Raman spectroscopic investigation of frozen and deparaffinized tissue sections of pediatric tumors: neuroblastoma and ganglioneuroma

Raman spectroscopy is a nondestructive technique that can provide information at the molecular level about the biochemicals in tissues. We have investigated the cellular regions in neuroblastoma and ganglioneuroma using Raman spectroscopy and compared their spectral characteristics with those of the corresponding normal adrenal gland. Thin sections from both the frozen and the corresponding formalin-fixed paraffin-processed (FFPP) tissues were studied in conjunction with the pathological examination of the tissues. Investigation of the spectral data shows that the normal adrenal gland tissues have higher levels of carotenoids,lipids,andcholesterolcomparedwith theneuroblastomaandganglioneuromafrozentissues.However, incomparisonwith the frozen tissues, the FFPP tissues show a significant alteration of several biochemicals, including the complete removal of carotenoids,lipids,and cholesterolin the adrenal tissues.A quantitative analysisusing chemometric methods of principalcomponent analysis and discriminant function analysis of the Raman spectral data obtained from the frozen tissues show a clear-cut classification among pathological groups with high sensitivity and specificity. We have validated the classification results of the FFPP tissues against a training set data obtained from the archived FFPP tissues of nine other patients. The validation process correctly identified and grouped the data with the training set of normal adrenal gland (>97% of the time) and neuroblastoma (100% of the time) tissues, whereas the validation was not so strong for ganglioneuroma. This study shows that Raman spectroscopycombinedwithchemometricmethodscanbesuccessfullyusedtodistinguishneuroblastomaandganglioneuromaatcellular levelinfrozentissuesections.Thisstudyalsoshowsthatformalin fixationandparaffinization/deparaffinizationoftissuescanalter their biochemical composition. Copyright © 2012 John Wiley & Sons, Ltd. Supporting information may be found in the online version of this article.