<p>The determination of the extent and margins of precancerous or cancerous lesions is still a problem in oncological surgeries today. This is more so with the ‘holy grail’ goal of optimal excision requiring total lesion removal and maximal neighboring tissue conservation especially for an important organ like the cervix. Although frozen section is the gold standard method available for aiding intraoperative diagnosis and surgery decision management, in dysplasia of the cervix where precancer cases are treated, it is not applicable due to various reasons, mainly to avoid over treatment. Still an optimal surgery is highly advocated and currently applying LUGOL dye to cervix is the accepted technique although not always accurate. Attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy can complement visual inspection after LUGOL application, and several efforts have shown its potential in this regard. Research efforts on the subject, however, have not explored the use of ATR-FTIR spectroscopy for determining the extent of precancer on fresh cervical biopsies. This work therefore sets out to investigate ATR-FTIR spectroscopy for estimating the extent of precancer on fresh cervical samples using discriminatory features from the fingerprint region, as well as entropy and bound water related markers. Our results show that posterior probabilities were statistically different for precancer and healthy sampling points, with an area under the receiver operating characteristic curve value of 0.678 ± 0.094. At a cut-off probability of 0.550, sensitivity and specificity were 70% and 59%, respectively. These results show that ATR-FTIR spectroscopy on fresh tissue can aid spatially resolved determination of the extent of precancer, and that bound water and entropy related spectral biomarkers can provide additional diagnostic information for the ultimate goal of aiding intraoperative decision management.</p>