■黑色素瘤的诊断传统上依赖于皮肤病理学家对苏木精和伊红(H&E)载玻片的显微镜检查以寻找特定的结构和细胞学特征。不幸的是,没有单一的分子标记物能够可靠地区分黑色素瘤和痣等良性病变。这项研究探索了组织内自发荧光分子的潜力,以提供指示黑素瘤中变性黑素细胞的分子指纹。
■使用高光谱成像(HSI)和光谱相量分析,我们调查了黑素瘤与皮内痣的自发荧光模式。使用紫外线激发和商用光谱共聚焦显微镜,我们从全切片样本中获得了无标签的恒生指数数据。
■我们的发现揭示了黑素瘤和皮内痣之间不同的光谱相量分布,黑色素瘤表现出更广泛的相相分布,表示更异质的自发荧光模式。值得注意的是,与较大相相关的较长波长与由专家皮肤病理学家使用H&E染色鉴定为黑色素瘤的区域相关。对五个黑素瘤(Breslow厚度范围为0.5mm至6mm)和五个皮内痣的相数簇内的相位和调制直方图的定量分析始终突出了两组之间的差异。我们进一步证明了使用相位和调制变量的质心比较来区分几种黑素细胞病变的潜力。值得注意的是,调制与相质心比较显示各组之间具有很强的统计学意义。此外,我们确定了负责组织自发荧光的分子内源性标记,包括胶原蛋白,弹性蛋白,NADH,FAD,还有黑色素.在黑色素瘤中,自发荧光的特征是相位贡献较高,表明黑素细胞巢中FAD和黑色素的增加。相比之下,NADH,弹性蛋白,胶原蛋白主导着痣的自发荧光。
■这项工作强调了自发荧光和HSI相量分析作为定量组织分子指纹图谱的有价值工具的潜力,从而支持更有效和定量的黑色素瘤诊断。
UNASSIGNED: Melanoma diagnosis traditionally relies on microscopic examination of hematoxylin and eosin (H&E) slides by dermatopathologists to search for specific architectural and cytological features. Unfortunately, no single molecular marker exists to reliably differentiate melanoma from benign lesions such as nevi. This study explored the potential of autofluorescent molecules within tissues to provide molecular fingerprints indicative of degenerated melanocytes in melanoma.
UNASSIGNED: Using hyperspectral imaging (HSI) and spectral phasor analysis, we investigated autofluorescence patterns in melanoma compared to intradermal nevi. Using UV excitation and a commercial spectral confocal microscope, we acquired label-free HSI data from the whole-slice samples.
UNASSIGNED: Our findings revealed distinct spectral phasor distributions between melanoma and intradermal nevi, with melanoma displaying a broader phasor phase distribution, signifying a more heterogeneous autofluorescence pattern. Notably, longer wavelengths associated with larger phases correlated with regions identified as melanoma by expert dermatopathologists using H&E staining. Quantitative analysis of phase and modulation histograms within the phasor clusters of five melanomas (with Breslow thicknesses ranging from 0.5 mm to 6 mm) and five intradermal nevi consistently highlighted differences between the two groups. We further demonstrated the potential for the discrimination of several melanocytic lesions using center-of-mass comparisons of phase and modulation variables. Remarkably, modulation versus phase center of mass comparisons revealed strong statistical significance among the groups. Additionally, we identified the molecular endogenous markers responsible for tissue autofluorescence, including collagen, elastin, NADH, FAD, and melanin. In melanoma, autofluorescence is characterized by a higher phase contribution, indicating an increase in FAD and melanin in melanocyte nests. In contrast, NADH, elastin, and collagen dominate the autofluorescence of the nevus.
UNASSIGNED: This work underscores the potential of autofluorescence and HSI-phasor analysis as valuable tools for quantifying tissue molecular fingerprints, thereby supporting more effective and quantitative melanoma diagnosis.