washed at 0 deg.C3(26.8mL, 282.5mmol) was added slowly to DMF (26mL, 339mmol) and dichloromethane (100 mL). After 60 minutes, a solution of cyclopentanone (10mL, 113mmol) in methylene chloride was added to the reaction mixture. After stirring for 16 hours at 25 ℃, the resulting red solution was poured into ice water, washed with NaOH solution and NaHCO3Adjusting pH of the solution to neutral, extracting with dichloromethane, and collecting organic layer with anhydrous Na2SO4Dried and concentrated. The crude yellow oil was of good purity and was used directly in the next reaction.
The reaction formula is as follows:
(3) synthesis of Compound 3
To a 100mL round bottom flask was added dry DMF, CS2CO3(6.90g,21.2mmol), Compound 1(1.57g,7.07mmol) and Compound 2(2.47g,14.1mmol) and the reaction mixture was stirred at 35 ℃ for 24 h. The insoluble material was then filtered and washed with ethanol, and the filtrate was concentrated under vacuum. The resulting residue was dissolved in silica gel (CH)2Cl2EtOAc, 9: 1, v/v) column chromatography gave compound 3 as a dark orange solid (1.03g, 49%);
characterization data for compound 4 are shown below:
1H NMR(400MHz,CDCl3)δ10.53(s,1H),7.08(d,J＝8.8Hz,1H),6.93(d,J＝2.4Hz,1H),6.83(dd,J＝8.4Hz&2.4Hz,1H),6.62(s,J＝3.2Hz,1H),5.47(t,1H),3.93-3.86(m,1H),3.69-3.64(m,1H),2.82-2.89(m,2H),2.81-2.71(m,4H),1.93-1.86(m,2H),1.61–1.76(m,2H).
13C NMR(100MHz，d6-DMSO)δ182.9,163.0,157.5,151.8,137.1,127.7,122.3,115.7,115.6,113.3,103.7,95.8,61.5,29.5,24.5,23.6,23.3,18.3.
ESI-HRMS m/z [ Compound 3+ H]+Calcd.for C18H18O4299.13,Found 299.1277.
Indicating the successful synthesis of compound 3;
(4) synthesis of Compound 4
A100 mL round-bottom flask was charged with a mixed solution of tetrahydrofuran and ethanol (10mL/10mL), compound 3(360mg,1.2mmol) was added, and after mixing well, a mixture of malononitrile (90mg,1.36mmol), 1 drop of tetrahydropyrrole and 1 drop of acetic acid was added. After the reaction solution reacts for 10h, TLC is used for tracking the reaction, after the raw material compound 3 disappears, trifluoroacetic acid (0.1mmol) is added, and then the mixture is stirred for 15-30min at room temperature. Tracking the reaction by TLC, and removing the solvent by reduced pressure distillation after trifluoroacetic acid disappears; separating and purifying by silica gel column chromatography to obtain compound 4(209mg, 66%);
characterization data for compound 4 are shown below:
1H NMR(400MHz,d6-DMSO)δ10.53(s,1H),7.80(s,1H),7.34(d,J＝8.4Hz,1H),7.23(s,1H),6.79(s,1H),6.75(dd,J＝8.4Hz&1.6Hz,1H),2.91-2.98(m,2H),2.82-2.89(m,2H).
13C NMR(100MHz，d6-DMSO)δ164.8,160.2,153.0,146.3,134.0,128.7,127.6,117.2,116.1,114.3,113.8,113.2,102.7,28.9,24.7.
ESI-HRMS m/z [ Compound 4+ H ]]+Calcd.for C16H10N2O2263.08,Found 263.0814.
This example illustrates the successful synthesis of compound 4;
(5) preparation of fluorescent Probe molecule CN
In a 50mL round-bottom flask was added tetrahydrofuran (10mL), compound 4(110mg,0.42mmol) and diisopropylethylamine (0.4 mL); under ice-water bath conditions, a solution of methacryloyl chloride (134mg,1.26mmol) in dichloromethane (10mL) was slowly added dropwise. After reacting for 1h in an ice water bath, the reaction mixture was transferred to room temperature and reacted for 10 h. The reaction was followed by TLC and was complete. Then, methylene chloride was added thereto to dissolve the compound, and after adding silica gel, the solvent was distilled off under reduced pressure, followed by separation and purification by silica gel column chromatography to obtain a near-infrared probe CN (128mg, 92%).
