2.10. Assessment of neurodegeneration in cell cultures

Neurodegeneration of the cultured cortical neurons was measured using FJ dye according to a previously published procedure [18], [54]. The cell culture plates were fixed using cold ethanol (−20 °C) for 30 min and washed three times with distilled water. The cells were treated with the dye (final concentration of 0.0004% FJ in 0.1% acetic acid), gently shaken for 30 min in the dark at room temperature and then washed twice with distilled water. Finally, the fluorescence intensity was determined using a FLUOstar Galaxy spectrophotometer (BMG, Lab Technologies, Ortenberg, Germany) at 485/530 nm.