CN115536669B - Electron donor-electron acceptor (D-A) near infrared luminous cell lipid drop fluorescence imaging probe and application thereof - Google Patents
Electron donor-electron acceptor (D-A) near infrared luminous cell lipid drop fluorescence imaging probe and application thereof Download PDFInfo
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Abstract
An electron donor-electron acceptor (D-A) near infrared luminous cell lipid drop fluorescent imaging probe and application thereof belong to the technical field of biological imaging. The structural formula of the fluorescent probe is shown as follows, and the triphenylamine group is electron donor, 1, 5-benzene oxide [1,2-b:4,5-b ]']The dithiophene and benzyl cyanide groups are electron acceptors, and the obtained fluorescent probe Lipi-Deep Red has very good imaging effect. The invention also discloses application of the fluorescent probe for specifically marking lipid droplets in cells under a structured light illumination microscope (SIM), tracking distribution and dynamic changes of lipid droplets in living cells and tracking relative dynamic changes of lipid droplets in living cells and mitochondria. Experiments prove that the fluorescent probe Lipi-Deep Red has the advantages of high brightness, imaging signal to noise ratio, ultrahigh light stability, lower cytotoxicity and the like, and has great application prospect.
Description
Technical Field
The invention belongs to the technical field of biological imaging, and particularly relates to an electron donor-electron acceptor (D-A) near infrared luminous cell lipid drop fluorescent imaging probe and a method for specifically marking lipid drops in cells under a structured light illumination microscope (SIM), tracking distribution and dynamic changes of lipid drops in living cells (shown in figure 7); the use of the relative kinetic changes of lipid droplets and mitochondria in living cells was followed (as shown in figure 8).
Background
Lipid droplets are a spherical organelle that is widely distributed in almost all organisms from prokaryotes to humans and whose constituent components are a neutral lipid core (triglycerides and cholesterol esters) and a phospholipid monolayer. With the recent intensive research into lipid droplets, many cell activities involving lipid droplets have been started, for example: membrane structure synthesis and transport, protein storage and degradation, inflammation, and other symptoms. Lipid droplet dysfunction can induce the development of a variety of diseases such as fatty liver, atherosclerosis, cancer, diabetes, and the like. Thus, research on lipid droplets has become the hottest area in cell biology in the last decade.
Confocal and wide-field fluorescence imaging techniques have been widely used in order to study the diverse functions of lipid droplets. However, due to the diffraction limit of light, the resolution limit achievable by conventional imaging techniques is only around 250nm, which is a significant concern for observing and studying the movement of small lipid droplets. While a structured light illumination microscope (SIM) with nanoscale resolution can observe and realize dynamic monitoring of small lipid droplets, which has strong requirements on the light stability of fluorescent probes, common lipid droplet commercial probes BODIPY and Nile Red (Materials, 2018, 11, 1768) have their own disadvantages, for example, BODIPY can cause overlapping of absorption and emission spectra due to smaller stokes shift, and Nile Red faces the problems of poor lipid droplet staining specificity and stability. In addition, the probe with light emission in the near infrared band can overcome the autofluorescence interference of biological tissues in the imaging process, and the development of various organelle probes with near infrared light emission is greatly attracting the attention of researchers, while lipid droplet fluorescent probes with near infrared light emission are more rare. Therefore, development of near infrared fluorescent probes with large stokes shift, high lipid droplet specificity and ultra-high light stability has become an urgent need to realize high quality lipid droplet fluorescent imaging.
Disclosure of Invention
In view of the shortcomings of the current cytosolic lipid drop probes in imaging, the invention aims to provide a novel electron donor-electron acceptor (D-A) near infrared luminescent cytosolic lipid drop fluorescent imaging probe and application of the same in specific labeling of lipid drops in cells, tracking of distribution and dynamics changes of lipid drops in living cells and tracking of relative dynamics changes of lipid drops in living cells and mitochondria under a structured light illumination microscope (SIM).
The invention relates to a novel electron donor-electron acceptor (D-A) near infrared luminous cell lipid drop fluorescent imaging probe, which is characterized in that: the molecular structure is a donor-acceptor structure, and the chemical structure is shown as a formula (I):
wherein the triphenylamine group is an electron donor, and the 1, 5-tetraoxide benzo [1,2-b:4,5-b' ] dithiophene and the benzyl cyanide group are electron acceptors.
