Keywords
- Cross-correlation
- E222Q mutant
- Fluorescence correlation spectroscopy
- Green fluorescent protein
- Photodynamics
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC (1994) Green fluorescent protein as a marker for gene expression. Science 263:802–805
Tsien RY (1998) The green fluorescent protein. Annu Rev Biochem 67:509–544
Jung G, Zumbusch A (2006) Improving autofluorescent proteins: comparative studies of the effective brightness of green fluorescent protein (GFP) mutants. Micr Res Tech 69:175–185
Hsu ST, Blaser G, Jackson SE (2009) The folding, stability and conformational dynamics of beta-barrel fluorescent proteins. Chem Soc Rev 38:2951–2965
Miyawaki A, Sawano A, Kogure T (2003) Lighting up cells: labelling proteins with fluorophores. Nat Cell Biol Suppl S1–7
Lippincott-Schwartz J, Patterson GH (2003) Development and use of fluorescent protein markers in living cells. Science 300:87–91
Chudakov DM, Belousov VV, Zaraisky AG, Novoselov VV, Staroverov DB, Zorov DB, Lukyanov S, Lukyanov KA (2003) Kindling fluorescent proteins for precise in vivo photolabeling. Nat Biotechnol 21:191–194
Post JN, Lidke KA, Rieger B, Anrdt-Jovin DJ (2005) One- and two-photon photoactivation of a paGFP-fusion protein in live Drosophila embryos. FEBS Lett 579:325–330
Kim PK, Mullen RT, Schumann U, Lippincott-Schwartz J (2006) The origin and maintenance of mammalian peroxisomes involves a de novo PEX16-dependent pathway from the ER. J Cell Biol 173:521–532
Dickson RM, Cubitt AB, Tsien RY, Moerner WE (1997) On/off blinking and switching behavior of single molecules of green fluorescent protein. Nature 388:355–358
Day RN, Davidson MW (2009) The fluorescent protein palette: tools for cellular imaging. Chem Soc Rev 38:2887–2921
Patterson GH, Lippincott-Schwartz J (2002) A photoactivatable GFP for selective photolabeling of proteins and cells. Science 297:1873–1877
Lippincott-Schwartz J, Patterson GH (2008) Fluorescent proteins for photoactivation experiments. Methods Cell Biol 85:45–61
Betzig E, Patterson GH, Sougrat R, Lindwasser OW, Olenych S, Bonifacino JS, Davidson MW, Lippincott-Schwartz J, Hess HF (2006) Imaging intracellular fluorescent proteins at nanometer resolution. Science 313:1642–1645
Ando R, Hama H, Yamamoto-Hino M, Mizuno H, Miyawaki A (2002) An optical marker based on the UV-induced greento- red photoconversion of a fluorescent protein. Proc Natl Acad Sci USA 99:12651–12656
Ando R, Mizuno H, Miyawaki A (2004) Regulated fast nucleocytoplasmic shuttling observed by reversible protein highlighting. Science 306:1370–1373
Lippincott-Schwartz J, Altan-Bonnet N, Patterson GH (2003) Photobleaching and photoactivation: following protein dynamics in living cells. Nat Cell Biol Suppl S7–14
Cinelli RAG, Pellegrini V, Ferrari A, Faraci P, Nifosi R, Tyagi M, Giacca M, Beltram F (2001) Green fluorescent proteins as optically controllable elements in bioelectronics. Appl Phys Lett 79:3353–3355
Bizzarri R, Serresi M, Cardarelli F, Abbruzzetti S, Campanini B, Viappiani C, Beltram F (2010) Single amino acid replacement makes Aequorea victoria fluorescent proteins reversibly photoswitchable. J Am Chem Soc 132:85–95
Habuchi S, Ando R, Dedecker P, Verheijen W, Mizuno H, Miyawaki A, Hofkens J (2005) Reversible single-molecule photoswitching in the GFP-like fluorescent protein Dronpa. Proc Natl Acad Sci USA 102:9511–9516
Hell SW, Jakobs S, Kastrup L (2003) Imaging and writing at the nanoscale with focused visible light through saturable optical transitions. Appl Phys A Mater Sci Process 77:859–860
Hofmann M, Eggeling C, Jakobs S, Hell SW (2005) Breaking the diffraction barrier in fluorescence microscopy at low light intensities using reversibly photoswitchable proteins. Proc Natl Acad Sci USA 102:17565–17569
Subach FV, Patterson GH, Manley S, Gillette JM, Lippincott-Schwartz J, Verkhusha VV (2009) Photoactivatable mCherry for high-resolution two-color fluorescence microscopy. Nat Methods 6:153–159
