2. Academic Validation
  3. ExoSloNano: multimodal nanogold labels for identification of macromolecules in live cells and cryo-electron tomograms

ExoSloNano: multimodal nanogold labels for identification of macromolecules in live cells and cryo-electron tomograms

  • Nat Methods. 2025 Nov 28. doi: 10.1038/s41592-025-02928-4.
Lindsey N Young # 1,  Alice Sherrard # 2,  Huabin Zhou 3,  Farhaz Shaikh 1 4,  Joshua Hutchings 1 5,  Margot Riggi 6,  Mythreyi Narasimhan 1,  W Alexander Flaherty 1,  Eric J Bennett 1,  Michael K Rosen 3 7,  Antonio J Giraldez 8,  Elizabeth Villa 9 10
Affiliations

Affiliations

  • 1 School of Biological Sciences, University of California, San Diego, La Jolla, CA, USA.
  • 2 Yale University, New Haven, CT, USA.
  • 3 University of Texas Southwestern Medical Center, Dallas, TX, USA.
  • 4 School of Medicine, University of California, San Francisco, San Francisco, CA, USA.
  • 5 Chan Zuckerberg Imaging Institute, Redwood City, CA, USA.
  • 6 Max Planck Institute for Biochemistry, Munich, Germany.
  • 7 Howard Hughes Medical Institute, Dallas, TX, USA.
  • 8 Yale University, New Haven, CT, USA. antonio.giraldez@yale.edu.
  • 9 School of Biological Sciences, University of California, San Diego, La Jolla, CA, USA. evilla@ucsd.edu.
  • 10 Howard Hughes Medical Institute, La Jolla, CA, USA. evilla@ucsd.edu.
  • # Contributed equally.
PMID: 41315814 DOI: 10.1038/s41592-025-02928-4
Abstract

In situ cryo-electron microscopy (cryo-EM) enables the direct interrogation of structure-function relationships by resolving macromolecular structures in their native cellular environment. Recent progress in sample preparation, imaging and data processing has enabled the identification and determination of large biomolecular complexes. However, the majority of proteins are of a size that still eludes identification in cellular cryo-EM data, and most proteins exist in low copy numbers. Therefore, novel tools are needed for cryo-EM to identify macromolecules across multiple size scales (from microns to nanometers). Here we introduce nanogold probes for detecting specific proteins using correlative light and electron microscopy, cryo-electron tomography (cryo-ET) and resin-embedded electron microscopy. These nanogold probes can be introduced into live cells, in a manner that preserves intact molecular networks and cell viability. We use this ExoSloNano system to identify both cytoplasmic and nuclear proteins by room-temperature electron microscopy, and resolve associated structures by cryo-ET. By providing high-efficiency protein labeling in live cells and molecular specificity within cryo-ET tomograms, ExoSloNano expands the proteome available to electron microscopy.

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