The chemical name of the cell lipid drop fluorescent probe is 4- (6- (4- (diphenylamino) phenyl) -4, 8-bis (2-methoxyethoxy) -1, 5-benzene oxide [1,2-b:4,5-b' ] dithiophene-2-yl) benzonitrile, which is abbreviated as Lipi-Deep Red. The probe is a novel fluorescent molecule synthesized by the invention, and the preparation reaction formula is as follows:
aiming at the defects of the current cell lipid drop probe in imaging, the invention selects D-A near infrared luminescent fluorescent molecules in the aspect of developing novel lipid drop imaging fluorescent probes. In the invention, triphenylamine groups are electron donors, and 1, 5-tetraoxide benzo [1,2-b:4,5-b' ] dithiophene and benzyl cyanide groups are electron acceptors. The prepared fluorescent molecule with the donor-acceptor structure shows stronger near infrared luminescence and larger Stokes shift. The introduction of bis (2-methoxyethoxy) further improves the hydrophilicity and hydrophobicity of the fluorescent probe, allowing the probe to better target cell lipid droplets. In the aspect of fluorescence imaging application, large Stokes displacement can effectively avoid cross overlapping of absorption and emission spectrums, and near infrared emission can effectively reduce the influence of cell autofluorescence background signals. Finally, smaller molecular structures can also be effective in increasing cell membrane permeability. Based on the advantages, the fluorescent probe Lipi-Deep Red prepared by the invention can be used as a lipid drop probe to realize high-quality fluorescent imaging (confocal imaging and structured light illumination microscope).
The invention relates to the use of a fluorescent probe Lipi-Deep Red for specifically labeling lipid droplets in cells, tracking distribution and dynamic changes of lipid droplets in living cells and tracking relative dynamic changes of lipid droplets in living cells and mitochondria under a structured light illumination microscope (SIM) (examples 7 and 8).
The cells of the invention are HeLa cells.
The fluorescent probe Lipi-Deep Red of the lipid droplets in the specific labeled cells prepared by the invention is a fluorescent probe with near infrared light emission, ultrahigh dyeing selectivity and light stability, which can be used for confocal imaging and structured light illumination microscope (SIM) imaging.
Experimental results prove that the fluorescent probe Lipi-Deep Red has ultrahigh dyeing selectivity, and under the same dyeing and imaging conditions, the Lipi-Deep Red shows more excellent dyeing selectivity than Nile Red. The MTT assay demonstrated that Lipi-Deep Red has very low cytotoxicity and is compatible with commercial mitochondrial probe MitoTracker Green. Most importantly, lipi-Deep Red has ultra-high light stability and can be used for SIM imaging to dynamically track lipid droplets and to dynamically track interactions between lipid droplets and mitochondria. Therefore, the fluorescent probe Lipi-Deep Red can be used as a powerful tool for tracking the distribution and dynamic process of lipid droplets in living cells by using the specific labeled lipid droplets, and is expected to become a commercial lipid droplet SIM super-resolution imaging fluorescent probe. More importantly, the method can provide a new field of view for the research of the lipid drop cell biology and promote the development of the lipid drop cell biology.
In a word, the probe Lipi-Deep Red is a brand-new fluorescent probe, and compared with other lipid droplet fluorescent probes, the Lipi-Deep Red has the advantages of high imaging signal to noise ratio, ultrahigh lipid droplet staining selectivity, light stability, lower cytotoxicity and the like. In view of the characteristics, the application of the fluorescent imaging method in the cell lipid drop fluorescence imaging has wide prospect.
Drawings
Fig. 1: nuclear magnetic hydrogen spectrum of fluorescent probe Lipi-Deep Red prepared in example 1 of the present invention;
fig. 2: mass spectrum of fluorescent probe Lipi-Deep Red prepared in example 1 of the present invention;
fig. 3: absorption-emission spectra of the fluorescent probe Lipi-Deep Red prepared in example 1 of the present invention in toluene (tolene);
wherein, the left dotted line part is an absorption spectrum, and the right solid line part is an emission spectrum.
Fig. 4: the fluorescent probe Lipi-Deep Red prepared in example 1 of the present invention was stained for HeLa cells for 24 hours under different concentrations, and a bar chart of cell viability was obtained;
fig. 5: the fluorescent probe Lipi-Deep Red prepared in example 1 of the present invention was co-localized with the commercial lipid droplet fluorescent probe BODIPY493/503 in HeLa cells;
wherein the first photograph (a) is a fluorescence photograph of BODIPY493/503 in a 488-510nm imaging channel under 470nm laser excitation; the second photograph (b) is a fluorescence photograph of Lipi-Deep Red in a 650-680nm imaging channel under 470nm laser excitation; the third photograph (c) is a superposition of the first two fluorescent photographs and the bright field photograph using imagej software; the fourth photograph (d) is the pearson correlation coefficient r=0.92 for the first two fluorescent channels. Ruler: 10 μm.
