Pancreata from mice were fixed with 10% zinc formalin overnight and paraffin embedded. 5-μm sections of the pancreata were generated, and staining was performed after blocking with 5% normal goat serum with anti-Insulin (Linco) and anti-Glucagon (Cell Signaling) antibodies using established protocols. Antigen retrieval was prepared by using citrate buffer pH 6.0. After staining, slides were mounted with antifade mounting medium containing 4,6-diamidino-2-phenylindole (DAPI) (Vector Laboratories). Immunofluorescent images of pancreatic sections were obtained using a Nikon Storm/Tirf/Epifluorescence. Images used for β-cell mass calculations were obtained with an EVOS FL Auto imaging system. The images of hematoxylin and eosin (H&E)-stained pancreatic sections were obtained using an AmScope light microscope. For analysis of β-cell mass and α:β cell ratio, the images were analyzed by using the Nikon Elements Advanced Research software program.
Storm tirf epifluorescence
Manufactured by Nikon
The Storm/Tirf/Epifluorescence is a versatile lab equipment that enables various fluorescence microscopy techniques. It provides the capability to perform Super-resolution Microscopy (STORM), Total Internal Reflection Fluorescence (TIRF) Microscopy, and Epifluorescence Microscopy. This equipment allows researchers to visualize and analyze samples at the nanoscale level with high spatial and temporal resolution.
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2 protocols using storm tirf epifluorescence
1
Pancreatic Immunofluorescence Histology Protocol
2
Pancreatic Immunofluorescence Histology Protocol
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Pancreata from mice were fixed with 10% zinc formalin overnight and paraffin embedded. 5-μm sections of the pancreata were generated, and staining was performed after blocking with 5% normal goat serum with anti-Insulin (Linco) and anti-Glucagon (Cell Signaling) antibodies using established protocols. Antigen retrieval was prepared by using citrate buffer pH 6.0. After staining, slides were mounted with antifade mounting medium containing 4,6-diamidino-2-phenylindole (DAPI) (Vector Laboratories). Immunofluorescent images of pancreatic sections were obtained using a Nikon Storm/Tirf/Epifluorescence. Images used for β-cell mass calculations were obtained with an EVOS FL Auto imaging system. The images of hematoxylin and eosin (H&E)-stained pancreatic sections were obtained using an AmScope light microscope. For analysis of β-cell mass and α:β cell ratio, the images were analyzed by using the Nikon Elements Advanced Research software program.
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