研究目的
Investigating the out-of-plane nanoscale reorganization of lipid molecules and nanoparticles using plasmonic spectroscopy.
研究成果
The study provides insights into the nanoscopic organization of lipid bilayers and their interactions with gold nanoparticles, revealing dynamic topographical information and sub-nanometer out-of-plane motion. The findings suggest that the HBL system with NPoM geometry is suitable for studying lipid motion and diffusion, though alternative models may be needed for more complex membrane components like proteins.
研究不足
The HBLs are supported on a solid surface, which may affect the mobility of the lipid layer and embedded components. The system's complexity increases with the introduction of large transmembrane proteins. The study suggests further development to include unsupported lipid membranes for more accurate modeling of cell membranes.
1：Experimental Design and Method Selection:
The study uses nanoparticle-on-mirror (NPoM) cavities to assemble hybrid lipid bilayers (HBLs) between a flat gold surface and gold nanoparticles (AuNPs). The lipid molecules are assembled in a flow-cell system, with the lower leaflet consisting of octadecanethiol (ODT) and the upper leaflet formed by lipid vesicles containing POPC and DOTAP.
2：Sample Selection and Data Sources:
The samples include HBLs assembled on template-stripped gold substrates with AuNPs. Data is collected using dark-field microscopy and spectroscopy.
3：List of Experimental Equipment and Materials:
Equipment includes a custom-built flow-cell, darkfield microscope with a 100× objective, QE65000 Ocean Optics spectrometer, and atomic force microscope (AFM). Materials include ODT, POPC, DOTAP lipids, and 100 nm diameter AuNPs.
4：Experimental Procedures and Operational Workflow:
The HBLs are prepared and incubated with AuNPs, then imaged using dark-field microscopy to track nanoparticle diffusion and spectroscopic changes. AFM is used to verify the lipid layer's presence and integrity.
5：Data Analysis Methods:
Single particle tracking is performed using FIJI software and TrackMate plugin to analyze diffusion dynamics. Dark-field spectra are analyzed to determine gap sizes and nanoparticle positions.
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