Determining the active site in real-life solid catalysts remains an intellectual challenge and is crucial for exploring the road towards their rational design. In recent years various micro-spectroscopic methods have revealed valuable structure-activity data at the level of a single catalyst particle, even under reaction conditions. Herein, we introduce Tip-Enhanced FLuorescence (TEFL) microscopy as a novel and versatile characterization tool for catalysis research. This has been achieved using a Fluid Catalytic Cracking (FCC) catalyst as showcase material. Thin sectioning of industrially used FCC particles together with selective staining of Brønsted acidity has enabled high-resolution TEFL mapping of different catalyst regions. Hyperspectral information gained via TEFL microscopy reveals a spatial distribution of Brønsted acidity within individual zeolite domains in different regions of the FCC catalyst particle. Comparison of TEFL measurements from different FCC particles showed significant intra- and inter-particle heterogeneities both in zeolite domain size and chemical reactivity.