Green fluorescent protein microscopy

Shimomura, O. (2005a). The discovery of aequorin and green fluorescent protein. J. Microscopy 217 3-15. 

Terry, B. R., Matthews, E. K., and Haseloff, J. (1995). Molecular characterization of recombinant green fluorescent protein by fluorescent correlation microscopy. Biochem. Biophys. Res. Commun. 217 21—27. 

Patterson GF1, Knobel SM, Sharif WD, Rain SR, Piston DW (1997) Use of the green fluorescent protein and its mutants in quantitative fluorescence microscopy. Biophys J 73 2782-2790... 

Squire, A., Verveer, P. J., Pocks, O. and Bastiaens, P. I. H. (2004). Red-edge anisotropy microscopy enables dynamic imaging of homo-FRET between green fluorescent proteins in cells. J. Struct. Biol. 147, 62-9. 

Since about the mid-1990s, the green fluorescent protein (GFP), and its spectral variants [1] have become some of the most exciting molecules in microscopy, biochemistry, and cell biology (see Chapter 5). 

Pepperkok, R., Squire, A., Geley, S. and Bastiaens, P. I. (1999). Simultaneous detection of multiple green fluorescent proteins in live cells by fluorescence lifetime imaging microscopy. Curr. Biol. 9, 269-72. 

Lansford, R., Bearman, G., and Fraser, S. E. 2001. Resolution of multiple green fluorescent protein color variants and dyes using two-photon microscopy and imaging spectroscopy. J. Biomed. Opt. 6(3) 311-18. 

Figure 9.17 Green fluorescent protein (GFP) synthesis in water-in-oil emulsion as visualized by fluorescence microscopy. (Adapted from Pietrini and Luisi, 2004). Shown are the compartments in which GFP has been expressed (green in the original), (a) Typical micrographs of the cell-free GFP synthesis in Span 80 (0.45% v/v)/Tween 80 (0.05% v/v)/aqueous solution (0.5% v/v) in mineral oil emulsion droplets, preparation at 4 °C incubation at 37°C (i) 0 min, (ii) 11 min, (iii) 23 min, (iv) 32 min, (v) 44 min, (vi) 57 min, (vii) 21 h. Negative control (viii) 0 min, (ix) 21 h. The bar represents 50 p.m. (b) Kinetics of the cell-free GFP synthesis in emulsion droplets, on average 10 droplets with diameters of 30-60 um are evaluated per time point, cell-free enhanced GFP synthesis in emulsion droplets (i, ii and iii are three independent experiments) and negative controi (iv and v are two independent experiments).

Green fluorescent protein (GFP) and related fluorescent proteins can be used to label practically any protein or subcellular compartment of living cells (49). Transfection of cells with plasmids that encode appropriately targeted fluorescent fusion proteins has been used to define the plasma membrane, early endosomes, late endosomes, caveolae, the golgi complex, the ER, and other subcellular locations. Several fluorescent small molecules are also available for labehng specific cellular organelles, including endosomes and lysosomes, for analysis by fluorescence microscopy. 

Patterson GH, Knobel SM, Sharif WD, Kain SR, Piston DW. Use of the green fluorescent protein and its mutants in quantitative fluorescence microscopy. Biophys. J. 1997 73 2782-2790. Kummer AD, Kompa C, Lossau H, POllinger-Dammer F, Michel-Beyerle ME, Silva CM, Bylina EJ, Coleman WJ, Yang MM, Youvan DC. Dramatic reduction in fluorescence quantum yield in mutants of green fluorescent protein due to fast internal conversion. Chem. Phys. 1998 237 183-193. 

Volkmer A, Subramaniam V, Birch DJS, Jovin TM. One- and two-photon excited fluorescence lifetimes and anisotropy decays of green fluorescent proteins. Biophys. J. 2000 78 1589-1598. Subramaniam V, Hanley QS, Clayton AHA, Jovin TM. Photophysics of green and red fluorescent proteins implications for quantitative microscopy. Methods Enzymol. 2003 360 178-201. Rizzo MA, Springer GH, Granada B, Fdston DW. An improved cyan fluorescent protein variant useful for FRET. Nature Biotechnol. 2004 22 445-449. 

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