研究目的
To develop super blinking and biocompatible nanoprobes based on dye-doped BSA nanoparticles for super-resolution imaging, specifically for single molecule localization microscopy (SMLM).
研究成果
The study successfully developed super blinking and biocompatible nanoprobes based on dye-doped BSA nanoparticles, which exhibited improved blinking properties and localization precision for SMLM applications. These nanoprobes were effectively used for high-resolution imaging of breast cancer cells and exosomes, demonstrating their potential as versatile fluorescent agents in super-resolution microscopy.
研究不足
The relatively large size of the SBNs (~50 nm) may induce a spatial resolution lower than the localization precision, especially when labeling small targets like exosomes. This could potentially be addressed by using smaller scaffolds or combining with expansion microscopy (ExM).
1：Experimental Design and Method Selection:
The study involved the fabrication of super blinking nanoprobes (SBNs) by doping organic dyes (Alexa Fluor 647 and Alexa Fluor 594) into BSA nanoparticles. The blinking properties of these nanoprobes were investigated for their application in SMLM.
2：Sample Selection and Data Sources:
SKBR3 and MCF7 breast cancer cells and exosomes derived from SKBR3 cells were used as model systems to test the application of SBNs in targeted imaging and super-resolution microscopy.
3：List of Experimental Equipment and Materials:
The study utilized a Zeiss Elyra P.1 microscope for SMLM and TIRF imaging, a Shimadzu UV-3600 PC spectrophotometer for extinction spectra, a Shimadzu RF-5301PC spectrofluorimeter for photoluminescence spectra, and a JEM-2100 electron microscope for TEM imaging.
4：Experimental Procedures and Operational Workflow:
The fabrication of SBNs involved doping organic dyes into BSA nanoparticles, followed by characterization of their optical properties, blinking behavior, and application in cell and exosome imaging using SMLM.
5：Data Analysis Methods:
The localization precision of the nanoprobes was analyzed using the Zeiss Zen 2012 software, and the hydrodynamic diameter and zeta potential of nanoparticles were measured by a Malvern Zetasizer Nano ZS90.
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