期刊
BIOMEDICAL OPTICS EXPRESS
卷 10, 期 6, 页码 2708-2718出版社
OPTICAL SOC AMER
DOI: 10.1364/BOE.10.002708
关键词
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资金
- European Research Council [ERC CoG-724489]
- EMBL Interdisciplinary Postdoc Programme (EIPOD) under Marie Curie Actions COFUND
- European Molecular Biology Laboratory
Three-dimensional single molecule localization microscopy relies on the fitting of the individual molecules with a point spread function (PSF) model. The reconstructed images often show local squeezing or expansion in z. A common cause is depth-induced aberrations in conjunction with an imperfect PSF model calibrated from beads on a coverslip, resulting in a mismatch between measured PSF and real PSF. Here, we developed a strategy for accurate z-localization in which we use the imperfect PSF model for fitting, determine the fitting errors and correct for them in a post-processing step. We present an open-source software tool and a simple experimental calibration procedure that allow retrieving accurate z-positions in any PSF engineering approach or fitting modality, even at large imaging depths. (C) 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement
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