Hi everyone, I'm a master's degree student and recently started working with endoplasmic reticulum stress (Er-stress). In the literature, I found that staining with Thioflavin-T (ThT) is one way to measure Er-stress because, during Er-stress, protein aggregates are formed. I tried the assay once, and it worked! But when I repeated it, there was no difference in fluorescence intensity between my control group and the positive control (cells treated with 1 µM of Thapsigargin, an Er-stress inducer). I'm trying to perform the staining using a fluorescence microscope. Does anyone have any tips on what can be done?