Correlation of Live Imaging and Fixed-Expansion Microscopy to study intracellular pathogens

LiFEx correlates live-cell dynamics with Ten-fold Robust Expansion Microscopy to show that membrane alterations develop within minutes after T6SS-5 assembly in Burkholderia thailandensis-infected epithelial cells.

• Why it matters: Expansion microscopy can resolve ultrastructure, but intracellular bacterial imaging is difficult because bacterial expansion is incomplete and sensitive to fixation, limiting links between structure and infection timing.
• What they did: The study adapted Ten-fold Robust Expansion Microscopy for Burkholderia thailandensis-infected epithelial cells, comparing live-to-fixed workflows and testing how fixation and peptidoglycan affect bacterial expansion.
• The result: Host cells expanded about 9-fold, while bacterial expansion was incomplete and heterogeneous; fixation conditions strongly influence bacterial expansion and peptidoglycan partially limits it, enabling minute-scale structure–dynamics correlation.

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