Authors
Basak S, Inamdar K, Pollack YG, Albert L, Jans DC, Jakobs S, Enderlein J, Tsukanov R, Opazo F
Journal
BioRxiv
Citation
bioRxiv 2025.12.17.694880
Abstract
Direct STORM relies on robust fluorophore blinking; observations suggest that AlexaFluor647 underperforms when coupled to nanobodies, thereby undermining the advantages of nanobodies. We show that the self-blinking dye JF635b preserves excellent blinking behavior upon nanobody conjugation, enabling sparse, stable localizations in saline buffers. Using these self-blinking nanobodies, we obtained robust dSTORM, FL-SMLM, and MINFLUX images, achieving localization precision of ∼1 nm. Its simple implementation improves reproducibility and makes single-molecule localization microscopy more accessible.
