CN112341374B - Compound, preparation method and application of compound as pH ratio detection type fluorescent probe - Google Patents
Compound, preparation method and application of compound as pH ratio detection type fluorescent probe Download PDFInfo
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- C07D209/56—Ring systems containing three or more rings
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- C07D209/60—Naphtho [b] pyrroles; Hydrogenated naphtho [b] pyrroles
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- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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- 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"
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Abstract
The invention discloses a compound, a preparation method and application thereof as a pH ratio detection type fluorescent probe, wherein the chemical structure of the compound is as follows:
the compound can be used as a fluorescent probe for pH in the endoplasmic reticulum, has endoplasmic reticulum targeting capability, and can realize sensitive detection of pH change of the endoplasmic reticulum by ratiometric fluorescence imaging.
Description
Technical Field
The invention relates to the technical field of fluorescence imaging detection, in particular to a compound, a preparation method and application thereof as a pH ratio detection type fluorescent probe.
Background
The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.
Intracellular pH plays an important role in many physiological processes, such as cell proliferation, apoptosis, regulation of enzymatic activity, and ion transport. The endoplasmic reticulum is a core intracellular organelle in the secretory pathway, is a base for the synthesis of a series of important biological macromolecules, such as proteins, lipids and saccharides, and is also an important place for maintaining calcium homeostasis and detoxifying toxic substances. The pH of the endoplasmic reticulum is an important parameter and is important for regulating the physiological functions of the endoplasmic reticulum, including protein sorting and targeting during secretion and recovery of resident chaperones. Under normal physiological conditions, the pH of the ER is generally considered to be the same as the pH of the cytoplasm (pH 7.2 ± 0.2). Any abnormal change in pH of the endoplasmic reticulum in the secretory pathway may disrupt enzyme activity, classification and processing of secreted proteins, and calcium storage. Therefore, the development of a detection means capable of targeting endoplasmic reticulum in situ and quantitatively determining the pH of the endoplasmic reticulum in real time has important significance for deeply disclosing the biological action of the endoplasmic reticulum in physiological events.
Fluorescence imaging has many advantages such as high sensitivity, noninvasive detection and in-situ detection, and is an attractive pH value detection method. In recent years, small molecule fluorescent probes have attracted attention because of their advantages such as easy modification of chemical structure and convenient adjustment of emission color, and are widely used in cell imaging research. However, the conventional small molecule fluorescent probe mostly depends on signal output of absolute intensity, and is easily interfered by various external factors, so that accurate quantification of an analyte is difficult. The imaging of the ratio-type fluorescent probe relies on the ratio of two or more detection signals as signal output, so that the interference of external factors can be effectively reduced, the accuracy of molecular detection and imaging is improved, and the quantitative detection of an analyte is realized. Currently, there are still few reports on ratiometric fluorescent probes for detecting pH in the endoplasmic reticulum.
Disclosure of Invention
In order to solve the defects of the prior art, the invention aims to provide a compound, a preparation method and application thereof as a pH ratio type fluorescent probe for detecting pH, wherein the compound can be used as a fluorescent probe for detecting pH in an endoplasmic reticulum, has an endoplasmic reticulum targeting capability and can realize sensitive detection of pH change of the endoplasmic reticulum by ratio fluorescence imaging.
In order to realize the purpose, the technical scheme of the invention is as follows:
in one aspect, a compound has the chemical structure shown in the following formula:
on the other hand, a method for preparing the above compound, 1,1, 2-trimethyl-1H-benzo [ e ] indole and iodopropionic acid are subjected to quaternization to obtain a product 1, N-tosylethylenediamine, the product 1 is subjected to amidation of carboxyl and amino groups to obtain a product 2, and the product 2 is subjected to condensation reaction of methyl and aldehyde groups with p-hydroxybenzaldehyde to obtain the above compound.
In a third aspect, use of a compound as described above for detecting pH.