The characterization data of the near infrared probe CN are as follows:
1H NMR(400MHz,d6-DMSO)δ7.79(s,1H),7.50(d,J＝8.4Hz,1H),7.27(d,J＝2.0Hz,1H),7.20(s,1H),7.12(dd,J＝8.4Hz&2.4Hz,1H),6.31(s,1H),5.94(s,1H),2.97–2.92(m,2H),2.89–2.85(m,2H),2.01(s,3H).
13C NMR(100MHz,d6-DMSO)δ166.7,165.4,153.6,153.4,149.0,139.9,136.8,130.2,130.0,127.3,121.8,121.0,118.4,117.3,116.1,111.9,26.7,26.5,19.8.
ESI-HRMS m/z: [ fluorescent probe CN + H]+Calcd.for C20H14N2O3331.11,Found 331.1077.。
This example illustrates the successful preparation of a near infrared fluorescent probe molecule CN.
Example 2: detection of cysteine by fluorescent probe molecule CN
(1) Preparation of PBS buffer solution: accurately weighing 1.70g of monopotassium phosphate and 395mg of sodium hydroxide in a 250mL volumetric flask, fixing the volume by using secondary distilled water, shaking up, standing for a moment, taking 1.0mL, measuring the pH value to be 7.40 by using a pH meter, and sealing for later use.
(2) Preparation of test solution for probe: the probe compound CN was dissolved in N, N-Dimethylformamide (DMF) to give 1.0X 10-3The probe mother liquor of mol/L is ready for use. The test was carried out using 0.1mL of 1.0X 10-3mol/L probe stock solution was added to 9ml of PBS buffer (pH 7.4) in a volume ratio of DMF to 1:1, then adding cysteine (0.01mol/L) in different volumes, and then adding the mixture in the volume ratio of 1: the volume of the mixed solution of the PBS buffer solution 1 and the DMF is up to 10mL, so that probe molecule CN test solutions with different cysteine concentrations (0-110 mu mol/L) are obtained.
When cysteine is added into the probe mother liquor to obtain a probe molecule CN test solution with the cysteine concentration of 80 mu mol/L, the color appearance change of the near-infrared fluorescent probe molecule CN solution before and after the cysteine is added is shown in figure 1, wherein the left figure in figure 1 is observed under a UV lamp; the right image in FIG. 1 is observed under sunlight, and it can be seen from FIG. 1 that the color of the probe mother liquor is changed after cysteine is added, the color is changed from weak color fluorescence to strong red fluorescence under UV, and the color is changed from light orange to purple under sunlight, which indicates that the probe molecule CN test solution of the invention can judge whether cysteine exists through color change.
The fluorescence emission spectrum of the probe molecule CN test solution with different cysteine concentrations (0-110 mu mol/L) is shown in FIG. 2, and it can be seen from FIG. 2 that the intensity of the fluorescence emission wavelength at 655nm is gradually increased along with the increase of the cysteine concentration.
The fluorescence intensity of probe molecule CN test solutions with different cysteine concentrations (0-110 mu mol/L) at the fluorescence emission wavelength of 655nm is tested, a linear relation graph of the cysteine (Cys) concentration (0-110 mol/L) and the fluorescence emission wavelength of the fluorescence probe CN at the intensity of 655nm is shown in figure 3, and the fluorescence intensity and the cysteine concentration are in a linear positive correlation as can be seen from figure 3.
Example 3: recognition selectivity of fluorescent probe molecule CN to cysteine
The selectivity is an important condition for determining the performance of the fluorescent probe, so this example also explores the fluorescent response of the probe molecule CN to other common amino acids and anions, including common amino acids such as cysteine (Cys), proline (Pro), tyrosine (Tyr), lysine (Lys) and NaHCO3、Na2S、CuSO4The influence of the inorganic salts on the probe molecule CN was examined by replacing cysteine in the probe test solution having a cysteine concentration of 50. mu. mol/L obtained in example 1 with a common amino acid such as proline (Pro), tyrosine (Tyr) and lysine (Lys) and NaHCO, respectively3、Na2S、CuSO4Inorganic salts were used, and the measurement results are shown in FIG. 4.
