Clarification and confocal imaging of the nonhuman primate placental micro-anatomy

J. A. Sargent, V. H.J. Roberts, J. E. Gaffney, A. E. Frias

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Geometry of the placental villous vasculature is a key determinant of maternal–fetal nutrient exchange for optimal fetal growth. Recent advances in tissue clarification techniques allow for deep high-resolution imaging with confocal microscopy; however, the methodology lacks a signal:noise ratio of sufficient magnitude to allow for quantitative analysis. Thus, we sought to develop a reproducible method to investigate the 3D vasculature of the nonhuman primate placenta for subsequent data analysis. Fresh placental tissue was dissected, formalin fixed, clarified using a modified Visikol® protocol and immunolabeled for CD31 (fetal endothelium) and cytokeratin-7 (villous trophoblast) for confocal imaging of the microanatomy. We present a detailed clarification and staining protocol augmented for imaging of nonhuman primate placental tissue. The image stacks generated by this refined staining method and our data acquisition parameters can be analyzed quantitatively to provide insights regarding the villous and vascular microanatomy of the placenta.

Original languageEnglish (US)
Pages (from-to)79-84
Number of pages6
JournalBioTechniques
Volume66
Issue number2
DOIs
StatePublished - Feb 2019

Keywords

  • Confocal microscopy
  • Immuno-histochemistry
  • Nonhuman primate
  • Placenta
  • Tissue clarification

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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