The compound provided by the invention utilizes phenolic hydroxyl as a response site of pH, and regulates and controls the fluorescence property of the probe through an intramolecular charge transfer effect. Under alkaline conditions, the phenolic hydroxyl group is deprotonated to convert to a phenolate ion, resulting in a red-shift in the fluorescence emission wavelength of the probe. As the basicity increases, the fluorescence emission wavelength of the probe gradually red-shifts from 545nm to 584 nm.
In a fourth aspect, the use of a compound as described above as a probe for detecting pH ratio fluorescence.
In a fifth aspect, the application of the compound in preparing a target endoplasmic reticulum pH fluorescent probe or a ratio type pH detection probe.
The invention has the beneficial effects that:
1. the compound provided by the invention can be used as a fluorescent probe for efficiently targeting endoplasmic reticulum.
2. The ratiometric fluorescent probe for detecting the pH of the endoplasmic reticulum of the compound provided by the invention shows good absorption and fluorescence spectrum response in buffer solutions with different pH values, and has good reversibility of acid-base response and photostability.
3. The compound of the invention has good biocompatibility and low cell toxicity, and can be used for pH detection of endoplasmic reticulum in living cells.
4. The compound of the invention can be applied to the detection of in-situ pH in an inflammation mouse model.
5. The compound disclosed by the invention is simple to synthesize, has obvious reversible color change in acidic and alkaline environments, and can be used for visual detection of the acid-base properties of a solution.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, are included to provide a further understanding of the invention, and are included to illustrate an exemplary embodiment of the invention and not to limit the invention.
FIG. 1 is a spectrum of a probe pH-ER prepared in example 1 of the present invention in buffers with different pH values, wherein a is an ultraviolet absorption spectrum and b is a fluorescence emission spectrum;
FIG. 2 is a graph showing reversibility of the probe ph-ER prepared in example 1 of the present invention in an environment where acidity and alkalinity are repeatedly changed;
FIG. 3 is a diagram of confocal fluorescence imaging of a probe ph-ER prepared in example 1 after co-staining with different subcellular organelle localization dyes in a normal human hepatocyte HL-7702, wherein a-c are diagrams of co-staining with endoplasmic reticulum dyes, d-e are diagrams of co-staining with mitochondrial dyes, f-h are diagrams of co-staining with lysosomal dyes, i is a diagram of fluorescence intensity trend of the probe ph-ER and Golgi dyes, j is a diagram of fluorescence intensity trend of the probe ph-ER and the mitochondrial dyes, and k is a diagram of fluorescence intensity trend of the probe ph-ER and the lysosomal dyes; l is a fluorescence intensity trend chart of the probe ph-ER and the Golgi dye;
fig. 4 is a representation of confocal fluorescence imaging of the probe pH-ER prepared in example 1 in human normal hepatocytes HL-7720 in buffer media of different pH values, a, e, m, i, q are images of green channels of the probe in the cell, b, f, j, n, r are images of red channels of the probe in the cell, c, g, k, o, s are images of bright fields of the cell, d, h, l, p, t are channels of ratios of red and green channels of the probe in the cell, u is an output graph of fluorescence intensity ratio data of the red and green channels, a-d is pH 6.0, e-h is pH 6.5, i-l is pH 7.0, m-p is pH 7.5, and q-t is pH 8.0;
FIG. 5 is a graph showing in situ fluorescence imaging characterization of probe pH-ER prepared in example 1 for detecting pH changes of endoplasmic reticulum at normal and inflammatory sites in an inflammatory mouse model, from left to right, respectively showing 520nm imaging window imaging graph, 620nm imaging window imaging graph, and fluorescence imaging intensity ratio (FI) of 620nm and 520nm windows620nm/FI520nm) And (6) outputting the data.
Detailed Description
It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
In order to realize the purpose of ratio fluorescence imaging for sensitively and quantitatively detecting the pH change of endoplasmic reticulum, the invention provides a compound, a preparation method and application thereof as a pH ratio type fluorescence probe for detecting the pH change.
