The presence of amyloid was confirmed by Congo red staining, showing apple-green birefringence under polarized light, as well as by a Thioflavin
S stain under fluorescence microscopy, which was positive (Figure 3).
In a recent work, the NP conjugating with thioflavin
T (ThT) has been reported, which enables ultrasensitive detection of amyloid protein .
T as an efficient inducer and selective fluorescent sensor for the human telomeric G-quadruplex DNA," Journal of the American Chemical Society, vol.
The mature [PrP.sup.Sc] aggregates are characterized by their resistance to proteinase K digestion, apple-green birefringence when stained with Congo red, and their ability to enhance the fluorescence emission of the dye thioflavin
In addition to Congo red, amyloid can also be stained by thioflavin
T and metachromatic dyes, such as crystal violet.
Fluorescent probes, such as benzothiazole dye thioflavin
T (ThT), are widely and efficiently used for diagnosing amyloid fibril formation and studying fibrillar structure (Figure 1(a)) [13-16].
To investigate the effects of FDE on the fibrillation and cytotoxicity of A[beta] protein, the inhibitory effects of FDE on A[beta]1-42 fibril formation were determined by using thioflavin
T fluorescence (Th-T) and Electron Microscopy (EM); the protective effects against cytotoxicity induced by A[beta]1-42 in SH-SY5Y cells were evaluated by MTT assay.
Application of thioflavin
T (ThT), a fluorescent dye that quantifies the presence of fibrillated aggregates or amyloids of misfolded protein, to the ASA, PAFA, or QuIC permits real-time measurement of the seeding activity of [PrP.sup.Sc] .
S (ThS) staining, sections were immersed in 0.5% ThS dissolved in phosphatebuffered saline (PBS; Sigma-Aldrich, St.
Aggregation is measured by fluorescence spectroscopy using the dye thioflavin
T (ThT), that selectively binds to sites formed by the cross-beta amyloid assembly.
Amyloid fibers are commonly detected by the fluorescent dye thioflavin
T (ThT), which becomes fluorescent when bound to the cross-[beta] spine of the amyloid fiber .
After completion of the experimental procedure, the area of no-reflow (ANR) was delineated by intra-atrial injection of 1 mL/kg of the fluorescent dye thioflavin
S (Sigma Chemical Co., USA) that had been dissolved in 0.9% saline and then centrifuged at 1500 rpm for 5 min.