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TitleSerialized on-grid lift-in sectioning for tomography (SOLIST) enables a biopsy at the nanoscale.
Journal, issue, pagesNat Methods, Vol. 21, Issue 9, Page 1693-1701, Year 2024
Publish dateSep 13, 2024
AuthorsHo Thuy Dung Nguyen / Gaia Perone / Nikolai Klena / Roberta Vazzana / Flaminia Kaluthantrige Don / Malan Silva / Simona Sorrentino / Paolo Swuec / Frederic Leroux / Nereo Kalebic / Francesca Coscia / Philipp S Erdmann /
PubMed AbstractCryo-focused ion beam milling has substantially advanced our understanding of molecular processes by opening windows into cells. However, applying this technique to complex samples, such as tissues, ...Cryo-focused ion beam milling has substantially advanced our understanding of molecular processes by opening windows into cells. However, applying this technique to complex samples, such as tissues, has presented considerable technical challenges. Here we introduce an innovative adaptation of the cryo-lift-out technique, serialized on-grid lift-in sectioning for tomography (SOLIST), addressing these limitations. SOLIST enhances throughput, minimizes ice contamination and improves sample stability for cryo-electron tomography. It thereby facilitates the high-resolution imaging of a wide range of specimens. We illustrate these advantages on reconstituted liquid-liquid phase-separated droplets, brain organoids and native tissues from the mouse brain, liver and heart. With SOLIST, cellular processes can now be investigated at molecular resolution directly in native tissue. Furthermore, our method has a throughput high enough to render cryo-lift-out a competitive tool for structural biology. This opens new avenues for unprecedented insights into cellular function and structure in health and disease, a 'biopsy at the nanoscale'.
External linksNat Methods / PubMed:39271806 / PubMed Central
MethodsEM (tomography) / EM (subtomogram averaging)
Resolution7.3 - 20.0 Å
Structure data

EMDB-50580: SOLIST cryo-tomogram of native left ventricle mouse heart muscle #1
Method: EM (tomography)

EMDB-50582: SOLIST native mouse heart muscle tomogram #2
Method: EM (tomography)

EMDB-50592: SOLIST native mouse heart muscle tomogram #3
Method: EM (tomography)

EMDB-50603: SOLIST cryo-tomogram of native mouse corpus callosum #1
Method: EM (tomography)

EMDB-50604: SOLIST cryo-tomogram of native mouse corpus callosum #2
Method: EM (tomography)

EMDB-50605: native mouse liver solist tomogram #1
Method: EM (tomography)

EMDB-50606: native mouse liver solist tomogram #2
Method: EM (tomography)

EMDB-50607: SOLIST cryo-tomogram of human brain organoid #1
Method: EM (tomography)

EMDB-50608: SOLIST cryo-tomogram of human brain organoid #2
Method: EM (tomography)

EMDB-50620: S. cerevisiae ribosome from SOLIST lamellas
Method: EM (subtomogram averaging) / Resolution: 7.3 Å

EMDB-50630: mus musculus ribosome from SOLIST lamellas
Method: EM (subtomogram averaging) / Resolution: 8.3 Å

EMDB-50646: mus musculus heart thin filament from SOLIST lamellas
Method: EM (subtomogram averaging) / Resolution: 20.0 Å

EMDB-50655: mus musculus heart thin filament with myosin heads from SOLIST lamellas
Method: EM (subtomogram averaging) / Resolution: 19.0 Å

Source
  • Mus musculus (house mouse)
  • Homo sapiens (human)
  • Saccharomyces cerevisiae (brewer's yeast)

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