In one exemplary embodiment of the present invention, there is provided a compound having a chemical structure represented by the following formula:
in another embodiment of the present invention, there is provided a method for preparing the above compound, wherein 1,1, 2-trimethyl-1H-benzo [ e ] indole and iodopropionic acid are subjected to quaternization to obtain a product 1, N-tosylethylenediamine, the product 1 is subjected to amidation of carboxyl and amino groups to obtain a product 2, and the product 2 is subjected to condensation reaction of methyl and aldehyde groups with p-hydroxybenzaldehyde to obtain the above compound.
1,1, 2-trimethyl-1H-benzo [ e ]]The chemical structural formula of the indole is
The chemical structural formula of the iodopropionic acid is
N-toluenesulfonylethyl esterThe chemical structural formula of the diamine is
The chemical structure of the parahydroxybenzaldehyde is
The synthetic route is as follows:
in some embodiments, the process of quaternization is: 1,1, 2-trimethyl-3H-indole was refluxed with iodopropionic acid in acetonitrile at 80 ℃ for 48H.
In some embodiments, the molar ratio of 1,1, 2-trimethyl-3H-indole to iodopropionic acid is 1: 1.5.
In this series of examples, the molar ratio of 1,1, 2-trimethyl-1H-benzo [ e ] indole to iodopropionic acid was 1: 1.5.
In some embodiments, the course of the amidation reaction is: under the alkaline condition, the product 1 and a condensing agent react at a temperature of-1-0 ℃, and then N-tosylethylenediamine is added to react at room temperature. The room temperature in the present invention is an indoor temperature, and is generally 15 to 30 ℃.
In this series of examples, triethylamine was used to provide basic conditions.
In this series of examples, the condensing agent was a mixed system of EDC (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide) and HOBt (1-hydroxybenzotriazole). The molar ratio of N-tosylethylenediamine, EDC and HOBt is 1: 0.9-1.1.
In the series of embodiments, the reaction time is 0.5 to 1 hour at-1 to 0 ℃. When the reaction time is 0.5h, the reaction can be ensured to be complete.
In the series of examples, the reaction time at room temperature is 12-24 h. When the reaction time is 24 hours, the completion of the reaction can be ensured.
In some embodiments, the molar ratio of N-tosylethylenediamine to product 1 is 1:0.9 to 1.1.
In some embodiments, the solvent for the amidation reaction is dichloromethane and N, N-Dimethylformamide (DMF).
In some embodiments, the conditions of the condensation reaction are: in an alkaline environment, the reaction temperature is 70-90 ℃, and the reaction time is 3-12 h.
In the series of embodiments, piperidine is used to provide an alkaline environment, and the pH value is 8-9.
In this series of examples, the condensation reaction was carried out at a reaction temperature of 80 ℃ for a reaction time of 6 hours.
In this series of examples, the solvent for the condensation reaction was absolute ethanol.
In the series of examples, the feeding ratio of the product 2 to the p-hydroxybenzaldehyde is 1: 1.1-1.3.
In some embodiments, the material after the condensation reaction is purified by column chromatography after the solvent is removed. The purity of the target compound can be ensured.
In this series of examples, the developing solvent for column chromatography was a mixture of dichloromethane and methanol. When the volume ratio of the dichloromethane to the methanol is 20:1, the purification effect is better.
In a third embodiment of the invention, there is provided the use of a compound as described above for detecting pH.
The compound provided by the invention utilizes phenolic hydroxyl as a response site of pH, and regulates and controls the fluorescence property of the probe through an intramolecular charge transfer effect. Under alkaline conditions, the phenolic hydroxyl group is deprotonated to convert to a phenolate ion, resulting in a red-shift in the fluorescence emission wavelength of the probe. As the basicity increases, the fluorescence emission wavelength of the probe gradually red-shifts from 545nm to 584 nm.
In some embodiments, detecting pH is detecting endoplasmic reticulum pH.
In a fourth embodiment of the present invention, there is provided a use of the above compound as a probe for detecting pH ratio type fluorescence.
In a fifth embodiment of the invention, the application of the compound in preparing a target endoplasmic reticulum pH fluorescent probe or a ratio type pH detection probe is provided.
In order to make the technical solutions of the present invention more clearly understood by those skilled in the art, the technical solutions of the present invention will be described in detail below with reference to specific embodiments.
