Quantum dot immunocytochemical localization of somatostatin in somatostatinoma by Widefield Epifluorescence, super-resolution light, and immunoelectron microscopy

J Histochem Cytochem. 2012 Nov;60(11):832-43. doi: 10.1369/0022155412459856. Epub 2012 Aug 16.

Abstract

Quantum dot nanocrystal probes (QDs) have been used for detection of somatostatin hormone in secretory granules of somatostatinoma tumor cells by immunofluorescence light microscopy, super-resolution light microscopy, and immunoelectron microscopy. Immunostaining for all modalities was done using sections taken from an epoxy resin-embedded tissue specimen and a similar labeling protocol. This approach allowed assessment of labeling at light microscopy level before examination at super-resolution and electron microscopy level and was a significant aid in interpretation. Etching of ultrathin sections with saturated sodium metaperiodate was a critical step presumably able to retrieve some tissue antigenicity masked by processing in epoxy resin. Immunofluorescence microscopy of QD-immunolabeled sections showed somatostatin hormone localization in cytoplasmic granules. Some variable staining of tumor gland-like structures appeared related to granule maturity and dispersal of granule contents within the tumor cell cytoplasm. Super-resolution light microscopy demonstrated localization of somatostatin within individual secretory granules to be heterogeneous, and this staining pattern was confirmed by immunoelectron microscopy.

MeSH terms

  • Adult
  • Ampulla of Vater / pathology*
  • Female
  • Humans
  • Immunohistochemistry / methods*
  • Microscopy, Fluorescence / methods
  • Microscopy, Immunoelectron / methods*
  • Quantum Dots*
  • Somatostatin / analysis*
  • Somatostatinoma / pathology*

Substances

  • Somatostatin