Confocal macromolecules

Confocal scanning Examination of cells in Cryoelectron Imaging of biological macromolecules in the... 

Eukaryotic cells utilize an efficient transport system that delivers macromolecules fast and secure to their destination. In the case of the small GTP binding proteins of the Ras family the modified C-terminus seems to be sufficient for addressing the polypeptide to its target membrane (in the case of Ras itself the plasma membrane). Lipopeptides with the C-terminal structure of N-Ras (either a pen-tamer with a C-terminal carboxymethylation and farnesylation or a heptapeptide with a palmitoyl thioester in addition) and a N-terminal 7-nitrobenz-2-oxa-l,3-diazolyl (NBD) fluorophore were microin-jected into NIH3T3 fibroblast cells and the distribution of the fluorophore was monitored by confocal laser fluorescence microscopy. Enrichment of the protein in the plasma membrane was efficient only for peptides with two hydrophobic modification sites, while the farnesylated but not palmitoylated peptide was distributed in the cytosol.1121... 

De Smedt S, Meyvis E, van Oostveldt P, Blonk J, Hennink W, Demeester J. The diffusion of macromolecules in dextran methacrylate solutions and gels as studied by confocal scanning laser microscopy. Macromolecules 1997, 30, 4863-4870. 

Watkins, A. W. Anseth, K. S. Investigation of molecular transport and distributions in polyfethylene glycol) hydrogels with confocal laser scanning microscopy. Macromolecules. 2005, 38, 1326-1334. 

FCS is a type of spectroscopy that is based on the measurement of fluorescence intensity and the analysis of its fluctuations, which result from diffusion of the observed fluorophore in the excitation volume or changes in the fluorescence quantum yield arising from chemical reactions. Measurements are usually made on only a few molecules at a time, of the order of 10 milliseconds. This is achieved by illuminating tiny sample volumes ( 1 femtoliter) by employing two-photon excitation microscopy (see section Two-Photon Confocal Microscopy above). FCS is used to measure translational diffusion coefficients of macromolecules, the number of fluorescent molecules under observation and the... 

Ma, Y Farinha, J. P. S. Winnik, M. A. Yaneff, P. V. Ryntz, R. A. Compatihihty of chlorinated polyolefin with the components of thermoplastic polyolefin A study by laser scaiming confocal fluorescence microscopy. Macromolecules 2004, 37, 6544—6552. 

MA2 Maeda, Y., Yamamoto, H., and Ikeda, L, Phase separation of aqueous solutions of poly(iV-isopropylacrylamide) investigated by confocal Raman microscopy, Macromolecules, 36, 5055, 2003. 

---