The use of remote optical feedback systems represents a promising approach for minimally invasive, nerve-sparing laser surgery. Autofluorescence properties can be exploited for a fast, robust identification of nervous tissue. With regard to the crucial step towards clinical application, the impact of laser ablation on optical properties in the vicinity of structures of the head and neck has not been investigated up to now. We acquired 24,298 autofluorescence spectra from 135 tissue samples (nine ex vivo tissue types from 15 bisected pig heads) both before and after ER:YAG laser ablation. Sensitivities, specificities, and area under curve(AUC) values for each tissue pair as well as the confusion matrix were statistically calculated for pre-ablation and post-ablation autofluorescence spectra using principal component analysis (PCA), quadratic discriminant analysis (QDA), and receiver operating characteristics (ROC). The confusion matrix indicated a highly successful tissue discrimination rate before laser exposure, with an average classification error of 5.2%. The clinically relevant tissue pairs nerve/cancellous bone and nerve/salivary gland yielded an AUC of 100% each. After laser ablation, tissue discrimination was feasible with an average classification accuracy of 92.1% (average classification error 7.9%). The identification of nerve versus cancellous bone and salivary gland performed very well with an AUC of 100 and 99%, respectively. Nerve-sparing laser surgery in the area of the head and neck by means of an autofluorescence-based feedback system is feasible even after ER-YAG laser-tissue interactions. These results represent a crucial step for the development of a clinically applicable feedback tool for laser surgery interventions in the oral and maxillofacial region.