研究目的
To develop sulfur-rich carbon dots as a fluorescent imaging probe for distinguishing pathological changes in mouse bone cells, specifically for detecting and distinguishing bone-related diseased cells from normal cells.
研究成果
Sulfur-rich carbon dots synthesized via one-pot hydrothermal method exhibit small size, high sulfur content, tunable fluorescence, low cytotoxicity, and good stability. They effectively distinguish bone-related diseased cells from normal cells through differential fluorescence emission, offering a novel non-invasive imaging probe for bone disease detection with potential for future biomedical applications.
研究不足
The study is limited to in vitro experiments with specific cell types (HeLa and mouse bone cells); in vivo applications and broader biological contexts are not explored. The synthesis method may require optimization for scalability, and the specificity of C-dots for other cell types or diseases is not investigated.
1:Experimental Design and Method Selection:
The study uses a one-pot hydrothermal method to synthesize sulfur-doped carbon dots (C-dots) from methionine and ethylene diamine precursors. Various characterization techniques (XRD, TEM, XPS, FT-IR, UV-vis absorption, fluorescence spectra) are employed to analyze structure and properties. Biological experiments involve cytotoxicity assessment and cell imaging.
2:Sample Selection and Data Sources:
Precursors include methionine and ethylene diamine. Biological samples include HeLa cells and mouse bone-related normal and diseased cells.
3:List of Experimental Equipment and Materials:
Equipment includes Bruker diffractometer (XRD), VG Thermo ESCALAB 250 spectrometer (XPS), JEOL 1200X TEM, Varian Cary 50 UV-vis spectrophotometer, Horiba Jobin Yvon Fluorolog-3 fluorescence spectrophotometer, Zetasizer Nano ZS System (zeta potential), Perkin-Elmer 100 FTIR spectrometer, confocal microscopy with 405 and 488 nm lasers. Materials include deionized water, methionine, ethylene diamine, dialysis membrane (MWCO = 3000 Da), sodium hydroxide, Tricaine, and embryo media.
4:Experimental Procedures and Operational Workflow:
Synthesis involves dissolving precursors in water, hydrothermal treatment at 250°C for 10 h, drying, dialysis, and neutralization. Characterization involves XRD, TEM, XPS, FT-IR, UV-vis, fluorescence measurements. Cytotoxicity is assessed using HeLa cells incubated with C-dots and viability monitored. Cell imaging is performed using confocal microscopy on HeLa and mouse bone cells incubated with C-dots.
5:Data Analysis Methods:
Data analysis includes particle size calculation from TEM, elemental analysis from XPS and EDS, spectral analysis for fluorescence properties, and statistical analysis for cell viability.
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JEOL 1200X TEM
1200X
JEOL
To perform transmission electron microscopy (TEM) on the sulfur C-dots for microstructural analysis.
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Perkin-Elmer 100 FTIR spectrometer
100
Perkin-Elmer
To record Fourier transform infrared (FTIR) spectra of the sulfur C-dots.
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Modulab 2015 water purification system
2015
San Antonio, TX
To obtain purified water with specific surface tension and resistivity for use in the synthesis of sulfur-doped carbon dots.
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Bruker diffractometer
Bruker
To record X-ray diffraction (XRD) patterns of the sulfur C-dots using Cu Kα radiation.
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VG Thermo ESCALAB 250 spectrometer
ESCALAB 250
VG Scientific
To perform X-ray photoelectron spectroscopy (XPS) on the sulfur C-dots.
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Varian Cary 50 UV–vis spectrophotometer
Cary 50
Varian
To record ultraviolet-visible (UV–vis) absorption spectra of the sulfur C-dots.
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Horiba Jobin Yvon Fluorolog-3 fluorescence spectrophotometer
Fluorolog-3
Horiba Jobin Yvon
To record fluorescence spectra of the sulfur C-dots and capture digital photographs for CIE chromaticity analysis.
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Zetasizer Nano ZS System
Nano ZS
Malvern, Inc.
To analyze the zeta potential of sulfur C-dots in aqueous solution using a 633 nm laser source.
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Confocal microscopy
To image sulfur C-dots incubated with cells using 405 nm and 488 nm lasers for fluorescence detection.
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Dialysis membrane
MWCO = 3000 Da
To purify the synthesized sulfur C-dots by dialysis.
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