研究目的
Developing a rapid and simple methodology for the biomolecular analysis of single cells and microenvironments using a stick-and-peel plasmonic sensing platform.
研究成果
The developed stick-and-peel plasmonic sensing platform provides a rapid, cost-effective, and label-free methodology for the biomolecular analysis of single cells and microenvironments. It enables the quantification and mapping of local target concentrations with high spatial resolution, offering new opportunities in clinical care such as on-site molecular pathology.
研究不足
The study is limited by the potential evaporation of the small volume of lysis buffer, which could concentrate the cell contents and affect the accuracy of ATP concentration measurements. Additionally, the methodology may require optimization for different cell types and biomolecules.
1：Experimental Design and Method Selection:
The study employed a stick-and-peel plasmonic sensing platform for the detection of ATP in single cells and microenvironments. The platform uses substrate-bound assemblies of plasmonic gold nanoparticles linked by reconfigurable oligonucleotides that disassemble upon target binding.
2：Sample Selection and Data Sources:
Two ovarian cancer cell lines, SKOV3.ip1 and HEY, were used to quantify intracellular ATP levels and elucidate differences and cellular distribution.
3：ip1 and HEY, were used to quantify intracellular ATP levels and elucidate differences and cellular distribution.
List of Experimental Equipment and Materials:
3. List of Experimental Equipment and Materials: Colloidal gold nanoparticle solutions, ATP-aptamer linked nanoparticle assemblies, lysis buffer, and darkfield microscopy were used.
4：Experimental Procedures and Operational Workflow:
The nanoparticle sensors were anchored on an elastomeric substrate and sandwiched with cells cultured on an adherent cover slip in the presence of microlitres of lysis buffer. Changes in the light scattering intensity were measured.
5：Data Analysis Methods:
The scattering intensity from individual nanostructures was used for the quantification and mapping of ATP levels. Image interpolation methods were applied to generate concentration maps.
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