Bibliographic Data
- Authors: Basak S., Inamdar K., Pollack Y. G., Albert L., Jans D. C., Jakobs S., Enderlein J., Tsukanov R., Opazo F.
- Title: Self-blinking dye restores efficient use of nanobodies in single-molecule localization microscopy
- Journal: bioRxiv
- DOI: 10.64898/2025.12.17.694880
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.