Randall Centre for Cell & Molecular Biophysics, Guy's Campus, King's College London, UK.
Randall Centre for Cell & Molecular Biophysics, Guy's Campus, King's College London, UK.
Int J Biochem Cell Biol. 2021 May;134:105931. doi: 10.1016/j.biocel.2021.105931. Epub 2021 Feb 17.
In single molecule localisation microscopy (SMLM) a super-resolution image of the distribution of fluorophores in the sample is built up from the localised positions of many individual molecules. It has become widely used due to its experimental simplicity and the high resolution that can be achieved. However, the factors which limit resolution in a reconstructed image, and the artefacts which can be present, are completely different to those present in standard fluorescent microscopy techniques. Artefacts may be difficult for users to identify, particularly as they can cause images to appear falsely sharp, an effect called artificial sharpening. Here we discuss the different sources of error and bias in SMLM, and the methods available for avoiding or detecting them.
在单分子定位显微镜（SMLM）中，通过对许多单个分子的局域化位置进行构建，可以得到样本中荧光团分布的超分辨率图像。由于其实验的简单性和可实现的高分辨率，它已被广泛应用。然而，限制重构图像分辨率的因素以及可能存在的伪影与标准荧光显微镜技术中存在的因素完全不同。对于用户来说，这些伪影可能难以识别，特别是因为它们可能会导致图像出现虚假的锐化效果，这种效果被称为人为锐化。在这里，我们讨论了 SMLM 中的不同误差源和偏差，以及可用于避免或检测它们的方法。
