Method for Estimating the Tip Geometry of Scanning Ion Conductance Microscope Pipets
Method for Estimating the Tip Geometry of Scanning Ion Conductance Microscope Pipets
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DOI：
10.1021/ac301851n
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发表时间：
2012-11-06
影响因子：
7.4
通讯作者：
Moss, Guy W. J.
中科院分区：
文献类型：
--
作者：
Caldwell, Matthew;Del Linz, Samantha J. L.;Moss, Guy W. J.
扫描离子电导显微镜（SICM）提供了在生理条件下对生物细胞和组织进行无接触、高分辨率成像的能力。然而，成像分辨率高度依赖于SICM探头的几何形状，这通常是未知的。小的，高分辨率的探针太精细，光学图像，到目前为止，几何估计通常需要电子显微镜（EM）。这是耗时的并且容易失败，并且不能提供关于探针的关键内部几何形状的信息。在这里，我们展示了一种新的方法来确定SICM尖端的几何形状，克服了EM成像的局限性。该方法涉及拟合的分析模型，以电流的变化，在准控制的吸管尖端的断裂。可以常规地使用SICM装置本身获得数据，因此我们的方法为在SICM成像和测量中更好地量化开辟了道路。
Scanning ion conductance microscopy (SICM) offers the ability to perform contact-free, high-resolution imaging of biological cells and tissues at physiological conditions. However, imaging resolution is highly dependent on the geometry of the SICM probe, which is generally not known. Small, high-resolution probes are too fine to image optically and, to date, geometry estimation has usually required electron microscopy (EM). This is time-consuming and prone to failure and cannot provide information about the crucial internal geometry of the probe. Here we demonstrate a new method for determining SICM tip geometry that overcomes the limitations of EM imaging. The method involves fitting an analytical model to current changes during quasi-controlled breakage of the pipet tip. The data can be routinely obtained using the SICM apparatus itself and our method thus opens the way for substantially better quantification in SICM imaging and measurement.