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TitleCorrelating cryo-super resolution radial fluctuations and dual-axis cryo-scanning transmission electron tomography to bridge the light-electron resolution gap.
Journal, issue, pagesJ Struct Biol, Vol. 215, Issue 3, Page 107982, Year 2023
Publish dateJun 1, 2023
AuthorsPeter Kirchweger / Debakshi Mullick / Prabhu Prasad Swain / Sharon G Wolf / Michael Elbaum /
PubMed AbstractVisualization of organelles and their interactions with other features in the native cell remains a challenge in modern biology. We have introduced cryo-scanning transmission electron tomography ...Visualization of organelles and their interactions with other features in the native cell remains a challenge in modern biology. We have introduced cryo-scanning transmission electron tomography (CSTET), which can access 3D volumes on the scale of 1 micron with a resolution of nanometers, making it ideal for this task. Here we introduce two relevant advances: (a) we demonstrate the utility of multi-color super-resolution radial fluctuation light microscopy under cryogenic conditions (cryo-SRRF), and (b) we extend the use of deconvolution processing for dual-axis CSTET data. We show that cryo-SRRF nanoscopy is able to reach resolutions in the range of 100 nm, using commonly available fluorophores and a conventional widefield microscope for cryo-correlative light-electron microscopy. Such resolution aids in precisely identifying regions of interest before tomographic acquisition and enhances precision in localizing features of interest within the 3D reconstruction. Dual-axis CSTET tilt series data and application of entropy regularized deconvolution during post-processing results in close-to-isotropic resolution in the reconstruction without averaging. The integration of cryo-SRRF with deconvolved dual-axis CSTET provides a versatile workflow for studying unique objects in a cell.
External linksJ Struct Biol / PubMed:37268154
MethodsEM (tomography)
Structure data

EMDB-16178: Dual-axis CSTET tomogram at 850 nm thick region, weighted back projection
Method: EM (tomography)

EMDB-16179: Deconvolved dual-axis CSTET tomogram of a WI-38 fibroblast cell at 850 nm thick part
Method: EM (tomography)

Source
  • Homo sapiens (human)

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