Background: Squash cytology is of significant importance in intraoperative consultation of central nervous system (CNS) pathology. There are several studies on squash cytology of CNS lesions, and only a few of them deal with spinal lesions alone. Aims: (1) To evaluate intraoperative squash cytology of spinal lesions. (2) To correlate cytological diagnosis with histopathological diagnosis and assess the diagnostic accuracy. (3) To study Ki67 expression on squash smears and determine whether it can assist in grading spinal tumours on cytology. Materials and Methods: A prospective study was conducted on 68 patients with clinico-radiologically diagnosed lesions of the spine. Intraoperative squash smears were stained with haematoxylin-eosin (H&E) stain, Papanicolaou (Pap) stain, and May-Grünwald-Giemsa (MGG) stain. Subsequently, histological diagnosis was made. Ki67 immunostaining was performed on squash smears and histology sections. Results: The sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy of squash cytology in spinal lesions were 84.6, 100, 100, 23.1, and 80.88%, respectively. On immunocytochemistry, the mean Ki67 labelling indices for grade I, II, and III tumours were 0, 0.33 and 9%, respectively. Conclusion: Squash smear cytology is a rapid intraoperative technique for diagnosing spinal lesions, with high specificity and high positive predictive value. It is more effective in diagnosing neoplasms than non-neoplastic lesions. Ki67 immunostaining can be done on cytology smears to effectively differentiate between WHO grade I and grade II spinal tumours.