1：Experimental Design and Method Selection:

The probe was synthesized via Knoevenagel condensation and Click reaction. Spectroscopic measurements (UV-Vis and fluorescence) were conducted in CH3CN/H2O (5:1 v/v) to study metal ion binding. Cell imaging used confocal microscopy.

2：Sample Selection and Data Sources:

Hela cells were cultured in DMEM with FBS. Metal ion solutions (perchlorate and nitrate salts) were prepared in CH3CN or deionized water.

3：List of Experimental Equipment and Materials:

NMR spectrometers (Bruker AVANCE III 400 or 500), mass spectrometer (AB SCIEX TripleTOF 4600), UV-Vis spectrophotometer (Lambda 750, PerkinElmer), spectrofluorometer (F-7000, Hitachi), confocal microscope (Zeiss LSM 710), silica gel (Qingdao Haiyang 300-400 mesh), Bio-Beads S-X1 beads, solvents (CH3CN, H2O, THF, CH2Cl2, etc.), reagents (piperidine, acetic acid, Mg(ClO4)2, CuSO4·5H2O, sodium ascorbate, metal salts).

4：Experimental Procedures and Operational Workflow:

Synthesis of probe via multi-step reactions, purification by chromatography. Spectroscopic titrations with metal ions in CH3CN/H2O. Cell culture, treatment with probe and metal ions, incubation, rinsing, and confocal imaging with excitation at 630 nm and emissions at 640-700 nm and 701-758 nm.

5：2O. Cell culture, treatment with probe and metal ions, incubation, rinsing, and confocal imaging with excitation at 630 nm and emissions at 640-700 nm and 701-758 nm. Data Analysis Methods:

5. Data Analysis Methods: Binding constants determined using 1:2 host-guest complex equation with linear least-squares fitting. Detection limits calculated as 3σ/S. Fluorescence intensity ratios (I673/I730, I640-700/I701-758) used for ratiometric analysis.