0605497 - ÚFCH JH 2026 RIV GB eng J - Journal Article
Hekrdla, Miroslav - Roesel, D. - Hansen, N. - Frederick, S. - Umar, K. - Petráková, Vladimíra
Optimized molecule detection in localization microscopy with selected false positive probability.
Nature Communications. Roč. 16, č. 1 (2025), č. článku 601. ISSN 2041-1723. E-ISSN 2041-1723
R&D Projects: GA ČR(CZ) GM21-17847M; GA MŠMT(CZ) LM2023066
Grant - others:AV ČR(CZ) LQ200402101
Program: Prémie Lumina quaeruntur
Institutional support: RVO:61388955
Keywords : Single-molecule localization microscopy * background estimation * tracking
OECD category: Physical chemistry
Impact factor: 14.7, year: 2023 ; AIS: 5.66, rok: 2023
Method of publishing: Open access
DOI: https://doi.org/10.1038/s41467-025-55952-5

Single-molecule localization microscopy (SMLM) allows imaging beyond the diffraction limit. Detection of molecules is a crucial initial step in SMLM. False positive detections, which are not quantitatively controlled in current methods, are a source of artifacts that affect the entire SMLM analysis pipeline. Furthermore, current methods lack standardization, which hinders reproducibility. Here, we present an optimized molecule detection method which combines probabilistic thresholding with theoretically optimal filtering. The probabilistic thresholding enables control over false positive detections while optimal filtering minimizes false negatives. A theoretically optimal Poisson matched filter is used as a performance benchmark to evaluate existing filtering methods. Overall, our approach allows the detection of molecules in a robust, single-parameter and user-unbiased manner. This will minimize artifacts and enable data reproducibility in SMLM.
Permanent Link: https://hdl.handle.net/11104/0363220