Assessing the efficacy of coherent anti-Stokes Raman scattering microscopy for the detection of infiltrating glioblastoma in fresh brain samples
Assessing the efficacy of coherent anti-Stokes Raman scattering microscopy for the detection of infiltrating glioblastoma in fresh brain samples
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DOI：
10.1002/jbio.201500323
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发表时间：
2017-03-01
影响因子：
2.8
通讯作者：
Kirsch, Matthias
中科院分区：
文献类型：
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作者：
Galli, Roberta;Uckermann, Ortrud;Kirsch, Matthias
相干反斯托克斯拉曼散射（汽车）显微镜是一种新兴的技术，用于识别脑肿瘤。然而，迄今为止，通过汽车显微镜对大量样品和体内的肿瘤识别已经在组织学切片上进行了回顾性验证，这仅为汽车图像的解释提供了粗略的参考，而没有在细胞水平上进行匹配。因此，利用荧光标记直接评估实体和浸润性肿瘤的汽车图像的解释。表达绿色荧光蛋白（GFP）的胶质母细胞瘤细胞用于诱导小鼠肿瘤（n = 7）。汽车显微镜成像的细胞的肿瘤性质通过在新鲜脑切片和体内处理GFP的双光子荧光而明确地验证。在人胶质母细胞瘤的新鲜的未固定的活组织检查（n = 10）中，5-氨基乙酰丙酸诱导的原卟啉IX的荧光被用于鉴定肿瘤组织。胶质母细胞瘤细胞的独特形态学特征，即较大的细胞核，明显的核膜和核仁，在小鼠和人类脑肿瘤的汽车图像中被识别。这种方法表明，由汽车提供的化学对比度允许在新鲜组织中定位浸润的肿瘤细胞，并且汽车图像中的细胞形态对于肿瘤识别是有用的。
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.