1：Experimental Design and Method Selection:

The study developed tapping AFM-IR, a hybrid technique combining atomic force microscopy (AFM) and infrared (IR) spectroscopy, to characterize soft polymeric nanoparticles. It involved theoretical modeling of the tapping mode interactions and comparison with contact mode.

2：Sample Selection and Data Sources:

Poly(lactic acid) (PLA) and poly(D,L-lactic-co-glycolic acid) (PLGA) nanoparticles were synthesized using nanoemulsion and nanoprecipitation methods, with and without the drug pipemidic acid (PIP). Samples were characterized using nanoparticle tracking analysis (NTA) and dynamic light scattering (DLS).

3：List of Experimental Equipment and Materials:

Equipment included an AFM-IR system (NanoIR2s, Anasys Instruments), QCL IR source (MIRcat, Daylight Solutions), FTIR spectrometer (Bruker Vertex 70), sonicator (Sonopuls HD 2070, Bandelin), NTA system (NanoSight LM10, Malvern), and DLS system (Zetasizer Nano ZS90, Malvern). Materials included PLGA, PLA, PVA, PIP, solvents like DCM, MeOH, acetone, and injectable water.

4：Experimental Procedures and Operational Workflow:

Nanoparticles were synthesized, diluted, deposited on Au-coated substrates, and dried. AFM-IR measurements were performed in tapping mode with specific IR wavenumbers for chemical mapping. Spectra and images were processed using MountainsMap software and Savitzky-Golay filtering.

5：Data Analysis Methods:

Data were analyzed to obtain topography, IR maps, and spectra, with comparisons to FTIR references. Statistical analysis included mean diameter calculations with standard deviations.