Protein localization in electron micrographs using fluorescence nanoscopy.
Nat Methods
; 8(1): 80-4, 2011 Jan.
Article
em En
| MEDLINE
| ID: mdl-21102453
ABSTRACT
A complete portrait of a cell requires a detailed description of its molecular topography proteins must be linked to particular organelles. Immunocytochemical electron microscopy can reveal locations of proteins with nanometer resolution but is limited by the quality of fixation, the paucity of antibodies and the inaccessibility of antigens. Here we describe correlative fluorescence electron microscopy for the nanoscopic localization of proteins in electron micrographs. We tagged proteins with the fluorescent proteins Citrine or tdEos and expressed them in Caenorhabditis elegans, fixed the worms and embedded them in plastic. We imaged the tagged proteins from ultrathin sections using stimulated emission depletion (STED) microscopy or photoactivated localization microscopy (PALM). Fluorescence correlated with organelles imaged in electron micrographs from the same sections. We used these methods to localize histones, a mitochondrial protein and a presynaptic dense projection protein in electron micrographs.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Microscopia Eletrônica
/
Nanotecnologia
/
Proteínas Luminescentes
/
Microscopia de Fluorescência
Limite:
Animals
Idioma:
En
Revista:
Nat Methods
Assunto da revista:
TECNICAS E PROCEDIMENTOS DE LABORATORIO
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Estados Unidos