Hess ST, Girirajan TP, Mason MD (2006) Ultra-high resolution imaging by fluorescence photoactivation localization microscopy. Biophys J 91:4258–4272
Rust MJ, Bates M, Zhuang X (2006) Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat Methods 3:793–795
Egner A, Geisler C, von Middendorff C, Bock H, Wenzel D, Medda R, Andresen M, Stiel AC, Jakobs S, Eggeling C, Schönle A, Hell SW (2007) Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters. Biophys J 93:3285–3290
Flors C, Hotta J, Uji-i H, Dedecker P, Ando R, Mizuno H, Miyawaki A, Hofkens J (2007) A stroboscopic approach for fast photoactivation-localization microscopy with Dronpa mutants. J Am Chem Soc 129:13970–13977
Hofmann M, Eggeling C, Jakobs S, Hell SW (2005) Breaking the diffraction barrier in fluorescence microscopy at low light intensities by using reversibly photoswitchable proteins. PNAS 102:17565–17569
Luin S, Voliani V, Lanza G, Bizzarri R, Amat P, Tozzini V, Serresi M, Beltram F (2009) Raman study of cromophore states in photochromic fluorescent proteins. J Am Chem Soc 131:96–103
Andresen M, Stiel AC, Trowitzsch S, Weber G, Eggeling C, Wahl MC, Hell SW, Jakobs S (2007) Structural basis for reversible photoswitching in Dronpa. Proc Natl Acad Sci USA 104:13005–13009
Andresen M, Wahl MC, Stiel AC, Gräter F, Schäfer LV, Trowitzsch S, Weber G, Eggeling C, Grubmüller H, Hell SW, Jakobs S (2005) Structure and mechanism of the reversible photoswitch of a fluorescent protein. Proc Natl Acad Sci USA 102:13070–13074
Stiel AC, Trowitzsch S, Weber G, Andresen M, Eggeling C, Hell SW, Jakobs S, Wahl MC (2007) 1.8 Å bright-state structure of the reversibly switchable fluorescent protein Dronpa guides the generation of fast switching variants. Biochem J 402:35–42
Moerner WE (2002) Single-molecule optical spectroscopy of autofluorescent proteins. J Chem Phys 117:10925–10937
Haupts U, Maiti S, Schwille P, Webb WW (1998) Dynamics of fluorescence fluctuations in green fluorescent protein observed by fluorescence correlation spectroscopy. Proc Natl Acad Sci USA 95:13573–13578
Moerner WE, Fromm DP (2003) Methods of single-molecule fluorescence spectroscopy and microscopy. Rev Sci Inst 74:3597–3619
Haustein E, Schwille P (2007) Fluorescence correlation spectroscopy: novel variations of an established technique. Annu Rev Biophys Biomol Struct 36:151–169
Marriott G, Mao S, Sakata T, Ran J, Jackson DK, Petchprayoon C, Gomez TJ, Warp E, Tulyathan O, Aaron HL, Isacoff EY, Yan Y (2008) Optical lock-in detection imaging microscopy for contrast-enhanced imaging in living cells. Proc Natl Acad Sci USA 105:17789–17794
Quercioli V, Bosisio C, Daglio SC, Rocca F, D'Alfonso L, Collini M, Baldini G, Chirico G, Bettati S, Raboni S, Campanini B (2010) Photoinduced millisecond switching kinetics in the GFPMut2 E222Q mutant. J Phys Chem B 114:4664–4677
Jung G, Brauchle C, Zumbusch A (2001) Two-color fluorescence correlation spectroscopy of one chromophore: application to the E222Q mutant of the green fluorescent protein. J Chem Phys 114:3149–3156
Zimmer M (2002) Green fluorescent protein (GFP): applications, structure, and related photophysical behavior. Chem Rev 102:759–781
Jung G, Wiehler J, Zumbusch A (2005) The photophysics of green fluorescent protein: influence of the key amino acids at positions 65, 203, and 222. Biophys J 88:1932–1947
Heilemann M, Margeat E, Kasper R, Sauer M, Tinnefeld P (2005) Carbocyanine dyes as efficient reversible single-molecule optical switch. J Am Chem Soc 127:3801–3806
Patterson GH, Piston DW (2000) Photobleaching in two-photon excitation microscopy. Biophys J 78:2159–2162
Seefeldt B, Kasper R, Seidel T, Tinnefeld P, Dietz K-J, Heilemann M, Sauer M (2008) Fluorescent proteins for single-molecule fluorescence applications. J Biophoton 1:74–82
