Molecule counts in localization microscopy with organic fluorophores
Single-molecule localization microscopy (SMLM) can be used to count fluorescently labeled molecules even when they are not individually resolved. We demonstrate SMLM molecule counting for nucleic acids labeled with the organic fluorophore Alexa Fluor 647 and imaged under photoswitching conditions. F...
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| Main Authors: | , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
April 19, 2017
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| In: |
ChemPhysChem
Year: 2017, Volume: 18, Issue: 8, Pages: 942-948 |
| ISSN: | 1439-7641 |
| DOI: | 10.1002/cphc.201601425 |
| Online Access: | Verlag, Volltext: http://dx.doi.org/10.1002/cphc.201601425 Verlag, Volltext: https://onlinelibrary.wiley.com/doi/abs/10.1002/cphc.201601425 
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| Author Notes: | Christos Karathanasis, Franziska Fricke, Gerhard Hummer, and Mike Heilemann |
| Summary: | Single-molecule localization microscopy (SMLM) can be used to count fluorescently labeled molecules even when they are not individually resolved. We demonstrate SMLM molecule counting for nucleic acids labeled with the organic fluorophore Alexa Fluor 647 and imaged under photoswitching conditions. From the observed distributions of the number of fluorophore blinking events, we extract the number of fluorophores per spot using a statistical model. We validate the molecule counting method for single Alexa Fluor 647 fluorophores, and for trimers of Alexa Fluor 647 constructed on a DNA origami structure. This simple counting strategy enables quantitative super-resolution imaging with organic fluorophores. |
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| Item Description: | Gesehen am 16.10.2018 First published: 14 February 2017 |
| Physical Description: | Online Resource |
| ISSN: | 1439-7641 |
| DOI: | 10.1002/cphc.201601425 |