Laser desorption followed by jet-cooling allows wavelength-selective as well as mass-selective detection of molecules desorbed from a surface without fragmentation. The cooling characteristics and detection sensitivity of laser desorption jet-cooling of organic molecules are investigated. From the rotational contour of the electronic origin of the S1 ← S0 transition of laser-desorbed anthracene, rotational cooling to 5-10 K is demonstrated. Vibrational cooling is studied for laser-desorbed diphenylamine, a molecule with low-energy vibrations, and a vibrational temperature below 15 K is found. The absolute detection sensitivity is determined for the perylene molecule. Using two-color (1+1) resonance enhanced multi-photon ionization (with a measured ionization efficiency of 0.25) for detection, it is found that one ion is produced in the detection region for every 2×105 perylene molecules evaporated from the desorption laser spot. A two-color (1+1) REMPI spectrum (400 points) of perylene is recorded using only 30 picogram of material. © 1990 Springer-Verlag.