In vivo plaque recognition may be important for safe and precise intra-arterial atheroma ablation during laser coronary angioplasty. This study examined the feasibility and sensitivity of utilizing quantitative fluorescence spectroscopy and video-enhanced fluorescence imaging for plaque identification in atherosclerotic human necropsy arterial wall before and after laser atheroma ablation. With wide-band (450 to 490 nm) blue light excitation, the 540 nm fluorescence intensity ratio of normal to diseased sites (n = 13) was 2.09 +/- 0.82 (p less than 0.001) and video fluorescence imaging provided enhanced delineation of atheroma surface characteristics. Continuous argon and pulsed excimer (308 nm) laser ablation of atheroma decreased fluorescence intensity ratios by 42 and 20% (p less than 0.001), respectively (that is, from abnormal to nearly normal). Low power 325 nm laser-excited fluorescence spectroscopy from normal (n = 115) and abnormal (n = 146) necropsy sites revealed an average 45% decrease in atheroma fluorescence intensity (p less than 0.0001) and changes in fluorescence spectra appearance that corresponded to plaque morphologic subtypes. Studies using a dual laser system combining 325 nm laser-excited fluorescence plaque recognition and a 480 nm pulsed dye laser for tissue ablation with common optical fibers demonstrated normalization of both fluorescence intensity and spectra appearance after laser atheroma ablation. Thus, in vitro analysis of surface arterial fluorescence by quantitative spectroscopy and video fluorescence imaging reliably differentiate plaque from normal tissue and may provide the feedback signal needed to activate a laser source for selective plaque removal.