Example 1:
synthesis of fluorescent probe:
1,1, 2-trimethyl-1H-benzo [ e ] indole (0.21g,1mmol) and iodopropionic acid (0.30g,1.5mmol) were weighed out, added to a two-necked flask under nitrogen, dissolved in acetonitrile (20mL) and refluxed at 80 ℃ for 48H. After completion of the reaction, most of the solvent was removed and the remaining concentrate was precipitated with ethyl acetate to give the product 1 as a pale yellow solid. The product 1 was used directly in the next reaction without further purification.
Product 1(0.56g,2mmol) was weighed out, taken in a two-necked flask under nitrogen, dissolved in DMF (1mL) and dichloromethane (10mL), added triethylamine (500. mu.L), HOBT (0.42g,2mmol) and EDC (0.38 g,2mmol), reacted at 0 ℃ for 0.5h, then N-toluenesulfonylethylenediamine (0.42g,2mmol) was added and the reaction was continued at room temperature for 24 h. After completion of the reaction, most of the solvent was removed, and the remaining concentrate was precipitated with ether. The product 2 was obtained as a dark blue color, and the product 2 was used in the next reaction without further purification.
P-hydroxybenzaldehyde (0.14g, 1.2mmol) and product 2(0.47g, 1mmol) were weighed out and added to a two-necked flask under nitrogen protection, 5mL of ethanol was added to dissolve the mixture, 10. mu.L of piperidine was added under stirring, the mixture was heated to 80 ℃ and refluxed for 6 hours, and then 10. mu.L of acetic acid was added and reacted for 10 minutes. After the reaction was stopped, the solvent was distilled off under reduced pressure, followed by reaction with dichloromethane: methanol 10:1 as a developing solvent, and separation and purification were performed by silica gel column chromatography to obtain ph-ER (0.41g, 70%) as a red solid.
Nuclear magnetic and mass spectral characterization of ph-ER:
1H NMR(400MHz,MeOD)δ8.42(d,J＝15.8Hz,1H),8.34(d,J＝8.5Hz,1H),8.15(d,J ＝8.9Hz,1H),8.10(d,J＝8.2Hz,1H),7.97(d,J＝8.8Hz,2H),7.89(d,J＝8.9Hz,1H),7.75(t, J＝7.2Hz,1H),7.64(t,J＝7.7Hz,1H),7.56(d,J＝8.3Hz,2H),7.40(d,J＝15.8Hz,1H),7.26 (d,J＝8.0Hz,2H),6.88(d,J＝8.7Hz,2H),4.90(t,J＝6.6Hz,2H),3.14(t,J＝6.1Hz,2H),2.90 (t,J＝6.5Hz,2H),2.80(t,J＝6.1Hz,2H),2.38(s,3H),2.05(s,6H).13C NMR(101MHz, MeOD)δ181.95,170.52,154.06,143.26,138.33,137.34,137.15,134.14,133.38,130.95,129.89, 129.33,127.96,127.44,126.52,126.45,125.11,122.52,117.91,111.90,105.91,63.01,53.25, 42.37,41.55,39.32,33.86,29.35,28.93,26.71,25.74,22.34,20.05,13.06.HRMS(ESI)m/z:[M+] calculated for C34H36N3O4S+,582.2427,found 582.2346.
effect test of ph-ER:
generally, the dye molecules can be dissolved in physiological saline, buffer solution or water-soluble organic solvent such as acetonitrile, dimethylsulfoxide, etc., and then added with appropriate buffer solution and other organic reagents for the test. The absorption and fluorescence spectral response of the probe pH-ER in PBS buffer solutions with different pH values along with the change of pH values are respectively explored and used for imaging experiments of living cells and inflammatory mice. The living cell staining method is to incubate the cultured cells in a culture solution containing probe molecules, remove the incubation solution after incubation for a certain time, and perform a confocal imaging experiment. The mouse staining method is to inject the probe into the normal part and inflammation part of mouse in situ, and after some time, to take the living fluorescence imaging of the mouse injected probe.
