1：Experimental Design and Method Selection:

The probe was synthesized via condensation of substituted formyl-1H-pyrrole and rhodamine 6G hydrazone under mild conditions. FRET was utilized for ratiometric sensing.

2：Sample Selection and Data Sources:

HeLa cells were used for bioimaging. Metal ion solutions were prepared from nitrate or chloride salts.

3：List of Experimental Equipment and Materials:

Instruments included Elemental Vario EL analyzer, Bruker AV400 NMR spectrometer, Purkinje General TU-1800 spectrophotometer, Varian CARY Eclipse spectrophotometer, Bruker SMART APEX II CCD diffractometer, Quantaurus Tau C11367 for fluorescence lifetime and quantum yield, Zeiss Leica inverted epifluorescence/reflectance laser scanning confocal microscope. Materials included solvents, starting materials, and salts.

4：Experimental Procedures and Operational Workflow:

Synthesis involved refluxing with acetic acid. UV-vis and fluorescence titrations were performed by adding aliquots of reagents to probe solutions. Bioimaging involved incubating HeLa cells with probe and Cu2+, followed by confocal microscopy.

5：Data Analysis Methods:

Spectra were analyzed for absorbance and emission changes. Binding constant was determined using Benesi-Hildebrand expression. DFT calculations were performed with Gaussian 09.