Fig. 6: quantification diagrams of light stability of the fluorescent probes Lipi-Deep Red and the lipid droplet fluorescent probe Nile Red prepared in the embodiment 1 of the present invention in HeLa cells;
wherein, the part a is 1 st photo and 50 th photo of 50 photos of the fluorescent probes Lipi-Deep Red and Nile Red which are continuously imaged in the same area after HeLa cells are stained; panel b shows the data processing of the relative fluorescence intensity trend with the number of images for 50 consecutive images of Lipi-Deep Red and Nile Red, respectively, using Origin software. Ruler: 10 μm.
Fig. 7: the fluorescent probe Lipi-Deep Red prepared in the embodiment 1 of the invention tracks the kinetics process of the lipid droplets in HeLa cells in a SIM mode;
wherein, from left to right, the three pictures are the 1 st, 1000 th and 2000 th SIM mode pictures in sequence. Ruler: 2 μm.
Fig. 8: fluorescent probes Lipi-Deep Red and commercial mitochondrial probe MitoTracker Green prepared in example 1 of the present invention track HeLa intracellular lipid droplets and mitochondrial dynamics in SIM mode:
wherein the three pictures from left to right are the 1 st, 50 th and 100 th SIM mode pictures in sequence. Ruler: 2 μm.
Detailed Description
Example 1:
1. synthesis of 4- (6- (4- (diphenylamino) phenyl) -4, 8-bis (2-methoxyethoxy) -1, 5-tetraoxide benzo [1,2-b:4,5-b' ] dithiophene-2-yl) benzonitrile
To a composition containing 2, 6-dibromo-4, 8-bis (2-methoxyethoxy) benzo [1,2-b:4,5-b ]']A reaction flask of dithiophene 1, 5-tetraoxide (0.222 g,0.397 mmol), triphenylamine boric acid (0.110 g,0.379 mmol), potassium carbonate (553 mg,4.00 mmol) and tetraphenylpalladium phosphate (0.058 g,0.050 mmol) was charged with 50mL of a mixed solvent of toluene, ethanol and water (V) Toluene (toluene) ：V Ethanol ：V Water and its preparation method =8: 1: 1). The reaction system was stirred at 90 ℃ for 1 hour, and the filtrate was extracted and spin-dried to give unpurified reaction intermediate 2. The resulting reaction intermediate 2, (4-cyanophenyl) boric acid (0.29 g,1.99 mmol), potassium carbonate (553 mg,4.00 mmol) and tetrakis triphenylphosphine palladium (0.058 g,0.050 mmol) were added to a reaction flask, and 50mL of a mixed solvent of toluene, ethanol and water, from which oxygen was removed (V Toluene (toluene) ：V Ethanol ：V Water and its preparation method =8: 1: 1). The reaction system was stirred at 90℃for 12 hours, and after extraction and purification by column chromatography, 23mg (0.032 mmol) was obtained9%) of Lipi-Deep Red in the form of a Red powder.
1 H NMR(500MHz,CDCl 3 ):δ7.92(d,J＝8.5Hz,2H),7.87(s,1H),7.78(d,J＝ 8.5Hz,2H),7.66(d,J＝8.8Hz,2H),7.54(s,1H),7.33(t,J＝7.9Hz,4H),7.18–7.13 (m,6H),7.07(d,J＝8.8Hz,2H),4.65–4.61(m,2H),4.59–4.56(m,2H),3.84–3.80 (m,4H),3.47(d,J＝1.8Hz,6H).
FIG. 1 shows nuclear magnetic resonance hydrogen spectra of the fluorescent probe Lipi-Deep Red synthesized in example 1, and FIG. 2 shows mass spectra of the fluorescent probe Lipi-Deep Red synthesized in example 1, indicating that the target product Lipi-Deep Red is prepared.
