Depth-dependent PSF calibration and aberration correction for 3D single-molecule localization
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 PS...
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| Main Authors: | , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
2 May 2019
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| In: |
Biomedical optics express
Year: 2019, Volume: 10, Issue: 6, Pages: 2708-2718 |
| ISSN: | 2156-7085 |
| DOI: | 10.1364/BOE.10.002708 |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1364/BOE.10.002708 Verlag, lizenzpflichtig, Volltext: https://www.osapublishing.org/boe/abstract.cfm?uri=boe-10-6-2708 
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| Author Notes: | Yiming Li, Yu-Le Wu, Philipp Hoess, Markus Mund, and Jonas Ries |
| Summary: | 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. |
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| Item Description: | Gesehen am 07.12.2020 |
| Physical Description: | Online Resource |
| ISSN: | 2156-7085 |
| DOI: | 10.1364/BOE.10.002708 |