Ultraviolet absorption, fluorescence emission and reversibility selection experiments of the probe pH-ER in buffers with different pH values:
the UV absorption and fluorescence response properties of probe pH-ER in PBS (0.1M) at different pH were explored. Buffers of different pH were prepared by adding as little sodium hydroxide or hydrochloric acid solution as possible to PBS at pH 7.4. The absorption spectrum of the probe pH-ER (20 μ M) shows a change with high sensitivity with increasing pH (4-11). As shown in FIG. 1a, the maximum absorption peak of the probe in a buffer medium with pH 5 is around 450nm, and the maximum absorption red of the probe is shifted to around 550nm as the pH value increases. FIG. 1b shows the fluorescence spectral response of probe pH-ER (10. mu.M) in buffers with different pH values (4-11). Under the excitation of light at 480nm, the fluorescence intensity of the probe at 545nm is gradually reduced along with the gradual increase of the pH, and the fluorescence intensity at 584nm is gradually increased, so that the fluorescence response of the probe with high sensitivity to the pH is proved, and the pH can be quantitatively detected through ratiometric fluorescence.
FIG. 2 shows the reversibility of the probe ph-ER in an environment with reciprocating acidic and basic properties. The probe (10. mu.M) was added to PBS at pH 4.5 for fluorescence detection, and the spectral response was recorded by changing the pH between 4.5 and 9.5 with constant adjustment. Ordinate FI584nm/FI545nmRepresents the ratio of the fluorescence intensity of the probe at 584nm and 545 nm. The results show that pH-ER consistently exhibits good, transient fluorescence response with good reversibility in pH 4.5 and 9.5 buffer.
Endoplasmic reticulum targeting experiments with ph-ER:
human normal hepatocyte HL-7702 was cultured in DMEM medium containing polyclonal antibody and fetal bovine serum albumin. After incubating the cells with 20 μ M probes separately and with commercial dyes (including endoplasmic reticulum, mitochondria, lysosomes) that localize different subcellular organelles for 30min, the co-localization imaging experiments were performed using confocal laser microscopy. Co-localization cell imaging experiments As shown in FIG. 3, the probe ph-ER in FIGS. 3a, b, c only overlaps well with the fluorescence of the endoplasmic reticulum commercial localization dye. FIG. 3d, e, f are graphs showing the trend of fluorescence intensity of probe Ph-ER and commercial dye in a certain linear region. The better the fluorescence intensity overlap, the more consistent the trend, indicating the better the co-localization effect, and thus the probe exhibits the ability to efficiently target the endoplasmic reticulum.
Confocal fluorescence imaging experiments of probes in living cells at different pH:
human hepatocyte HL-7702 is cultured in DMEM culture solution containing double antibody and fetal bovine serum albumin. The cells were then incubated with pH-ER (20. mu.M) for 10 min at 37 ℃ and after washing off the probe, incubation was continued for 10 min with PBS buffer (0.01M) containing 10. mu.M nigericin, an ionophore, at various pH's (6.0,6.5,7.0,7.5,8.0) followed by confocal imaging. Due to the intervention of nigericin, the intracellular pH value will change according to the change of the PBS buffer pH. As shown in FIG. 4, after the cells were incubated with PBS at pH 6.00, the green channel and the red channel of the probe showed similar fluorescence in the cells, and as the pH of the cell incubation solution increased from 6.00 to 8.00, the green channel fluorescence of Ph-ER in the cells gradually decreased, and the red channel fluorescence gradually increased. The above results indicate that the probe Ph-ER can be used with high sensitivity to monitor pH changes in living cells.
