Micropatterning Transmission Electron Microscopy Grids to Direct Cell Positioning within Whole-Cell Cryo-Electron...

Area of Science:

• Cell biology
• Structural biology
• Microscopy

Background:

• Whole-cell cryo-electron tomography (cryo-ET) offers high-resolution structural insights of macromolecules within cells.
• Preparing cells on transmission electron microscopy (TEM) grids for cryo-ET while maintaining physiological states presents significant challenges.

Purpose of the Study:

• To present a detailed protocol for using micropatterning to guide eukaryotic cell growth on TEM grids.
• To enable improved sample preparation for whole-cell cryo-ET.

Main Methods:

• Micropatterning involves depositing extracellular matrix (ECM) proteins in specific patterns on TEM grid foils.
• Anti-fouling coatings are applied to non-patterned areas to control cell adhesion.
• This technique supports diverse cell types and experimental systems.

Main Results:

• Micropatterning successfully directs and promotes eukaryotic cell growth on TEM grids.
• The protocol allows cells to be maintained in a near-native, physiologically relevant state.
• The method is adaptable to various cell types and complex cellular communities.

Conclusions:

• Micropatterning is a versatile technique for preparing samples for whole-cell cryo-ET.
• It facilitates studies of intracellular structures and complex biological systems.
• This approach integrates well with downstream cryo-ET workflows like cryo-CLEM and cryo-FIB.