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A fluorescent silicon nano-dot and its preparation method and application

A silicon nanometer and fluorescence technology, applied in the fields of nanomaterials and biology, can solve the problems of poor stability, high cost, short imaging time, etc., and achieve the effects of good cleaning resistance, cheap raw materials, and simple preparation method.

Active Publication Date: 2018-11-09
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved in the present invention is to provide a fluorescent silicon nano-dot (SiNDs) to realize ultra-long-time, cleaning-resistant, fix-resistant, and permeabilization-resistant lysosome imaging, which solves the high cost of the prior art, Problems such as poor stability and short imaging time

Method used

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  • A fluorescent silicon nano-dot and its preparation method and application
  • A fluorescent silicon nano-dot and its preparation method and application
  • A fluorescent silicon nano-dot and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Taking AEEA as an example, the preparation of fluorescent silicon nanodots includes the following steps:

[0049] (1) Weigh Rose Bengal and fully dissolve it in ultrapure water, add an appropriate volume of AEEA so that the mass ratio of AEEA to Rose Bengal is 100:3. The two are fully mixed and transferred to a hydrothermal reaction kettle;

[0050] (2) Reaction: React at 160°C for 3 hours in a hydrothermal reactor to form a silicon nanodot solution;

[0051] (3) Purification: the target ultra-bright fluorescent silicon nano-dot solution is obtained by dialysis.

[0052] For a schematic diagram of the reaction, see figure 1 , the transmission electron microscopy results of the prepared SiNDs are shown in figure 2 , the statistical results of particle size distribution are shown in image 3 ,, its UV-Vis absorption spectrum see Figure 4 , and its fluorescence excitation emission spectrum is shown in Figure 5 , its extinction at 650nm in different pH environments...

Embodiment 2

[0054] The lysosome imaging effect of SiNDs prepared in Example 1 on A549 cells was tested as follows:

[0055] (1) Cell culture: A549 cells were cultured at 5×10 3 The density of cells / well will be seeded in a 96-well plate at 37°C, 5% CO 2 Cultivate in the environment for 24h;

[0056] (2) Cell staining: replace the culture medium in each well with fresh culture medium containing 20 g / mL SiNDs, and incubate at 37°C, 5% CO 2 After culturing in the environment for 2, 4, 18 and 24 hours respectively, they were washed twice with phosphate buffered saline (PBS). Then, stain with commercial red lysosome fluorescent dye (LT-Red) and cell nucleus staining reagent (Hoechst 33342) for 30 and 10 minutes, respectively. Finally, wash the cells twice with PBS;

[0057] (3) Confocal fluorescence microscope imaging observation: using lasers with wavelengths of 405, 488 and 552nm as excitation light, Hoechst 33342 dye emits blue fluorescence under excitation of 405nm excitation light, wh...

Embodiment 3 and 4

[0060] The lysosome imaging effect of commercial lysosomal green fluorescent dye (LT-Green) and red fluorescent dye (LT-Red) on A549 cells was tested.

[0061] The operation process of Examples 3 and 4 is basically the same as that of Example 1, only need to replace SiNDs in step (2) with 1 LT-Green and LT-Red for 30 min, 4 h and 8 h respectively.

[0062] Lysosome imaging effect see Figure 8 and 9 . Observed Figure 8 and Figure 9 It can be seen that the change of dosing time has little effect on the green lysosomal fluorescent dye LT-Green, but it greatly changes the staining site of LT-Red. As the dosing time was extended to 4h, LT-Red was transferred to the cytoplasm and nuclear membrane other than the lysosome, and even at 8h, some dyes had entered the nucleus. It can be seen that, compared with LT-Red, the SiNDs synthesized in the present invention are more suitable for long-term tracking of lysosome imaging.

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Abstract

The invention discloses fluorescent SiNDs (silicon nanodots). The fluorescent SiNDs are prepared from silane and Bengal rose with a hydrothermal method through one step. Compared with the prior art, the prepared SiNDs have ultrahigh fluorescent quantum yield (100%) and can realize long-time specific imaging of mammal cytolysosome. Besides, the imaging effect of lysosome of the SiNDs is not affected by cell cleaning, immobilization, permeabilization and the like and the fluorescent SiNDs have the advantages of cleaning resistance, immobilization resistance and permeabilization resistance. Meanwhile, the SiNDs also have the advantages of low preparation cost, simple synthesis method, good water dispersity, narrow fluorescent emission peak width, good photostability, good cytocompatibility, low photocytotoxicity and the like and are expected to become novel lysosome fluorescent probes.

Description

technical field [0001] The invention belongs to the field of nano-materials and biotechnology, and in particular relates to a fluorescent silicon nano-dot and its preparation method and application. Background technique [0002] Fluorescent materials are widely used in biomedicine. Compared with traditional organic fluorescent small molecules, fluorescent nanomaterials have attracted extensive attention in recent years because of their advantages such as better photostability and tunable excitation and emission wavelengths. At present, there are many types of fluorescent nanomaterials, but they all have certain shortcomings. For example, traditional semiconductor quantum dots (such as cadmium selenide, lead sulfide, etc.) generally contain toxic heavy metal elements, so they are widely used in biomedicine. The use dose is greatly limited; although the biocompatibility of noble metal nanoclusters (such as gold nanoclusters, silver nanoclusters) and upconversion nanoparticles...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C09K11/59B82Y20/00B82Y40/00G01N21/64
CPCB82Y20/00B82Y40/00C09K11/59G01N21/6458
Inventor 吴富根陈晓凯张晓东
Owner SOUTHEAST UNIV
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