研究目的
To design and synthesize a bioorthogonally applicable, tetrazine functionalized rhodaphenothiazine probe for light-assisted photooxidation-based fluorogenic labeling schemes, suitable for STED super-resolution microscopy.
研究成果
The developed tetrazine-rhodaphenothiazine probe exhibits a novel photooxidation-based fluorogenic mechanism, enabling selective activation of specifically conjugated probes with minimal background fluorescence. It is suitable for STED super-resolution microscopy, offering potential for advanced imaging applications.
研究不足
The study is limited to in vitro and fixed cell imaging. The applicability in live cells and tissues, as well as the potential phototoxicity of the photooxidation process, were not explored.
1：Experimental Design and Method Selection:
The study involved the synthesis of a photoactivatable fluorogenic tetrazine-rhodaphenothiazine probe and its evaluation in light-assisted, bioorthogonal labeling schemes. The probe's ability to sensitize 1O2 generation upon illumination and undergo self-oxidation to a fluorescent sulfoxide product was examined.
2：Sample Selection and Data Sources:
The probes were tested in solution and in fixed COS7 cells tagged with cyclooctynylated phalloidin for actin filament labeling.
3：List of Experimental Equipment and Materials:
A confocal microscope with a green laser (552 nm) and a STED microscope with a 660 nm depletion laser were used for imaging.
4：Experimental Procedures and Operational Workflow:
The probes were illuminated with green or orange LEDs to study photooxidation. The specifically conjugated probes were selectively photooxidized and imaged under a confocal microscope.
5：Data Analysis Methods:
LC-MS and emission spectra were used to monitor the formation of oxidized products and their spectral characteristics.
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