1：Experimental Design and Method Selection:

The study involved synthesizing a rhodamine-deoxylactam-based fluorescent probe (dRB-OPD) with o-phenylenediamino as the recognition group for NO detection. Fluorescence and UV-vis spectroscopy were used to characterize the probe's response.

2：Sample Selection and Data Sources:

The probe was tested with various concentrations of NO and other potential interferents in phosphate-buffered saline (PBS) solution. Cell imaging was performed using HepG2 and RAW

3：7 cells. List of Experimental Equipment and Materials:

2 Equipment included a Bruker Avance II 400 spectrometer for NMR, Bruker Autoflex MALDI-TOF mass spectrometer for MS, Perkin-Elmer 2400 elemental analyzer, Hitachi F-4600 spectrophotometer for fluorescence, Agilent CARY 60 UV-Vis spectrophotometer, Mettler-Toledo Delta 320 pH meter, and Nikon A1 plus confocal microscope. Materials included reagents from Sigma-Aldrich and Aladdin.

4：Experimental Procedures and Operational Workflow:

The probe was synthesized via chemical reactions, purified, and characterized. Fluorescence measurements were conducted with excitation at 550 nm and emission at 560-640 nm. Cell cultures were incubated and imaged under specific conditions.

5：Data Analysis Methods:

Data were analyzed using fluorescence intensity measurements, NMR and MS for structural confirmation, and statistical methods for detection limits and selectivity.