研究目的
Developing an efficient method for cancer cell labeling using silica fluorescent dye-doped nanoparticles and second antibody, focusing on the recognition of MUC1 protein in human breast carcinoma MCF-7 cells.
研究成果
The developed RBITC-doped silica nanoparticles conjugated with goat anti-rabbit IgG serve as effective, photostable probes for cancer cell recognition, offering advantages over traditional dye labeling methods. This approach can be adapted for various biomolecules and pathogens by changing the antibody.
研究不足
The study focuses on MCF-7 cells and MUC1 protein recognition, potentially limiting its applicability to other cell types or biomarkers. The photostability comparison is with a specific dye, which may not represent all organic dyes.
1:Experimental Design and Method Selection:
The study employed the reverse microemulsion method for synthesizing dye-doped silica nanoparticles and the EDC/sulfo-NHS method for antibody conjugation.
2:Sample Selection and Data Sources:
Human breast carcinoma MCF-7 cells were used, cultured in DMEM medium.
3:List of Experimental Equipment and Materials:
Included tetraethyl orthosilicate (TEOS), TritonX-100, cyclohexane, n-hexanol, ammonium hydroxide, rhodamine B isothiocynate (RBITC), 3-aminopropyltriethoxysilane (APTES), and others.
4:Experimental Procedures and Operational Workflow:
Detailed steps included nanoparticle synthesis, PEG introduction, antibody immobilization, and cell labeling.
5:Data Analysis Methods:
Fluorescence intensity was measured using a spectrofluorometer, and imaging was done with a fluorescence microscope.
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Transmission electron microscope
JEM100CXII
JEOL
Used to characterize the size and uniformity of nanoparticles.
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Spectrofluorometer
F-4600
Hitachi
Used to record excitation and emission spectra.
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Zetasizer
Nano-ZS
Malvern Instruments
Used for dynamic light scattering and ζ-potential measurements.
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Tetraethyl orthosilicate
TEOS
Aldrich Chemical Co. Inc
Used in the synthesis of silica nanoparticles.
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TritonX-100
Aldrich Chemical Co. Inc
Used in the reverse microemulsion method for nanoparticle synthesis.
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Rhodamine B isothiocynate
RBITC
Aldrich Chemical Co. Inc
Fluorescent dye used in the synthesis of dye-doped silica nanoparticles.
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3-aminopropyltriethoxysilane
APTES
Aldrich Chemical Co. Inc
Used to form a conjugate with RBITC for nanoparticle synthesis.
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Fluorescence microscope
ECLIPSE TE2000-U
Nikon
Used for fluorescence imaging of labeled cells.
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IR spectrum
Nicolet-5700
USA
Used to confirm the modified carboxyl on the surface of nanoparticles.
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