BL-induced cell wall expansion is paralleled by changes in the BRI1-GFP fluorescence lifetime.Kirstin ElgassKatharina CaesarFrank SchleifenbaumYorkDieter StierhofAlfred J. MeixnerKlaus Harter
GFP has a rel- atively short fluorescence lifetime, but significant differences exist among mu- tants. Lifetimes vary from 1.3 ns for cyan to 2.6 ns for green S65T and 3.7 ns for yel- low fluorescent proteins, although these values are known to vary with tempera- ture, pH and fusion ...
Fluorescence lifetime measurements were performed using a MicroTime200 optical setup. GFP samples were placed as drops on top of 175 µm glass slides (Precision Cover Glass No:1.5H, Marienfeld), mounted on an inverted microscope (IX83 inverted, Olympus) with a 60× water immersion objective ...
Lifetime intensity images from cells expressing either of the constructs showed a clear difference (Fig. 1b, left panel). The corresponding time constant τof the sEcGFP fluorescence lifetime decay was determined by fitting the respective TCSPC diagram (Fig. 1b, right panel). For ...
The identified mutations in 尾-strand 7 render GFP fluorescence intensity and lifetime sensitive to conformational changes of the PKA-sensing domain. In addition, sensors engineered from the GCaMP2 calcium indicator to incorporate a conformation-sensitive GFP (csGFP) exhibited calcium-dependent ...
FluorescenceLifetimeCorrelation SpectroscopytoQuantifyProtein–Protein InteractionsinLiveCell.《MICROSCOPY RESEARCHANDTECHNIQUE》.2011,第74卷第788- 793页. Lan-YingLee等.Vectorsformulti- colorbimolecularfluorescence complementationtoinvestigateprotein- proteininteractionsinlivingplant ...
Indeed, GFPmut2 is a fast folding mutant of the original protein with a high stability, and it is generally assumed that the lifetime of this protein is longer than the cell cycle [51]. In our case, segregation phenomena, i.e. strong fluorescence decreases for a major part of the ...
Acceptor photobleaching (APB) and one- and two-photon fluorescence lifetime imaging microscopy (FLIM) are used to quantitatively determine FRET efficiency values. A FRET standard is introduced based on a tandem construct comprising donor and acceptor together with a 20 amino acid long cleavable ...
it should be emphasized that at latter times the cells continue to produce GFP and that fluorescence intensity increases over time. The transfection efficiency in the monolayer exposed to 10 min of PA waves generated with a picosecond laser was 5.0% for plasmid vector concentrations of 0.5 ...
Elgass K, Caesar K, Schleifenbaum F, Meixner AJ, Harter K. The fluorescence lifetime of BRI1-GFP as probe for the noninvasive determination of the membrane potential in living cells. Proc SPIE. 2010:7568.Elgass, K. , Caesar, K. , Schleifenbaum, F. , Meixner, A.J. and Harter...