Spectrally coded optical nanosectioning (SpecON) with biocompatible metal-dielectric-coated substrates
Spectrally coded optical nanosectioning (SpecON) with biocompatible metal-dielectric-coated substrates
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DOI：
10.1073/pnas.1307222110
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发表时间：
2013-12-10
影响因子：
11.1
通讯作者：
Heinze, Katrin G.
中科院分区：
文献类型：
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作者：
Elsayad, Kareem;Urich, Alexander;Heinze, Katrin G.
荧光纳米切片在亚微米区域以上的界面是理想的许多学科在生命科学。然而，目前大多数方法的缺点是先验地需要在轴向尺寸上对雕刻的点扩散函数进行物理扫描，这对于光学敏感或高度动态的样品可能是不希望的。在这里，我们展示了一种荧光成像方法，可以通过利用简单的生物相容性纳米结构上方的荧光团的位置相关发射光谱来克服扫描的需要。为了实现这一目标，我们设计了一种薄的金属介电涂层衬底，其中对总测量荧光的光谱修饰可用于估计衬底上方10-150 nm距离内的轴向荧光团分布，精度可达5-10 nm。验证了该方法的建模和可行性，并成功地应用于迁移细胞中纳米级粘附蛋白和丝状足动力学的研究。一般原理可能会在单分子研究、生物传感和研究快速动态过程中找到更广泛的应用。
Fluorescence nanosectioning within a submicron region above an interface is desirable for many disciplines in the life sciences. A drawback, however, to most current approaches is the a priori need to physically scan a sculptured point spread function in the axial dimension, which can be undesirable for optically sensitive or highly dynamic samples. Here we demonstrate a fluorescence imaging approach that can overcome the need for scanning by exploiting the position-dependent emission spectrum of fluorophores above a simple biocompatible nanostructure. To achieve this we have designed a thin metal-dielectric-coated substrate, where the spectral modification to the total measured fluorescence can be used to estimate the axial fluorophore distribution within distances of 10-150 nm above the substrate with an accuracy of up to 5-10 nm. The modeling and feasibility of the approach are verified and successfully applied to elucidate nanoscale adhesion protein and filopodia dynamics in migrating cells. It is likely that the general principle can find broader applications in, for example, single-molecule studies, biosensing, and studying fast dynamic processes.