@inproceedings{ee8967147d104eb7b80de9ede353be0d,
title = "Cryogenic single-molecule active control microscopy with a photoactivatable fluorescent protein",
abstract = "Single-molecule superresolution methods enable imaging of specifically-labeled biological samples with structures on length scales below the diffraction limit of visible light. Imaging samples at cryogenic temperatures (77 K) significantly reduces photobleaching, allowing more photons to be collected per emitter and thus improving the localization precision. Cryogenic single-molecule imaging also facilitates correlative imaging with cryogenic electron tomography (cryoET), which provides images of whole biological cells with high-resolution cellular contrast. Combining these two techniques by performing optical imaging under conditions that do not damage the sample for cryoET allows the combination of the high sensitivity and specificity from single-molecule fluorescence with the cellular context from cryoET. In this work, we use PAmKate, a red photoactivatable fluorescent protein, to perform cryogenic single-molecule imaging of proteins in the model organism Caulobacter crescentus at 77 K with sufficiently low illumination powers to prevent damage of the cryogenic sample. The enhanced number of photons detected allows localization precision to be improved to values below 10 nm.",
keywords = "Correlative imaging, Cryogenic, Cryopalm, Fluorescent protein, Photoactivation, Photophysics",
author = "Sartor, {Annina M.} and Dahlberg, {Peter D.} and Jiarui Wang and Saumya Saurabh and Lucy Shapiro and Moerner, {W. E.}",
note = "Funding Information: This work was supported in part by the National Institute of General Medical Sciences Grant Nos. R35GM118067 (to W.E.M.) and R35GM118071 (to L.S.). A.M.S is a William and Lynda Steere Bio-X Stanford Interdisciplinary Graduate Fellow, and J.W. is a Mona M. Burgess Bio-X Stanford Interdisciplinary Graduate Fellow. L.S. is a Chan Zuckerberg Biohub Investigator. Publisher Copyright: {\textcopyright} COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only.; Single Molecule Spectroscopy and Superresolution Imaging XIII 2020 ; Conference date: 01-02-2020 Through 02-02-2020",
year = "2020",
doi = "10.1117/12.2546333",
language = "English (US)",
series = "Progress in Biomedical Optics and Imaging - Proceedings of SPIE",
publisher = "SPIE",
editor = "Ingo Gregor and Felix Koberling and Rainer Erdmann",
booktitle = "Single Molecule Spectroscopy and Superresolution Imaging XIII",
}