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Pulse-chase methods offer powerful tools for following the evolution of a biological system over time, but are usually limited to ensemble measurements of the average behavior of very large numbers of cells. Here we describe three methods ranging from a true pulse-chase, through selective regional photoactivation, to pharmacological induction of an altered protein state, which can be applied to time-dependent studies at the single-cell level. These methods are exemplified by experimental protocols to follow region-selective nuclear envelope targeting of nascent phospholipids, a nascent nuclear lamin protein (lamin B1), and an immature lamin precursor (prelamin A).

Original publication




Journal article


Methods Mol Biol

Publication Date





159 - 176


Nucleoplasmic reticulum, Photoactivatable fluorescent proteins, Pulse-chase, Secondary ion mass spectrometry, Single-cell assays, Stable isotope labeling, Super-resolution light microscopy, Animals, Cell Nucleus, Cells, Cultured, Gene Expression, Genes, Reporter, Image Processing, Computer-Assisted, Mice, Microscopy, Electron, Microscopy, Fluorescence, Molecular Imaging, Nuclear Proteins, Protein Transport, Single-Cell Analysis, Staining and Labeling