Enhanced fluorescence cell imaging with metal-coated slides
Enhanced fluorescence cell imaging with metal-coated slides
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DOI：
10.1117/12.728545
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发表时间：
2007-03
期刊：
影响因子：
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通讯作者：
E. Le Moal;E. Fort;S. Lévêque-Fort;A. Janin;H. Murata;F. Cordelières
中科院分区：
文献类型：
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作者：
E. Le Moal;E. Fort;S. Lévêque-Fort;A. Janin;H. Murata;F. Cordelières
在过去的十年里，荧光显微镜取得了重大进展。新型荧光标记的引入，以及原始显微镜技术的发展，使得研究活细胞中的生物分子相互作用和检查活组织的结构和功能成为可能。这些创新技术的出现对所有生命科学都产生了巨大的影响。然而，许多生物和医学应用涉及检测微量的生物分子，并且受到常见观察条件下信号微弱的限制。在这里，我们表明，银和金涂层的显微镜载玻片可以用作反射镜基板，以有效地提高检测灵敏度时，荧光显微镜应用于微米厚的生物样品。我们报告了四倍增强的荧光信号和显着加强的图像对比度，当使用标准的空气显微镜物镜的镜子基板。我们证明，金属涂层基板提供了一种手段，以获得灵敏度增强的荧光检测与干光学，同时保持宽视场观察和大景深。这对于细胞和组织诊断分析的自动化和高通量应用是至关重要的优势。
The last decade has witnessed momentous advances in fluorescence microscopy. The introduction of novel fluorescent markers, together with the development of original microscopy techniques, made it possible to study biomolecular interactions in living cells and to examine the structure and function of living tissues. The emergence of these innovative techniques had a remarkable impact on all the life sciences. However, many biological and medical applications involve the detection of minute quantities of biomolecules, and are limited by the signal weakness in common observation conditions. Here, we show that silver and gold-coated microscope slides can be used as mirror substrates to efficiently improve detection sensitivity when fluorescence microscopy is applied to micrometer-thick biological samples. We report a fourfold enhancement of the fluorescence signal and a noticeable strengthening of the image contrast, when mirror substrates are used with standard air microscope objectives. We demonstrate that metal-coated substrates provide the means to get sensitivity-enhanced fluorescence detection with dry optics, while keeping a wide field observation and a large depth of field. This is a crucial advantage for automated and high-throughput applications to cell and tissue diagnostic analysis.