The Pathogen-Host Interface in Three Dimensions: Correlative FIB/SEM Applications

Area of Science:

• Microbiology
• Cell Biology
• Microscopy

Background:

• Intracellular pathogens employ complex interactions to survive and replicate within host cells.
• Multidimensional fluorescence microscopy is a key tool for studying these molecular and cellular events.
• Advancements in 3D electron microscopy offer high-resolution visualization of pathogens within their cellular context.

Purpose of the Study:

• To introduce correlative focused ion beam/scanning electron microscopy (c-FIB/SEM) tomography.
• To illustrate the utility of c-FIB/SEM for intracellular pathogen research.
• To discuss the limitations and future prospects of this correlative imaging approach.

Main Methods:

• Correlative focused ion beam/scanning electron microscopy (c-FIB/SEM) tomography.
• Integration of multidimensional fluorescence microscopy with volume electron microscopy.
• Case studies on cellular invasion by Shigella, Salmonella, and Brucella.

Main Results:

• c-FIB/SEM tomography enables detailed visualization of pathogen-host interactions.
• Combining fluorescence and electron microscopy provides both molecular and ultrastructural information.
• The technique has been successfully applied to study the invasion mechanisms of key intracellular pathogens.

Conclusions:

• Correlative microscopy, particularly c-FIB/SEM, is a powerful approach for studying intracellular pathogens.
• This technique bridges the gap between molecular/functional data and ultrastructural context.
• Further development holds promise for advancing our understanding of host-pathogen dynamics.