3D imaging of the early embryonic chicken heart with focused ion beam scanning electron microscopy

Monique Y. Rennie, Curran G. Gahan, Claudia Lopez, Kent Thornburg, Sandra Rugonyi

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Early embryonic heart development is a period of dynamic growth and remodeling, with rapid changes occurring at the tissue, cell, and subcellular levels. A detailed understanding of the events that establish the components of the heart wall has been hampered by a lack of methodologies for three-dimensional (3D), high-resolution imaging. Focused ion beam scanning electron microscopy (FIB-SEM) is a novel technology for imaging 3D tissue volumes at the subcellular level. FIB-SEM alternates between imaging the block face with a scanning electron beam and milling away thin sections of tissue with a FIB, allowing for collection and analysis of 3D data. FIB-SEM was used to image the three layers of the day 4 chicken embryo heart: myocardium, cardiac jelly, and endocardium. Individual images obtained with FIB-SEM were comparable in quality and resolution to those obtained with transmission electron microscopy. Up to 1,100 serial images were obtained in 4 nm increments at 4.88 nm resolution, and image stacks were aligned to create volumes 800-1,500 μm3 in size. Segmentation of organelles revealed their organization and distinct volume fractions between cardiac wall layers. We conclude that FIB-SEM is a powerful modality for 3D subcellular imaging of the embryonic heart wall.

Original languageEnglish (US)
Pages (from-to)1111-1119
Number of pages9
JournalMicroscopy and Microanalysis
Volume20
Issue number4
DOIs
StatePublished - 2014

Fingerprint

chickens
Focused ion beams
ion beams
Imaging techniques
Scanning electron microscopy
scanning electron microscopy
Tissue
myocardium
organelles
embryos
Electron beams
Volume fraction
electron beams
methodology
Transmission electron microscopy
Scanning
transmission electron microscopy
scanning
high resolution
cells

Keywords

  • cardiac outflow tract
  • embryonic heart development
  • FIB-SEM
  • focused ion beam
  • scanning electron microscopy

ASJC Scopus subject areas

  • Instrumentation

Cite this

3D imaging of the early embryonic chicken heart with focused ion beam scanning electron microscopy. / Rennie, Monique Y.; Gahan, Curran G.; Lopez, Claudia; Thornburg, Kent; Rugonyi, Sandra.

In: Microscopy and Microanalysis, Vol. 20, No. 4, 2014, p. 1111-1119.

Research output: Contribution to journalArticle

@article{7beb086d9486486185db2c1b25e90c81,
title = "3D imaging of the early embryonic chicken heart with focused ion beam scanning electron microscopy",
abstract = "Early embryonic heart development is a period of dynamic growth and remodeling, with rapid changes occurring at the tissue, cell, and subcellular levels. A detailed understanding of the events that establish the components of the heart wall has been hampered by a lack of methodologies for three-dimensional (3D), high-resolution imaging. Focused ion beam scanning electron microscopy (FIB-SEM) is a novel technology for imaging 3D tissue volumes at the subcellular level. FIB-SEM alternates between imaging the block face with a scanning electron beam and milling away thin sections of tissue with a FIB, allowing for collection and analysis of 3D data. FIB-SEM was used to image the three layers of the day 4 chicken embryo heart: myocardium, cardiac jelly, and endocardium. Individual images obtained with FIB-SEM were comparable in quality and resolution to those obtained with transmission electron microscopy. Up to 1,100 serial images were obtained in 4 nm increments at 4.88 nm resolution, and image stacks were aligned to create volumes 800-1,500 μm3 in size. Segmentation of organelles revealed their organization and distinct volume fractions between cardiac wall layers. We conclude that FIB-SEM is a powerful modality for 3D subcellular imaging of the embryonic heart wall.",
keywords = "cardiac outflow tract, embryonic heart development, FIB-SEM, focused ion beam, scanning electron microscopy",
author = "Rennie, {Monique Y.} and Gahan, {Curran G.} and Claudia Lopez and Kent Thornburg and Sandra Rugonyi",
year = "2014",
doi = "10.1017/S1431927614000828",
language = "English (US)",
volume = "20",
pages = "1111--1119",
journal = "Microscopy and Microanalysis",
issn = "1431-9276",
publisher = "Cambridge University Press",
number = "4",

}

TY - JOUR

T1 - 3D imaging of the early embryonic chicken heart with focused ion beam scanning electron microscopy

AU - Rennie, Monique Y.

AU - Gahan, Curran G.

AU - Lopez, Claudia

AU - Thornburg, Kent

AU - Rugonyi, Sandra

PY - 2014

Y1 - 2014

N2 - Early embryonic heart development is a period of dynamic growth and remodeling, with rapid changes occurring at the tissue, cell, and subcellular levels. A detailed understanding of the events that establish the components of the heart wall has been hampered by a lack of methodologies for three-dimensional (3D), high-resolution imaging. Focused ion beam scanning electron microscopy (FIB-SEM) is a novel technology for imaging 3D tissue volumes at the subcellular level. FIB-SEM alternates between imaging the block face with a scanning electron beam and milling away thin sections of tissue with a FIB, allowing for collection and analysis of 3D data. FIB-SEM was used to image the three layers of the day 4 chicken embryo heart: myocardium, cardiac jelly, and endocardium. Individual images obtained with FIB-SEM were comparable in quality and resolution to those obtained with transmission electron microscopy. Up to 1,100 serial images were obtained in 4 nm increments at 4.88 nm resolution, and image stacks were aligned to create volumes 800-1,500 μm3 in size. Segmentation of organelles revealed their organization and distinct volume fractions between cardiac wall layers. We conclude that FIB-SEM is a powerful modality for 3D subcellular imaging of the embryonic heart wall.

AB - Early embryonic heart development is a period of dynamic growth and remodeling, with rapid changes occurring at the tissue, cell, and subcellular levels. A detailed understanding of the events that establish the components of the heart wall has been hampered by a lack of methodologies for three-dimensional (3D), high-resolution imaging. Focused ion beam scanning electron microscopy (FIB-SEM) is a novel technology for imaging 3D tissue volumes at the subcellular level. FIB-SEM alternates between imaging the block face with a scanning electron beam and milling away thin sections of tissue with a FIB, allowing for collection and analysis of 3D data. FIB-SEM was used to image the three layers of the day 4 chicken embryo heart: myocardium, cardiac jelly, and endocardium. Individual images obtained with FIB-SEM were comparable in quality and resolution to those obtained with transmission electron microscopy. Up to 1,100 serial images were obtained in 4 nm increments at 4.88 nm resolution, and image stacks were aligned to create volumes 800-1,500 μm3 in size. Segmentation of organelles revealed their organization and distinct volume fractions between cardiac wall layers. We conclude that FIB-SEM is a powerful modality for 3D subcellular imaging of the embryonic heart wall.

KW - cardiac outflow tract

KW - embryonic heart development

KW - FIB-SEM

KW - focused ion beam

KW - scanning electron microscopy

UR - http://www.scopus.com/inward/record.url?scp=84905236118&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84905236118&partnerID=8YFLogxK

U2 - 10.1017/S1431927614000828

DO - 10.1017/S1431927614000828

M3 - Article

VL - 20

SP - 1111

EP - 1119

JO - Microscopy and Microanalysis

JF - Microscopy and Microanalysis

SN - 1431-9276

IS - 4

ER -