Measuring true localization accuracy in super resolution microscopy with DNA-origami nanostructures
A common method to assess the performance of (super resolution) microscopes is to use the localization precision of emitters as an estimate for the achieved resolution. Naturally, this is widely used in super resolution methods based on single molecule stochastic switching. This concept suffers from...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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IOP Publishing
2017-01-01
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| Series: | New Journal of Physics |
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| Online Access: | https://doi.org/10.1088/1367-2630/aa5f74 |
| _version_ | 1827873613056835584 |
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| author | Matthias Reuss Ferenc Fördős Hans Blom Ozan Öktem Björn Högberg Hjalmar Brismar |
| author_facet | Matthias Reuss Ferenc Fördős Hans Blom Ozan Öktem Björn Högberg Hjalmar Brismar |
| author_sort | Matthias Reuss |
| collection | DOAJ |
| description | A common method to assess the performance of (super resolution) microscopes is to use the localization precision of emitters as an estimate for the achieved resolution. Naturally, this is widely used in super resolution methods based on single molecule stochastic switching. This concept suffers from the fact that it is hard to calibrate measures against a real sample (a phantom), because true absolute positions of emitters are almost always unknown. For this reason, resolution estimates are potentially biased in an image since one is blind to true position accuracy, i.e. deviation in position measurement from true positions. We have solved this issue by imaging nanorods fabricated with DNA-origami. The nanorods used are designed to have emitters attached at each end in a well-defined and highly conserved distance. These structures are widely used to gauge localization precision. Here, we additionally determined the true achievable localization accuracy and compared this figure of merit to localization precision values for two common super resolution microscope methods STED and STORM. |
| first_indexed | 2024-03-12T16:39:35Z |
| format | Article |
| id | doaj.art-8a5b13662f59455aa7d06fbec4bdb145 |
| institution | Directory Open Access Journal |
| issn | 1367-2630 |
| language | English |
| last_indexed | 2024-03-12T16:39:35Z |
| publishDate | 2017-01-01 |
| publisher | IOP Publishing |
| record_format | Article |
| series | New Journal of Physics |
| spelling | doaj.art-8a5b13662f59455aa7d06fbec4bdb1452023-08-08T14:36:02ZengIOP PublishingNew Journal of Physics1367-26302017-01-0119202501310.1088/1367-2630/aa5f74Measuring true localization accuracy in super resolution microscopy with DNA-origami nanostructuresMatthias Reuss0https://orcid.org/0000-0001-8914-3830Ferenc Fördős1Hans Blom2Ozan Öktem3https://orcid.org/0000-0002-1118-6483Björn Högberg4https://orcid.org/0000-0003-2715-7887Hjalmar Brismar5https://orcid.org/0000-0003-0578-4003Science for Life Laboratory, Department of Applied Physics, Royal Institute of Technology, Solna, SwedenDepartment of Medical Biochemistry and Biophysics, Division of Biomaterials and Regenerative Medicine, Karolinska Institutet, Solna, SwedenScience for Life Laboratory, Department of Applied Physics, Royal Institute of Technology, Solna, SwedenDepartment of Mathematics, Royal Institute of Technology, Stockholm, SwedenDepartment of Medical Biochemistry and Biophysics, Division of Biomaterials and Regenerative Medicine, Karolinska Institutet, Solna, SwedenScience for Life Laboratory, Department of Applied Physics, Royal Institute of Technology, Solna, Sweden; Department of Women’s and Children’s Health, Karolinska Institutet, Solna, SwedenA common method to assess the performance of (super resolution) microscopes is to use the localization precision of emitters as an estimate for the achieved resolution. Naturally, this is widely used in super resolution methods based on single molecule stochastic switching. This concept suffers from the fact that it is hard to calibrate measures against a real sample (a phantom), because true absolute positions of emitters are almost always unknown. For this reason, resolution estimates are potentially biased in an image since one is blind to true position accuracy, i.e. deviation in position measurement from true positions. We have solved this issue by imaging nanorods fabricated with DNA-origami. The nanorods used are designed to have emitters attached at each end in a well-defined and highly conserved distance. These structures are widely used to gauge localization precision. Here, we additionally determined the true achievable localization accuracy and compared this figure of merit to localization precision values for two common super resolution microscope methods STED and STORM.https://doi.org/10.1088/1367-2630/aa5f74superresolution microscopylocalization accuracyDNA origami |
| spellingShingle | Matthias Reuss Ferenc Fördős Hans Blom Ozan Öktem Björn Högberg Hjalmar Brismar Measuring true localization accuracy in super resolution microscopy with DNA-origami nanostructures New Journal of Physics superresolution microscopy localization accuracy DNA origami |
| title | Measuring true localization accuracy in super resolution microscopy with DNA-origami nanostructures |
| title_full | Measuring true localization accuracy in super resolution microscopy with DNA-origami nanostructures |
| title_fullStr | Measuring true localization accuracy in super resolution microscopy with DNA-origami nanostructures |
| title_full_unstemmed | Measuring true localization accuracy in super resolution microscopy with DNA-origami nanostructures |
| title_short | Measuring true localization accuracy in super resolution microscopy with DNA-origami nanostructures |
| title_sort | measuring true localization accuracy in super resolution microscopy with dna origami nanostructures |
| topic | superresolution microscopy localization accuracy DNA origami |
| url | https://doi.org/10.1088/1367-2630/aa5f74 |
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