Correlative cryo-imaging of the cellular universe with soft X-rays and laser light used to track F-actin structures in mammalian cells

Mohamed Koronfel, Ilias Kounatidis, Dennis M. Mwangangi, Nina Vyas, Chidinma Okolo, Archana Jadhav, Tom Fish, Phatcharin Chotchuang, Albert Schulte, Robert C. Robinson, Maria Harkiolaki

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

Imaging of actin filaments is crucial due to the integral role that they play in many cellular functions such as intracellular transport, membrane remodelling and cell motility. Visualizing actin filaments has so far relied on fluorescence microscopy and electron microscopy/tomography. The former lacks the capacity to capture the overall local ultrastructure, while the latter requires rigorous sample preparation that can lead to potential artefacts, and only delivers relatively small volumes of imaging data at the thinnest areas of a cell. In this work, a correlative approach utilizing in situ super-resolution fluorescence imaging and cryo X-ray tomography was used to image bundles of actin filaments deep inside cells under near-native conditions. In this case, fluorescence 3D imaging localized the actin bundles within the intracellular space, while X-ray tomograms of the same areas provided detailed views of the local ultrastructure. Using this new approach, actin trails connecting vesicles in the perinuclear area and hotspots of actin presence within and around multivesicular bodies were observed. The characteristic prevalence of filamentous actin in cytoplasmic extensions was also documented.

Original languageEnglish
Pages (from-to)1479-1485
Number of pages7
JournalActa Crystallographica Section D: Structural Biology
Volume77
DOIs
Publication statusPublished - Dec 1 2021

Keywords

  • Actin filaments
  • Correlative imaging
  • Soft X-ray tomography
  • Structured illumination
  • Super-resolution microscopy

ASJC Scopus subject areas

  • Structural Biology

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