Fluorescence lifetime imaging of endogenous fluorophores in histopathology sections reveals differences between normal and tumor epithelium in carcinoma in situ of the breast.
Fluorescence lifetime imaging of endogenous fluorophores in histopathology sections reveals differences between normal and tumor epithelium in carcinoma in situ of the breast.
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在组织病理学切片中内源性荧光团的荧光寿命成像揭示了乳腺癌原位癌中正常和肿瘤上皮之间的差异。
DOI：
10.1007/s12013-009-9046-7
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发表时间：
2009
影响因子：
2.6
通讯作者：
Keely PJ
中科院分区：
文献类型：
--
作者：
Conklin MW;Provenzano PP;Eliceiri KW;Sullivan R;Keely PJ
在这项研究中，从乳腺癌小鼠模型的组织学切片的经典检查已被扩展到现代多光子激发和光子计数技术。这种方法的优点是从荧光发射信号中量化潜在的诊断参数，从而通过测量荧光强度、寿命和光谱来补充传统的描述性分级过程。我们探讨了伊红和苏木精染色的组织学切片的临床“金标准”是否会提供具有诊断价值的光学生物标志物特征。或者，我们检查了未染色的载玻片的强度和/或相关内源性荧光团的荧光寿命的变化。虽然曙红提供了一个强的发射信号，并具有独特的光谱和寿命，我们发现，它是不是有用的荧光生物标记物，特别是当与苏木精组合。相反，我们发现来自内源性荧光团NADH和FAD的荧光的性质指示组织的病理状态。比较原位癌区域与邻近的组织学正常区域，我们发现肿瘤细胞产生更高的强度并且具有更长的荧光寿命。通过在780 nm和890 nm激发下成像，我们能够通过分离发射光谱来区分FAD与NADH的荧光。在肿瘤细胞中向更长寿命的转变与FAD和NADH的自由或结合状态以及激发波长无关。大多数形式的癌症具有改变的代谢和氧化还原比;在这里，我们提出了一种具有早期检测这些变化的潜力的方法，这些变化保存在固定的组织样本中，例如经典的组织病理学切片。
The classical examination of histology slides from a mouse model of breast cancer has been extended in this study to incorporate modern multiphoton excitation and photon-counting techniques. The advantage of such approaches is quantification of potential diagnostic parameters from the fluorescence emission signal, whereby the traditional descriptive staging process is complemented by measurements of fluorescence intensity, lifetime, and spectra. We explored whether the clinical “gold standard” of eosin and hematoxylin stained histology slides would provide optical biomarker signatures of diagnostic value. Alternatively, we examined unstained slides for changes in intensity and/or fluorescence lifetime of relevant endogenous fluorophores. Although eosin provided a strong emission signal and had distinct spectra and lifetime, we found that it was not useful as a fluorescent biological marker, particularly when combined with hematoxylin. Instead, we found that the properties of the fluorescence from the endogenous fluorophores NADH and FAD were indicative of the pathological state of the tissue. Comparing regions of carcinoma in situ to adjacent histologically normal regions, we found that tumor cells produced higher intensity and had a longer fluorescence lifetime. By imaging at 780 nm and 890 nm excitation, we were able to differentiate the fluorescence of FAD from NADH by separating the emission spectra. The shift to a longer lifetime in tumor cells was independent of the free or bound state of FAD and NADH, and of the excitation wavelength. Most forms of cancer have altered metabolism and redox ratios; here we present a method that has potential for early detection of these changes, which are preserved in fixed tissue samples such as classic histopathology slides.
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影响因子：
2.4
作者：
Gannot, I;Ron, I;Gandjbakhche, A
通讯作者：
Gandjbakhche, A
影响因子：
3.8
作者：
Kirkpatrick, Nathaniel D.;Brewer, Molly A.;Utzinger, Urs
通讯作者：
Utzinger, Urs
影响因子：
5.3
作者：
GUY, CT;CARDIFF, RD;MULLER, WJ
通讯作者：
MULLER, WJ
影响因子：
11.2
作者：
Chaiswing, Luksana;Zhong, Weixiong;Oberley, Terry D.
通讯作者：
Oberley, Terry D.
影响因子：
11.2
作者：
Bird, DK;Yan, L;Ramanujam, N
通讯作者：
Ramanujam, N