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  • Title: A tridimensional view of the organization of actin filaments in the central nervous system by use of fluorescent photooxidation.
    Author: Capani F, Saraceno E, Boti VR, Aon-Bertolino L, Fernández JC, Gato F, Kruse MS, Giraldez L, Ellisman MH, Coirini H.
    Journal: Biocell; 2008 Apr; 32(1):1-8. PubMed ID: 18669318.
    Abstract:
    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.
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