Camera-based localization microscopy optimized with calibrated structured illumination

Martin Schmidt, Adam C. Hundahl, Henrik Flyvbjerg, Rodolphe Marie, Kim I. Mortensen*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Until very recently, super-resolution localization and tracking of fluorescent particles used camera-based wide-field imaging with uniform illumination. Then it was demonstrated that structured illuminations encode additional localization information in images. The first demonstration of this uses scanning and hence suffers from limited throughput. This limitation was mitigated by fusing camera-based localization with wide-field structured illumination. Current implementations, however, use effectively only half the localization information that they encode in images. Here we demonstrate how all of this information may be exploited by careful calibration of the structured illumination. Our approach achieves maximal resolution for given structured illumination, has a simple data analysis, and applies to any structured illumination in principle. We demonstrate this with an only slightly modified wide-field microscope. Our protocol should boost the emerging field of high-precision localization with structured illumination.

Original languageEnglish
Article number41
JournalCommunications Physics
Issue number1
Number of pages9
Publication statusPublished - 2021

Bibliographical note

Funding Information:
This work was funded by the Novo Nordisk Foundation Challenge Programme (NNF16OC0022166). We thank A. Kristensen for generously sharing microscopy components with us.


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