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Exploring shaped focused ion beams for lamella preparation.

Johann Brenner1, Jürgen M Plitzko2, Sven Klumpe2

  • 1Research Group CryoEM Technology, Max Planck Institute of Biochemistry, Martinsried, Germany; Research Group Mechanisms of Cellular Quality Control, Max Planck Institute of Biophysics, Frankfurt am Main, Germany.

Ultramicroscopy
|January 13, 2026
PubMed
Summary

Focused ion beam (FIB) milling for transmission electron microscopy (TEM) now uses an 'ion knife' probe. This astigmatic probe enables improved material removal and feasibility for cryogenic lamella preparation.

Keywords:
Beam shapingBiologyCryo-FIBCryo-electron tomographyFocused ion beamsLamella preparationTEMion knifepatterning

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Area of Science:

  • Materials Science and Engineering
  • Microscopy and Imaging
  • Nanotechnology

Background:

  • Focused ion beam (FIB) milling is a standard technique for transmission electron microscopy (TEM) sample preparation.
  • Conventional FIB milling typically produces non-isometric lamellae with dimensions significantly different in thickness versus width and length.
  • The standard Gaussian-like ion probe limits precise control over material ablation geometry.

Purpose of the Study:

  • To investigate an alternative ion probe shape for improved FIB milling.
  • To characterize the 'ion knife' probe for enhanced material removal and lamella geometry control.
  • To demonstrate the utility of the 'ion knife' for cryogenic sample preparation.

Main Methods:

  • Utilized the stigmator as a quasi-cylindrical lens to create a highly astigmatic ion probe, termed 'ion knife'.
  • Characterized probe dimensions and material removal patterns using spot burn cross-sections.
  • Developed a method for rapid imaging-based approximation of probe shapes for parameter alignment.
  • Applied the 'ion knife' for milling cellular lamellae and sectioning cryo-lift-out volumes.

Main Results:

  • The 'ion knife' probe spreads ion current over a larger area, altering material removal dimensions compared to Gaussian probes.
  • A method for rapid probe shape approximation via imaging was successfully demonstrated.
  • The 'ion knife' proved feasible for milling cellular lamellae and sectioning cryo-lift-out volumes in cryogenic preparation.

Conclusions:

  • The astigmatic 'ion knife' probe offers a novel approach to FIB milling for TEM sample preparation.
  • This technique allows for modified material removal characteristics and improved control over lamella geometry.
  • The 'ion knife' shows significant potential for advancing cryogenic sample preparation in microscopy.