研究目的
Investigating the effect of surface coverage on the cellular interaction mechanisms of ultrasmall luminescent gold nanoparticles, including membrane-binding, uptake, and their use as optical probes for imaging.
研究成果
Surface coverage is a crucial factor in regulating the cellular interaction of ultrasmall luminescent AuNPs, with lower coverage favoring membrane-binding and higher coverage favoring uptake. The pH-responsive dual emissions enable effective subcellular imaging and tracking. This provides a strategy for designing tunable nanoparticles for biomedical applications.
研究不足
The study is limited to HeLa cells and specific AuNP formulations; generalizability to other cell types or nanoparticles may require further investigation. The experimental conditions (e.g., pH range, incubation times) might not cover all biological scenarios. Potential optimizations include exploring a wider range of surface coverages and additional functional ligands.
1：Experimental Design and Method Selection:
Designed and synthesized ultrasmall luminescent gold nanoparticles (AuNPs) with different surface coverages (29% to 47%) by co-introducing glutathione and a cell-penetrating peptide (CR8). Used these AuNPs as fluorescent probes to study their interaction with living cells, including membrane-binding, uptake, and subcellular imaging. Methods included synthesis, characterization (e.g., emission spectra, zeta potential), cell culture, fluorescent imaging, and inhibitor studies.
2：8). Used these AuNPs as fluorescent probes to study their interaction with living cells, including membrane-binding, uptake, and subcellular imaging. Methods included synthesis, characterization (e.g., emission spectra, zeta potential), cell culture, fluorescent imaging, and inhibitor studies.
Sample Selection and Data Sources:
2. Sample Selection and Data Sources: Used HeLa cells as the model cell line. AuNPs were synthesized with varying ligand densities (07CR-AuNPs, 09CR-AuNPs, 13CR-AuNPs). Data were acquired through spectroscopic measurements and microscopic imaging.
3：List of Experimental Equipment and Materials:
Equipment included spectrometers for emission and absorption spectra, zeta potential analyzer, gel electrophoresis apparatus, fluorescent microscope for imaging. Materials included gold nanoparticles, glutathione, CR8 peptide, cell culture reagents, endocytic inhibitors (chlorpromazine, methyl-β-cyclodextrin, amiloride, sodium azide), and Lysotracker Green dye.
4：Experimental Procedures and Operational Workflow:
Synthesized AuNPs with different surface coverages. Characterized their physicochemical properties. Incubated cells with AuNPs at various pH levels and times. Performed fluorescent imaging to monitor membrane-binding and uptake. Used inhibitors to study endocytosis pathways. Conducted 3D imaging and colocalization studies with Lysotracker Green.
5：Data Analysis Methods:
Analyzed luminescence intensities and ratios (e.g., I615 nm/I810 nm) from images. Used statistical methods to quantify membrane binding and uptake percentages. Colocalization analysis was performed to determine lysosomal involvement.
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