Correlative light and electron microscopy (CLEM) is a collection of methods that allows us to locate a molecule of interest within a high-resolution cellular map.

This e-learning video focuses on a specific method where we preserve the fluorescence throughout the electron microscopy (EM) sample preparation. This allows us to image the same section with the light and electron microscope, so that we can overlay the information coming from both imaging modalities with very high precision.

Target audience

This video is targeted at scientists interested in subcellular localisation of a protein of interest by combining light and electron microscopy.

This video takes you step-by-step through the method. However, because of its complexity, a basic level of knowledge in sample preparation for EM and ultramicrotomy would be advantageous.

Learning objectives

• Prepare sample used for on-section CLEM
• Evaluate tricks for light and electron microscopy imaging
• Use methods to obtain high precision correlation of the light and electron microscopy images using special beads/fiducials and dedicated software solutions

CLEM protocols

• Precise, correlated fluorescence microscopy and electron tomography of lowicryl sections using fluorescent fiducial markers. Kukulski W, et al. (2012). Methods Cell Biol. 111. 235-57. Europe PMC link

• eC-CLEM: A multidimension, multimodel software to correlate intermodal images with a focus on light and electron microscopy. Heiligenstein X, et al. (2017) Methods Cell Biol. 140. 335-52. Europe PMC link

Feedback

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Project team

Project manager: Jacqueline Dreyer-Lamm

Executive producer, scientific advisor and voice over: Rachel Mellwig

Technical guidance: Pedro Machado & Paolo Ronchi

Editor and scientific oversight: Yannick Schwab

Video producer and motion graphics: Claudiu Grozea