Correlative video light/electron microscopy.
This unit describes newly developed methods that allow the examination of living cells by time-lapse analysis with the subsequent identification of the just-observed organelle under an electron microscope. To understand how such cellular functions, such as intracellular traffic, cytokinesis, and cell migration, are organized and executed in vivo, it is most useful to observe living cells in real time with the spatial resolution afforded by electron microscopy (EM). Most suitable for this is a conceptually simple, yet powerful, method called correlative video light/electron microscopy (CVLEM), by which observations of the in vivo dynamics and the ultrastructure of intracellular objects can indeed be combined to achieve the above-mentioned result. This unit describes this methodology, illustrates the type of questions that the CVLEM approach was designed to address, and discusses the expertise required for successful application of the technique.
Polishchuk
R S
RS
Consorzio Mario Negri Sud, S. Maria Imbara (Chieti), Italy.
Mironov
A A
AA
D000818:Animals; D002477:Cells / Q000648:ultrastructure*; D003199:Computer Systems; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D021621:Imaging, Three-Dimensional; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D018613:Microscopy, Confocal / Q000379:methods; D008854:Microscopy, Electron / Q000379:methods*; D016707:Tissue Fixation / Q000379:methods; D014162:Transfection; D014741:Video Recording
D049452:Green Fluorescent Proteins