Relocation is the key to successful correlative fluorescence and scanning electron microscopy

Area of Science:

• Cell Biology
• Microscopy Techniques
• Biophysics

Background:

• Correlative microscopy combines multiple imaging modalities to provide comprehensive sample analysis.
• Current methods can be time-consuming and lack high-throughput capabilities for nanoscale investigations.
• Automated solutions are needed to bridge length scales and improve efficiency in biological imaging.

Purpose of the Study:

• To develop and validate an automated hardware and software solution for swift correlative sample array mapping.
• To enable high-throughput, high-resolution imaging of fluorescently labeled molecules in cells and tissues.
• To demonstrate the system's utility in analyzing molecular expression differences and tissue composition.

Main Methods:

• Development of an automated sample-relocation holder for precise sample transfer between imaging platforms.
• Integration of wide-field optical/fluorescence microscopy with scanning electron microscopy.
• Application to in vitro HeLa cells labeled for monosialoganglioside one (GM-1) and zebrafish (Danio rerio) tissues.

Main Results:

• Achieved swift correlative fluorescence and electron microscopy analysis of HeLa cells within minutes of preparation.
• Enabled high-throughput, spatially resolved nanoscopic analysis of molecular composition in distinct cell populations.
• Demonstrated accurate relocation of fluorescent macromolecular complexes in zebrafish tissue sections for albumin distribution analysis.

Conclusions:

• The automated sample-relocation holder significantly enhances the efficiency and throughput of correlative microscopy.
• This approach facilitates answering complex biological questions by integrating multi-scale imaging data.
• The system supports rigorous, statistically relevant, results-driven research in cell biology and tissue analysis.