Pancreatic Tissue Sectioning and Immunostaining

The pancreas was divided into consecutive tissue blocks (~5 mm) with alternating collection between paraffin-embedding and snap frozen18 (link). In this study, paraffin-embedded sections (5 μm) were stained with the following primary antibodies (all 1:500): polyclonal guinea pig anti-porcine insulin (DAKO, Carpinteria, CA), mouse monoclonal anti-human glucagon (Sigma-Aldrich, St. Louis, MO), polyclonal goat anti-somatostatin (Santa Cruz, Santa Cruz, CA), polyclonal goat anti-human PP (DAKO), and DAPI (Invitrogen, Carlsbad, CA). The primary antibodies were detected using a combination of DyLight 488, 549, and 649-conjugated secondary antibodies (1:200, Jackson ImmunoResearch Laboratory, West Grove, PA).

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Poudel A., Fowler J.L., Zielinski M.C., Kilimnik G, & Hara M. (2016). Stereological analyses of the whole human pancreas. Scientific Reports, 6, 34049.

Publication 2016

polyclonal guinea pig anti-porcine insulin mouse monoclonal anti-human glucagon polyclonal goat anti-somatostatin DAPI DyLight 488 649-conjugated secondary antibodies

Antibodies Dapi Glucagon Goat Guinea pig Human Mouse Pancreas Paraffin Porcine insulin Somatostatin Tissue

Corresponding Organization :

Other organizations : University of Chicago

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Variable analysis

independent variables

Paraffin-embedding vs. snap freezing of pancreatic tissue blocks

dependent variables

Insulin expression
Glucagon expression
Somatostatin expression
Pancreatic polypeptide (PP) expression

control variables

Tissue block size (~5 mm)
Paraffin-embedded section thickness (5 μm)
Primary antibody dilution (1:500)
Secondary antibody dilution (1:200)
DAPI staining for nuclear visualization

positive controls

None specified

negative controls

None specified

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