hFTAA Staining of Amyloid Deposits: A Technique to Visualize Amyloid Fibrils Using Oligothiophene-Based Dye for Fluorescence Imaging in Tissue Sections

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.

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.
2. On the day of staining, prepare a working solution by diluting the stock 1:10,000 in Phosphate-buffered saline (PBS).

2. Preparation of Tissue Samples

NOTE: Many tissue types can be imaged using hFTAA as an amyloi....

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-I.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
hFTAA/Amytracker545 	Ebba Biotech
Dako fluorescene mounting medium	Agilent technologies 	GM304
LeicaDM6000 	Leica
Lumen 200 	Prior
Spectraview system	 ASI spectral imaging