Correlated fluorescence and 3D electron microscopy with high sensitivity and spatial precision.

Area of Science:

• Cell Biology
• Microscopy Techniques
• Biophysics

Background:

• Correlative electron and fluorescence microscopy offers potential for ultrastructural analysis of cellular events.
• Existing methods suffer from low precision and sensitivity, limiting their application.

Purpose of the Study:

• To develop a novel method for high-precision mapping of fluorescence signals to 3D electron tomograms.
• To enable visualization of dynamic and rare cellular processes at the ultrastructural scale.

Main Methods:

• Direct mapping of signals from approximately 20 fluorescent protein molecules to 3D electron tomograms.
• Achieving a mapping precision of less than 100 nm.

Main Results:

• Successfully identified individual HIV particles bound to mammalian cell surfaces.
• Visualized flared growing microtubule plus-ends in fission yeast.
• Localized Rvs167 to endocytic sites in budding yeast and characterized vesicle scission dynamics.

Conclusions:

• The developed method significantly enhances the precision and sensitivity of correlative microscopy.
• This technique facilitates direct correlation of fluorescence and electron microscopy for ultrastructural visualization of cellular processes.