Eggeling C, Hilbert M, Bock H, Ringemann C, Hofmann M, Stiel AC, Andresen M, Jakobs S, Egner A, Schoenle A, Hell SW (2007) Reversible photoswitching enables single-molecule fluorescence fluctuation spectroscopy at high molecular concentration. Micr Res Tech 70:1003–1009
Bizzarri R, Arcangeli C, Arosio D, Ricci F, Faraci P, Cardarelli F, Beltram F (2006) Development of a novel GFP-based ratiometric excitation and emission pH indicator for intracellular studies. Biophys J 90:3300–3314
Bizzarri R, Serresi M, Luin S, Beltram F (2009) Green fluorescent protein based pH indicators for in vivo use: a review. Anal Bioanal Chem 393:1107–1122
Schwille P, Haupts U, Maiti S, Webb WW (1999) Molecular dynamics in living cells observed by fluorescence correlation spectroscopy with one- and two-photon excitation. Biophys J 77:2251–2265
Bosisio C, Quercioli V, Collini M, D’Alfonso L, Baldini G, Bettati S, Campanini B, Raboni S, Chirico G (2008) Protonation and conformational dynamics of GFP mutants by two-photon excitation fluorescence correlation spectroscopy. J Phys Chem B 112:8806–8814
Malvezzi-Campeggi F, Jahnz M, Heinze KG, Dittrich P, Schwille P (2001) Light-induced flickering of DsRed provides evidence for distinct and interconvertible fluorescent states. Biophys J 81:1776–1785
Wang ZF, Shah JV, Chen ZP, Sun CH, Berns MW (2004) Fluorescence correlation spectroscopy investigation of a GFP mutant-enhanced cyan fluorescent protein and its tubulin fusion in living cells with two-photon excitation. J Biomed Opt 9:395–403
Jung G, Werner M, Schneider M (2008) Efficient photoconversion distorts the fluorescence lifetime of GFP in confocal microscopy: a model kinetic study on mutant Thr203Val. ChemPhysChem 9:1867–1874
Chirico G, Cannone F, Baldini G, Diaspro A (2003) Two-photon thermal bleaching of single fluorescent molecules. Biophys J 84:588–598
Kim SA, Heinze KG, Bacia K, Waxham MN, Schwille P (2005) Two-photon cross-correlation analysis of intracellular reactions with variable stoichiometry. Biophys J 88:4319–4336
Nifosì R, Tozzini V (2006) Cis–trans photoisomerization of the chromophore in the green fluorescent protein variant E2GFP: a molecular dynamics study. Chem Phys 323:358–368
Wong FHC, Banks DS, Abu-Arish A, Fradin C (2007) A molecular thermometer based on fluorescent protein blinking. J Am Chem Soc 129:10302–10303
Cannone F, Milani R, Chirico G, Diaspro A, Krol S, Campanini B (2007) Voltage regulation of single green fluorescent protein mutants. Biophys Chem 125:368–374
Paradise A, Levin MK, Korza G, Carson JH (2007) Significant proportions of with reduced intracellular nuclear transport proteins mobilities resolved by fluorescence correlation spectroscopy. J Mol Biol 365:50–65
Wang Z, Shah JV, Berns MW, Cleveland DW (2006) In vivo quantitative studies of dynamic intracellular processes using fluorescence correlation spectroscopy. Biophys J 91:343–351
Reiff DF, Ihring A, Guerrero G, Isacoff EY, Joesch M, Nakai J, Borst A (2005) In vivo performance of genetically encoded indicators of neural activity in flies. J Neurosci 25:4766–4778
Cormack BP, Valdivia RH, Falkow S (1996) FACS-optimized mutants of the green fluorescent protein (GFP). Gene 173:33–38
Abbruzzetti S, Grandi E, Viappiani C, Bologna S, Campanini B, Raboni S, Bettati S, Mozzarelli A (2005) Kinetics of acid-induced spectral changes in the GFPmut2 chromophore. J Am Chem Soc 127:626–635
Chirico G, Diaspro A, Cannone F, Collini M, Bologna S, Pellegrini V, Beltram F (2005) Selective fluorescence recovery after bleaching of single E(2)GFP proteins induced by two-photon excitation. ChemPhysChem 6:328–335
Chattoraj M, King BA, Bublitz GU, Boxer SG (1996) Ultra-fast excited state dynamics in green fluorescent protein: multiple states and proton transfer. Proc Natl Acad Sci USA 93:8362–8367
