Pancreatic Islet Embedding for Paraffin Sections.

TitlePancreatic Islet Embedding for Paraffin Sections.
Publication TypeJournal Article
Year of Publication2018
AuthorsKong Y, Ebrahimpour P, Liu Y, Yang C, Alonso LC
JournalJ Vis Exp
Issue136
Date Published2018 06 29
ISSN1940-087X
KeywordsAnimals, Histological Techniques, Humans, Islets of Langerhans, Mice, Paraffin, Rats
Abstract

Experiments using isolated pancreatic islets are important for diabetes research, but islets are expensive and of limited abundance. Islets contain a mixed cell population in a structured architecture that impacts function, and human islets are widely variable in cell type composition. Current frequently used methods to study cultured islets include molecular studies performed on whole islets, lumping disparate islet cell types together, or microscopy or molecular studies on dispersed islet cells, disrupting islet architecture. For in vivo islet studies, paraffin-embedded pancreas sectioning is a powerful technique to assess cell-specific outcomes in the native pancreatic environment. Studying post-culture islets by paraffin sectioning would offer several advantages: detection of multiple outcomes on the same islets (potentially even the exact-same islets, using serial sections), cell-type-specific measurements, and maintaining native islet cell-cell and cell-substratum interactions both during experimental exposure and for analysis. However, existing techniques for embedding isolated islets post-culture are inefficient, time consuming, prone to loss of material, and generally produce sections with inadequate islet numbers to be useful for quantifying outcomes. Clinical pathology laboratory cell block preparation facilities are inaccessible and impractical for basic research laboratories. We have developed an improved, simplified bench-top method that generates sections with robust yield and distribution of islets. Fixed islets are resuspended in warm histological agarose gel and pipetted into a flat disc on a standard glass slide, such that the islets are distributed in a plane. After standard dehydration and embedding, multiple (10+) 4 - 5 Āµm sections can be cut from the same islet block. Using this method, histologicalĀ and immunofluorescent analyses can be performed on mouse, rat, and human islets. This is an effective, inexpensive, time-saving approach to assess cell-type-specific, intact-architecture outcomes from cultured islets.

DOI10.3791/57931
Alternate JournalJ Vis Exp
PubMed ID30010652
PubMed Central IDPMC6102017
Grant ListR01 DK105837 / DK / NIDDK NIH HHS / United States
R01 DK114686 / DK / NIDDK NIH HHS / United States
UC4 DK098085 / DK / NIDDK NIH HHS / United States