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A preparation method of nano fluorescent probe and its application in detecting the activation status of bv2 cells

A nano-fluorescence probe, activated state technology, applied in nanotechnology, nanotechnology, fluorescence/phosphorescence, etc., can solve problems such as toxicity, and achieve the effects of easy operation, good biocompatibility, and easy promotion

Active Publication Date: 2022-06-14
LIAONING UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Furthermore, neurostimulants have toxic effects on dopamine neurons in the presence of BV2 cells

Method used

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  • A preparation method of nano fluorescent probe and its application in detecting the activation status of bv2 cells
  • A preparation method of nano fluorescent probe and its application in detecting the activation status of bv2 cells
  • A preparation method of nano fluorescent probe and its application in detecting the activation status of bv2 cells

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Experimental program
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Embodiment 1

[0027] The preparation of embodiment 1 nanometer fluorescent probe

[0028] Weigh 2g (11.5mmol) of arginine, 0.09g of alanine (1mmol), dissolve in 20mL of ultrapure water, add 1.5g (25mmoL) of urea, after complete ultrasonic dissolution, heat in the reaction kettle at 200°C for 10h , to obtain nano-fluorescent probes.

[0029] The above-mentioned prepared nano fluorescent probes were scanned by electron microscope, and the results were as follows: figure 1 shown, from figure 1 It can be seen that the average particle size of the prepared nano fluorescent probe is 4nm, and the distribution is uniform, and it is uniformly distributed spherical particles.

[0030] figure 2 It is the nano-fluorescent probe sample prepared in Example 1, and the fluorescence spectrum is obtained by using a fluorescence spectrophotometer to perform fluorescence detection at the optimal excitation wavelength. It can be seen from the figure that the optimum excitation wavelength of the nano fluore...

Embodiment 2

[0032] After the nano fluorescent probe prepared in Example 1 was filtered, dialyzed, and freeze-dried, 0.004 g was dissolved in 200 mL of PBS buffer solution, so that the concentration of the nano fluorescent probe solution was 0.02 mg / mL, and each well of the 96-well plate 100 μL of cell suspension was added to the well, and the density of the cell suspension was 10,000 cells / mL, and 100 μL of nano fluorescent probe solution was added to each well, and incubated at 20°C for 5 min.

[0033] Detect the fluorescence intensity value on a fluorescent microplate reader, repeat 8 times, such as image 3 shown. The fluorescence intensity of the nano fluorescent probe under the microplate reader is 2.2×10 7 . Normal BV2 cells are not activated, and the concentration of specific components of membrane proteins and cytokines is low. According to the principle of non-specific adsorption, the combination of nano-fluorescent probes with biothiols and polypeptides produces a slight fluor...

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Abstract

The invention belongs to the technical field of fluorescent nanomaterials, and specifically provides a method for preparing a nanofluorescent probe, comprising the following steps: completely dissolving a carbon source in water, adding a small molecule ligand to the solution for doping; ultrasonically dissolving, and heating obtain the target product. The carbon source is one or more of natural amino acids. The small molecule ligand is urea or ammonia water. In molar ratio, carbon source: small molecule ligand = 1:1‑5. The nano fluorescent probe prepared in the invention is a spherical nano particle with a fluorescent performance, which uses carbon atoms as a skeleton structure. The nano fluorescent probe can not only identify BV2 cells in different activation states, but also has the characteristics of good biocompatibility, safety, non-toxicity, stable effect and the like. It mainly specifically recognizes BV2 cell membrane proteins and cytokines; its preparation method is simple in process, easy to operate, low in preparation cost and easy to popularize.

Description

technical field [0001] The invention belongs to the technical field of fluorescent nanometer materials, and in particular relates to a preparation method of a nanometer fluorescent probe and its application in detecting the activation state of BV2 cells. Background technique [0002] Microglia (BV2) participate in a series of neurodegenerative diseases, and its activation and neuroinflammation are the main features of neuropathology. It induces endogenous immune responses to central nervous system injury and disease, thereby exerting neuroprotective or neurotoxic effects. Any neurological disorder typically results in inflammation and activation of microglial cells with increased numbers and altered phenotypes of glial cells, a phenomenon termed "reactive gliosis". In acute neurodegenerative diseases (stroke, cerebral hypoxia, traumatic brain injury), the phenotype of microglia changes and releases inflammatory mediators, mainly cytokines and chemokines. These acute inflam...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C09K11/65C01B21/082B82Y30/00B82Y40/00G01N21/64
CPCC09K11/65C09K11/0883C01B21/0605B82Y30/00B82Y40/00G01N21/6428C01P2004/04
Inventor 刘学毕秀丽刘禹霖陈扬
Owner LIAONING UNIVERSITY
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