Work

CLEM with Commercially Available Reagents to Facilitate Immunolocalization

Year: 2020

Type: article

Abstract: There is often a need to locate the same cellular structure of interest in light and electron microscopy, which can be a difficult task. Here we present a method that uses only commercially available reagents and standard epi-fluorescence and transmission electron microscopy (TEM) technology to make correlative light and electron microscopy (CLEM) available to a large group of researchers without specialized CLEM hardware. This was achieved by seeding cells on photo-etched gridded cover slips and staining the protein to be localized with a secondary antibody coupled to both a fluorophore and 10 nm gold. The presence of the grid allowed for the alignment of light microscopy images with TEM images and the double-labeled antibody revealed co-localization of the fluorophore with gold particles.

Source: Microscopy Today

DOI: 10.1017/s1551929520001078

Authors: Leslie Cummins, Vincent Tu, Joshua Mayoral, Louis M. Weiss, Frank P. Macaluso, Vera DesMarais

Institution: Albert Einstein College of Medicine

Language: English

Topic: Advanced Electron Microscopy Techniques and Applications

Subfield: Structural Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: bronze

Funder: National Cancer Institute

Award: Cancer Center Support Grant