Towards Multiparametric Fluorescent Imaging of Amyloid Formation: Studies of a YFP Model of α-Synuclein Aggregation

Protein Solubility and Protein Homeostasis: A Generic View of Protein Misfolding Disorders

“…This theoretical analysis can be combined with a range of biophysical experiments that allow key properties governing protein aggregation to be defined in vitro and in vivo in a highly quantitative manner by using a variety of sophisticated fluorescence methods (van Ham et al 2010), as well as label-free Figure 2. Schematic illustration of the link between solubility and homeostasis of proteins.…”

Superresolution Imaging of Amyloid Fibrils with Binding-Activated Probes

“…Since fluorescent proteins are considerably larger than the aggregating protein or peptide, in many cases this induces artifacts. 6,7 Moreover, covalent linking of fluorophores to monomeric proteins can interfere with the packing of amyloids. 6 To avoid such artifacts and to enable visualization of amyloid structures in their natural environment, amyloid reactive dyes such as congo red or Thioflavin T have been used.…”

“…6,7 Moreover, covalent linking of fluorophores to monomeric proteins can interfere with the packing of amyloids. 6 To avoid such artifacts and to enable visualization of amyloid structures in their natural environment, amyloid reactive dyes such as congo red or Thioflavin T have been used. Recently, luminescent conjugated polyelectrolyte probes, also called luminescent conjugated oligothiophenes (LCOs), have been utilized to identify and image amyloid deposits.…”

Design and application of a confocal microscope for spectrally resolved anisotropy imaging

“…FRET causes depolarization of the sensitized emission because of the transfer of excitation energy to a second fluorophore the orientation of which is not constrained by the photoselection rules of the directly excited fluorophores. A decrease in the anisotropy can be measured when identical fluorophores are in close proximity (homo-FRET) allowing protein oligomerization to be detected [7,26]. Recently, Rizzo and Piston [25] demonstrated that mapping the FRET-dependent depolarization in hetero-FRET experiments could be also useful to measure protein-protein interactions.…”

“…The most important applications that we foresee for hyper-dimensional imaging microscopy include (i) unmixing of multiple FRET-based biosensors [31], (ii) detection of interactions in multi-molecular complexes and (iii) enhanced contrast for tissue imaging. In our laboratories, HDIM will complement existing techniques that we are using for probing the molecular mechanisms underlying neurodegeneration [26] and cancer [32]. However, it is likely that HDIM will find numerous applications not only in biology and biophysics but also in material sciences and spectroscopy.…”