Fluorescing the electron: strategies in correlative experimental design

Area of Science:

• Cell Biology
• Microscopy
• Biophysics

Background:

• Correlative light and electron microscopy (CLEM) combines live-cell imaging and fluorescence microscopy with high-resolution electron microscopy (EM).
• CLEM enables visualization of molecular localization and cellular ultrastructure simultaneously.
• Advancements in CLEM are crucial for understanding complex cellular processes.

Purpose of the Study:

• To review common strategies employed in CLEM.
• To detail sample preparation and imaging workflows for CLEM applications.
• To highlight the benefits and applications of different CLEM approaches.

Main Methods:

• Review of three primary CLEM strategies: time-resolved CLEM, cell sorting for EM, and spatial CLEM.
• Discussion of sample preparation techniques tailored for each strategy.
• Illustration of workflows for combining light and electron microscopy.

Main Results:

• Time-resolved CLEM allows live-cell imaging integrated with EM resolution.
• Cell sorting enables targeted EM analysis of fluorescently labeled cells.
• Spatial CLEM facilitates correlation of fluorescent features with ultrastructure.

Conclusions:

• CLEM offers powerful approaches for bridging light and electron microscopy.
• The choice of CLEM strategy depends on the specific research question and sample.
• Future directions include integrating super-resolution microscopy with EM for enhanced CLEM capabilities.