Photostability of green and yellow fluorescent proteins with fluorinated chromophores, investigated by fluorescence correlation spectroscopy

2008 ◽  
Vol 136 (1) ◽  
pp. 38-43 ◽  
Author(s):  
S. Veettil ◽  
N. Budisa ◽  
G. Jung
Keyword(s):  
Fluorescence Correlation Spectroscopy ◽  
Fluorescent Proteins ◽  
Correlation Spectroscopy ◽  
Fluorescence Correlation

scholarly journals Photodynamics of Red Fluorescent Proteins Studied by Fluorescence Correlation Spectroscopy

2004 ◽  
Vol 86 (1) ◽  
pp. 384-394 ◽  
Author(s):  
Andreas Schenk ◽  
Sergey Ivanchenko ◽  
Carlheinz Röcker ◽  
Jörg Wiedenmann ◽  
G. Ulrich Nienhaus
Keyword(s):  
Fluorescence Correlation Spectroscopy ◽  
Fluorescent Proteins ◽  
Correlation Spectroscopy ◽  
Fluorescence Correlation ◽  
Red Fluorescent Proteins

Properties of Baculovirus Particles Displaying GFP Analyzed by Fluorescence Correlation Spectroscopy

2002 ◽  
Vol 383 (12) ◽  
pp. 1941-1946 ◽  
Author(s):  
J. Toivola ◽  
K. Ojala ◽  
P.O. Michel ◽  
M. Vuento ◽  
C. Oker-Blom
Keyword(s):  
Fluorescence Correlation Spectroscopy ◽  
Fusion Proteins ◽  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Sodium Dodecyl ◽  
Recombinant Baculovirus ◽  
Mean Value ◽  
Correlation Spectroscopy ◽  
Viral Envelope ◽  
Fluorescence Correlation

Abstract Recombinant baculovirus particles displaying green fluorescent protein (GFP) fused to the major envelope glycoprotein gp64 of the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) were characterized by fluorescence correlation spectroscopy (FCS). FCS detected Brownian motion of single, intact recombinant baculovirus display particles with a diffusion coefficient (D) of (2.89±0.74)10 8 cm2s 1 and an apparent hydrodynamic radius of 83.35±21.22 nm. In the presence of sodium dodecyl sulfate (SDS), Triton X-100, and octylglucoside, the diffusion time was reduced to the 0.2 ms range (D = 7.5710 7 cm2s 1), showing that the fusion proteins were anchored in the viral envelope. This allowed for a calculation of the number of single gp64 fusion proteins incorporated in the viral membrane. A mean value of 3.2 fluorescent proteins per virus particle was obtained. Our results show that FCS is the method of choice for studying enveloped viruses such as a display virus with one component being GFP.

scholarly journals Diffusion behavior of the fluorescent proteins eGFP and Dreiklang in solvents of different viscosity monitored by fluorescence correlation spectroscopy

2016 ◽  
Vol 3 (1) ◽  
Author(s):  
Cornelia Junghans ◽  
Franz-Josef Schmitt ◽  
Vladana Vukojević ◽  
Thomas Friedrich
Keyword(s):  
Fluorescence Correlation Spectroscopy ◽  
Diffusion Processes ◽  
Fluorescent Proteins ◽  
Correlation Spectroscopy ◽  
Multiple Time Scales ◽  
Multiple Time ◽  
Fluorescence Excitation ◽  
Dark State ◽  
Fluorescence Correlation ◽  
Observation Volume

AbstractFluorescence correlation spectroscopy relies on temporal autocorrelation analysis of fluorescence intensity fluctuations that spontaneously arise in systems at equilibrium due to molecular motion and changes of state that cause changes in fluorescence, such as triplet state transition, photoisomerization and other photophysical transformations, to determine the rates of these processes. The stability of a fluorescent molecule against dark state conversion is of particular concern for chromophores intended to be used as reference tags for comparing diffusion processes on multiple time scales. In this work, we analyzed properties of two fluorescent proteins, the photoswitchable Dreiklang and its parental eGFP, in solvents of different viscosity to vary the diffusion time through the observation volume element by several orders of magnitude. In contrast to eGFP, Dreiklang undergoes a dark-state conversion on the time scale of tens to hundreds of microseconds under conditions of intense fluorescence excitation, which results in artificially shortened diffusion times if the diffusional motion through the observation volume is sufficiently slowed down. Such photophysical quenching processes have also been observed in FCS studies on other photoswitchable fluorescent proteins including Citrine, from which Dreiklang was derived by genetic engineering. This property readily explains the discrepancies observed previously between the diffusion times of eGFP- and Dreiklang-labeled plasma membrane protein complexes.

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