Document Type
Conference Proceeding
Publication Date
3-12-2024
Abstract
We develop six-dimensional single-molecule orientation-localization microscopy (SMOLM) to measure the 3D positions and 3D orientations simultaneously of single fluorophores. We show how careful optimization of phase and polarization modulation components can encode phase, polarization, and angular spectrum information from each fluorescence photon into a microscope’s dipole-spread function. We used the transient binding and blinking of Nile red (NR) to characterize the helical structure of fibrils formed by designed amphipathic peptides, KFE8L and KFE8D, and the pathological amyloid-beta peptide Aβ42. We also deployed merocyanine 540 to uncover the interfacial architectures of biomolecular condensates.
Recommended Citation
Copyright 2024 Society of Photo‑Optical Instrumentation Engineers (SPIE). One print or electronic copy may be made for personal use only. Systematic reproduction and distribution, duplication of any material in this publication for a fee or for commercial purposes, and modification of the contents of the publication are prohibited. Tingting Wu, Weiyan Zhou, Jai S. Rudra, Rohit V. Pappu, and Matthew D. Lew "6D single-fluorogen orientation-localization microscopy for elucidating the architecture of beta-sheet assemblies and biomolecular condensates", Proc. SPIE 12853, High-Speed Biomedical Imaging and Spectroscopy IX, 1285302 (12 March 2024); https://doi.org/10.1117/12.3005992
Included in
Bioimaging and Biomedical Optics Commons, Biological and Chemical Physics Commons, Electromagnetics and Photonics Commons, Nanoscience and Nanotechnology Commons, Optics Commons