Technical Abstract: Fluorescence responses of wholesome and unwholesome chicken carcasses were characterized and further evaluated for potential on-line applications for detection and classification of wholesome and unwholesome chicken carcasses. For this study, unwholesome chicken carcasses included cadaver and those diseased, such as airsacculitis and septicemia. Fluorescence characteristics from chicken carcasses were dynamic in nature. Emission peaks and ridges (maxima) were observed at 386, 444, 472, 512, and 554 nm and valleys (minima) were observed at 410, 460, 484, 538 nm. One of the major factors affecting the line shapes of fluorescence responses on chicken carcass skin layers was absorption by hemoglobin. With the use of normalized ratio spectra (NRS) approach, oxyhemoglobin was shown to be the major constituent in chicken carcasses affecting the fluorescence emission line shapes. A subtle line shape changes in the NRS also provided a qualitative means to assess the minute differences in oxy- and deoxyhemoglobin compositions perturbed by the poultry diseases such as septicemia and airsacculitis. With the use of simple fluorescence band ratios as multivariate model, wholesome and unwholesome chicken carcasses were correctly classified with 97.1 % and 94.8 % accuracies, respectively. On-line implementation of fluorescence techniques for the assessment of chicken carcass wholesomeness appears promising.