As can be seen from FIG. 4, the fluorescence intensity of the fluorescent probe molecule CN at 655nm is significantly enhanced only after the addition of cysteine, and other substances are not changed. This result indicates that the selectivity of the fluorescent probe molecule A for cysteine is relatively good.
Example 5: application of near-infrared fluorescent probe molecule in detecting cysteine in living cells
HeLa cells (purchased from ATCC, USA) were planted on 14mm glass cover slips and attached for 24h before fluorescence imaging experiments. Then incubated with 10. mu. mol/L of probe CN (solvent DMSO) at 37 ℃ for 30 minutes and washed with PBS buffer (3 times); and then collecting fluorescence signals at 645-690 nm by using a 635nm semiconductor laser as an excitation light source. The bioimaging graph of the fluorescent probe molecule CN in the detection of cysteine in living cells is shown in FIG. 5. As can be seen in FIG. 5, incubation of HeLa cells with 10. mu. mol/L probe CN at 37 ℃ for 30min observed weak fluorescence, indicating that probe CN can penetrate into the cells and detect endogenous Cys by emitting near infrared fluorescence. To verify the specificity of intracellular cysteines, two control experiments were performed. Viable cells were first incubated with an aqueous solution (10. mu. mol/L) of the known thiol blocker N-ethylmaleimide (NEM) at 37 ℃ for 60min, and then with 10. mu. mol/L of probe CN at 37 ℃ for 30 min. As expected, the fluorescence intensity decreased significantly. In another control experiment, cells were incubated with 10. mu. mol/L NEM and 10. mu. mol/L Cys in sequence for 60min, respectively, followed by 10. mu. mol/L CN probe at 37 ℃ for 30min with significant time-dependent fluorescence enhancement. The three experimental results clearly show that the fluorescent probe CN has strong responsiveness and selectivity to cysteine in a cell body, and has good application value in the aspect of biological test.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A near-infrared fluorescent probe molecule CN is characterized by having the following structure:
2. the method for synthesizing the near-infrared fluorescent probe molecule CN according to claim 1, which is characterized by comprising the following steps:
(1) in an organic solvent 1, 2, 4-dihydroxy benzaldehyde, pyridinium p-toluenesulfonate and 3, 4-dihydro-2H-pyran react to generate a compound 1;
(2) PBr is prepared from3Adding the mixture into an organic solvent 2, and then adding cyclopentanone to react to generate a compound 2;
(3) in organic solvent 3, compound 1, compound 2 and CS2CO3Reacting to generate a compound 3;
(4) adding the compound 3 into an organic solvent 4, uniformly mixing, adding malononitrile, tetrahydropyrrole and acetic acid for reaction, tracking the reaction by TCL, adding trifluoroacetic acid for continuous reaction after the raw materials disappear, and obtaining a compound 4 after the trifluoroacetic acid disappears;
(5) adding the compound 4 and diisopropylethylamine into an organic solvent 5, and adding methacryloyl chloride for reaction to obtain a target product, namely the near-infrared fluorescent probe molecule CN.
3. The method for synthesizing the near-infrared fluorescent probe molecule CN according to claim 2, characterized in that:
the organic solvent 1 in the step (1) is dichloromethane;
the molar ratio of the 2, 4-dihydroxy benzaldehyde to the 3, 4-dihydro-2H-pyran in the step (1) is 1: 2-3; the dosage of the pyridinium p-toluenesulfonate in the step (1) meets the condition that the molar ratio of the 2, 4-dihydroxybenzaldehyde to the pyridinium p-toluenesulfonate is 1: 0.01 to 0.1;
the reaction in the step (1) is stirring reaction at room temperature for 3-5 h.
4. The method for synthesizing the near-infrared fluorescent probe molecule CN according to claim 2, characterized in that:
the organic solvent 2 in the step (2) is at least one of N, N-dimethylformamide and dichloromethane;
PBr described in step (2)3And cyclopentanone in a molar ratio of 2-3: 1;
the reaction in the step (2) is carried out at 25 ℃ for 12-20 h.
