Fluorescence probe for detecting lipid droplets and production method and application thereof

A fluorescent probe and lipid droplet technology, applied in the field of analytical chemistry, can solve the problems of poor photostability, easy to be bleached, and unstable imaging process, etc., and achieve the effects of good photostability, easy purification, and fast response speed

Inactive Publication Date: 2019-08-23
UNIV OF JINAN
View PDF5 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, Nile Red, as the first commercialized fluorescent dye, is unstable and easily bleached during imaging.
[0003] Most of the existing lipid droplet probes are small molecule fluorescent probes, which are not photostable and easy to be bleached, while there are relatively few reports on polymer-based lipid droplet probes, especially biocompatible polymers. There are fewer reports on lipid droplet probes as substrates

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Fluorescence probe for detecting lipid droplets and production method and application thereof
  • Fluorescence probe for detecting lipid droplets and production method and application thereof
  • Fluorescence probe for detecting lipid droplets and production method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Synthesis of Fluorescent Probe PEG-NBD

[0026]

[0027] In a 25 mL eggplant-shaped flask, add 0.2 g PEG-200 and 10 mL dichloromethane, then add 0.5 g 4-chloro-7-nitrobenzo-2-oxa-1,3-oxadiazole (NBD- Cl), finally add a few drops of triethylamine (TEA), and react at room temperature for 24 hours under the protection of nitrogen. The obtained crude product was rotary evaporated, and petroleum ether / dichloro (V / V) = 2:1 was used as the eluent, and the product PEG-NBD was purified through a silica gel column. That 1 H NMR spectrum such as figure 1 .

Embodiment 2

[0028] Embodiment 2 The selectivity of fluorescent probe PEG-NBD

[0029] Prepare 5 mL of PBS (pH=7.4) aqueous solution of amino acids, metal ions, and active oxygen at a concentration of 100 mM and the fluorescent probe PEG-NBD mother solution obtained in Example 1 at a concentration of 1 mM for future use.

[0030] Add 20 μL probe mother solution, 200 μL DMSO and 100 equivalents of amino acids, ions, thiols and reactive oxygen species, dilute to 2 mL with phosphate buffer solution PBS, shake well and perform fluorescence detection (λ ex = 470 nm), establish a histogram of fluorescence intensity and each ion (amino acid or active oxygen), the results are as follows figure 2 , 1-28 added detection substances were PBS solution, arginine, serine, alanine, aspartic acid, glutamic acid, histidine, isoleucine, threonine, glutamine, N -Acetyl-L-cysteine, N-acetylglycine, aluminum chloride, calcium chloride, copper chloride, ferrous sulfate, potassium iodide, magnesium chloride, so...

Embodiment 3

[0031] Example 3 Fluorescence spectra of fluorescent probe PEG-NBD in different polar solvents

[0032] Choose eight solvents with different polarities, including water, methanol, acetonitrile, dimethyl sulfoxide, dichloromethane, tetrahydrofuran, dioxane and toluene; prepare 5 mL of the fluorescent probe obtained in Example 1 with a concentration of 1 mM The PEG-NBD mother solution was used as a backup. The final concentration of the prepared probe was 5 µg / mL, which were added to solvents of different polarities, and the fluorescence detection (λ ex = 460 nm), and its fluorescence spectrum is shown in image 3 shown. Depend on image 3 It can be seen that the fluorescence intensity varies greatly depending on the polarity of the solution, and the greater the polarity, the lower the fluorescence, indicating that the fluorescent probe PEG-NBD has a polar response.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a fluorescence probe for detecting lipid droplets in biological cells. A chemical structural formula of the fluorescence probe for detecting the lipid droplets in the biologicalcells is as shown in the description. The fluorescence probe is a polyethylene glycol fluorescence probe based on biocompatibility, and can well position the lipid droplets in the cells, the responsespeed is high, and the anti-interference capability is strong. The fluorescence probe is produced from polyethylene glycol and 4-chloro-7-nitro-1,2,3-benzoxadiazole through reaction at room temperature, synthesis steps of the probe are simple, and purification is easy.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry, and in particular relates to a polyethylene glycol-based fluorescent probe for detecting intracellular lipid droplets and an application thereof. Background technique [0002] Lipid droplets are spherical organelles whose core is composed of neutral lipids, including triglycerides and cholesteryl esters. The main function of lipid droplets is to regulate the energy dynamic balance of cells. In white adipose tissue, lipid droplets mainly store triglyceride and store energy; in brown adipose tissue, lipid droplets are closely connected with mitochondria to decompose triglyceride and provide cells and bio-organism energy. Lipid droplets are the most hydrophobic organelles in cells and widely exist in animal, plant, bacterial, and yeast cells; their sizes also vary greatly, ranging from 40 nm to 100 μm in diameter. Studies have shown that lipid droplet is not just an energy storage, bu...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C07D271/12C09K11/06G01N21/64C08G65/333
CPCC07D271/12C08G65/33396C08G2650/04C08G2650/50C09K11/06C09K2211/1048C09K2211/1475G01N21/6428G01N21/6486
Inventor 林伟英于法祺景新颖
Owner UNIV OF JINAN
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap