An enzyme-responsive FRET nanoprobe was designed and developed based on AIE-driven fluorescent polysaccharide polymersomes to study the real-time delivery aspects in intracellular compartments live cancer cells.

Molecular sensors based on intramolecular Förster resonance energy transfer (FRET) have become versatile tools to monitor regulatory molecules in living tissue. However, their use is often compromised by low signal strength and excessive noise. We analyzed signal/noise (SNR) aspects of spectral FRET analysis methods, with the following conclusions: The most commonly used method (measurement of ...

Methods The FRET donor, CFP, and FRET acceptor, YFP, were fused to aminoand carboxy-terminal ends of sGC subunits. After generation of recombinant baculovirus strains fluorescent tagged sGC subunits were co-expressed in Sf9cells. Fluorescent variants of sGC were analyzed in vitro in cytosolic fractions by sensitized emission FRET. In addition, fluorescent tagged sGC subunits were analyzed in vi...

We show that FRET between Pacific Blue (PB) and Alexa488 (A488) covalently attached to a DNA scaffold can be reversibly controlled by photochromic switching of a spiropyran derivative. With the spiropyran in the closed spiro isomeric form, FRET occurs freely between PB and A488. UV-induced isomerization to the open merocyanine form shuts down the FRET process by efficient quenching of the PB ex...

Förster resonance energy transfer (FRET) microscopy is a powerful technique routinely used to monitor interactions between biomolecules. Here, we focus on the techniques that are used for investigating the structure and interactions of nucleic acids (NAs). We present a brief overview of the most commonly used FRET microscopy techniques, their advantages and drawbacks. We list experimental appro...

BACKGROUND While the static structure of the intracellular Ca(2+) release channel, the ryanodine receptor type 1 (RyR1) has been determined using cryo electron microscopy, relatively little is known concerning changes in RyR1 structure that accompany channel gating. Förster resonance energy transfer (FRET) methods can resolve small changes in protein structure although FRET measurements of RyR1...

Förster resonant energy transfer (FRET) can be an efficient mechanism between densely packed fluorescent emitters. It plays a key role in photosynthesis but may also detrimental. In optoelectronic devices for instance, FRET funnels to quenching sites and favors losses. Here, we image individual self-assembled chains of stacked CdSe nanoplatelets demonstrate fluorescence intermittency (blinking)...

Fluorescence resonance energy transfer (FRET) analysis and the recently developed proximity ligation assay (PLA) are widely used to study protein-protein interactions in situ. We have developed correlative FRET-PLA to monitor interactions between membrane proteins that frequently cause problems in confirmatory co-immunoprecipitation assays. Correlative FRET-PLA is particularly aimed at deliveri...

Cryoelectron microscopy studies have identified distinct locations of tropomyosin (Tm) within the Ca(2+)-free, Ca(2+)-saturated, and myosin-S1-saturated states of the thin filament. On the other hand, steady-state Förster resonance energy transfer (FRET) studies using functional, reconstituted thin filaments under physiological conditions of temperature and solvent have failed to detect any mov...

Conformational states of the metastable drkN SH3 domain were characterized using single-molecule fluorescence techniques. Under nondenaturing conditions, two Förster resonance energy transfer (FRET) populations were observed that corresponded to a folded and an unfolded state. FRET-estimated radii of gyration and hydrodynamic radii estimated by fluorescence correlation spectroscopy of the two c...