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TitleUltrastructure of human brain tissue vitrified from autopsy revealed by cryo-ET with cryo-plasma FIB milling.
Journal, issue, pagesNat Commun, Vol. 15, Issue 1, Page 2660, Year 2024
Publish dateMar 26, 2024
AuthorsBenjamin C Creekmore / Kathryn Kixmoeller / Ben E Black / Edward B Lee / Yi-Wei Chang /
PubMed AbstractUltrastructure of human brain tissue has traditionally been examined using electron microscopy (EM) following fixation, staining, and sectioning, which limit resolution and introduce artifacts. ...Ultrastructure of human brain tissue has traditionally been examined using electron microscopy (EM) following fixation, staining, and sectioning, which limit resolution and introduce artifacts. Alternatively, cryo-electron tomography (cryo-ET) allows higher resolution imaging of unfixed cellular samples while preserving architecture, but it requires samples to be vitreous and thin enough for transmission EM. Due to these requirements, cryo-ET has yet to be employed to investigate unfixed, never previously frozen human brain tissue. Here we present a method for generating lamellae in human brain tissue obtained at time of autopsy that can be imaged via cryo-ET. We vitrify the tissue via plunge-freezing and use xenon plasma focused ion beam (FIB) milling to generate lamellae directly on-grid at variable depth inside the tissue. Lamellae generated in Alzheimer's disease brain tissue reveal intact subcellular structures including components of autophagy and potential pathologic tau fibrils. Furthermore, we reveal intact compact myelin and functional cytoplasmic expansions. These images indicate that plasma FIB milling with cryo-ET may be used to elucidate nanoscale structures within the human brain.
External linksNat Commun / PubMed:38531877 / PubMed Central
MethodsEM (tomography)
Structure data

EMDB-43330: Tomogram of human Alzheimer's disease brain tissue showing variety of membrane vesicles and axonal cross-sections
Method: EM (tomography)

EMDB-43331: Tomogram of human Alzheimer's disease brain tissue showing longitudinal axonal cross-section
Method: EM (tomography)

EMDB-43334: Tomogram of human Alzheimer's disease brain tissue showing variety of membrane vesicles
Method: EM (tomography)

Source
  • Homo sapiens (human)

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