Co-staining with Thioflavin S & Carboxyfluorescein

1.) Prepare 0.01% thioflavin S (T1892 , Sigma) solution in F12+ [0.01g / 100 ml F12+]

2.) Filter the staining solution with a 0.22 µm Millipore filter.

3.) Incubate the staining solution and F12+ wash for approximately 30 minutes to adjust solution temperature to 37°C

4.) Stain with thioflavin S for 15 minutes

5.) Wash with F12+ once

6.) Stain with carboxyfluorescein 1:200 (C1354 Molecular Probes,1mM w/DMSO) for 30 minutes

7.) Wash with F12+ twice

8.) View immediately under the fluorescent microscope using FITC filter.

9.) Fix with 2% formalin for 20 to 30 minutes

10.) Coverslip with water-soluble mounting media