A kind of lysosome super-resolution fluorescence imaging dye and its synthesis method and application

A technology of super-resolution fluorescence and synthesis method, applied in the field of lysosome super-resolution fluorescence imaging dye and its synthesis, can solve the problems of photoactivation performance failure, limited application, fluorescence interference, etc., and achieve the effect of good imaging effect

Active Publication Date: 2020-12-04
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, when the rhodamine spiramide dye is exposed to the acidic environment in the lysosome, the fluorescence generated by its acid activation will seriously interfere or even lead to the complete failure of the photoactivation performance. Applications in volumetric super-resolution imaging

Method used

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  • A kind of lysosome super-resolution fluorescence imaging dye and its synthesis method and application
  • A kind of lysosome super-resolution fluorescence imaging dye and its synthesis method and application
  • A kind of lysosome super-resolution fluorescence imaging dye and its synthesis method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 3-Acetamido-substituted rhodamine ethylmorpholine spiroamide (P1) synthetic route and product structure are as follows:

[0025]

[0026] Synthesis steps and characterization: 3-nitrorhodamine (2mmol, 0.974g) and 2-ethylaminomorpholine (2mmol, 0.146g) were dissolved in absolute ethanol (35mL). The temperature was raised to 78°C for reflux, and after stirring for 4 hours, the solvent was evaporated under reduced pressure, and the product was separated and purified by column chromatography (silica gel, petroleum ether / ethyl acetate, 4:1v / v) to obtain a light yellow powder (1.14g, 95 %). Then all the powder was dissolved in methanol (5mL), stirred under hydrogen atmosphere and palladium carbon (10%wt) catalysis for 1 hour, the filtrate was taken by suction filtration, and the final white powdery intermediate (1.07g , 99%). The white powder intermediate (0.57g, 1mmol) and acetyl chloride (0.12g, 1.5mmol) were mixed in dichloromethane (15mL), stirred for 2 hours, and th...

Embodiment 2

[0029] The product P1 in Example 1 was dissolved in methylene chloride / methanol (9 / 1, v / v) mixed solvent (concentration is 10 - 5 M), trifluoroacetic acid (2.3 μL, 1000 eq) was added to the mixed solution. Time-resolved UV-Vis absorption spectra were measured before and after acid addition ( Figure 4 ), the results showed that the absorbance at the maximum absorption wavelength of P1 did not increase with the increase of acidification time, indicating that the P1 molecule has the property of acid resistance.

[0030] MCF-7 cells were co-stained with the product 3-acetylamino-substituted rhodamine ethylmorpholine spiroamide P1 (10 μM) in Example 1 and a commercial lysosome labeling dye (LTG, 0.1 μM), and the MCF-7 cells were co-stained by laser co-staining. A focusing inverted microscope was used to observe the fluorescent staining in the two channels in real time. The excitation light wavelength of the green channel is 488nm, and the fluorescence signal in the 500-550nm ban...

Embodiment 3

[0032] (1) 3-nitrorhodamine and 2-ethylaminomorpholine are dissolved in absolute ethanol according to the molar ratio of substances (1:2.5), and the temperature is raised to reflux, and after stirring for 3 hours, the solvent is evaporated to obtain a colorless solid, and further Separation and purification by silica gel column chromatography to obtain 3-nitrorhodamine ethyl morpholine amide.

[0033] (2) Take the product 3-nitrorhodamine ethyl morpholine amide in the above step (1), dissolve it in an appropriate amount of methanol, stir for 3 hours under a hydrogen atmosphere and palladium carbon (5%wt) catalysis, and take the filtrate by suction filtration , After distilling off the solvent under reduced pressure, it was separated and purified by silica gel column chromatography to obtain 3-aminorhodamine ethyl morpholine amide.

[0034] (3) Get the product 3-aminorhodamine ethyl morpholine amide product in the above step (2) and acetyl chloride according to a certain substa...

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Abstract

The invention provides a lysosome super-resolution fluorescent imaging dye and its synthesis method and application. The specific molecular structure of the dye is rhodamine ethyl morpholine amide substituted by 3-acetylamino, and its structural formula is shown in the following formula (1): The lysosome super-resolution fluorescent imaging dye of the present invention not only has acid-resistant performance, but also retains light-activated performance. Therefore, this kind of acid-resistant light-activated dye can be used in super-resolution imaging technology in lysosome without interference from the pH of the biological environment, so as to achieve good imaging effect. In addition, the lysosome super-resolution fluorescent imaging dye of the present invention can also be used as a molecular fluorescent probe in the field of sensing and detection.

Description

technical field [0001] The invention belongs to the field of fluorescent dyes and super-resolution fluorescent imaging, and in particular relates to a lysosome super-resolution fluorescent imaging dye and its synthesis method and application. Background technique [0002] As an important acidic organelle in eukaryotic cells, lysosomes contain more than 60 kinds of acid hydrolases, cathepsins, and various specific membrane proteins. It not only has the function of intracellular digestion, but also has the function of regulating secretion in some cases. When the number and distribution of lysosomes are abnormal due to their own variation or external influence, it may cause lung diseases (silicosis, tuberculosis, etc.) ), lysosomal storage diseases (Taiwan-Sarritz syndrome, type II glycogen storage disease, cellular inclusion disease, etc.), tumors and other diseases. As the "digestive organ" in the cell, the research on lysosome has always been a hot spot in life science. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D491/107C09B11/20C09K11/06G01N21/64
CPCC09B11/20C09K11/06C09K2211/1029C09K2211/1033C09K2211/1088G01N21/6486
Inventor 徐兆超陈婕祁清凯
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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