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.
Elements advanced research software program
Manufactured by Nikon
Nikon Elements Advanced Research software program is a comprehensive image analysis and visualization tool developed for scientific and research applications. It provides advanced features for acquiring, processing, analyzing, and managing high-quality digital images and data from various imaging techniques.
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2 protocols using elements advanced research software program
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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