CLEM with Commercially Available Reagents to Facilitate Immunolocalization. (July 2020)

• Record Type: Journal Article
• Authors: Cummins, Leslie; Tu, Vincent; Mayoral, Joshua; Weiss, Louis M.; Macaluso, Frank P.; DesMarais, Vera
• Is Part Of: Microscopy today. Volume 28:Number 4(2020)
• Page Start: 20
• Page End: 23
• Publication Date: 2020-07
• Subjects: transmission electron microscopy, -- immunoelectron microscopy, -- fluorescence microscopy, -- correlative microscopy, -- Toxoplasma gondii; Microscope and microscopy -- Periodicals; 502.8
• DOI: 10.1017/S1551929520001078
• Languages: English
• ISSNs: 1551-9295

Abstract

Abstract: 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.