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The main focus of this chapter is the theoretical and instrumental processes that underpin densitometric methods widely used in thin-layer chromatography (TLC). Densitometric methods include UV–vis, luminescence and fluorescence optical measurements as well as infrared and Raman spectroscopic measurements. The chapter is divided in two general parts: a theoretical part and a practical part. The systems for direct radioactivity measurements and the combination of TLC with mass spectrometry are also discussed. All these systems allow measuring an intensity distribution directly on a TLC plate. We call this “in situ detection” because no analyte is removed from the plate.