Imaging of biofilms using cryo-scanning electron microscopy.

Area of Science:

• Microscopy
• Microbiology
• Materials Science

Background:

• Biofilms present significant microscopy challenges due to size, fragility, and substrate variability.
• Cryo-scanning electron microscopy (cryo-SEM) preserves native structures, unlike room temperature SEM.
• Cryo-SEM is established for environmental and food biofilms but underutilized for medical biofilms.

Purpose of the Study:

• To provide an overview of the cryo-scanning electron microscopy (cryo-SEM) workflow for biofilm imaging.
• To highlight equipment options and discuss the advantages and limitations of each step.
• To guide researchers in applying cryo-SEM to various biofilm types, including medically relevant ones.

Main Methods:

• Cryo-scanning electron microscopy (cryo-SEM) workflow involves sample freezing (high-pressure, plunge, or slush nitrogen).
• Subsequent steps include fracturing, sublimation, and coating before imaging.
• Review of cryo-SEM equipment and methodologies for biofilm analysis.

Main Results:

• Cryo-SEM provides rapid sample preparation, yielding native-like biofilm structures.
• The technique is suitable for imaging microbiological components in various contexts.
• Discussion of specific workflow steps' suitability for different biofilm research.

Conclusions:

• Cryo-SEM is a powerful technique for visualizing biofilms with minimal structural distortion.
• Understanding the cryo-SEM workflow is crucial for successful biofilm imaging.
• Further application of cryo-SEM to medically relevant biofilms is encouraged.