Bz x700 microscope

Fluorescent images were obtained using a Keyence BZ-X700 microscope and associated software. Bright-field and fluorescent images were obtained using a Keyence BZ-X700 microscope and associated software. Fluorescent images of 3D samples were taken using a Leica TCS FLIM SP8 (Wetzlar, Germany) confocal microscope. Scanning electron microscopy (SEM) was performed using a Zeiss Evo MA10 (Carl Zeiss Microscopy, Germany) to visualize the morphological properties of 3D brain-like tissue samples. Constructs were thoroughly dried and cross-sectioned and then gold-sputter–coated to prepare for subsequent SEM analysis.

For mouse islets, following stress treatment, 50–100 intact islets were transferred to a polysterene round bottom tube and CellROX Deep Red Reagent (Invitrogen) was added at 1:500 dilution to culture media followed by incubation at 37 °C for 45 min. Islets were washed 2X with PBS, dispersed to single cell suspension, attached to slides using cytospin, and imaged using fluorescence microscopy (Keyence BZ-X700 microscope). GFP signal was used to identify transduced cells and the CellROX signal from each cell was quantified and normalized by cell area using BZ-X Advanced Analysis Software. Signal from at least 60 GFP positive cells was assessed for each condition per experiment.
For Min6 cells, following stress treatment, CellROX Deep Red Reagent (Invitrogen) was added at 1:500 dilution directly to culture media followed by incubation at 37 °C for 30 min. Cells were washed 2X with PBS and imaged using fluorescence microscopy (Keyence BZ-X700 microscope). Bright field images were used to determine cell areas and CellROX signal was quantified from at least 6 imaging fields for each group using BZ-X Advanced Analysis Software.