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Biocell ; 32(1): 1-8, Apr. 2008. ilus
Article Dans Anglais | LILACS | ID: lil-541117

Résumé

Cellular and subcellular organization and distribution of actin filaments have been studied with various techniques. The use of fluorescence photo-oxidation combined with phalloidin conjugates with eosin has allowed the examination of the precise cellular and subcellular location of F-actin. Correlative fluorescence light microscopy and transmission electron microscopy studies of F-actin distribution are facilitated with this method for morphological and physiological studies. Because phalloidin-eosin is smaller than other markers, this method allows the analysis of the three-dimensional location of F-actin with high-resolution light microscopy, three-d serial sections reconstructions, and electron tomography. The combination of selective staining and three-dimensional reconstructions provide a valuable tool for revealing aspects of the synaptic morphology that are not available when conventional electron microscopy is used. By applying this selective staining technique and three-dimensional imaging, we uncovered the structural organization of actin in the postsynaptic densities in physiological and pathological conditions.


Sujets)
Humains , Animaux , Actines/métabolisme , Éosine jaunâtre/pharmacologie , Éosine jaunâtre/métabolisme , Photooxidation , Système nerveux central/métabolisme , Système nerveux central/ultrastructure , Coloration et marquage/méthodes , Colorants fluorescents/pharmacologie , Phalloïdine/pharmacologie , Imagerie tridimensionnelle/méthodes , Modèles moléculaires , Cytosquelette d'actine/métabolisme , Cytosquelette d'actine/ultrastructure , Microscopie de fluorescence/méthodes , Oxydoréduction , Photons
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