Print Email Facebook Twitter Polarized stimulated-emission depletion and dark-state lifetime at vacuum and cryogenic temperature conditions Title Polarized stimulated-emission depletion and dark-state lifetime at vacuum and cryogenic temperature conditions Author Hulleman, C.N. (TU Delft ImPhys/Computational Imaging) Moerland, R.J. (TU Delft ImPhys/Computational Imaging) Stallinga, S. (TU Delft ImPhys/Imaging Physics) Rieger, B. (TU Delft ImPhys/Computational Imaging) Department ImPhys/Imaging Physics Date 2021 Abstract With the growing popularity of cryogenic correlative light and electron microscopy, it is becoming increasingly important to bridge the resolution gap between these two modalities. At cryogenic temperatures, the photon yield of fluorophores is a few orders of magnitude higher than at room temperature, enabling localization precisions on the Ångström scale. The current challenge is to induce sparsity at cryogenic temperatures such that individual fluorescent molecules can be localized. In this paper, we demonstrate the progress of using polarized stimulated-emission depletion (STED) to induce sparsity at cryogenic temperatures and in vacuum. We generate linear polarization of arbitrary in-plane orientations to achieve polarized STED with a sparsity of 3.3:1. Furthermore, we have probed the dark-state lifetime of ATTO 647N at cryogenic temperatures and in vacuum at room temperature. This dark state in vacuum is long-lived (τ=38 ms) and could be the cause for reduced photostability of fluorophores under STED illumination in vacuum. The experiments were done on an in-house designed and built liquid nitrogen cryostat, enabling 30 hours of stable cryogenic fluorescence microscopy. To reference this document use: http://resolver.tudelft.nl/uuid:defdf77e-25c6-44eb-9570-22379e537eee DOI https://doi.org/10.1103/PhysRevA.104.063516 ISSN 2469-9926 Source Physical Review A: covering atomic, molecular, and optical physics and quantum information, 104 (6) Part of collection Institutional Repository Document type journal article Rights © 2021 C.N. Hulleman, R.J. Moerland, S. Stallinga, B. Rieger Files PDF PhysRevA.104.063516.pdf 3.45 MB Close viewer /islandora/object/uuid:defdf77e-25c6-44eb-9570-22379e537eee/datastream/OBJ/view