Background: Tuberous sclerosis complex (TSC) is an autosomal dominant multisystem disorder with two gene loci located on chromosomes 9q34 (TSC1) and 16p13 (TSC2). Brain abnormalities in TSC include cortical tubers, subependymal nodules, giant cell astrocytomas, and white matter lesions. Cortical tubers present disordered focal neocortical formation. However, their biology remains to be elucidated. Recently, proton magnetic resonance spectroscopy has been clinically applied to the differential diagnosis of brain changes as a noninvasive neuroimaging tool. The purpose of this study was to investigate cortical tubers by single-voxel proton spectroscopy. Material/Methods: Twenty-four children with TSC were examined using a 1.5T scanner with a standard head coil. The group of patients consisted of 12 girls and 12 boys aged 3 weeks to 28 years (median: 8.66 years). Ten healthy children (examined for other reasons, with normal MR images) were the control group. Integrated MR/MRS examinations were performed. Proton MR spectroscopy images were obtained using single-voxel point resolved spectroscopy, the PRESS technique with TE=35 ms and TR=1500 ms. Results: Proton MR spectroscopy of cortical tubers revealed increased mI/Cr ratio (1.023 versus 0.553 in healthy children) and slightly decreased NAA/Cr (0.952 vs. 1.268) and NAA/Cho ratios (0.948 vs. 1.208) in all the spectra of TSC patients. The Cho/Cr ratio was almost the same as in the control group (1.079 vs. 1.058). Lactate peaks were present in ten cortical tubers. Conclusions: Proton spectroscopy can be useful in the examination of cortical tubers in TSC as a noninvasive method to investigate neurochemistry of the brain.