Hybrid fluorescence and electron cryo-microscopy for simultaneous electron and photon imaging

Area of Science:

• Correlative microscopy
• Integrated optical and electron imaging systems
• Advanced microscopy techniques

Background:

• Correlative microscopy traditionally requires separate fluorescence and electron microscopes.
• Integrating both into a single device presents challenges in objective lens arrangement for simultaneous specimen focusing.
• Previous methods involved specimen movement (lateral displacement or rotation) for sequential imaging, preventing simultaneous acquisition.

Purpose of the Study:

• To present a versatile approach for integrating fluorescence and transmission electron microscopy into a single device.
• To enable simultaneous imaging capabilities in correlative microscopy.
• To overcome the limitations of sequential imaging in existing correlative microscopy setups.

Main Methods:

• Development of an integrated microscopy device combining fluorescence light and transmission electron microscopy.
• Utilized a reflection mirror to facilitate simultaneous imaging.
• Demonstrated the system's potential through imaging experiments.

Main Results:

• Successfully demonstrated simultaneous hybrid microscopy.
• Achieved sequential imaging of fluorescent proteins in cells using integrated fluorescence and electron microscopy.
• Performed cathodoluminescence imaging of fluorescent beads within the same integrated system.

Conclusions:

• The developed integrated device offers a versatile solution for simultaneous correlative microscopy.
• This approach advances the field by enabling combined light and electron imaging in a single instrument.
• The system has potential applications in various biological and materials science imaging studies.