Tumor removal typically involves electrocautery, but no studies to date have quantified the effect of electrocautery on fluorescence emission. Electrocautery was applied to N=4 locations of the oral cavity and striated leg muscle of a live Yorkshire pig. Autofluorescence of cauterized tissues and surrounding regions was measured at distinct time points up to 120 minutes following cauterization. The fluorescence lifetime was spectrally resolved in four spectral detection channels that maximized the signal emanating from endogenous fluorophores of interest. The autofluorescence emission (355 nm excitation) was temporally resolved using a high-speed digitizer; resulting fluorescence decay characteristics were retrieved using the Laguerre deconvolution technique. Histology was performed and co-registered with the autofluorescence data. Results show that cauterized tissue presents a distinct autofluorescence signature from surrounding regions immediately after cauterization. Differences become less evident with time. The autofluorescence-derived parameters suggest altered metabolism in peripheral regions compared to the region of maximal damage. Within the time-frame of this study, tissues investigated show variable degrees of recovery from the effects of electrocautery that can be monitored by changes in fluorescence lifetime characteristics. Our findings suggest delineation of pathologic conditions could be affected by tissue cauterization and that future studies in this area will be necessary.