title: Fluorescence photoconversion of biarsenical-labeled cells for correlated electron microscopy (EM).
abstract: Correlated light microscopy (LM)/electron microscopy (EM) analysis can be achieved by using biarsenical dyes to fluorescently label tetracysteine-tagged proteins. Once live cell imaging using LM is complete, cellular activity can be halted promptly using a glutaraldehyde-based fixative. Rapid fixation preserves cellular ultrastructure and limits diffusion of reaction products. This protocol provides details on rapid fixation of cells, followed by fluorescence photoconversion of 3,3'-diaminobenzidine tetrahydrochloride (DAB) and sample processing for EM that can be correlated with the live cell LM images.
authors.forename: Guido M
authors.initials: GM
authors.lastname: Gaietta
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Deerinck
authors.forename: Mark H
authors.initials: MH
authors.lastname: Ellisman
mesh_terms: D001152:Arsenicals; D002460:Cell Line; D005456:Fluorescent Dyes; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008854:Microscopy, Electron / Q000379:methods; D011994:Recombinant Proteins / Q000032:analysis*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D001152:Arsenicals; D005456:Fluorescent Dyes; D011994:Recombinant Proteins