Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York, USA.
Biophys J. 2011 Apr 6;100(7):L34-6. doi: 10.1016/j.bpj.2011.02.034.
We have developed a method of performing near-field fluorescence correlation spectroscopy via an array of planarized circular apertures of 50 nm diameter. This technique provides 1 μs and 60 nm resolution on proximal samples, including live cells, without incorporating a scanning probe or pulsed lasers or requiring penetration of the sample into the aperture. Millions of apertures are created in an array within a thin film of aluminum on a coverslip and planarized to achieve no height distinction between the apertures and the surrounding metal. Supported lipid bilayers and plasma membranes from live cells adhere to the top of this substrate. We performed fluorescence correlation spectroscopy to demonstrate the sub-diffraction-limited illumination with these devices.
我们开发了一种通过 50nm 直径的平面化圆形孔阵列进行近场荧光相关光谱学的方法。该技术在不引入扫描探针或脉冲激光器或要求样品穿透孔径的情况下,为近距样品(包括活细胞)提供了 1μs 和 60nm 的分辨率。在盖玻片上的铝薄膜内的阵列中创建了数百万个孔,并进行了平面化处理,以使孔和周围金属之间没有高度差异。来自活细胞的支持脂双层和质膜附着在该基底的顶部。我们进行了荧光相关光谱学实验,以证明使用这些器件的亚衍射限制照明。