Robles Francisco E, Deb Sanghamitra, Wilson Jesse W, Gainey Christina S, Selim M Angelica, Mosca Paul J, Tyler Douglas S, Fischer Martin C, Warren Warren S
Department of Chemistry, Duke University, Durham, North Carolina 27708, USA.
Department of Pathology, Duke University Medical Center, Durham, NC 27705, USA.
Biomed Opt Express. 2015 Aug 27;6(9):3631-45. doi: 10.1364/BOE.6.003631. eCollection 2015 Sep 1.
Metastatic melanoma is associated with a poor prognosis, but no method reliably predicts which melanomas of a given stage will ultimately metastasize and which will not. While sentinel lymph node biopsy (SLNB) has emerged as the most powerful predictor of metastatic disease, the majority of people dying from metastatic melanoma still have a negative SLNB. Here we analyze pump-probe microscopy images of thin biopsy slides of primary melanomas to assess their metastatic potential. Pump-probe microscopy reveals detailed chemical information of melanin with subcellular spatial resolution. Quantification of the molecular signatures without reference standards is achieved using a geometrical representation of principal component analysis. Melanin structure is analyzed in unison with the chemical information by applying principles of mathematical morphology. Results show that melanin in metastatic primary lesions has lower chemical diversity than non-metastatic primary lesions, and contains two distinct phenotypes that are indicative of aggressive disease. Further, the mathematical morphology analysis reveals melanin in metastatic primary lesions has a distinct "dusty" quality. Finally, a statistical analysis shows that the combination of the chemical information with spatial structures predicts metastatic potential with much better sensitivity than SLNB and high specificity, suggesting pump-probe microscopy can be an important tool to help predict the metastatic potential of melanomas.
转移性黑色素瘤预后较差，但尚无方法能可靠预测特定分期的黑色素瘤哪些最终会发生转移，哪些不会。尽管前哨淋巴结活检（SLNB）已成为转移性疾病最有力的预测指标，但大多数死于转移性黑色素瘤的患者前哨淋巴结活检结果仍为阴性。在此，我们分析原发性黑色素瘤薄活检切片的泵浦 - 探测显微镜图像，以评估其转移潜能。泵浦 - 探测显微镜能以亚细胞空间分辨率揭示黑色素的详细化学信息。利用主成分分析的几何表示法，在无参考标准的情况下实现分子特征的量化。通过应用数学形态学原理，将黑色素结构与化学信息结合起来进行分析。结果显示，转移性原发性病变中的黑色素化学多样性低于非转移性原发性病变，且包含两种不同的表型，提示疾病具有侵袭性。此外，数学形态学分析表明转移性原发性病变中的黑色素具有独特的“尘状”特征。最后，统计分析表明，化学信息与空间结构相结合预测转移潜能的敏感性远高于SLNB，且特异性高，这表明泵浦 - 探测显微镜可成为帮助预测黑色素瘤转移潜能的重要工具。
