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

Monique Y. Rennie, Curran G. Gahan, Claudia S. López, Kent L. Thornburg, Sandra Rugonyi

Research output: Contribution to journalArticle

4 Scopus citations

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 - Aug 2014

Keywords

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

ASJC Scopus subject areas

  • Instrumentation

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