Multifocal two-photon laser scanning microscopy combined with photo-activatable GFP for in vivo monitoring of intracellular protein dynamics in real time

Martini J, Schmied K, Palmisano R, Tönsing K, Anselmetti D, Merkle T (2007)
Journal of Structural Biology 158(3): 401-409.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
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Abstract / Bemerkung
We used multifocal two-photon laser scanning microscopy for local and selective protein activation and quantitative investigation of intracellular protein dynamics. The localized activation was realized with photo-activatable green-fluorescent-proteins (pa-GFP) and optical two-photon excitation in order to investigate the real-time intracellular dynamics in vivo. Such processes are of crucial importance for a deep understanding and modelling of regulatory and metabolic processes in living cells. Exemplarily, the intracellular dynamics of the Arabidopsis MYB transcription factor LHY/CCA1-like 1 (LCL1) that contains both a nuclear import and a nuclear export signal was quantitatively investigated. We used tobacco BY-2 protoplasts co-transfected with plasmids encoding photo-activatable green fluorescent protein (pa-GFP) fusion proteins and a red fluorescing transfection marker and measured the rapid nuclear export of pa-GFP-LCL1 I after its photo-activation in the nucleus. In contrast, an export-negative mutant of LCL1 remained trapped inside the nucleus. We detemined average time constants of 51s and 125s for the decrease of fluorescence in the nucleus due to active bi-directional nuclear transport of pa-GFP-LCL1 and diffusion of pa-GFP, respectively. (c) 2007 Elsevier Inc. All rights reserved.
Erscheinungsjahr
Zeitschriftentitel
Journal of Structural Biology
Band
158
Zeitschriftennummer
3
Seite
401-409
ISSN
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Martini J, Schmied K, Palmisano R, Tönsing K, Anselmetti D, Merkle T. Multifocal two-photon laser scanning microscopy combined with photo-activatable GFP for in vivo monitoring of intracellular protein dynamics in real time. Journal of Structural Biology. 2007;158(3):401-409.
Martini, J., Schmied, K., Palmisano, R., Tönsing, K., Anselmetti, D., & Merkle, T. (2007). Multifocal two-photon laser scanning microscopy combined with photo-activatable GFP for in vivo monitoring of intracellular protein dynamics in real time. Journal of Structural Biology, 158(3), 401-409. doi:10.1016/j.jsb.2006.12.012
Martini, J., Schmied, K., Palmisano, R., Tönsing, K., Anselmetti, D., and Merkle, T. (2007). Multifocal two-photon laser scanning microscopy combined with photo-activatable GFP for in vivo monitoring of intracellular protein dynamics in real time. Journal of Structural Biology 158, 401-409.
Martini, J., et al., 2007. Multifocal two-photon laser scanning microscopy combined with photo-activatable GFP for in vivo monitoring of intracellular protein dynamics in real time. Journal of Structural Biology, 158(3), p 401-409.
J. Martini, et al., “Multifocal two-photon laser scanning microscopy combined with photo-activatable GFP for in vivo monitoring of intracellular protein dynamics in real time”, Journal of Structural Biology, vol. 158, 2007, pp. 401-409.
Martini, J., Schmied, K., Palmisano, R., Tönsing, K., Anselmetti, D., Merkle, T.: Multifocal two-photon laser scanning microscopy combined with photo-activatable GFP for in vivo monitoring of intracellular protein dynamics in real time. Journal of Structural Biology. 158, 401-409 (2007).
Martini, Joerg, Schmied, Katja, Palmisano, Ralf, Tönsing, Katja, Anselmetti, Dario, and Merkle, Thomas. “Multifocal two-photon laser scanning microscopy combined with photo-activatable GFP for in vivo monitoring of intracellular protein dynamics in real time”. Journal of Structural Biology 158.3 (2007): 401-409.

14 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Identification of 30 MYB transcription factor genes and analysis of their expression during abiotic stress in peanut (Arachis hypogaea L.).
Chen N, Yang Q, Pan L, Chi X, Chen M, Hu D, Yang Z, Wang T, Wang M, Yu S., Gene 533(1), 2014
PMID: 24013078
Fluorescent proteins for live-cell imaging with super-resolution.
Nienhaus K, Nienhaus GU., Chem Soc Rev 43(4), 2014
PMID: 24056711
Advances in multiphoton microscopy technology.
Hoover EE, Squier JA., Nat Photonics 7(2), 2013
PMID: 24307915
A new set of reversibly photoswitchable fluorescent proteins for use in transgenic plants.
Lummer M, Humpert F, Wiedenlübbert M, Sauer M, Schüttpelz M, Staiger D., Mol Plant 6(5), 2013
PMID: 23434876
Characterization and drug resistance patterns of Ewing's sarcoma family tumor cell lines.
May WA, Grigoryan RS, Keshelava N, Cabral DJ, Christensen LL, Jenabi J, Ji L, Triche TJ, Lawlor ER, Reynolds CP., PLoS One 8(12), 2013
PMID: 24312454
Nucleo-cytoplasmic transport of proteins and RNA in plants.
Merkle T., Plant Cell Rep 30(2), 2011
PMID: 20960203
Combining ocFLIM and FIDSAM reveals fast and dynamic physiological responses at subcellular resolution in living plant cells.
Elgass K, Caesar K, Harter K, Meixner AJ, Schleifenbaum F., J Microsc 242(2), 2011
PMID: 21118238
Patterned two-photon photoactivation illuminates spatial reorganization in live cells.
Smith AW, Smoligovets AA, Groves JT., J Phys Chem A 115(16), 2011
PMID: 21391691
Fluorescent proteins and their applications in imaging living cells and tissues.
Chudakov DM, Matz MV, Lukyanov S, Lukyanov KA., Physiol Rev 90(3), 2010
PMID: 20664080
Nucleocytoplasmic distribution of the Arabidopsis chromatin-associated HMGB2/3 and HMGB4 proteins.
Pedersen DS, Merkle T, Marktl B, Lildballe DL, Antosch M, Bergmann T, Tönsing K, Anselmetti D, Grasser KD., Plant Physiol 154(4), 2010
PMID: 20940346
High-resolution mosaic imaging with multifocal, multiphoton photon-counting microscopy.
Chandler E, Hoover E, Field J, Sheetz K, Amir W, Carriles R, Ding SY, Squier J., Appl Opt 48(11), 2009
PMID: 19363544
Novel application of fluorescence lifetime and fluorescence microscopy enables quantitative access to subcellular dynamics in plant cells.
Elgass K, Caesar K, Schleifenbaum F, Stierhof YD, Meixner AJ, Harter K., PLoS One 4(5), 2009
PMID: 19492078
Chromosomal high mobility group (HMG) proteins of the HMGB-type occurring in the moss Physcomitrella patens.
Kiilerich B, Stemmer C, Merkle T, Launholt D, Gorr G, Grasser KD., Gene 407(1-2), 2008
PMID: 17980517

27 References

Daten bereitgestellt von Europe PubMed Central.

Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation.
Zipfel WR, Williams RM, Christie R, Nikitin AY, Hyman BT, Webb WW., Proc. Natl. Acad. Sci. U.S.A. 100(12), 2003
PMID: 12756303
Nonlinear magic: multiphoton microscopy in the biosciences.
Zipfel WR, Williams RM, Webb WW., Nat. Biotechnol. 21(11), 2003
PMID: 14595365

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