1：Experimental Design and Method Selection:

The study involved synthesizing a pyrene-based Schiff base (5-DP) via a single-step condensation reaction and investigating its aggregation-induced emission (AIE) properties in THF/water mixtures. Methods included photophysical studies, fluorescence quenching assays for nitroaromatic explosives, and encapsulation with Pluronic P123 for nanoparticle formation and cell imaging.

2：Sample Selection and Data Sources:

Samples included 5-DP synthesized from pyrene-1-carboxaldehyde and dimethyl 5-aminoisophthalate, nitroaromatic explosives (e.g., TNP, DNP, TNB), and HeLa cells for cytotoxicity and imaging studies. Data were sourced from spectroscopic measurements, microscopy, and biological assays.

3：List of Experimental Equipment and Materials:

Equipment included NMR spectrometers (Bruker Advance (III) 400 MHz), mass spectrometer (Brucker–Daltonics micro TOF–QII), FT-IR spectrometer (Bruker Tensor-27), UV-Vis spectrophotometer (Varian Carry 100 Bio), spectrofluorometer (Horiba Jobin Yvon Fluoromax-4p), DLS instrument (NanoPlus 3), TCSPC system (Horiba Jobin Yvon Fluorocube-01-NL), FE-SEM (Zeiss Supra55), elemental analyzer (Thermo Scientific FLASH 2000), electrochemical workstation (CHI620D), and confocal microscope (Olympus FV1200MPE). Materials included solvents (THF, water, etc.), explosives (Sigma-Aldrich, Loba Chemie), and Pluronic P

4：Experimental Procedures and Operational Workflow:

1 Synthesis of 5-DP via condensation in ethanol; preparation of nanoaggregates in THF/water mixtures; fluorescence studies with varying water fractions; quenching titrations with nitroaromatics; encapsulation with Pluronic P123 for nanoparticle formation; cytotoxicity assessment via MTT assay; cell imaging with confocal microscopy.

5：Data Analysis Methods:

Data analysis included calculation of quantum yields using quinine sulfate as standard, Stern-Volmer plots for quenching constants, lifetime measurements from TCSPC, particle size analysis from DLS, and statistical analysis for cytotoxicity (two-way ANOVA).