Aperio scanscope digital slide scanner

The maxillae were decalcified in 10% ethylenediaminetetraacetic acid (EDTA) solution (pH 7.2) with weekly changes of solution over 8 weeks. The samples were subsequently processed and embedded in paraffin wax allowing the sectioning along the parasagittal plane. Sections (4 µm thick) were obtained, stained with H&E, and three representative sections were selected per maxilla specifically to include buccal and mid-socket regions that was determined based on the morphology of the remaining molar in the maxillae. The slides were scanned using the Aperio Scanscope Digital Slide Scanner (Leica Biosystems Inc, Buffalo Grove, IL, USA) and histomorphometric analysis was performed. All quantification was performed by a single observer and repeated twice.

Pancreata were fixed in 10% formalin (pH 6.90–7.10) and embedded in paraffin. Tissue sections (2 µm) were cut and selected 50 µm apart to avoid overlapping of islets for hematoxylin and eosin staining, which was accomplished except for 1 big islet. Five slides and a minimum of 14 islets per mouse were analyzed in aged-matched ctrl and spd diabetic mice with an age from 14 to 29 weeks. Images were captured with an Aperio ScanScope digital slide scanner (Leica Biosystems, Germany) at 40-fold magnification. Islets were counted and classified in a blinded fashion by two independent observers. Immune cell infiltration in islets was classified as published before^{24 (link)}: grade 0: no infiltration, grade 1: peri-insulitis to 10% infiltration, grade 2: 10–50% infiltration, grade 3: 50–75% infiltration, grade 4: 75–100% infiltration.