Example 2: determination of the absorption-emission Spectrum of the fluorescent Probe Lipi-Deep Red prepared in example 1
The fluorescent probe Lipi-Deep Red synthesized in example 1 was prepared as a 10. Mu.M solution in 10mL of toluene (tolene) solvent. The absorption spectrum is obtained by scanning with an ultraviolet-visible spectrophotometer within the wavelength range of 300-800 nm, the fluorescence emission spectrum is obtained by collecting with an optical fiber type fluorescence spectrometer under 470nm excitation, the absorption-emission spectrum (the left broken line part is the absorption spectrum and the right solid line part is the emission spectrum) of the fluorescence probe Lipi-Deep Red in toluene solution as shown in figure 3 is obtained by processing data with Origin software, and the absorption-emission peak position of the fluorescence probe Lipi-Deep Red and the large Stokes shift are described.
Example 3: culture of HeLa cells
All percentages in this example are volume fractions.
HeLa cell lines were grown at 37℃and CO 2 The culture was performed in an incubator at a concentration of 5%, and the culture medium was high-sugar DMEM containing 10% fetal bovine serum and 1% diabody (a mixture of green streptomycin). Among them, fetal bovine serum, diabody and high-sugar DMEM were purchased directly from biological reagent companies.
After the cells grew to log phase, we passaged the cells: sucking 5mL of culture solution in a cell culture bottle, cleaning the cell surface with 2mL of high-sugar DMEM culture solution (containing 1% double antibody) without fetal calf serum, sucking the culture solution, and digesting the treated cells with 0.5mL of pancreatin for 2 min until the cell surface is largeAfter partial cell wall removal, adding 2mL of high-sugar DMEM culture solution containing 10% of fetal calf serum and 1% of double antibodies, blowing uniformly, taking a proper amount of the cell dispersion solution, respectively transferring into a new cell culture bottle and a new culture dish, and putting CO 2 The cells are cultured in a cell incubator, and the cells in the culture dish are used for confocal or SIM imaging experiments after the concentration is proper.
Example 4: test of cytotoxicity of fluorescent Probe Lipi-Deep Red prepared in example 1
We performed cytotoxicity tests on the fluorescent probe Lipi-Deep Red using 3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide (MTT). HeLa cells were seeded in 96-well plates (1.times.10 per well 4 Individual cells), put into CO 2 The cells were cultured in a cell incubator for 24 hours. The medium of the middle 60 wells was then changed to a medium containing different concentrations (0, 0.5, 1.0, 2.0, 5.0 and 10.0. Mu.M) of fluorescent probe Lipi-Deep Red and 1% (volume fraction) DMSO (10 sets of parallel assays were set for each concentration), and after incubation for a further 24 hours, MTT reagent (10. Mu.L per well) was added to these wells and the incubation was continued for 4 hours in a cell incubator. The original culture solution in these wells was removed, and the formazan crystals formed were dissolved in DMSO (100 μl per well) and then allowed to stand at room temperature for 30 minutes, and the absorbance of each well was measured at 490nm using an enzyme-labeled instrument. Since only living cells can react with MTT reagent to generate formazan crystals, we can calculate the cell survival rate by comparing the average absorbance value of each group of wells with different concentrations with the average absorbance value of the control group (10 wells with probe concentration of 0), and the result shows that the fluorescent probe Lipi-Deep Red has very small cytotoxicity and the fluorescent probe Lipi-Deep Red with concentration of 10.0 mu M does not influence the normal growth of HeLa cells within 24 hours as shown in FIG. 4.
Example 5: co-staining experiment of the fluorescent Probe Lipi-Deep Red prepared in example 1 with the lipid drop fluorescent Probe BODIPY493/503 in HeLa cells
We cultured HeLa cells in 20mm diameter glass bottom dishes in CO 2 Propagation was carried out in an incubator for 2 days. After removal from the incubator, the original DMEM broth in the dish was removed and 1mL of a culture medium containing Lipi-Deep Red (2. Mu.M), BODIPY493/503 (2. Mu.M) and 1% (volume fraction) was added) DMEM medium in DMSO was placed in a cell incubator for further incubation for 2 hours. After removal, the sample was washed 3 times with HBSS solution, and fluorescence imaging was performed in HBSS solution. As shown in FIG. 5, we can clearly observe that the fluorescent probe Lipi-Deep Red prepared in example 1 and the lipid droplet fluorescent probe BODIPY493/503 can well realize co-localization in HeLa cells, demonstrating that the fluorescent probe Lipi-Deep Red prepared in example 1 has excellent cell lipid droplet specificity.
Example 6: light stability test of fluorescent Probe Lipi-Deep Red prepared in example 1
After 2 dishes of HeLa cells of example 5 were removed from the cell incubator, the original DMEM medium was removed, DMEM medium (containing 1% DMSO) containing 2. Mu.M Lipi-Deep Red and 2. Mu.M Nile Red was added, and the dishes were returned to the incubator for 2 hours, and after 2 dishes were removed, washed 3 times with HBSS solution, respectively, and fluorescence imaging was performed. After 50 consecutive images were formed, respectively, as shown in FIG. 6, we found that Nile Red was photo-bleached very quickly, and the relative fluorescence intensity of the fluorescent probe Lipi-DSB prepared in example 1 was maintained at 90% or more after 50 images were formed, demonstrating its excellent photo-stability.
Example 7: fluorescent probe Lipi-Deep Red prepared in example 1 follows the kinetics of HeLa intracellular lipid droplets in the SIM mode
After taking out the culture dish fully covered with HeLa cells in example 5 from the incubator, the original DMEM culture solution was removed, DMEM culture solution containing 2. Mu.M Lipi-Deep Red and 1% DMSO was added, and after being put back into the incubator for 2 hours, the culture dish was taken out, washed 3 times with HBSS solution, and then SIM imaging was performed. As shown in FIG. 7, after selecting a region with higher lipid drop content, after continuously imaging 2000 images in the SIM mode, the images still can maintain significant fluorescence intensity, and from the continuous 2000 SIM images, we find the rapid movement of the small lipid drops, which illustrates the unprecedented ultra-high light stability of the fluorescent probe Lipi-Deep Red and the practicability of the fluorescent probe for super-resolution imaging of lipid drops in tracking the dynamics of lipid drops.
Example 8: fluorescent probes Lipi-Deep Red and commercial mitochondrial Probe MitoTracker Green prepared in example 1 track HeLa intracellular lipid droplets and mitochondrial dynamics in the SIM mode
We removed the HeLa cell-plated petri dish from the incubator in example 5, removed the original DMEM broth, added DMEM broth containing 2. Mu.M Lipi-Deep Red, 1. Mu.M commercial mitochondrial probe MitoTracker Green and 1% (volume fraction) DMSO, returned to the incubator for 2 hours, removed, washed 3 times with HBSS solution, and then SIM imaged. After selecting a region where lipid droplets and mitochondria coexist, as shown in fig. 8, 100 continuous images can be formed in the SIM mode, the lipid droplets can still maintain significant fluorescence intensity while the mitochondrial probe can hardly detect fluorescence signals, which highlights the high light stability of Lipi-Deep Red from the side. Furthermore, from this succession of 100 SIM photographs we found a change in the relative positions of lipid droplets and mitochondria. The unprecedented ultra-high light stability of the fluorescent probe Lipi-Deep Red and the practicability of the fluorescent probe as a lipid drop super-resolution imaging fluorescent probe in the aspect of double-color imaging are demonstrated.
Claims (2)
1. An electron donor-electron acceptor type near infrared luminous cell lipid drop fluorescent imaging probe has a structural formula shown as follows,
2. use of an electron donor-electron acceptor type near infrared luminescent cell lipid droplet fluorescence imaging probe according to claim 1 for specific labeling of lipid droplets in cells, tracking distribution and kinetic changes of lipid droplets in living cells or tracking relative kinetic changes of lipid droplets in living cells and mitochondria under a structured light illumination microscope, characterized in that: the cells are HeLa cells.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109722237A (en) * | 2019-01-07 | 2019-05-07 | 华南理工大学 | A kind of fluorescence probe based on condensed ring oxidation of thiophene and its application in cell imaging |
| CN113651834A (en) * | 2021-08-25 | 2021-11-16 | 吉林大学 | Fluorescent probe based on dithienobenzene derivative and application of fluorescent probe in cell lipid drop imaging |
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| CN109722237A (en) * | 2019-01-07 | 2019-05-07 | 华南理工大学 | A kind of fluorescence probe based on condensed ring oxidation of thiophene and its application in cell imaging |
| CN113651834A (en) * | 2021-08-25 | 2021-11-16 | 吉林大学 | Fluorescent probe based on dithienobenzene derivative and application of fluorescent probe in cell lipid drop imaging |
Non-Patent Citations (1)
| Title |
|---|
| Zhen, Shijie.Efficient Red/Near-Infrared Fluorophores Based on Benzo[1,2-b:4,5-b']dithiophene 1,1,5,5-Tetraoxide for Targeted Photodynamic Therapy and In Vivo Two-Photon Fluorescence Bioimaging.《Advanced Functional Materials》.2018,第28卷(第13期),1-15页. * |
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