Shedding light onto live molecular targets

Weissleder, Ralph; Ntziachristos, Vasilis

doi:10.1038/nm0103-123

Cited by 1,773 publications

(1,352 citation statements)

References 83 publications

(86 reference statements)

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“…Likewise, it can be expected that functional imaging will further develop to image molecular processes within cells of complete organisms (Weissleder and Ntziachristos 2003). Functional Xuorescence microscopy is also suitable in a highthroughput setting and it can be expected that this will play a signiWcant role in elucidating the causality in biochemical network structure in the near future by directly probing protein reaction states in living cells.…”

Section: Resultsmentioning

confidence: 99%

Quantitative microscopy and systems biology: seeing the whole picture

Verveer

Bastiaens

2008

Histochem Cell Biol

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Understanding cellular function requires studying the spatially resolved dynamics of protein networks. From the isolated proteins we can only learn about their individual properties, but by investigating their behavior in their natural environment, the cell, we obtain information about the overall response properties of the network module in which they operate. Fluorescence microscopy methods provide currently the only tools to study the dynamics of molecular processes in living cells with high temporal and spatial resolution. Combined with computational approaches they allow us to obtain insights in the reactiondiVusion processes that determine biological function on the scale of cells.

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Section: Resultsmentioning

confidence: 99%

Quantitative microscopy and systems biology: seeing the whole picture

Verveer

Bastiaens

2008

Histochem Cell Biol

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“…For biological applications, it is preferably that such complexes allow for sensitization of the metal using the lowest possible energy (longer wavelength) in order to minimize autofluorescence and to increase the penetration depth of excitation into the sample. 12 Another recently described Eu(III) chelate, which has the advantage of red shifted excitation at 335 nm, was reported by Matsumoto et. al.…”

Section: Introductionmentioning

confidence: 99%

Highly Luminescent Lanthanide Complexes of 1-Hydroxy-2-pyridinones

et al. 2008

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The synthesis, X-ray structure, stability, and photophysical properties of several trivalent lanthanide complexes formed from two differing bis-bidentate ligands incorporating either alkyl or alkyl ether linkages and featuring the 1-hydroxy-2-pyridinone (1,2-HOPO) chelate group in complex with Eu(III), Sm(III) and Gd(III) are reported. The Eu(III) complexes are among some of the best examples, pairing highly efficient emission ( Eu tot Φ ~ 21.5 %,) with high stability (pEu ~ 18.6) in aqueous solution, and are excellent candidates for use in biological assays. A comparison of the observed behavior of the complexes with differing backbone linkages shows remarkable similarities, both in stability and photophysical properties. Low temperature photophysical measurements for a Gd(III) complex were also used to gain insight into the electronic structure, and were found to agree with corresponding TD-DFT calculations for a model complex. A comparison of the high resolution Eu(III) emission spectra in solution and from single crystals also revealed a more symmetric coordination geometry about the metal ion in solution due to dynamic rotation of the observed solid state structure.

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“…1 So far, three far-red fluorescent proteins have been discovered, all with emission maximum (k max em ) of 650 nm: HcRed (k max em ¼ 645 nm), 2 aeFP595 AQ143 (k max em ¼ 655 nm) 3 and mPlum (k max em ¼ 649 nm). 4 Of these, only mPlum is monomeric and it also exhibits the highest quantum yield, about 0.10.…”

Section: Introductionmentioning

confidence: 99%

Unique interactions between the chromophore and glutamate 16 lead to far‐red emission in a red fluorescent protein

et al. 2009

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mPlum is a far-red fluorescent protein with emission maximum at~650 nm and was derived by directed evolution from DsRed. Two residues near the chromophore, Glu16 and Ile65, were previously revealed to be indispensable for the far-red emission. Ultrafast time-resolved fluorescence emission studies revealed a time dependent shift in the emission maximum, initially about 625 nm, to about 650 nm over a period of 500 ps. This observation was attributed to rapid reorganization of the residues solvating the chromophore within mPlum. Here, the crystal structure of mPlum is described and compared with those of two blue shifted mutants mPlum-E16Q and -I65L. The results suggest that both the identity and precise orientation of residue 16, which forms a unique hydrogen bond with the chromophore, are required for far-red emission. Both the far-red emission and the time dependent shift in emission maximum are proposed to result from the interaction between the chromophore and Glu16. Our findings suggest that significant red shifts might be achieved in other fluorescent proteins using the strategy that led to the discovery of mPlum.

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Shedding light onto live molecular targets

Cited by 1,773 publications

References 83 publications

Quantitative microscopy and systems biology: seeing the whole picture

Quantitative microscopy and systems biology: seeing the whole picture

Highly Luminescent Lanthanide Complexes of 1-Hydroxy-2-pyridinones

Unique interactions between the chromophore and glutamate 16 lead to far‐red emission in a red fluorescent protein

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