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TitleExoSloNano: multimodal nanogold labels for identification of macromolecules in live cells and cryo-electron tomograms.
Journal, issue, pagesNat Methods, Vol. 23, Issue 1, Page 131-142, Year 2026
Publish dateNov 28, 2025
AuthorsLindsey N Young / Alice Sherrard / Huabin Zhou / Farhaz Shaikh / Joshua Hutchings / Margot Riggi / Mythreyi Narasimhan / W Alexander Flaherty / Eric J Bennett / Michael K Rosen / Antonio J Giraldez / Elizabeth Villa /
PubMed AbstractIn situ cryo-electron microscopy (cryo-EM) enables the direct interrogation of structure-function relationships by resolving macromolecular structures in their native cellular environment. Recent ...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.
External linksNat Methods / PubMed:41315814 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution10.5 - 18.0 Å
Structure data

EMDB-71113: ExoSloNano: STA on nucleosomes from cryo-FIB-ET
Method: EM (subtomogram averaging) / Resolution: 18.0 Å

EMDB-71202: ExoSloNano, STA of 1.4 nm NG labeling of the ribosome from vitreous cells
Method: EM (subtomogram averaging) / Resolution: 10.5 Å

EMDB-71205: ExoSloNano proof of principle labeling the ribosome in intact and vitreous cells with 5 nm NG
Method: EM (subtomogram averaging) / Resolution: 16.0 Å

EMDB-71211: ExoSloNano: labeling macroH2A nucleosomes with 1.4 nm NG in intact cells.
Method: EM (subtomogram averaging) / Resolution: 18.0 Å

Source
  • Homo sapiens (human)

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