A direct fluorescence-based technique for cellular localization of amylin

Amylin has been implicated in type II diabetes because of its inherent property to misfold into toxic aggregates. Although it has been shown that amylin interacts with cell membranes, no study to date has monitored the association process using a direct approach. The present study uses confocal microscopy to identify the localization of carboxyfluorescein-labeled amylin in RIN-5F cells. In addition, the size of the aggregates that are formed was evaluated using nanoparticle tracking analysis. In support of previous findings, amylin was observed to interact with and remain associated with the cell membrane. The cell membrane-associated aggregates spanned a size range of 130-800nm.