Cryo-FIB-SEM serial milling and block face imaging: large volume structural analysis of biological tissues preserved close to their native state

N. Vidavsky, A. Akiva, I. Kaplan-Ashiri, K. Rechav, L. Addadi, S. Weiner, A. Schertel

Research output: Contribution to journalArticleAcademicpeer-review

25 Citations (Scopus)
1 Downloads (Pure)

Abstract

Many important biological questions can be addressed by studying in 3D large volumes of intact, cryo fixed hydrated tissues (⩾10,000 μm3) at high resolution (5–20 nm). This can be achieved using serial FIB milling and block face surface imaging under cryo conditions. Here we demonstrate the unique potential of the cryo-FIB-SEM approach using two extensively studied model systems; sea urchin embryos and the tail fin of zebrafish larvae. We focus in particular on the environment of mineral deposition sites. The cellular organelles, including mitochondria, Golgi, ER, nuclei and nuclear pores are made visible by the image contrast created by differences in surface potential of different biochemical components. Auto segmentation and/or volume rendering of the image stacks and 3D reconstruction of the skeleton and the cellular environment, provides a detailed view of the relative distribution in space of the tissue/cellular components, and thus of their interactions. Simultaneous acquisition of secondary and back-scattered electron images adds additional information. For example, a serial view of the zebrafish tail reveals the presence of electron dense mineral particles inside mitochondrial networks extending more than 20 μm in depth in the block. Large volume imaging using cryo FIB SEM, as demonstrated here, can contribute significantly to the understanding of the structures and functions of diverse biological tissues.

Original languageEnglish
Pages (from-to)487-495
Number of pages9
JournalJournal of Structural Biology
Volume196
Issue number3
DOIs
Publication statusPublished - 1 Dec 2016
Externally publishedYes

Keywords

  • 3D imaging
  • Biomineralization
  • Cryo-FIB-SEM
  • Sea urchin embryo
  • Zebrafish

Fingerprint Dive into the research topics of 'Cryo-FIB-SEM serial milling and block face imaging: large volume structural analysis of biological tissues preserved close to their native state'. Together they form a unique fingerprint.

  • Cite this