研究目的
To develop a highly selective and specific fluorescent probe for imaging lipid droplets in liver cells, enabling distinction between normal and cancer cells based on lipid droplet differences.
研究成果
TPA-SD is an effective probe for specific imaging of lipid droplets, showing no cytotoxicity and enabling clear distinction between normal and cancer liver cells based on lipid droplet abundance. It holds promise for diagnostic applications in lipid-related diseases.
研究不足
The probe's specificity and performance were tested only in selected cell lines; further validation in vivo or with other cell types may be needed. Potential interference in complex biological environments was not fully explored.
1:Experimental Design and Method Selection:
The study involved designing a fluorescent probe TPA-SD with AIE and ESIPT properties for specific imaging of lipid droplets. Methods included synthesis, spectroscopic characterization, and cellular imaging.
2:Sample Selection and Data Sources:
Human cell lines (HeLa, L02, HepG2) were used. Lipid droplets were induced using oleic acid.
3:List of Experimental Equipment and Materials:
Equipment included NMR spectrometer, mass spectrometer, spectrophotometer, fluorescence spectrometer, confocal microscope, and MTT assay kit. Materials included chemical reagents for synthesis and cell culture.
4:Experimental Procedures and Operational Workflow:
Synthesis of TPA-SD, characterization of optical properties, cytotoxicity testing, and confocal microscopy imaging of cells stained with the probe.
5:Data Analysis Methods:
Fluorescence intensity measurements, statistical analysis of lipid droplet numbers, and Pearson's correlation coefficient for colocalization.
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NMR Spectrometer
Bruker Advance III
Bruker
Recording NMR spectra for chemical characterization
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Mass Spectrometer
Bruker Apex IV FTMS
Bruker
Measuring mass spectra using electrospray ionization
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Fluorescence Spectrometer
Hitachi F-7000
Hitachi
Measuring fluorescence spectra
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Confocal Fluorescence Microscope
Olympus IX81
Olympus
Fluorescence imaging of cells
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Spectrophotometer
Purkinje TU-1901
Purkinje
Recording absorption spectra
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pH Meter
Mettler Toledo FE-30
Mettler Toledo
pH measurements
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Photonic Multi-Channel Analyzer
PMA-12
Measuring absolute fluorescence quantum yield
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ELISA Reader
Bio-Rad 680
Bio-Rad
Measuring absorbance for MTT assay
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BODIPY 493/503
Commercially available marker for lipid droplet imaging
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