Cell nucleus targeting carbon dot, preparation and application
A cell nucleus, carbon dot technology, applied in nano-carbon, fluorescence/phosphorescence, instruments, etc., can solve the problems of low fluorescence quantum yield, unclear imaging, long time into cells, etc., to achieve high chemical stability, photostability Good, the effect of fast infiltration time
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Embodiment 1
[0041] The preparation method of carbon dots, concrete steps are as follows:
[0042] Dissolve semicarbazide hydrochloride (0.0270 g) and m-phenylenediamine (0.0170 g) in absolute ethanol (3 ml) to prepare a precursor mixture. After sonicating for five minutes, the mixture was transferred to a Teflon-lined In a high pressure reactor. Subsequently, the autoclave was placed in an oven and heated to reflux at 180° C. for 12 hours. Wait for the reaction kettle to cool down to room temperature naturally to obtain a brown solution, and then use dichloromethane and methanol as eluents to purify the obtained solution by silica gel column chromatography. The organic solvent in the obtained carbon dot solution was removed with a rotary evaporator to obtain a carbon dot solid. Dissolve the carbon dots solid in DMSO to obtain a stock solution of carbon dots at a concentration of 2 mg / mL.
Embodiment 2
[0044] The stability research of carbon dots on common metal ions and amino acids in cells, the specific steps are as follows:
[0045] A certain amount of carbon dot stock solution was added to the PBS buffer solution to make the concentration 10 μg / mL, and a certain amount of metal ions and amino acids were added to make the concentrations of metal ions and amino acids 100 μM and 1 mM. The resulting solution was transferred to a fluorescence cuvette, and the fluorescence intensity was measured with a fluorescence spectrophotometer. Such as figure 1 As shown, after adding metal ions or amino acids, the fluorescence intensity of carbon dots did not change significantly, indicating that carbon dots have good stability to common metal ions and amino acids in cells.
[0046] The stability research of carbon dots on the common ROS in the process of cell ferroptosis, the specific steps are as follows:
[0047]A certain amount of carbon dot stock solution was added to the PBS buff...
Embodiment 3
[0049] The rapid cell imaging research of carbon dots, the specific steps are as follows:
[0050] HepG2 cells were seeded in confocal culture dishes at 37 °C, 5% CO 2 , in a cell culture incubator with saturated humidity for 24 h (the cell culture medium contains DMEM high-glucose medium, 10% volume fraction of gibco serum, 100 μg / mL penicillin and 100 μg / mL streptomycin); the carbon dot solution Add it to a confocal culture dish to make the final concentration 4 μg / mL, excite it with a light source with an excitation wavelength of 458nm, perform confocal imaging, and observe the time when carbon dots enter the cells. Such as image 3 As shown, after the carbon dots were co-incubated with the cells for 5 minutes, the entire nucleus showed green fluorescence, which indicated that the carbon dots had easily penetrated the cell membrane and entered the nucleus. As time increased, the fluorescence of the nucleus gradually increased and reached 25 It reaches a plateau in about 1...
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