Laitenberger Oskar, Aspelmeier Timo, Staudt Thomas, Geisler Claudia, Munk Axel, Egner Alexander
Department of Optical Nanoscopy, Institut für Nanophotonik e.V., 37077 Göttingen, Germany.
Institute for Mathematical Stochastics, Georg-August-University of Göttingen, 37073 Göttingen, Germany.
Nanomaterials (Basel). 2023 Jan 23;13(3):459. doi: 10.3390/nano13030459.
With the advent of fluorescence superresolution microscopy, nano-sized structures can be imaged with a previously unprecedented accuracy. Therefore, it is rapidly gaining importance as an analytical tool in the life sciences and beyond. However, the images obtained so far lack an absolute scale in terms of fluorophore numbers. Here, we use, for the first time, a detailed statistical model of the temporal imaging process which relies on a hidden Markov model operating on two timescales. This allows us to extract this information from the raw data without additional calibration measurements. We show this on the basis of added data from experiments on single Alexa 647 molecules as well as GSDIM/dSTORM measurements on DNA origami structures with a known number of labeling positions.
随着荧光超分辨率显微镜的出现，纳米级结构能够以前所未有的精度进行成像。因此，它作为一种分析工具在生命科学及其他领域正迅速变得重要起来。然而，到目前为止所获得的图像在荧光团数量方面缺乏绝对尺度。在此，我们首次使用了时间成像过程的详细统计模型，该模型依赖于在两个时间尺度上运行的隐马尔可夫模型。这使我们能够从原始数据中提取此信息，而无需额外的校准测量。我们基于单个Alexa 647分子实验的添加数据以及对具有已知标记位置数量的DNA折纸结构的GSDIM/dSTORM测量结果展示了这一点。
