Correlative cryo-fluorescence and cryo-scanning electron microscopy as a straightforward tool to study host-pathogen...

Area of Science:

• Microscopy
• Cell Biology
• Infectious Diseases

Background:

• Correlative light and electron microscopy (CLEM) allows high-resolution imaging of fluorescently tagged structures.
• Conventional fixation methods can introduce artifacts, hindering the study of biological processes in near-native states.
• Understanding pathogen-host interactions is crucial for developing effective treatments.

Purpose of the Study:

• To establish a novel correlative cryo-fluorescence microscopy and cryo-scanning electron microscopy (cryo-CLEM) workflow.
• To investigate the early cell-association mechanisms of Borrelia burgdorferi with mammalian neural cell lines.
• To provide a rapid and reliable method for studying pathogen-host interactions at high resolution.

Main Methods:

• Development of a correlative cryo-fluorescence microscopy and cryo-scanning electron microscopy workflow.
• Imaging of Borrelia burgdorferi interacting with neural cell lines using the established cryo-CLEM technique.
• Analysis of bacterial cell association and interaction with host cells in a near-native state.

Main Results:

• The novel cryo-CLEM workflow enables imaging of biological samples in a near-native state, minimizing fixation artifacts.
• The method was successfully applied to study the interaction between Borrelia burgdorferi and mammalian neural cell lines.
• The workflow demonstrated high speed (<3 hours), straightforwardness, and reliability in visualizing pathogen-host cell dynamics.

Conclusions:

• The developed cryo-CLEM workflow is an unprecedentedly fast and reliable solution for studying pathogen-host cell interactions.
• This technique provides valuable insights into the finer details of bacterial association with host cells prior to entry.
• The findings advance our understanding of the complex relationship between pathogens and host cells, particularly in the context of zoonotic diseases.