1：Experimental Design and Method Selection:

The study involved the design and synthesis of a deep-red fluorogenic probe (BT-NH) for NO detection, utilizing a benzo-bis(1,2,5-thiadiazol) fluorophore. The probe's sensitivity and selectivity to NO were evaluated through absorption and fluorescence spectra.

2：Sample Selection and Data Sources:

HepG2, SH-SY5Y, and 293T cells were used for in vitro studies, and Parkin null Drosophila were used as a PD model for in vivo studies.

3：List of Experimental Equipment and Materials:

Instruments included a Bruker Ultra Shield Plus for NMR, Synergy HTX microplate reader for absorption spectra, and Cytation 5 microplate reader for photoluminescence spectra. Confocal laser scanning microscopy (CLSM) was used for imaging.

4：Experimental Procedures and Operational Workflow:

Cells were treated with NO donors or inhibitors, incubated with BT-NH, and then imaged. Drosophila brains were dissected, incubated with BT-NH, and imaged.

5：Data Analysis Methods:

Fluorescence intensity was quantified to assess NO levels, with statistical analysis performed to determine sensitivity and selectivity.