Formalin-fixed paraffin-embedded pancreases were sectioned and immunostained using the antibodies indicated (ESM Table 1 ). Slides were imaged using a Leica DM5500 fluorescent microscope or Leica DM6000 Power Mosaic microscope (Leica Microsystems, Wetzlar, Germany). Immunostaining was quantified using ImageJ/FIJI (ImageJ Developers; version 1.52a; https://imageJ.nih.gov/ij/ ; and FIJI Contributors; version 2.0.0-rc-69/1.52n; https://fiji.sc/ ).
Dm 6000 power mosaic microscope
Manufactured by Leica
Sourced in Germany
The Leica DM 6000 Power Mosaic microscope is a high-performance laboratory equipment designed for advanced microscopy applications. It features a motorized stage and objective turret, enabling precise and efficient sample navigation and observation. The microscope is equipped with state-of-the-art optics and illumination systems to provide users with clear and detailed images.
Automatically generated - may contain errors
3 protocols using dm 6000 power mosaic microscope
1
Immunohistochemistry of Pancreatic Tissue
2
Pancreatic Beta Cell Quantification
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In brief, and as described previously [12 (link)], parallel pancreatic tissue sections were stained using standard protocols, buffers and diluents for insulin (rabbit anti-mouse insulin polyclonal antibody; 4590, Cell Signaling Technology), followed by incubation with horseradish peroxidase (HRP)-labelled polymer-conjugated goat anti-rabbit IgG (Dako EnVision+ System) and counterstaining with haematoxylin. Beta cell area was quantified from the total area (insulin-positive cells compared with non-positive tissue) using ImageJ (v1.53i; https://imagej.nih.gov ) on consecutive pancreatic serial sections cut at 200 µm intervals. Beta cell mass (per mg) was calculated by multiplying the relative insulin-positive area by the mass of the isolated pancreas before fixation. Images were captured using a Leica DM 4000 or Leica DM 6000 Power Mosaic microscope (Leica Microsystems).
3
Comet Assay for DNA Damage Assessment
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The comet assay was performed using the CometAssay Reagent Kit for Single Cell Gel Electrophoresis Assay following the manufacturer’s instructions (4250-050-01; Trevigen, Gaithersburg, MD). Exponentially growing cells were treated with 10 nmol/L to 1 μmol/L SN38 (Selleck Chemicals, Houston, TX) for 2 hours. High-resolution images of the comets formed were generated using the Leica DM 6000 Power Mosaic microscope (Leica Microsystems, Wetzlar, Germany). Tail length was quantified using the OpenComet v1.3 ImageJ (National Institutes of Health, Bethesda, MD) plug-in from comets generated from 3 independent experiments.
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