Probe in situ photoacoustic imaging experiments on normal and inflammatory sites in a mouse model of inflammation:
mice were treated with saline intraperitoneally on the left side as a control, and LPS (2mg/mL, 400. mu.L) was injected at the same position on the right side to induce the production of abdominal inflammation. After 4 hours, the mice were anesthetized with chloral hydrate and the experiment was performed. The probe ph-ER (50. mu.M, 50. mu.L) was intraperitoneally injected at the site where LPS (right) and physiological saline (left) were injected, and the in vivo fluorescence imaging detection was performed under a laser of 480nm, as shown in FIG. 5. As a result, it was found that, under a laser of 480nm, the fluorescence intensity of the LPS-stimulated side of the mouse was slightly higher than that of the side injected with physiological saline in the 520nm window, whereas the fluorescence intensity of the LPS-stimulated side of the mouse was significantly lower than that of the side injected with physiological saline in the 620nm window, and the LPS side had a lower FI620nm/FI520nmThe ratio shows that the LPS stimulation site presents weak acidity and the pH value is lower. Indicating that pH-ER can be used as a fluorescence ratio imaging probe to detect pH in vivo.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (14)
1. A compound characterized by the chemical structure represented by the formula:
2. a process for the preparation of the compound of claim 1, wherein 1,1, 2-trimethyl-1H-benzo [ e ] indole is quaternized with iodopropionic acid to obtain 1, N-tosylethylenediamine, amidation of carboxyl and amino groups is performed on product 1 to obtain 2, and condensation of methyl and aldehyde groups is performed on product 2 and p-hydroxybenzaldehyde to obtain the compound, the synthetic route is as follows:
3. a process for the preparation of a compound according to claim 2, characterized in that the amidation reaction is carried out by: under the alkaline condition, the product 1 and a condensing agent react at a temperature of-1-0 ℃, and then N-tosylethylenediamine is added to react at room temperature.
4. A process for the preparation of a compound according to claim 3, characterized in that basic conditions are provided by triethylamine.
5. The method for preparing the compound according to claim 3, wherein the condensing agent is a mixed system of EDC and HOBt.
6. The method according to claim 5, wherein the molar ratio of N-toluenesulfonylethylenediamine, EDC and HOBt is 1:0.9 to 1.1.
7. The method for preparing the compound according to claim 3, wherein the reaction is carried out at-1 to 0 ℃ for 0.5 to 1 hour.
8. The method for preparing a compound according to claim 3, wherein the reaction is carried out at room temperature for 12 to 24 hours.
9. The method according to claim 2, wherein the molar ratio of N-toluenesulfonylethylenediamine to product 1 is 1:0.9 to 1.1.
10. A process for the preparation of a compound according to claim 2, characterized in that the conditions of the condensation reaction are: in an alkaline environment, the reaction temperature is 70-90 ℃, and the reaction time is 3-12 h.
11. The method of claim 10, wherein piperidine is used to provide a basic environment at a pH of 8 to 9.
12. The process according to claim 2, wherein the solvent for the condensation reaction is absolute ethanol.
13. The method for preparing the compound according to claim 2, wherein the feed ratio of the product 2 to the p-hydroxybenzaldehyde is 1: 1.1-1.3.
14. Use of a compound of claim 1 for the preparation of a target endoplasmic reticulum pH fluorescent probe or a ratiometric pH detection probe.
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| CN109096189A (en) * | 2018-09-14 | 2018-12-28 | 济南大学 | The two-photon fluorescence probe of pH in a kind of detection endocytoplasmic reticulum |
| CN109651249A (en) * | 2019-01-08 | 2019-04-19 | 济南大学 | A kind of fluorescence probe detecting endocytoplasmic reticulum cysteine and its synthesis and application |
| CN110078665A (en) * | 2019-05-16 | 2019-08-02 | 济南大学 | A kind of hypochlorous fluorescence probe of detection of endoplasmic reticulum targeting and application |
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| CN109096189A (en) * | 2018-09-14 | 2018-12-28 | 济南大学 | The two-photon fluorescence probe of pH in a kind of detection endocytoplasmic reticulum |
| CN109651249A (en) * | 2019-01-08 | 2019-04-19 | 济南大学 | A kind of fluorescence probe detecting endocytoplasmic reticulum cysteine and its synthesis and application |
| CN110078665A (en) * | 2019-05-16 | 2019-08-02 | 济南大学 | A kind of hypochlorous fluorescence probe of detection of endoplasmic reticulum targeting and application |
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