Cell metabolism, tumour diagnosis and multispectral FLIM
Cell metabolism, tumour diagnosis and multispectral FLIM
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细胞代谢、肿瘤诊断和多光谱 FLIM
DOI：
10.1117/12.2003729
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Kalinina S
中科院分区：
文献类型：
--
作者：
Rueck A;Hauser C;Lorenz S;Mosch S;Rotte S;Kessler M;Kalinina S
在许多情况下，肿瘤组织的荧光引导诊断是不够的，因为假阳性结果干扰了结果。因此，肿瘤和炎症之间的区别可能是困难的。通过观察细胞代谢来改善荧光诊断可能是解决方案，这需要详细了解自发荧光的起源。然而，荧光团的复杂组合产生发射信号。同样在PDD（光动力学诊断）中，不同的光敏剂代谢物有助于荧光信号。因此，在许多情况下，荧光衰减不显示简单的单指数曲线。在这些情况下，当时间分辨和光谱分辨技术同时结合时，可以实现相当大的改进。重点讨论了卟啉类化合物的检测方法。用多光谱荧光显微镜（FLIM）研究了正常口腔角质形成细胞和不同来源的鳞癌细胞中蛋白结合型和游离型辅酶的区别。氧化还原比，这可以与荧光寿命的NADH和FAD的变化取决于细胞的状态。大多数研究在单层细胞培养中进行。然而，为了从更真实的体内情况中获得信息，还使用了受精卵的绒毛尿囊膜（CAM），其中允许肿瘤细胞或活检生长。这些测量的结果也将被讨论。
Fluorescence guided diagnosis of tumour tissue is in many cases insufficient, because false positive results are interfering with the outcome. Discrimination between tumour and inflammation could be therefore difficult. Improvement of fluorescence diagnosis through observation of cell metabolism could be the solution, which needs a detailed understanding of the origin of autofluorescence. However, a complex combination of fluorophores give rise to the emission signal. Also in PDD (photodynamic diagnosis) different photosensitizer metabolites contribute to the fluorescence signal. Therefore, the fluorescence decay in many cases does not show a simple monoexponential profile. In those cases a considerable improvement could be achieved when time-resolved and spectral-resolved techniques are simultaneously incorporated. The discussion will focus on the detection of NADH, FAD and 5-ALA induced porphyrins. With respect to NADH and FAD the discrimination between protein bound and free coenzyme was investigated with multispectral FLIM in normal oral keratinocytes and squamous carcinoma cells from different origin. The redox ratio, which can be correlated with the fluorescence lifetimes of NADH and FAD changed depending on the state of the cells. Most of the investigations were done in monolayer cell cultures. However, in order to get information from a more realistic in vivo situation additionally the chorioallantoismembrane (CAM) of fertilized eggs was used where tumour cells or biopsies were allowed to grow. The results of theses measurements will be discussed as well.
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