5. The method for synthesizing the near-infrared fluorescent probe molecule CN according to claim 2, characterized in that:
the organic solvent 3 in the step (3) is N, N-dimethylformamide;
the reaction in the step (3) is stirred at the temperature of 30-40 ℃ for 16-30 h;
the mol ratio of the compound 1 to the compound 2 in the step (3) is 1: 1-3; CS as described in step (3)2CO3The amount of (A) satisfies CS2CO3And the molar ratio of the compound 1 is 2-4: 1.
6. the method for synthesizing the near-infrared fluorescent probe molecule CN according to claim 2, characterized in that:
the organic solvent 4 in the step (4) is at least one of tetrahydrofuran and ethanol;
the trifluoroacetic acid, the compound 3 and the malononitrile in the step (4) are used in the following amounts: the molar ratio of trifluoroacetic acid, compound 3 and malononitrile is 1: 10-15: 10-15; the dosages of the pyrrolidine and the acetic acid in the step (4) meet the following requirements: in the step (4), 1 drop of pyrrolidine and 1 drop of acetic acid are correspondingly dropped every 0.1mmol of trifluoroacetic acid;
the reactions described in step (4) all refer to reactions carried out at room temperature.
7. The method for synthesizing the near-infrared fluorescent probe molecule CN according to claim 2, characterized in that:
the organic solvent 5 in the step (5) is tetrahydrofuran;
the step (5) of adding methacryloyl chloride refers to adding under the ice bath condition;
the reaction in the step (5) is carried out for 0.5-2 h in ice-water bath, and then the reaction is carried out for 8-12 h at room temperature;
the dosage of the compound 4 and the diisopropylethylamine in the step (5) meets the condition that 4-5 mmol of the compound 4 is correspondingly added into every 4mL of diisopropylethylamine; the dosage of the methacrylic chloride in the step (5) meets the following requirements: the mol ratio of the compound 4 to the methacryloyl chloride is 2-4: 1.
8. the near-infrared fluorescent probe molecule CN according to claim 1, used for cysteine detection.
9. The application of the near-infrared fluorescent probe molecule CN in the cysteine detection according to claim 8, wherein the near-infrared fluorescent probe molecule CN is characterized in that:
the range of the required excitation wavelength is 500-650 nm during detection, and the range of the fluorescence emission wavelength is 620-750 nm.
10. The application of the near-infrared fluorescent probe molecule CN in the cysteine detection according to claim 8, wherein the near-infrared fluorescent probe molecule CN is characterized in that:
the required excitation wavelength range is 500-650 nm during detection, and the wavelength range of fluorescence emission light is 655 nm.
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| CN113214285A (en) * | 2021-04-27 | 2021-08-06 | 宁波南大光电材料有限公司 | Preparation method of acid-sensitive monomer in ArF photoresist |
| CN114605376A (en) * | 2022-04-21 | 2022-06-10 | 济南大学 | Difunctional fluorescent probe for detecting cysteine and viscosity and preparation thereof |
| CN115925664A (en) * | 2023-01-06 | 2023-04-07 | 山东省华泰营养健康产业技术研究院有限公司 | Near-infrared fluorescent probe with large Stokes displacement and preparation method and application thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113214285A (en) * | 2021-04-27 | 2021-08-06 | 宁波南大光电材料有限公司 | Preparation method of acid-sensitive monomer in ArF photoresist |
| CN114605376A (en) * | 2022-04-21 | 2022-06-10 | 济南大学 | Difunctional fluorescent probe for detecting cysteine and viscosity and preparation thereof |
| CN115925664A (en) * | 2023-01-06 | 2023-04-07 | 山东省华泰营养健康产业技术研究院有限公司 | Near-infrared fluorescent probe with large Stokes displacement and preparation method and application thereof |
| CN115925664B (en) * | 2023-01-06 | 2023-05-09 | 山东省华泰营养健康产业技术研究院有限公司 | Near infrared fluorescent probe with large Stokes displacement and preparation method and application thereof |
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Application publication date: 20200616 |