Visualizing membrane traffic in vivo by combined video fluorescence and 3D electron microscopy.
In studies of dynamic cellular processes, it would be ideal to be able to combine the capability of in vivo fluorescence video microscopy with the power of resolution of electron microscopy (EM). This article describes an approach based on the association of these two techniques, by which an individual intracellular structure can be monitored in vivo, typically through the use of markers fused with green-fluorescent protein, and then analysed by EM and three-dimensional reconstruction methods, resulting in a 'snapshot' of its fine structure at any chosen time in its life cycle. The potential of this approach is discussed in relation to various aspects of cell biology and especially to the question of the morpho-functional organization of the intracellular membrane trafficking pathways.
Mironov
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AA
Dept of Cell Biology and Oncology, Istituto di Ricerche Farmacologiche 'Mario Negri', Consorzio Mario Negri Sud, Maria Imbaro (Chieti), Italy.
Polishchuk
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RS
Luini
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A
D000818:Animals; D001692:Biological Transport; D005453:Fluorescence; D049452:Green Fluorescent Proteins; D006801:Humans; D007425:Intracellular Membranes / Q000648:ultrastructure*; D008164:Luminescent Proteins; D008854:Microscopy, Electron / Q000379:methods*; D018715:Microscopy, Video / Q000379:methods*
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins