期刊
JOURNAL OF BIOPHOTONICS
卷 10, 期 3, 页码 404-414出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/jbio.201500323
关键词
coherent anti-Stokes Raman scattering; two-photon fluorescence; brain tumors; glioblastoma; green fluorescent protein; native tissue; human biopsies
资金
- German Federal Ministry of Education and Research (MediCARS project) [AZ. 13N10777]
- Medical Faculty of the Technische Universitat Dresden (Habilitationsforderung fur Frauen)
Coherent anti-Stokes Raman scattering (CARS) microscopy is an emerging technique for identification of brain tumors. However, tumor identification by CARS microscopy on bulk samples and in vivo has been so far verified retrospectively on histological sections, which only provide a gross reference for the interpretation of CARS images without matching at cellular level. Therefore, fluorescent labels were exploited for direct assessment of the interpretation of CARS images of solid and infiltrative tumors. Glioblastoma cells expressing green fluorescent protein (GFP) were used for induction of tumors in mice (n = 7). The neoplastic nature of cells imaged by CARS microscopy was unequivocally verified by addressing two-photon fluorescence of GFP on fresh brain slices and in vivo. In fresh unfixed biopsies of human glioblastoma (n = 10), the fluorescence of 5-aminolevulinic acid-induced protoporphyrin IX was used for identification of tumorous tissue. Distinctive morphological features of glioblastoma cells, i.e. larger nuclei, evident nuclear membrane and nucleolus, were identified in the CARS images of both mouse and human brain tumors. This approach demonstrates that the chemical contrast provided by CARS allows the localization of infiltrating tumor cells in fresh tissue and that the cell morphology in CARS images is useful for tumor recognition.
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