Thermochemical scanning probe lithography of protein gradients at the nanoscale
Thermochemical scanning probe lithography of protein gradients at the nanoscale
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DOI：
10.1088/0957-4484/27/31/315302
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发表时间：
2016-08-05
期刊：
影响因子：
3.5
通讯作者：
Riedo, E.
中科院分区：
文献类型：
--
作者：
Albisetti, E.;Carroll, K. M.;Riedo, E.
图案化纳米蛋白梯度是至关重要的研究各种细胞过程中的体外。尽管最近在纳米制造技术的发展，结合纳米分辨率和蛋白质浓度的精细控制仍然是一个开放的挑战。在这里，我们展示了使用热化学扫描探针光刻（tc-SPL）定义微米和纳米尺寸的图案与精确控制的蛋白质浓度。首先，通过在聚合物基底上扫描可加热的原子力显微镜尖端来进行tc-SPL，用于局部暴露表面上的反应性氨基，然后用链霉亲和素和层粘连蛋白将基底功能化。我们表明，通过荧光显微镜上的图案化梯度，它是可以精确地调整固定化蛋白质的浓度，通过改变图案化参数在tc-SPL。这为使用tc-SPL在纳米级定义蛋白质梯度铺平了道路，可用作化学线索。G.用于研究和调节体外细胞过程。
Patterning nanoscale protein gradients is crucial for studying a variety of cellular processes in vitro. Despite the recent development in nano-fabrication technology, combining nanometric resolution and fine control of protein concentrations is still an open challenge. Here, we demonstrate the use of thermochemical scanning probe lithography (tc-SPL) for defining micro- and nano-sized patterns with precisely controlled protein concentration. First, tc-SPL is performed by scanning a heatable atomic force microscopy tip on a polymeric substrate, for locally exposing reactive amino groups on the surface, then the substrate is functionalized with streptavidin and laminin proteins. We show, by fluorescence microscopy on the patterned gradients, that it is possible to precisely tune the concentration of the immobilized proteins by varying the patterning parameters during tc-SPL. This paves the way to the use of tc-SPL for defining protein gradients at the nanoscale, to be used as chemical cues e. g. for studying and regulating cellular processes in vitro.