hFTAA Staining of Amyloid Deposits: A Technique to Visualize Amyloid Fibrils Using Oligothiophene-Based Dye for Fluorescence Imaging in Tissue Sections
Published: April 30, 2023
Abstract
Source: Nyström, S. et al., Imaging Amyloid Tissues Stained with Luminescent Conjugated Oligothiophenes by Hyperspectral Confocal Microscopy and Fluorescence Lifetime Imaging. J. Vis. Exp. (2017).
This video describes a technique to stain amyloids in tissue sections using a luminescent-conjugated oligothiophene dye – heptamer-formyl thiophene acetic acid, or hFTAA – for fluorescence imaging. Amyloids are β-sheet-rich abnormal protein aggregates. Their detection in tissue samples is helpful in the diagnosis and treatment of amyloid-associated diseases or proteinopathies.
Protocol
1. LCO Staining Solution NOTE: If hFTAA is purchased from a commercial vendor, please follow the vendor's instructions instead of section 1. Resuspend the lyophilized hFTAA in 2 mM NaOH to prepare a stock solution of 1 mg/mL. Keep the stock in a glass vial at 4 °C. The stock can be stored for one year. On the day of staining, prepare a working solution by diluting the stock 1:10,000 in Phosphate-buffered saline (PBS). <p c…
Representative Results
Figure 1: Various tissue types and proteins aggregates showing h-FTAA staining of: (a) Mallory-Denk bodies consisting of keratin aggregates in a liver (counterstained with DAPI), (b) p62-positive r inclusions in sporadic inclusion-body myositis (s-IBM) muscle tissue, (c) Amyloid of islet amyloid polypeptide in human pancreas, (d) Amyloid of Immunoglobulin light …
Disclosures
The authors have nothing to disclose.
Materials
| hFTAA/Amytracker545 | Ebba Biotech | ||
| Dako fluorescene mounting medium | Agilent technologies | GM304 | |
| LeicaDM6000 | Leica | ||
| Lumen 200 | Prior | ||
| Spectraview system | ASI spectral imaging |
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Cite This Article
hFTAA Staining of Amyloid Deposits: A Technique to Visualize Amyloid Fibrils Using Oligothiophene-Based Dye for Fluorescence Imaging in Tissue Sections. J. Vis. Exp. (Pending Publication), e21045, doi: (2023).