Bz x810 all in one fluorescence microscope

Briefly, paraffin tissues were deparaffinized and dehydrated using Histo-Clear and a gradient of ethanol washes, respectively. Antigen retrieval was done by immersing slides in antigen unmasking solutions (Vector 21202), placing the slides in a microwave oven for 1 min and boiling the slides at 95–99 °C for 15 min. The sections were cooled to 22 °C (RT) and washed with distilled water and PBS. Then, 5% BSA was added to block non-specific reactions. The slices were incubated with indicated antibodies, followed by appropriate secondary antibodies. The slices were exposed with DAB chromogen, hematoxylin, and Periodic Acid-Schiff (PAS), based on the manufacturer’s instructions. The images were captured on an Olympus microscope or an All-in-One Fluorescence Microscope BZ-X810 (Keyence, Itasca, IL, USA).
For Immunofluorescence, briefly, pNETs cell lines were fixed with 4% PFA, and permeabilized in 0.3% Triton-100. Then, the cell samples were blocked in 3% BSA for 1 h and probed with appropriate primary and secondary antibodies. The immunofluorescence images were captured on an All-in-One Fluorescence Microscope BZ-X810 (Keyence).

Lipid peroxidation was examined using the fluorescent dye C11-BODIPY581/591 (No. D3861; Thermo Fisher Scientific, Waltham, MA, USA)^{29 (link)}. Fibroblasts were cultured to semi-confluence and then plated at 5,000 cells per well in a 96-well culture plate in normal medium. After incubation for 24 h, BSO and several other compounds (e.g. apomorphine and Fer-1) were added to each well. After incubation for 24 h, the cells were labeled with 5 μM C11-BODIPY581/591 for 30 min in the assay medium. The nuclei were stained with Hoechst 33,342. Representative images were obtained using a Keyence All-in-One Fluorescence BZ-X810 microscope (Keyence Co., Itasca, IL, USA).