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，样品中荧光团分布的超分辨率图像是根据许多单个分子的局部位置构建的。由于其实验简单和可以实现的高分辨率，它已被广泛使用。然而，限制重建图像分辨率的因素以及可能存在的人为因素与标准荧光显微镜技术中存在的因素完全不同。用户可能难以识别人工制品，特别是因为它们可能导致图像显得虚假清晰，这种效果称为人工锐化。在这里，我们讨论 SMLM 中不同的错误和偏差来源，以及可用于避免或检测它们的方法。