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

J. A. Sargent, Victoria Roberts, J. E. Gaffney, Antonio Frias

Research output: Contribution to journalArticle

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 1 2019

Fingerprint

Primates
Anatomy
Tissue
Imaging techniques
Placenta
Staining and Labeling
Keratin-7
Confocal microscopy
Trophoblasts
Fetal Development
Confocal Microscopy
Formaldehyde
Nutrients
Endothelium
Blood Vessels
Data acquisition
Food
Geometry
Chemical analysis

Keywords

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

ASJC Scopus subject areas

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

Cite this

Clarification and confocal imaging of the nonhuman primate placental micro-anatomy. / Sargent, J. A.; Roberts, Victoria; Gaffney, J. E.; Frias, Antonio.

In: BioTechniques, Vol. 66, No. 2, 01.02.2019, p. 79-84.

Research output: Contribution to journalArticle

Sargent, J. A. ; Roberts, Victoria ; Gaffney, J. E. ; Frias, Antonio. / Clarification and confocal imaging of the nonhuman primate placental micro-anatomy. In: BioTechniques. 2019 ; Vol. 66, No. 2. pp. 79-84.
@article{4f4b88a51027422fb96885266fb14011,
title = "Clarification and confocal imaging of the nonhuman primate placental micro-anatomy",
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{\circledR} 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.",
keywords = "Confocal microscopy, Immuno-histochemistry, Nonhuman primate, Placenta, Tissue clarification",
author = "Sargent, {J. A.} and Victoria Roberts and Gaffney, {J. E.} and Antonio Frias",
year = "2019",
month = "2",
day = "1",
doi = "10.2144/btn-2018-0110",
language = "English (US)",
volume = "66",
pages = "79--84",
journal = "BioTechniques",
issn = "0736-6205",
publisher = "Eaton Publishing Company",
number = "2",

}

TY - JOUR

T1 - Clarification and confocal imaging of the nonhuman primate placental micro-anatomy

AU - Sargent, J. A.

AU - Roberts, Victoria

AU - Gaffney, J. E.

AU - Frias, Antonio

PY - 2019/2/1

Y1 - 2019/2/1

N2 - 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.

AB - 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.

KW - Confocal microscopy

KW - Immuno-histochemistry

KW - Nonhuman primate

KW - Placenta

KW - Tissue clarification

UR - http://www.scopus.com/inward/record.url?scp=85061390203&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85061390203&partnerID=8YFLogxK

U2 - 10.2144/btn-2018-0110

DO - 10.2144/btn-2018-0110

M3 - Article

VL - 66

SP - 79

EP - 84

JO - BioTechniques

JF - BioTechniques

SN - 0736-6205

IS - 2

ER -