Direct optical measurement of intramolecular distances with angstrom precision
Optical investigations of nanometer distances between proteins, their subunits, or other biomolecules have been the exclusive prerogative of Förster resonance energy transfer (FRET) microscopy for decades. In this work, we show that MINFLUX fluorescence nanoscopy measures intramolecular distances down to 1 nanometer-and in planar projections down to 1 angstrom-directly, linearly, and with angstrom precision. Our method was validated by quantifying well-characterized 1- to 10-nanometer distances in polypeptides and proteins. Moreover, we visualized the orientations of immunoglobulin subunits, applied the method in human cells, and revealed specific configurations of a histidine kinase PAS domain dimer. Our results open the door for examining proximities and interactions by direct position measurements at the intramacromolecular scale.
Authors
- Kaushik Inamdar (ORCID: https://orcid.org/0000-0001-5959-6409)
- Stefan Jakobs (ORCID: https://orcid.org/0000-0002-8028-3121)
- Michael Weber (ORCID: https://orcid.org/0000-0001-9680-7543)
- Johannes Broichhagen (ORCID: https://orcid.org/0000-0003-3084-6595)
- Taukeer A. Khan (ORCID: https://orcid.org/0000-0001-5483-5837)
- Christian Griesinger (ORCID: https://orcid.org/0000-0002-1266-4344)
- Stefan Becker (ORCID: https://orcid.org/0000-0003-2041-5740)
- Steffen J. Sahl (ORCID: https://orcid.org/0000-0003-2917-0778)
- Christian Brüser (ORCID: https://orcid.org/0000-0003-1873-5120)
- Jessica Matthias (ORCID: https://orcid.org/0000-0001-6560-2174)
- Stefan W. Hell (ORCID: https://orcid.org/0000-0002-9638-5077)
Institutions
- Max Planck Institute for Medical Research (DE)
- Fraunhofer Institute for Translational Medicine and Pharmacology (DE)
- Universitätsmedizin Göttingen (DE)
- Nanoscale Microscopy and Molecular Physiology of the Brain Cluster of Excellence 171 — DFG Research Center 103 (DE)
- Max Planck Institute for Multidisciplinary Sciences (DE)
- University of Göttingen (DE)
Publication Details
- Journal
- Science
- Published
- 2024-10-10
- DOI
- https://doi.org/10.1126/science.adj7368
- Citations
- 58
- Primary Topic
- Advanced Fluorescence Microscopy Techniques
- Type
- article
- Field-Weighted Citation Impact
- 38.42