Progres c14 digital camera

For paraffin sections, pancreata were processed as previously described (Lattke et al. 2012 (link), Salem et al. 2014 (link)). Sections were incubated with primary antibodies overnight (Supplementary Table 2). Secondary antibodies were coupled with Alexa Fluor (Invitrogen) for immunofluorescence or with horseradish peroxidase that was developed by 3-amino-9-ethylcarbazole (DakoCytomation) for immune-histochemistry. Fibrosis was assessed using Pikro-Siriusrot solution (Morphisto, Frankfurt, Germany). Immunofluorescent stainings were visualized as before (Lattke et al. 2012 (link)), and other stainings were analysed on a Leica DM IRB microscope (Leica Microsystems) equipped with ProgRes C14 digital camera (Jenoptik).

Alkaline phosphatase (ALPL) activity was detected following previously published protocols [67 (link)], with modifications. Briefly, frozen uterine sections were fixed in 10% NBF for 10 min, and then washed with 1x phosphate-buffered saline (PBS) three times for 5 min each. The uterine sections were then incubated in the dark at 37°C for 30 min in a solution containing 0.5 mM naphthol AS-MX phosphate (ALPL substrate) and 1.5 mM Fast Blue RR in 0.1 M Tris-HCl, pH 8.5. Alkaline phosphatase activity releases orthophosphate and naphthol derivatives from the ALPL substrate. The naphthol derivatives are simultaneously coupled with the diazonium salt (Fast Blue RR) to form a dark dye marking the site of enzyme action. The slides were rinsed in tap water to terminate the enzymatic reaction. Stained uterine sections were visualized under an Olympus BX51 microscope equipped for light imaging and connected to a Jenoptik ProgRes C14 digital camera with c-mount interface containing a 1.4 Megapixel CCD sensor.