Brejc K, Sixma TK, Kitts PA, Kain SR, Tsien RY, Ormo M, Remington SJ (1997) Structural basis for dual excitation and photoisomerization of the Aequorea victoria green fluorescent protein. Proc Natl Acad Sci USA 94:2306–2311
Lossau H, Kummer A, Heinecke R, Pollinger-Dammer F, Kompa C, Bieser G, Jonsson T, Silva CM, Yang MM, Youvan DC, Michel-Beyerle ME (1996) Time-resolved spectroscopy of wild-type and mutant green fluorescent proteins reveals excited state deprotonation consistent with fluorophore-protein interactions. Chem Phys 213:1–16
Stoner-Ma D, Jaye AA, Ronayne KL, Nappa J, Meech SR, Tonge PJ (2008) An alternate proton acceptor for excited-state proton transfer in green fluorescent protein: rewiring GFP. J Am Chem Soc 130:1227–1235
Jung G, Wiehler J, Steipe B, Bräuchle C, Zumbusch A (2001) Single-molecule microscopy of the green fluorescent protein using simultaneous two-color excitation. ChemPhysChem 6:392–396
Cinelli RAG, Pellegrini V, Ferrari A, Faraci P, Nifosì R, Tyagi M, Giacca M, Beltram F (2001) Green fluorescent proteins as optically-controllable elements in bioelectronics. Appl Phys Lett 79:3353–3355
Yokoe H, Meyer T (1996) Spatial dynamics of GFP-tagged proteins investigated by local fluorescence enhancement. Nat Biotech 14:1252–1256
Redmond RW, Kochevar IE, Krieg M, Smith G, McGimpsey WG (1997) Excited state relaxation in cyanine dyes: a remarkably efficient reverse intersystem crossing from upper triplet levels. J Phys Chem A 101:2773–2777
Widengren J, Seidel CAM (2000) Manipulation and characterization of photo-induced transient states of merocyanine 540 by fluorescence correlation spectroscopy. Phys Chem Chem Phys 2:3435–3441
Huang Z, Ji D, Xia A (2005) Fluorescence intensity and lifetime fluctuations of single Cy5 molecules immobilized on the glass surface. Colloids Surf A Physicochem Eng Asp 257–258:203–209
Larkin JM, Donaldson WR, Foster TH, Knox RS (1999) Reverse intersystem crossing from a triplet state of rose Bengal populated by sequential 532- + 1064-nm laser excitation. Chem Phys 244:319–330
English DS, Harbron EJ, Barbara PF (2000) Probing photoinduced intersystem crossing by two-color. Double resonance single molecule spectroscopy. J Phys Chem A 104:9057–9061
Ringemann C, Schönle A, Giske A, von Middendorff C, Hell SW, Eggeling C (2008) Enhancing fluorescence brightness: effect of reverse intersystem crossing studied by fluorescence fluctuation spectroscopy. ChemPhysChem 9:612–624
Palm GJ, Zadnov A, Gaitanaris GA, Stauber R, Pavlakis GN, Wlodawer A (1997) The structural basis for spectral variations in green fluorescent protein. Nat Struct Biol 4:361–365
Weber W, Helms V, McCammon JA, Langhoff PW (1999) Shedding light on the dark and weakly fluorescent states of green fluorescent proteins. Proc Natl Acad Sci USA 96:6177–6182
Agmon N (2007) Kinetics of switchable proton escape from a proton-wire within green fluorescence protein. J Phys Chem B 111:7870–7878
Heilemann M, Dedecker P, Hofkens J, Sauer M (2009) Photoswitches: key molecules for subdiffraction-resolution fluorescence imaging and molecular quantification. Laser Photon Rev 3:180–202
Acknowledgments
This work has been also funded by the PRIN MIUR fund 2008 to M.C. (2008JZ4MLB) and by the FP7 program ENCITE (contract no. 201842) to G.C.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer-Verlag Berlin Heidelberg
About this chapter
Cite this chapter
Chirico, G., Collini, M., D’Alfonso, L., Caccia, M., Daglio, S.C., Campanini, B. (2011). Green Fluorescent Protein Photodynamics as a Tool for Fluorescence Correlative Studies and Applications. In: Jung, G. (eds) Fluorescent Proteins II. Springer Series on Fluorescence, vol 12. Springer, Berlin, Heidelberg. https://doi.org/10.1007/4243_2011_25
Download citation
DOI: https://doi.org/10.1007/4243_2011_25
Published:
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-23376-0
Online ISBN: 978-3-642-23377-7
eBook Packages: Chemistry and Materials ScienceChemistry and Material Science (R0)